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  • Genetics
  • 101
    ISSN: 0749-503X
    Keywords: Threonine metabolism ; amino acid biosynthesis ; homologous domains ; chromosome III ; gene organisation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The gene encoding theonine synthase (THR4) from the yeast Saccharomyces cerevisiae was cloned by complementation of a thr4 mutant. This gene was also found on a lambda clone (5239) consisting of a fragment of chromosome III inserted in the vector lambda MG3. The THR4 gene encodes a protein of 514 amino acids (M.W. 58 kDa), which has extensive homologies with E. coli threonine synthase (thrC) and B. subtilis threonine synthase. The 5′ flanking region of the gene contains three regulatory sequences. [TGACT(C)] for the general amino acid control (GCN).About 130 bp downstream of the THR4 gene another open reading frame (563 amino acids) is found in the opposite orientation. This may imply that this open reading frame, called CTR86, shares a terminator region with THR4. The function of the protein encoded by CTR86 is not yet clear, but the fact that the upstream region contains a GCN4 responsive site that the gene product may also be involved in amino acid biosynthesis.
    Additional Material: 4 Ill.
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  • 102
    ISSN: 0749-503X
    Keywords: Alcohol toxicity ; ion permeability ; plasma membrane ; H+-ATPase ; yeast ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The effect on n-alcohols on ATP-dependent generation of ΔpH and Em across the plasma membrane vesicles of the yeast Saccharomyces carlsbergenesis was investigated. The alcohols were shown to collapse ΔpH and Em in the order C2〈C3〈C4〈C5≤C6≥C7〉C8〉C11, the dissipation of Em being more pronounced. Inhibition of the plasmalemma H+-ATPase was insignificannt: at low alcohol concentrations its activity even increased. The basic reason for the toxic effect of the alcohols on the yeast cells was suggested to be due to the increase in the anion and proton permeability of the plasma membrane. Mg2+ partially prevented the increase in the plasmalemma ion permeability by the alcohols investigated.
    Additional Material: 5 Ill.
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  • 103
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990) 
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 104
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 367-382 
    ISSN: 0749-503X
    Keywords: Yeast genetics ; wine yeast ; Saccharomyces cerevisiae ; chromosomes ; karyotyping ; aneuploidy ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A general procedure is described for determining the chromosomal constitution of industrial strains of Saccharomyces cerevisiae based on analysis of segregation frequencies for input markers among random spore progeny of industrial-laboratory strain hybrids. The multiple auxotrophic haploid testers used carried a dominant erythromycin-resistance marker, allowing hybrids to be selected in mass matings with produced by the wild-type industrial strains. Analysis a number of independent crosses between the haploid testers and an unselected population of spores of each wine strain distinguished between disomic, trisomic and tetrasomic chromosomal complements in the parents. Possible explanations for a significant class of aberrant segregation frequencies are discussed.Results of the analysis indicate that UCD Enology 522 (Montrachet) is diployed and possibly trisomic for chromosome VII; 522X is diploid; UCD Enology 505 (California Champagne) is disomic for chromosome XVI, trisomic for chromosomes I, II, III, VI, VIII, IX, X, XII, XV, tetrasomic for chromosomes IV, XI, XIII, XIV and either trisomic or tetrasomic for chromosomes V and VII; and that UCD Enology 595 (Pasteur Champagne) is disomic for chromosomes I, II, III, IX, XVI, trisomic for chromosomes IV, VI, X, XII, XIV, XV, tetrasomic for chromosomes V, VIII, XI, XIII, and either disomic or tetrasomic for chromosome VII.
    Additional Material: 2 Ill.
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  • 105
    ISSN: 0749-503X
    Keywords: Multifunctional FAS2 gene ; Chromosome hybridisation ; spo11 mapping ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The trifunctional FAS2 gene encoding subunits α of the Saccharomyces cerevisiae fatty acid synthetase complex was mapped on the left arm of chromosome XVI 24 centinorgans proximal to GAL4 and 39 centimorgans distal and PEP4 relative to the centromere. Mapping was achieved by three-independent methods: meiotic co-segragation of FAS2 and ARO7 in recombination-deficient spo11-mutants; tetrad analysis of crosses between FAS2, GAL4 and PEP4; and Southern hybridization of purified FAS2 DBA with individual yeast chromosomes separated by pulsed-field gel electrophoresis.
    Additional Material: 2 Ill.
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  • 106
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 6 (1990), S. 451-454 
    ISSN: 0749-503X
    Keywords: DNA-binding ; dimerization ; helix-loop-helix ; PHO4 activator ; Saccharomyces cerevisiae ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Evidence is presented, based on sequence comparisons and secondary structure prediction, of the presence of a DNA-binding and dimerization helix-loop-helix motif in the yeast transcription activator PHO4. Interest in the existence of this first known motif in yeast is discussed.
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  • 107
    ISSN: 0749-503X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A cytochrome c protein gene, CYC10, of the dextran- and starch-fermenting yeast, Schwanniomyces occidentalis was cloned and characterized. The DNA sequence was determined, and the predicted amino acid sequence of the protein-coding region shares close homologies to the cytochrome c genes. A. S. occidentalis strain with a disruption of the gene revealed that CYC10 was the only functional cytochrome c protein-encoding gene in S. occidentalis, unlike the two cytochrome c protein genes (CYC1 and CYC7) in Saccharomyces cerevisiaear The CYC10 gene was oxygen-induced but not subject to catabolite repression.The expression of the CYC10 gene was studied in the heterologous yeast. S. cerevisiae. The oxygen induction of the gene was found to be identical to that of the CYC1 gene, indecating these two genes share similar or closely related cis- and trans- acting oxygen regulatory elements. However, the CYC10 gene was glucose repressed in S. cerevisiae strains; a phenomenon which was not observed in the native S. accidentalis cells. Search in the 5′ unstranslated region of the CYC10 gene revealed some homologies at -425 to -405 to UAS1 of the S. cerevisiae CYC1 gene. A deletion of a segment of upstream region including this sequence abolished expression in S. cerevisiae.Finally the phylogenetic relationships of different yeasts and fungi were determined based upon the amino acid sequences of the cytochrome c proteins. These relationships do not completely agree with classical divisions.
    Additional Material: 8 Ill.
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  • 108
    ISSN: 0749-503X
    Keywords: Cell wall porosity ; permeability ; polycation assay ; cell wall structure ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have developed a new assay to determine relative cell wall porosity in yeasts, which is based on polycation-induced leakage of UV-absorbing compounds. Polycations with a small hydrodynamic radius as measured by gel filtration (poly-L-lysine) caused cell leakage independent of cell wall porosity whereas polycations with a large hydrodynamic radius (DEAE-dextrans) caused only limited cell leakage due to limited passage through the cell wall. This allowed the ratio between DEAE-dextran- and poly-L-lysine-induced cell leakage to be used as a measure of cell wall porosity in Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe. Using this assay, we found that the composition of the growth medium affected cell wall porosity in S. cerevisiae. In addition, we could show that cell wall porosity is limited by the number of disulphide bridges in the wall and is dependent on cell turgor. It is argued that earlier methods to estimate cell wall porosity in S. cerevisiae resulted in large underestimations.
    Additional Material: 4 Ill.
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  • 109
    ISSN: 0749-503X
    Keywords: Chromosome III ; sequencing ; gene disruption ; ribokinase ; ARS ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We report here the DNA sequence of a segment of chromosome III extending over 8·2 kb. The sequence was determined using the random clone strategy followed by oligonucleotide-directed sequencing. The segments contains five long open reading frames, YCR521, 522, 523, 524 and 526, with only short distances between them. YCR523 (333 codons) endodes a ribokinase, a new function for yeast. YCR526 originates inside the MAT cassette, which is in continuity with the present segment, and extends over 358 codons outside of MAT. YCR524 (923 codons) codes for a putative membrane protein. YCR521, 522 and 524, have each been disrupted by insertion of a URA3 cassette and are non-essential genes. An active ARS element is located within YCR523 or its vicinity.
    Additional Material: 8 Ill.
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  • 110
    ISSN: 0749-503X
    Keywords: Peroxisomes ; oleic acid ; β-oxidation ; membrane proliferation ; Hansenula polymorpha ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We studied the physiological responses of Hansenula polymorpha during adaptation of cells to oleic acid-containing media. Growth experiments indicated that the organism was unable to use oleic acid as the sole source of carbon and energy. However, upon incubation of glucose-grown cells in mineral media containing oleic acid, activities of various enzymes of the β-oxidation pathway were induced. These enzymes were localized in microbodies together with alcohol oxidase. Furthermore, a drastic increase in phospholipid content of the cells was observed; this was due to a rapid proliferation of membranes. These consisted of a variable number of membranous layers which were continuous with the peroxisomal membrane. Upon continued incubation, the membrane proliferations extended and large compartments were formed. This process was dependent on the presence of peroxisomes in the cells since it was not observed in peroxisome-deficient mutant strains of H. polymorpha. The newly formed membranous compartments differed from peroxisomes since they did not contain peroxisomal matrix proteins; these were confined to the single enlarged organelle which was incorporated in the membranous structure and characterized by a large alcohol oxidase crystalloid. The membranous compartments are considered to be whole entities since they could not be separated from the peroxisomes by common cell fraction methods; also they were degraded entirely after a shift of cells to glucose-excess condition.Freeze fracturing reveled that the substructure of the membranes greatly resembled that of normal peroxisomal membranes. Since a distinct enhancement of different peroxisomal membrane proteins was observed during the initial hours after the shift, we assume that exposure of H. polymorpha to acid lead to a drastic overproduction of peroxisomal membranes.
    Additional Material: 12 Ill.
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  • 111
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 327-327 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 112
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 341-353 
    ISSN: 0192-253X
    Keywords: Vesicle movement ; myosin II ; cAMP ; Dictyostelium ; actin ; computer-assisted motion analysis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Dictyostelium amoebae were analyzed before and after rapid addition of 10-6 M cAMP for cellular motility, dynamic shape changes, and intracellular particle movement. Before cAMP addition, amoebae moved in a persistent anterior fashion and were elongate with F-actin localized predominantly in the anterior pseudopod. Intracellular particles moved rapidly and anteriorly. Within seconds after 10-6 M cAMP addition, cells stopped translocating, pseudopod formation ceased, intra-cellular particle movement was depressed, and F-actin was lost from the pseudopod and concomitantly relocalized in the cell cortex After 10 seconds, expansion zones reappeared but were small and no longer anteriorly localized. Vesicle movement partially rebounded but was no longer anteriorly directed. The myosin II null mutant HS2215 exhibited both depressed cellular translocation and vesicle movement. The addition of cAMP to HS2215 cells did not result in any detectable change in the random, depressed movement of particles. The results with HS2215 suggest that myosin II is essential for (1) rapid cellular translocation, (2) cellular polarity, (3) rapid particle movement, (4) anteriorly directed particle movement, and (5) the cAMP response. Electron micrographs suggest that at least half of the particles examined in this study contain in turn smaller membrane bound vesicles or multilameilar membrane bodies. The possible role of these vesicles is discussed.
    Additional Material: 10 Ill.
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  • 113
    ISSN: 0192-253X
    Keywords: Amoeboid movement ; cell adhesion ; cytoskeleton ; cell motility ; Dictyostelium ; microfilaments ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Ponticulin is a 17,000-dalton transmembrane glycoprotein that is involved in the binding and nucleation of actin filaments by Dictyostelium discoideum plasma membranes. The major actin-binding protein isolated from these membranes by F-actin affinity chromatography, ponticulin also binds F-actin on blot overlays. The actin-binding activity of ponticulin in vitro is identical to that observed for purified plasma membranes: it resists extraction with 0.1 N NaOH, is sensitive to high salt concentrations, and is destroyed by heat, proteolysis, and thiol reduction and alkylation. A cytoplasmic domain of ponticulin mediates binding to actin because univalent antibody fragments directed against the cytoplasmic surface of this protein inhibit 96% of the actin-membrane binding in sedimenlation assays. Antibody specific for ponticulin emoves both ponticu-lin and the ability to reconstitute actin nucleation activity from detergent extracts of solubilized plasma membranes. Levels of plasma membrane ponticulin increase 2- to 3-fold during aggregation streaming, when cells adhere to each other and are highly motile. Although present throughout the plasma membrane, ponticulin is preferentially localized to some actin-rich membrane structures, including sites of cell-cell adhesion and arched regions of the plasma membrane reminiscent of the early stages of pseudopod formation. Ponticulin also is present but not obviously enriched at phagocytic cups of log-phase amebae. These results indicate that ponticulin may function in vivo to attach and nucleate actin filaments at the cytoplas-mic surface of the plasma membrane. A 17,000-dalton analogue of ponticulin has been identified in human polymorphonuclear leukocyte plasma membranes by immunoblotting and immunofluo-rescence microscopy. These findings suggest that the structure and function of ponticulin in motile cells has been evolutionarily conserved.
    Additional Material: 8 Ill.
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  • 114
    ISSN: 0192-253X
    Keywords: Mutants ; proteolytic processing ; N-linked oligosaccharide ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Dictyostelium discoideum is a useful system to study the biosynthesis of lysosomal enzymes because of the relative ease with which it can be manipulated genetically and biochemically. Previous studies have revealed that lysosomal enzymes are synthesized in vegetatively growing amoebae as glycosylated precursor polypeptides that are phosphorylated and sulfated on their N-linked oligosaccharide side-chains upon arrival in the Golgi complex. The precursor polypeptiaes are membrane associated until they are proteolytically processed and deposited as soluble mature enzymes in lysosomes. In this paper we review biochemical experiments designed to determine the roles of post-translational modification, acidic pH compartments, and proteolytic processing in the transport and sorting of lysosomal enzymes. We also describe molecular genetic approaches that are being employed to study the biosynthesis of these enzymes. Mutants altered in the sorting and secretion of lysosomal enzymes are being analyzed biochemically, and we describe recent efforts to clone the genes coding for three lysosomal enzymes in order to better understand the molecular mechanisms involved in the targeting of these enzymes.
    Additional Material: 6 Ill.
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  • 115
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 473-483 
    ISSN: 0192-253X
    Keywords: N-glycosylation ; glycoprotein sorting ; oligosaccharide processing ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have used metabolic radiolabeling with oligosaccharide precursors, coupled with subcellular fractionation, to examine the distribution of several classes of asparagine-linked oligosaccharides during early development. In Dictyostelium, we have observed endoglycosidase H (endo H)-sensitive structures with sizes corresponding to 10 (Hex10) and 11 (Hex11) hexose residues on the chitobiose core. Only Hex11 was detected as the major structure on fucosylated endo H-resistant species. All Hex11 species cofractionated with plasma membrane and secreted glycoproteins, whereas Hex10 appeared to be confined to intracellular membrane and soluble glycoproteins. Sulfated species correlated with lysosomal and secreted fractions, and glucose residues were markedly depressed in Hex11 of secreted glycoproteins. Outer branch structural studies have revealed several components of the endo H-sensitive species. Usingα-mannosidase and β-hexosaminidase as diagnostic tools, species elucidated thus far are: a structure with 10 mannoses, a structure with nine mannoses and an intersecting N-acetylglucosamine, structures with three glucoses and seven or eight mannoses and several larger species with multiple blocks to digestion.
    Additional Material: 8 Ill.
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  • 116
    ISSN: 0192-253X
    Keywords: Dictyostelium ; glycosylation ; cell adhesion ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The monoclonal antibodies MUD1 and MUD50 recognize peptide and carbohydrate epitopes, respectively, on the Dictyoste-lium discoideum developmentally regulated cell surface glycoprotein PsA. PsA is a putative pre-spore cell adhesion molecule during the slug stage and is anchored to the cell membrane via a phos-phatidylinositol glycan. Here we report on the presence of a PsA-like homolog in several Dictyo-stelium species. The MUD1 epitope is mostly confined to the D. discoideum complex, and only one non-D. discoideum strain reacted strongly with this antibody. By contrast, the mAb MUD50 recognises a homologous molecule in several Dictyostelium species that do not react with MUD1. With mAb MUD50 immunoaffinity chromatography, it was possible to purify and obtain an amino-terminal sequence for the PsA homolog from the Dictvoste-lium sp. strain ZA3A and from one strain of D. pur-pureum.
    Additional Material: 5 Ill.
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  • 117
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 463-472 
    ISSN: 0192-253X
    Keywords: Sulfated/phosphorylated oligosaccharides ; mutants ; lysosomal enzymes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Dictyostelium discoideum synthesizes many highly immunogenic carbohydrates of unknown structure and function. We have used monoclonal antibodies prepared against one of these called CA1 to investigate its structure and the consequences of its loss. CA1 is preferentially expressed on lysosomal enzymes as a specific arrangement of mannose-6-SO4 residues on N-linked oligosaccharides. Mutant strains HL241 and HL243 do not express CA1, and synthesize a truncated lipid-linked oligosaccharide (LLO) precursor that lacks the critical mannose residues needed for expression. The lesion appears to result from the loss of mannosyl transferase activity involved in LLO biosynthesis. The truncated LLO is poorly transferred to an artificial peptide acceptor in a cell-free N-glycosylation assay, and this appears to result from improper topological localization of the LLO or to a lower affinity of the LLO for the oligosaccharyl transferase. Although both mutants share these lesions, they are biochemically and genetically distinct. Only HL243 is lower in N-glycosylation in intact cells, and this is not a result of an altered structure of the LLO. There are other differences between the strains. HL241 can form fruiting bodies at a slower rate than normal while HL243 cannot aggregate. Genetic analysis of defects shows that the CA1 lesion in HL241 is recessive, while the lesions in both CA1 and in development are dominant and co-segregate in HL243 and are, therefore, likely to be in the same gene. Lysosomal enzyme targeting is normal but enzyme processing proceeds at a 2-3 fold slower rate in HL241 and HL243 compared to wild-type. Strain HL244 does not express CA1 since it completely lacks protein sulfation, but lysosomal enzyme targeting and processing proceeds at a normal rate, showing that sulfate is not essential for these processes. Alterations in oligosaccharide structure can have individualized effects on the biosynthesis of lysosomal enzymes. The results presented here illustrate how this approach can be used to study both the structure and function of carbohydrate epitopes.
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  • 118
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 512-512 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 119
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 492-506 
    ISSN: 0192-253X
    Keywords: Prespore vesicle ; cellulose ; polysac-charide ; acid hydrolase spore coat protein ; secretion ; flow cytometry ; confocal microscopy ; macrocyst ; cellular slime mold ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The spore coat forms as a rigid extracellular wall around each spore cell during culmination. Coats purified from germinated spores contain multiple protein species and an approximately equal mass of polysaccharide, consisting mostly of cellulose and a galactose/N-acetyl-galactosamine polysaccharide (GPS). All but the cellulose are prepackaged during prespore cell differentiation in a regulated secretory compartment, the prespore vesicle. The morphology of this compartment resembles an anastomosing, tubular network rather than a spherical vesicle. The molecules of the prespore vesicle are not uniformly mixed but are segregated into partially overlapping domains. Although lysosomal enzymes have been found in the prespore vesicle, this compartment does not function as a lysosome because it is not acidic, and a common antigen associated with acid hydrolases is found in another, acidic vesicle population. All the prespore vesicle profiles disappear at the time of appearance of their contents outside of the cell; this constitutes an early stage in spore coat formation, which can be detected both by microscopy and flow cytometry. As an electron-dense layer, the future outer layer of the coat, condenses, cellulose can be found and is located immediately beneath this outer layer Certain proteins and the GPS become associated with either the outer or inner layers surrounding this middle cellulose layer. Assembly of the inner and outer layers occurs in part from a pool of glycoproteins that is shared between spores, and unincorporated molecules loosely reside in the interspore matrix, a location from which they can be easily washed away. When the glycosylation of several major protein species is disrupted by mutation, the coat is assembled, but differences are found in its porosity and the extractibility of certain proteins. In addition, the retention or loss of proteolytic fragments in the mutants indicates regions of spore coat proteins that are required for association with the coat. Comparative examination of the macrocyst demonstrates that patterns of molecular distributions are not conserved between the macrocyst and spore coats. Thus spore coat assembly is characterized by highly specific intermolecular interactions, leading to saturable associations of individual glycoproteins with specific layers and the exclusion of excess copies to the interspore space.
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  • 120
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 121
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 1-1 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 122
    Electronic Resource
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 27-40 
    ISSN: 0192-253X
    Keywords: Maternal effect ; Drosophila embryogenesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The Drosophila mutation, quartet, affects development at points in the life cycle that require intense mitotic activity. Examination of embryos affected by the maternal effect of quartet has revealed defects that can be attributed to incomplete chromosome separation at mitosis. These defects include uneven spacing of nuclei, strands of DNA creating bridges between nuclei, and abnormal amounts of DNA per nucleus. Nuclei in quartet-affected embryos also have a greater-than-normal number of centrosomes. Immunofluorescent examination of the spindles in quartet-affected embryos has revealed tripolar spindles and adjacent spindles that share a common spindle pole. Finally, chromosome separation distance was measured in anaphase and telophase spindles in quartet-affected embryos and found to be blocked in anaphase. Examination of mitotic figures in quartet larvae revealed a reduced mitotic index and an elevated frequency of abnormal mitotic figures. quartet could encode a function necessary for the disengagement of chromosomes in mitosis, for kinetochore function or for function of a spindle motor. Mutations in quartet prevent the post-translational modification of three abundant proteins. These proteins may be involved in chromosome separation in mitosis.
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  • 123
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 2-14 
    ISSN: 0192-253X
    Keywords: Gene expression ; mRNA localization ; ascidian embryos ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have cloned and characterized the temporal and spatial expression of ScCAl5, a cDNA clone encoding an actin gene in the ascidian Styela clava. The partial nucleotide and derived amino acid sequences of this single- copy gene suggest that it is a cytoskeletal actin. Northern analysis shows that ScCAl5 corresponds to a 1.8-kb mRNA that is transcribed during oogenesis, during embryonic development, and in the adult. In situ hybridization shows that maternal ScCA15 mRNA is distributed uniformly in the cyto- plasm of the oocyte and unfertilized egg. During the period of ooplasmic segregation following fertilization, however, ScCAl5 mRNA appears to be translocated into the ectoplasm, a specialized cytoplasmic region of the egg. During the early cleavages, the ectoplasmic transcripts are partitioned to ectodermal cells in the animal hemisphere, which are precursors of the epidermis and nervous system of the larva. Maternal ScCA15 mRNA is degraded just before gastrulation and replaced by zygotic transcripts which begin to accumulate between the neurula and mid-tailbud stages. Zygotic ScCAl5 mRNA accumulates primarily in the epidermal and neural cells, although lower levels of these transcripts may also be present in tail muscle cells. These results show that two mechanisms are used to concentrate ScCA15mRNA in the ectodermal cells during development: (1) localization and differential segregation of maternal transcripts and (2) specific expression of the ScCA15 gene. ScCAl5 mRNA is detected by in situ hybridization in the testes, ovaries, alimentary tract, and endostyle of adults. In the testes, ScCA15 mRNA is present in developing sperm, whereas in the ovary, these transcripts are present in the germinal epithelium and developing oocytes. In the alimentary tract, ScCAl5 mRNA is confined to the gastric epithelium of the esophagus, stomach, and intestine. Since the ScCA15 gene is expressed in embryonic and adult tissues that are undergoing rapid cell division, this actin is likely to function in some aspect of cell proliferation.
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  • 124
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    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 11 (1990), S. 15-26 
    ISSN: 0192-253X
    Keywords: Development ; gene regulation ; multigene family ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The temporal and spatial patterns of accumulation of transcripts from individual actin genes during Drosophila embryogenesis have been determined by in situ hybridization. We describe the subcloning into transcription vectors of unique DNA fragments derived from the 3′ transcribed, but nontranslated region of each actin gene. These fragments then served as templates for the synthesis in vitro of single-stranded, radioactive gene-specific RNA probes. Probe characterization and hybridization to developmental RNA blots are presented, demonstrated the independent developmental accumulation of actin transcripts from each gene. Each gene-specific probe has been hybridized in situ to the transcripts present in embryonic frozen sections. The results of these experiments have demonstrated that transcripts from each actin gene accumulate differentially in developing Drosophila tissues. The 5C and 42A actin genes are cytoplasmic actin genes, with transcripts distributed in all cells and tissues of the developing embryo. Therefore these genes presumably encode the cytoplasmic actins used for functions common to all cells. Transcripts from both cytoplasmic actin genes are evenly distributed in preblastoderm embryos, becoming localized to the periphery at blastoderm formation [5C: Burn et al.: Dev Biol 131:345-355, 1989]. Later in development, levels of these cytoplasmic transcripts vary in specific tissues. While the patterns of localization of 5C actin transcripts have been published [Burn et al.: Dev Biol 131:345-355, 1989], differential neurological localization is presented here; 42A transcripts are localized at higher concentrations in the midgut, the brain, nerve cord, and gonad. Both 87E and 57B transcripts accumulated in the developing larval body wall musculature, but at differing levels and in differing patterns. Transcripts of the 79B and the 88F actin genes were undetectable in embryos. The results of these experiments suggest dedicated contributions of individual actin genes to complex developmental processes.
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  • 125
    ISSN: 0192-253X
    Keywords: Embryogenesis ; cavitation ; Na ; K-ATPase ; mRNA ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: NaK-ATPase is a plasma membrane enzyme that plays a critical role in eutherian blastocoel formation (cavitation) by pumping Na+ into the extracellular space enclosed by the trophectoderm. Previous experiments with the mouse had shown that the α (catalytic) subunit of the enzyme becomes detectable by immunocyto-chemistry in the late morula, just prior to the onset of cavitation. In the present study we have used cDNAs corresponding to three mRNA isoforms of the α subunit and a β subunit to determine which genes are expressed during preimplantation development and to explore the timing of their expression. Of the three α subunit cDNAs tested by Northern blot hybridization with blastocyst RNA, only α1 produced a hybridization signal, recognizing a single mRNA about 4 kb in length. This mRNA is relatively abundant in zygotes but barely detectable by the 2-cell stage and then accumulates steadily thereafter to reach its preimplantation maximum in blastocysts. The β1 cDNA detected mRNA of about 2.6-2.8 kb. This mRNA is present in zygotes but could not be detected in 2-, 4-, or 8- cell stages; it is present at a low level in late morulae and is abundant in blastocysts. The temporal profile of accumulation of β1 mRNA thus matches more closely than does α1 the timing of appearance of the catalytic subunit. This suggests that the β subunit may regulate production of the holoenzyme and hence the timing of cavitation.
    Additional Material: 5 Ill.
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  • 126
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    Developmental Genetics 11 (1990), S. 77-87 
    ISSN: 0192-253X
    Keywords: Divergent homeobox ; embryonic transcription ; homeobox evolution ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We describe two homeobox sequences, TgHbox5 and TgHboxó, isolated from the Hawaiian sea urchin Tripneustes gratilla using a Drosophila Sex combs reduced probe. Sequence analysis shows that the encoded TgHbox5 home-odomain shares only 30-52% amino acid identity with homeodomains encoded by previously characterized genes, establishing that it is a divergent homeobox that is not in any known class of homeoboxes. TgHbox5 is expressed in the embryo as two major developmentally regulated transcripts. one at 5.0 kilobase (kb) appearing by blastula stage and the other at 2.7 kb appearing at pluteus stage. Multiple transcripts from TgHbox5 are present at a much lower level in adult tissues and are predominantly expressed in small and large intestines. The TgHbox6 homeobox is an Antennapedia-class homeobox, which appears not to be expressed during embryogenesis but produces abundant 3.6 and 3.2 kb transcripts in the six adult tissues examined.
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  • 127
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    Developmental Genetics 11 (1990), S. 49-64 
    ISSN: 0192-253X
    Keywords: Cell migration ; morphogenesis ; mutant analysis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Four recessive mutations that affect long-range embryonic migration of the two canal-associated neurons (CANs) in C. elegans were isolated and characterized with the goal of identifying genes involved in control of directed cell movement. Mutant animals were identified initially by their “withered” tails, a phenotype associated with abnormal CAN migration; the mutants were then analyzed for abnormal cell migrations by Nomarski microscopy. Based on genetic complementation tests, the mutations were assigned to four different loci, two new (mig-10 III, mig-11 III) and two previously identified (unc-39 V, vab-8 V). Mutations at all four loci affect CAN migration with high to moderate penetrance (the percentage of mutant animals that exhibit the phenotype). In addition, two other bilaterally symmetric pairs of neurons (ALM and HSN), the mesoblast M, and a pair of coelomocyte mother cells are affected by one or more of the mutations, generally with lower penetrance. With the exceptions of HSN and the right coelomocyte mother cell, which occasionally migrate beyond their normal destinations, the cells affected appear to migrate either incompletely or not at all. All the migration phenotypes show incomplete penetrance and variable expressivity, although genetic tests suggest that mutations at mig-10 and vab-8 result in complete or nearly complete loss of gene function. The variability in mutant phenotypes allowed tests for interdependence of several of the affected migrations; all those analyzed appeared independent of one another. The possible nature of the mutant defects and possible roles of these four loci in cell migration are discussed.
    Additional Material: 4 Ill.
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  • 128
    ISSN: 0192-253X
    Keywords: Somatic embryogenesis ; gene regulation ; transgenic plants ; β-glucuronidase ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Extensive studies of gene expression programs in carrot somatic embryos identified a gene, designated Dc3, that serves as a reliable molecular marker for the acquisition of embryogenic potential by carrot cells in culture. The complete sequence of a carrot genomic region, DcG3, encoding a Dc3-like mRNA, was determined. The DcG3 transcription unit contains a single intron and encodes mRNA that is expressed at high levels in embryonic tissue but is undetectable in somatic tissue of carrot. The predicted protein sequence of DcG3 is 163 amino acids and includes two approximately 50 amino acid direct repeats which in turn include additional repetitive elements with an unusual distribution of charged amino acids. Dc3 and Dc3-like mRNAs are encoded by a small divergent gene family. Furthermore, similarities of the Dc3 gene family with genes from other plant species that are expressed in response to environmental and developmental cues suggest a possible role in seed desiccation and possibly in more general water-stiess responses in plants. Analysis of transgenic tobacco containing a β-glucuronidase (GUS) reporter gene fused to a 1.7 kb 5′ upstream element of DcG3 defined a promoter/enhancer complex that confers developmentally and environmentally regulated expression of GUS activity. Thus, DcG3 is phylogenetically conserved together with the trans-acting factors required for its regulated expression in transgenic tobacco.
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  • 129
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    Developmental Genetics 11 (1990), S. 133-140 
    ISSN: 0192-253X
    Keywords: Morphogenesis ; heat shock ; phenocopies ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The construction of cell hairs (trichomes) on the wings of Drosophila occurs in synchrony on 30,000 cells over a period of about 20 hr. Changes in both morphology and patterns of protein synthesis occur rapidly during this time period. In this report we describe the use of stressinduced (heat shock) abnormalities in morphogenesis to provide further details on the stepwise processes of differentiation within single wing cells. A cartoon summary of the overall process and a discussion of some possible mechanisms is included.
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  • 130
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    Developmental Genetics 11 (1990), S. 141-148 
    ISSN: 0192-253X
    Keywords: Aging ; longevity ; genes and aging ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The first step in a genetic analysis of aging is to identify and characterize the genetic mutants and their controls that will be used. Such mutants or strains are initially identified by their effect on the life span. Yet many genetic interventions are known to have some effect on the life span without necessarily affecting the aging process. It is therefore necessary to prove that one is actually dealing with an aging mutant before one draws strong inferences from the data. Casarett's rules provide an operational test for doing so, relying as they do on the comparison of aging biomarkers in the experimental and reference strains. We show that our previously described genetically based long-lived NDC-L strain and its normal-lived NDC-R control strain differ only in the chronological age of expression of two behavioral and three physiological functional age biomarkers. They do not differ in the sequence or the physiological age of expression of these biomarkers. These two strains comply with the Casarett rules and thereby comprise a valid tool with which to conduct a comparative genetic analysis of aging. The implications of the available data are discussed, including the possibility that aging in these strains of Drosophila melanogaster may be the result of a multiphasic developmental process.
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  • 131
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    Developmental Genetics 11 (1990), S. 149-159 
    ISSN: 0192-253X
    Keywords: Superoxide dismutase ; glutathione peroxidase ; catalase ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The interaction of reactive oxygen metabolites with DNA is well characterized and may result in mutagenesis, chromosome aberrations, and modulation of gene expression. Superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) catalyze enzymatic reactions to remove oxidant stresses, particularly O2- and H2O2. The role of these enzymes during in utero development of the embryo and the developmental pattern of expression of the embryonic genes encoding them is not known. We examined the in utero developmental expression and activity of the three free-radical-metabolizing enzymes in mice. We collected mouse fetuses at different stages of development and examined total RNA populations by Northern and slot blots using gene-specific cDNA probes. In addition to quantifying the probe-specific RNAs, activities of the three enzymes were also evaluated on the same tissue samples.The gene-specific RNAs and the associated enzyme activities are detectable with somite formation (day 8 postcoitus [p.c.]) in mice. The relative RNA values for each of the genes studied are higher in in utero stages as compared with the adult. The specific activities of these enzymes, on the other hand, follow a characteristic increase with development and growth. The relative RNA levels for each of the genes studied are higher during in utero growth and development than the relative enzyme activity values (between day 8 and day 18, third trimester) in the liver and carcass. This may suggest that the mRNA specific to these genes may accumulate in utero and are not translated immediately. Such accumulating transcripts are translated efficiently after birth, when these enzyme are particularly needed with the advent of aerobic respiration.
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  • 132
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    Developmental Genetics 11 (1990) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 133
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    Developmental Genetics 11 (1990), S. 160-167 
    ISSN: 0192-253X
    Keywords: Flanking sequences ; insertion events ; gene expression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A series of constructs containing the developmentally regulated soybean lectin gene (Le1) were used to transform tobacco plants in order to assess developmental and quantitative regulation conferred by flanking sequences. The largest of the lectin constructs contained approximately 3,00 base pairs (bp) of Le1 5′ flanking region and 1,500 bp of the 3′ flanking region. The smallest construct contained no 5′ flanking region and 194 bp of the 3′ flanking region. ELISA assays of lectin in individual tobacco seeds and Southern blot analyses confirmed that most constructs were inherited as unique insertion events. Maximal expression of Le1 required more than 338 bp of 5′ sequence, indicating that far upstream factors are involved in quantitative control of lectin expression. Lectin expression declined more than 80% between deletions with 1,700 versus 338 bp of 5′ flanking sequence. In contrast, developmental control of lectin expression was maintained by Le1 inserts with only 190 bp of 5′ sequence. The lectin promoter offers a potential means to target high levels of gene expression to the developing seeds of soybean or other dicotyledonous plants.
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  • 134
    ISSN: 0192-253X
    Keywords: Human fetal globin gene ; hemoglobin switching ; mouse erythroleukemia cells ; erythroid differentiation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have analyzed the expression of endogenous murine genes and of transfected human fetal Aγ globin gene in GM 979, a mouse erythroleukemia line which produces adult as well as embryonic globins. Optimal induction of the endogenous murine adult globin genes was obtained with DMSO or HMBA while the ∊y and βh1 embryonic genes were preferentially induced by butyrate. Similarly, the transferred human Aγ globin gene was preferentially induced by butyrate. These results as well as previous observations in vivo or in erythroid cell cultures suggest that butyrate preferentially induces the expression of fetal globin genes.
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  • 135
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    Developmental Genetics 11 (1990), S. 175-175 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 136
    ISSN: 0192-253X
    Keywords: Video-image analysis ; contact inhibition ; collision theory ; neural crest ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Chimeras provide unique opportunities to study interactions between the phenotypically similar but genotypically allogeneic cell populations during embryogenesis in vivo. From the quantitative analysis of coat-color patterns in C3H/HeN↔BALB/cA chimeras, a model was proposed stating that the aggregability of the C3H/ HeN-derived melanoblasts in the chimeras was inversely related to the ratio between the mean free path of the epidermal melanoblasts in the normal C3H/He N mouse and that in the chimeras. As a corollary, the possibility was suggested that during the migration of melanoblasts, mechanisms identical with or similar to contact inhibition of movement might operate after collision between the isogeneic, but not between the allogeneic melanoblasts. With regard to the number of melanoblast clones in the trunk region of the mouse, the present series of analyses yielded the value of 24-28 arranged unilaterally; the value closely approximated the number of the somites in that region and provided further support for the proposition made earlier by Tachi [Dev Genet 9:121-154, 1988; “Development of Preimplantation Embryos and Their Environment.” New York: Alan R. Liss, Inc., 1989, pp 263-274].
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  • 137
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    Developmental Genetics 11 (1990), S. 270-279 
    ISSN: 0192-253X
    Keywords: Phenocopy ; developmental arrest ; heat lability ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Heat shock has a dramatic effect on the organization of the cytoplasm, causing the intermediate filament cytoskeleton to aggregate of the nucleus. This has previously been shown in cultured Drosophila and mammalian cells. In this paper we analyze the heat lability of the intermediate filament cytoskeleton in early Drosophila embryos by indirect immunofluorescence. At all stages of embryogenesis tested, the intermediatefilamentcytoskeleton, which is maternally provided, is severely disturbed by 30 min heat shock at 37°C. After the nuclei have migrated to the subcortical cytoplasm, it collapses around them. Nuclei in all heat-shocked embryos are considerably enlarged and become displaced. Embryos before cellular blastoderm stage, in which heat shock protein synthesis is not inducible, are irreversibly arrested in development by heat shock. Embryos at or after cellular blastoderm, which do synthesize heat shock proteins in response to stress, are also immediately arrested in development but continue development when returned to 25°C. We discuss the possibility that cytoplasmic events such as the intermediate filament cytoskeleton rearrangement may be involved in heat shock-mediated phenocopy induction.
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  • 138
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    Developmental Genetics 11 (1990), S. 280-288 
    ISSN: 0192-253X
    Keywords: Seed germination ; transcription control ; compartmentalization ; polysomes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The temporal expression pattern of the CAT-2 catalase isozyme in scutella of Zea mays seedlings normally coincides with that of other major glyoxysomal enzymes. In standard genetic lines (e.g., W64A), the CAT-2 enzyme is synthesized de novo after imbibition, reaches a peak at approximately 4 days later, and then declines steadily. In a high CAT-2 genetic line, R6-67, the enzyme accumulates in a linear fashion for at least 8 days after imbibition and reaches a level 3-fold higher than in W64A. During the first 9 days of early seedling growth in W64A, the correlation between Cat2 mRNA levels and CAT-2 protein suggests that pretranslational control governs Cat2 gene expression. In R6-67, the steady rise in CAT-2 protein appears to result from a pretranslational control mechanism in which Cat-2 mRNA apparently never declines to levels which would limit the rate of accumulation of CAT-2 protein. In addition, the amount of Cat2 mRNA bound to polysomes is 3-fold higher in R6-67 at day 9 relative to W64A at day 9, reflecting a much greater capacity to synthesize CAT-2 later in development. Despite substantial differences in Cat2 mRNA levels between genetic lines, early CAT-2 protein accumulation is similar until day 5, when other glyoxysomal enzymes also attain maximal activity levels. The early increase in CAT-2, between day 2 and day 5 post-imbibition, occurs despite a sharp decline in polysomal Cat2 mRNA. This is related to a transient decline in total extractable polysomes which paradoxically coincides with the peak in glyoxysomal enzyme activities. Early Cat2 gene expression is likely controlled by the compartmentalization of CAT-2 in glyoxysomes.
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  • 139
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    Developmental Genetics 11 (1990), S. 289-293 
    ISSN: 0192-253X
    Keywords: Cell or tissue specificity ; Glycine max ; Transformation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: A number of independently derived transgenic soybean plants expressing a chimeric β-glucuronidase (GUS) gene under the control of the 355 CaMV promoter and a nopaline synthase polyadenylation signal were recovered using direct DNA transfer via electric discharge particle acceleration. Expression of GUS in R, plants was localized using thin tissue sections. Many tissue types expressed GUS at various levels. Pericycle cells in root, parenchyma cells in xylem, and phloem tissues of stem and leaf had high levels of enzyme activity. Procambium, phloem, and cortex cells in root, vascular cambium cells in stem, and the majority of cortex cells in leaf midrib, expressed low or no GUS activity. Intermediate levels of GUS activity were detected in leaf mesophyll cells, certain ground tissue cells in stem and leaf midrib, and in trichome and epidermal guard cells. Thus, we conclude that the 35S CaMV promoter is cell-type specific and is developmentally regulated in soybean.
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  • 140
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    Developmental Genetics 11 (1990), S. 299-309 
    ISSN: 0192-253X
    Keywords: Ecdysone ; gene regulation ; imaginal discs ; morphogenesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The steroid hormone 20-hydroxyecdysone (20-HE) induces imcainol discs to form adult appendages in Drosophila. We have isolated a set of six ecdysone-responsive genes that apparently encode disc cell-surface or secreted proteins. Transcripts from one of these genes, IMP-E3, accumulate rapidly within 1-2 h in response to hormone. Developmentally,IMP-E3 transcripts reach maximum levels during the first stages of metamorphosis (white prepupae, WPP) and are primarily limited to imaginal tissues. Transcripts are also present during embryogenesis (0-3 h and 12-18 h). Two different-sized transcripts (1.2 and 1.4 kb) result from differential polyadenylation, with the larger transcript predominating in WPP. The conceptual IMP-E3 protein contains a signal peptide, an RGD sequence, and a potential glycosylphosphatidylinositol anchor. We speculate that the protein provides a transient cue important for imaginal disc morphogenesis.
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  • 141
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    Developmental Genetics 11 (1990), S. 294-298 
    ISSN: 0192-253X
    Keywords: Soybean ; Heat sensitive genotype ; heat shock proteins ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Arabidopsis thaliana seedlings as measured by an electrolyte leakage assay, have been found to be extremely sensitive to high temperature stress as compared to a high temperature tolerant variety (Tracy) of soybean. Over 50% ion leakage occurred in Arabidopsis leaves during a 15-minute exposure to 50°C, indicating a heat killing time of less than 15 minutes. In contrast, the heat killing time for soybean at 50°C was over five times longer. When soybean or Arabidopsis seedlings in culture plates were exposed to 37°C for 2 hours and then returned to 23°C, they suffered no apparent short-term or long-term damage. Soybean seedlings given a 42°C, treatment for 2 hours also showed no damage. Arabidopsis seedlings after a 42°C treatment for 2 hours showed no apparent immediate damage, but 48 hours after return to 23°C severe damage symptoms were visible and after 96 hours all the seedlings were dead. Both soybean and Arabidopsis seedlings synthesize heat shock proteins (hsps) when exposed to 42°C for 2 hours. The hsps synthesized are of similar molecular weights, although the relative abundances of the different size classes are very different in the two plants. Even though hsps are produced in Arabidopsis seedlings after a 2 hour exposure to 42°C their presence is not sufficient for the seedlings to recover from the effects of rhe heat shock when returned to 23°C. Our results show that Arabidopsis has a heat sensitive genotype. This along with its other characteristics should make it a good model system in which to assay in transgenic plants, the functions of homologous and heterologous genes that might be candidates for determining heat tolerance in plants.
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  • 142
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    Developmental Genetics 11 (1990), S. 310-317 
    ISSN: 0192-253X
    Keywords: Drosophila acidic protein ; leader sequence ; tubular structures ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The Drosophila gene ectodermal (ect, located at 67D8-10 on chromosome 3) is expressed for a short period at mid-embryogenesis in all ectodermally derived tissues except the nervous system. During this stage the tissues involved form tubular structures by a process of invagination followed by cell fusion. Here we report the sequence of the ect protein as deduced from the longest ORF (280 codons) of an ect cDNA. The principal molecular features of the ect protein are: (1) a consensus leader sequence for targeting to the rough ER; (2) a central domain containing a remarkably high density of acidic residues arranged in large clusters separated by smaller clusters of hydrophobic residues; (3) a consensus nuclear-targeting sequence near the C-terminus; (4) a single tyrosine residue located at a potential tyrosine-sulfation site. The antibody staining pattern of the ect protein corresponds to the in situ hybridization pattern of the transcript. A possible role for the ect protein in the complex process of fubular formation that occurs in embryonic ectodermal tissues is discussed.
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  • 143
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    Developmental Genetics 11 (1990) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 144
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    Journal of molecular evolution 17 (1981), S. 167-181 
    ISSN: 1432-1432
    Keywords: Evolution ; Genetics ; REH theory ; Mutations ; Natural selection ; Nucleic acids ; Proteins ; Paleogenetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have independently repeated the computer simulations on which Nei and Tateno (1978) base their criticism of REH theory and have extended the analysis to include mRNAs as well as proteins. The simulation data confirm the correctness of the REH method. The high average value of the fixation intensity μ2 found by Nei and Tateno is due to two factors: 1) they reported only the five replications in which μ2 was high, excluding the forty-five replications containing the more representative data;and 2) the lack of information, inherent to protein sequence data, about fixed mutations at the third nucleotide position within codons, as the values are lower when the estimate is made from the mRNAs that code for the proteins. REH values calculated from protein or nucleic acid data on the basis of the equiprobability of genetic events underestimate, not overestimate, the total fixed mutations. In REH theory the experimental data determine the estimate T2 of the time average number of codons that have been free to fix mutations during a given period of divergence. In the method of Nei and Tateno it is assumed, despite evidence to the contrary, that every amino acid position may fix a mutation. Under the latter assumption, the measure X2 of genetic divergence suggested by Nei and Tateno is not tenable: values of X2 for theα hemoglobin divergences are less than the minimum number of fixed substitutions known to have occurred. Within the context of REH theory, a paradox, first posed by Zuckerkandl, with respect to the high rate of covarion turnover and the nature of general function sites in proteins is resolved.
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  • 145
    ISSN: 1432-069X
    Keywords: Genetics ; Hair dysplasia ; Scanning electron microscopy ; Uncombable hair syndrome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
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  • 146
    ISSN: 1432-0983
    Keywords: Protoplast ; Fusion ; Mitochondrial ; Genetics
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Using a protoplast fusion technique we have been able to locate to the mitochondrial genome of the asporogenous yeast Torulopsis glabrata mutations conferring resistance to oligomycin, antimycin and diuron. When two strains differing in the size of their mtDNAs were fused the mitochondrial markers from the parent with the larger mtDNA (71–91) were transmitted predominantly among the fusion products. Both genetical and physical evidence support the occurrence of recombination in T. glabrata mitochondrial genome. Segregation of the mitochondrial genome appears to take place before the separation of the first bud from the fusion product.
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    Current genetics 4 (1981), S. 177-180 
    ISSN: 1432-0983
    Keywords: Genetics ; Yeasts, protoplasts ; Saccharomyces ; Hansenula
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protoplasts of petites of strains 625-CI of Saccharomyces diastaticus and NCYC 1085 of Saccharomyces cerevisiae, originally obtained from the National Collection of Yeast Cultures, England, were fused with protoplasts of Candida pseudotropicalis, Saccharomyces rosei, Yaccharbmycesmontanus, Pichiamembranefaciens, Hansenula anomala, Hansenula capsulata, and Schizosac-charomyces pombe. The respiratory-competent products of the fusions were selected on the basis of using at least one of the carbon sources utilized by the petite parent and not by the other. The products of the fusion of C. pseudotropicalis x 1085(p−) consisted of two cell types; an oval cell which utilized both lactose and maltose and fermented lactose vigorously, and a cylindrical form which fermented maltose slowly. The S. rosei x 1085(p−) hybrids had acquired the ability to metabolize and ferment galactose, and to ferment maltose, from the petite parent. The P. membranaefaciens x 625(p−) hybrids acquired the ability to metabolize galactose, sucrose and maltose, but fermented only glucose, weakly, like the P. membranaefaciens parent strain. The H. capsulate x 625(p−) hybrids, unlike the hybrids with P. membranaefaciens or S. rosei, resembled the petite parent morphologically and also had the fermentative abilities of this strain (galactose, maltose, sucrose and starch), and the ability to ferment starch was considerably enhanced. The S. montanus x 625(p−) hybrids acquired the ability to utilize starch. Schizosaccharomyces pombe x 625(p−) hybrids resembled S. pombe morphologically, but had the ability to metabolize galactose and starch. Some of the asci produced by these hybrids contained abnormal numbers of spores. H. anomala x 624 x(p−) hybrids fermented starch, though weakly.
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  • 148
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    Theoretical and applied genetics 60 (1981), S. 229-236 
    ISSN: 1432-2242
    Keywords: Glutenin ; Triticum ; Genetics ; SDS ; Polyacrylamide-gel-electrophoresis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The high-molecular-weight (HMW) subunits of glutenin from about 185 varieties were fractionated by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). About 20 different, major subunits were distinguished by this technique although each variety contained, with only a few exceptions, between 3 and 5 subunits. Further inter-varietal substitution lines to those already described (Payne et al. 1980) were analysed and the results indicate that all the HMW subunits are controlled by the homoeologous group 1 chromosomes. All hexaploid varieties studied except ‘NapHal’ contained two major subunits controlled by chromosome 1D. Their genes were shown to be tightly linked genetically for only four different types of banding patterns were observed. The nominal molecular weights determined after fractionation in 10% polyacrylamide gels were between 110,000 and 115,000 for the larger of the two subunits and between 82,000 and 84,000 for the smaller. One quarter of the varieties contained only one major HMW subunit controlled by chromosome 1B whereas the rest had two. The chromosome 1B subunits were the most varied and nine different banding patterns were detected. All the subunits had mobilities which were intermediate between those of the two chromosome 1D-controlled subunits. Only two types of HMW subunit controlled by chromosome 1A were detected in all the varieties examined; a single variety never contained both of these subunits and 40% of varieties contained neither. The chromosome 1A-controlled subunits had slightly slower mobilities in 10% gels than the largest HMW subunit controlled by chromosome 1D. About 100 single grains were analysed from each of five different crosses of the type (F1 of variety A × variety B) × variety C. The results indicate that the genes on chromosome 1B which control the synthesis of subunits 6, 7, 13, 14 and 17 are allelic, as are the genes of the chromosome 1A-controlled subunits, 1 and 2.
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  • 149
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    Theoretical and applied genetics 60 (1981), S. 265-268 
    ISSN: 1432-2242
    Keywords: Agrocybe ; Genetics ; Fruiting ; Biomass
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In the edible white rot fungus Agrocybe aegerita the threshold from mycelial growth to fruit body formation is under control of a single gene in both monokaryons and dikaryons. The allele su opens the pathway for fruiting and allows the subsequent expression of the fruiter genes fi+ (fruit body initials) and fb + (fruit bodies). Its allele, su +, suppresses monokaryotic fruiting completely and restricts dikaryotic fruiting drastically. The detection of this threshold gene su +/su and its action and interactions has practical implication in that an opportunity for concerted breeding is created. First results indicate that the fruiter genes are involved in two essential parameters of productivity. Both time of fruiting and biomass production depend on the two fruiter genes fi + and fb +. Comparable results obtained with two other basidiomycetes suggest that the genetic control of fruiting in Agrocybe aegerita is a general mechanism which may be made use of in breeding work with other basidiomycetes of economic value.
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  • 150
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    Environmental biology of fishes 6 (1981), S. 357-360 
    ISSN: 1573-5133
    Keywords: Behaviour ; Esterase ; Evolution ; Genetics ; Isozymes ; Stock structure ; Schools ; Starch gel
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Synopsis Heterogeneous gene frequencies of Est-1 across groups ofNotropis cornutus provide evidence of behaviourally imposed restrictions on stock structuring. Positive fixation indices (F1S = 0.056 and F1T = 0.085) were reflected by a deficiency of heterozygotes for pooled groups. The degree of subdivision ofN. cornutus stocks cannot be evaluated with the present evidence. but it is likely that their schooling behaviour is associated with significant genotypic structuring of the species.
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    Developmental Genetics 2 (1981) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 152
    ISSN: 0192-253X
    Keywords: nuclear differentiation ; cytoplasmic inheritance ; Paramecium tetraurelia ; mating type ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In P. tetraurelia each cell is determined to express only one of the two complementary mating types, O and E. This determination is under cytoplasmic control and seems to be achieved only by the commitment or noncommitment to the expression of mating type E. All the previously known mutations affecting the differentiation of mating type prevent the expression of the E mating type (O-restricted mutations) without affecting the determination process. An E-restricted mutation was obtained: mtFE. Its phenotypic properties indicate that the mutation affects the determination process itself. When an O cell becomes mtFE/mtFE it acquires the E mating type and an E-determining cytoplasm. We propose that this constitutive determination for the E mating type is due to the inefficiency of a factor which is normally active in an O cell. This factor would act like a repressor and stabilize the E functions under an inactive state.
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  • 153
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    Developmental Genetics 2 (1981), S. 279-290 
    ISSN: 0192-253X
    Keywords: temperature-sensitive mutant ; cytokinin ; hormonal metabolism ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Developmental controls of morphological mutants of Phaseolus vulgaris L. conditioned by two independent loci, DL1 and DL2, were examined through grafting experiments and hydroponic studies. Phenotypes of mutant classes were duplicated by unions of scions and stocks derived from different genotypes. Results indicate that DL1 and DL2 regulate a root and shoot factor respectively, contributing to the mutant types. The allelic dosages of DL1 in the root and DL2 in the shoot rather than the genotype of the whole plant per se determine the severity of the mutant expression. Plants heterozygous for both loci with a temperature-sensitive expression of the mutant phenotype were used to determine physiological components involved. The primary abnormal developmental event associated with the appearance of mutant phenotypes, the restricted root growth at high temperature, could be overcome by the addition of cytokinin in hydroponic solution. These observations suggest that DL1 and DL2 may be related to the regulation of hormonal function or metabolism.
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  • 154
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    Developmental Genetics 2 (1981) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 155
    ISSN: 0192-253X
    Keywords: suspensive and adhesive teratoma cells ; teratoma embryoid bodies ; cell differentiation ; endogenous prostanoid biosynthesis ; long-chain fatty acyl CoA derivatives ; mass fragmentography ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Attachment of the cell surface to a substratum may play a critical role in initiating some cellular developmental commitments and in sustaining differentiation of cells that have already been specialized. Embryoid bodies of teratoma OTT6050 were divided, on day 10 of initial culture, into myogenic adhesive cells which were already (at day 6) characterized by endogenous prostaglandin (PG)I2 formation and little-specialized suspensive cells which formed only thromboxane (TX)B2 in the same culture system. Since at day 10 both cell types reached a stationary phase in which the nature of each cell was mature enough for the analyses with mass fragmentographic technique and gas chromatography- mass spectrometry (GCMS), the total levels of predominant long-chain fatty acyl CoA (acyl CoA) derivatives could be measured comparatively as methyl esters after methanolysis. It was found as a result of major differentiation that adhesive cells had a rather low ratio of arachidonyl CoA to stearyl CoA, although adhesive cells accumulated a larger total amount of acyl CoA derivatives than that accumulated in suspensive cells.
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  • 156
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    Developmental Genetics 2 (1981), S. 99-111 
    ISSN: 0192-253X
    Keywords: CIPC as a mitotic inhibitor of Dictyostelium ; inhibition of Dictyostelium development by CIPC ; CIPC-resistant mutants ; Dictyostelium ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The anti-mitotic herbicide isopropyl N-(3-chlorophenyl) carbamate (CIPC) prevents the growth of amoebae of Dictyostelium discoideum without killing the cells for a period of time equivalent to one generation. During in-hibition, amoebae accumulate in prophase and metaphase of mitosis. After removal of CIPC, they continue through mitosis and then divide.The addition of CIPC to amoebae under starvation conditions prevents aggregation and concomitant cell elongation. The cells, however, do not lose their ability to adhere to a surface, and they remain viable. When CIPC is added to amoebae which have formed streams, it leads to the disintegration of streams into small clusters of cells and to a loss of cell elongation.Post-aggregation stages of development can be inhibited by CIPC at the mound, slug, or Mexican hat stages. Slugs break apart into distinct aggregates.Mutants resistant to CIPC can be obtained easily. Among these mutants, many become temperature sensititive for growth (27°C) or development (27°C or 15.5°C). Others show various abnormalities at the normal temperature (22°C). Most mutants are cross resistant to the microtubule inhibitors nocodazole and thiabendazole, and some are also resistant to CIPC during development.
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  • 157
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    Developmental Genetics 2 (1981), S. 147-158 
    ISSN: 0192-253X
    Keywords: triplo-lethal locus ; Tpl ; gene-dosage ; X chromosome ; Drosophila melanogaster ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Only a single locus (Tpl) is known in the Drosophila melanogaster genome that leads to early lethality when present as a heterozygous duplication (three doses) or deficiency (one dose). We report the recovery of third instar larvae (and of occasional adults) carrying a duplication for the triplo-lethal locus, Dp(Tpl). Karyotype analysis of the larvae showed that the individuals surviving were almost entirely 3X;2A metafemales. We examined the question of whether the entire X or a single X locus was a major factor permitting survival. X-Y translocations were used to produce females hyperploid for different portions of the X and carrying Dp(Tpl). Analysis of metaphase chromosomes by quinacrine fluorescence pattern indicates that the X chromosome region between 6D and 7DE must be present in an extra copy to enhance the survival of Tpl duplication-bearing females. Another type of experiment suggests that it is the region between 7C and 7DE which is essential.
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  • 158
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    Developmental Genetics 2 (1981), S. 131-146 
    ISSN: 0192-253X
    Keywords: rat chimeras ; hooded pigmentation ; melanocyte ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: New, improved media and procedures for making rat chimeric embryos and culturing them in vitro have been developed. We have produced 27 rat chimeras: 20 males and 7 females. This ratio of males to females is consistent with that seen in mouse chimeras, suggesting that rat sex chimeras develop as phenotypic males. By aggregating embryos containing appropriate genetic markers for pigment cell differentiation, it is possible to produce chimeras that elucidate the site of action of the hooded gene. The coat color patterns of black ↔ black hooded chimeras display a white belly spot. In black ↔ albino hooded chimeras, small patches of white hair appear on the head and a large white spot occurs on the belly. Black ↔ agouti hooded chimeras display both agouti and nonagouti pigmentation over the entire surface of the chimera. These animals are fully pigmented with no white spots. In black ↔ albino non-hooded chimeras, rather small irregular patches of black and white hairs are distributed throughout the pelage. Histological examination of sections of hair follicles obtained from the white areas in the head of black ↔ albino hooded chimeras revealed amelanotic melanocytes. On the other hand, hair bulbs from the white belly spots do not contain any such melanocytes. Thus the white hairs of the head are due to the presence of albino melanocytes, but the white hairs of the belly are due to the total absence of melanocytes. All these observations are consistent with the conclusion that the hooded gene acts within melanoblasts, probably to retard their migration from the neural crest and/or to prevent their entrance into the hair follicles of the white areas of hooded rats.
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  • 159
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    Developmental Genetics 2 (1981), S. 171-183 
    ISSN: 0192-253X
    Keywords: sperm ; F9 antigen ; T/t-complex ; immunolabeling ; scanning electron microscopy ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The antigens defined by conventional syngeneic antiserum against F9 embryonal carcinoma cells were localized on mature sperm using immunolabeling and scanning electron microscopy. Labeling patterns were compared for normal (+ / +) mice and mice bearing recessive t-haplotypes. The results showed that antigens detected by intact anti-F9 antiserum are expressed similarly in all genotypes, except for sperm from mice bearing the t12-haplotype where the frequency of labeled cells was reduced. Labeling with the IgM fraction of anti-F9 antiserum was lower on sperm from all t-genotypes examined, with sperm from + /t12 males showing the most marked reduction. In all cases, the labeling patterns were similar, and included a labeling of the whole sperm head with complete anti-F9 antiserum and a restriction of the label to the postacrosomal region when the IgM fraction was used.
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    Developmental Genetics 2 (1981), S. 219-222 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 161
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    Developmental Genetics 2 (1981), S. 49-73 
    ISSN: 0192-253X
    Keywords: determination ; Drosophila ; haltere disc ; homeotic mutation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Mutations at the bithorax locus transform anterior haltere tissue into anterior wing. These transformations could in principle be due to the mutations altering either the expression or cell heredity functions of determination. I have studied two alleles of the bithorax locus bx3 and bx34e using disc culture techniques and found that both produce their transformations by altering the expression of the determined state. I have also found that the expression of the temperature-sensitive allele, bx34e, can be altered by temperature shifts during the culture period. Evidence has been obtained that suggests that such changes in expression do not require growth or cell division.
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  • 162
    ISSN: 0192-253X
    Keywords: temporal genes ; GPDH isozymes ; regulation ; development, Drosophila ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The complete developmental program of glycerol-3-phosphate dehydrogenase in wild type Drosophila is described with respect to activity, isozyme expression, and GPDH-specific CRM. Variants of this developmental program have been isolated from natural populations which affect the rate of accumulation of only the GPDH-3 isozyme in both the larval and adult stages of development. This activity variation segregates as a single gene which is tightly linked to the structural element on Chromosome II, exhibits cis-control, and is tissue specific in expression. This gene meets all the criteria for temporal regulatory genes.
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  • 163
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    Developmental Genetics 2 (1981) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 164
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    Developmental Genetics 2 (1981), S. 159-170 
    ISSN: 0192-253X
    Keywords: Tetrahymena hegewischi ; timing of maturity ; cellular differentiation ; genetic ; environmental variation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The development of sexual maturity has been studied in Tetrahymena hegewischi. Progeny lines do not typically change from immaturity to mating with all different mating types during a single test interval, but about 30% do mature abruptly. Some testers are more likely than others to participate in the earliest mating reactions of progeny lines which do not mature abruptly. Subcaryonidal vegetative pedigrees of 10 pairs from 4 crosses revealed considerable intrapair variation in the time, measured in fissions, of maturity. The average intrapair coefficient of variation was 20%. A nested ANOVA revealed significant genomic effects on the immaturity interval, but no significant cytoplasmic or caryonidal effects; 56% of the total variation was non-genomic. Growth in different environments had highly significant effects on the immaturity interval. Subclones grown at 27°C with alternate day transfers took on the average 2 to 3 times as many fissions to mature as sister subclones grown at 27°C with daily transfers. Subclones grown at 18°C or 34°C and transferred on alternate days had intermediate maturation times. The greatest range in the immaturity interval among lines of the same genotype was from 34 to 143 fissions. The development of maturity in this species involves genetic control of timing, but the genetic differences are obscured by a large amount of intraclonal variation and sensitivity to the environment.
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  • 165
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    Developmental Genetics 2 (1981) 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 166
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    Developmental Genetics 2 (1981), S. 369-383 
    ISSN: 0192-253X
    Keywords: Dictyostelium discoideum ; aggregation-deficient mutants ; intracellular cAMP ; adenylate cyclase ; defective spore differentiation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Forty aggregation-deficient mutants of Dictyostelium discoideum were screened for changes in intracellular cAMP during the first 10 hr of starvation. The pools in 39 of the mutants remained low and relatively static during this period. However, amoebae of one mutant, strain HC151, exhibited significantly elevated levels of intracellular cAMP during vegetative growth and for several hours after starvation. A more detailed analysis of this mutant indicated that the elevated cAMP pools in these cells are a consequence of the premature appearance and partial activation of an adenylate cyclase. The mutation(s) altering adenylate cyclase regulation in this strain appears to map in linkage group IV. Complementation tests between strain HC151 and another mutant, HH201, which has recently been shown to produce an adenylate cyclase activity precociously [1], indicated that the mutations affecting adenylate cyclase activity in these strains map at different loci. Although both of these mutations behave recessively in heterozygous diploids with respect to gross development, an examination of early cAMP metabolism and terminal spore differentiation in these diploids suggest that these mutations are at least partially expressed during some stage(s) of the developmental cycle.
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  • 167
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    Developmental Genetics 2 (1981), S. 35-48 
    ISSN: 0192-253X
    Keywords: proteoglycan ; micromelia ; avian embryo ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Embryos homozygous for the recessive lethal gene, micromelia-Abbott, develop a severe form of micromelia, a parrot-like beak, and hemorrhagic skin. Feather development is also retarded. The reduction in length of the long bones of the leg can be traced to their cartilaginous stage Quantitative analysis of sulfated proteoglycan (PGS), a major macromolecular component of cartilage matrix, reveals that mutant tibiae, femora, and sterna contain significantly less uronic acid per μg of DNA than normal rudiments, indicating reduced accumulation of PGS in the mutant. Incorporation of radioactive precursors into cartilage PGS is severely reduced in relatively early developmental stages of a particular organ, but this reduction becomes less severe in cartilage taken from the same rudiment at a later developmental stage. Analysis of the sedimentation rate of PGS in sucrose gradients reveals no difference between normal and mutant in all cartilaginous types at all ages. These results suggest that the quantity and not the quality of PGS is affected in this mutant. The observation that the addition of para-nitrophenyl-β-D-xyloside to the culture medium can stimulate glycosaminoglycan synthesis to normal levels is interpreted to mean that the reduced levels of PGS may be the result of a reduced availability of the xylosylated protein backbone for PGS.
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  • 168
    ISSN: 0192-253X
    Keywords: alcohol dehydrogenase C2 ; isozymes ; temporal locus ; genetics ; chromosome 3 ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The tissue specificity of a proposed cis-acting temporal locus (Adh-3t), which regulates alcohol dehydrogenase C2 (ADH-C2) activity in mouse reproductive tissue extracts, has been examined in C5 7BL/6J, SM/J, F1 (SM/J × C5 7BL/6J) mice as well as in progeny of an (F1 [SM/J × C5 7BL/6J] × C5 7BL/6J) back-cross. Electrophoretic variants for ADH-C2, previously used to localize the gene (Adh-3) encoding this enzyme on chromosome 3, enabled the relative parental contributions to ADH-C2 phenotype in F1 and backcross mouse tissues to be determined. These analyses demonstrated that (1) stomach, kidney, lung, adrenals, seminal vesicles, epididymis, uterus, and ovary ADH-C2 is encoded by a single locus (Adh-3); Adh-3t is differentially active in various tissues, eg, lung exhibits no apparent activity whereas the temporal locus is fully active in seminal vesicles; (3) Adh-3t is probably differentically active in different cells of some tissues, eg, adrenals. Specific activity profiles of stomach and epididymal ADH-C2 during the neonatal development of C5 7BL/6J, SM/J, and F1 (SM/J × C5 7BL/6J) male mice supported the proposal for a cis-acting temporal locus for this enzyme. Genetic analyses examining segregation of Adh-3 and Adh-3t among backcross progeny suggested that these are distinct but closely linked loci, since one recombinant among 256 progeny was observed. Linkage data of Adh-3 with Va (varitint-waddler) and de (droopy ear) was also obtained, which suggested that Adh-3 is localized on chromosome 3 between Va and de.
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    Developmental Genetics 2 (1981), S. 357-367 
    ISSN: 0192-253X
    Keywords: amphibian hybrids ; exogastrulation ; hybrid lethality ; nucleocytoplasmic interactions ; triploidy ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Hybrids between species frequently arrest early in development. In the frog hybrid Rana catesbeiana female × Rana clamitans male, the embryo shows a characteristic development to an exogastrula which dies. This hybrid can be rescued by pressure suppression of the second polar body, which results in the addition of another haploid set of R catesbeiana chromosomes to the embryo. The triploid hybrid expresses genes from both species and can develop normally through metamorphosis. The results show that an R catesbeiana egg containing a full haploid set of R clamitans chromosomes is capable of development and that the usual developmental arrest caused by the R clamitans genome responds to chromosomal dosage.
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  • 170
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    Developmental Genetics 2 (1981), S. 203-218 
    ISSN: 0192-253X
    Keywords: cytokinesis ; fusome ; intercellular bridges ; oogenesis ; sterility ; tumor mutations ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The ethyl methane sulfonate-induced mutation, fs(1)1621, resides at 11.7 on the genetic map and within segment 4F1-5A1 of the cytological map of the X chromosome. When homozygous, fs(1)1621 renders females semisterile but has no effect on their viability; nor does it affect the viability or fertility of hemizygous males. Heterozygous females are fertile and have cytologically normal ovaries. The ovaries of homozygous females first produce normal oocytes, which, if fertilized, can develop into adult males or females. After this period, ovarian chambers containing only pseudonurse cells are formed, and finally mutant germaria produce only tumors. These contain hundreds to thousands of cells that appear to be derived from germarial cystocytes, because they occasionally form clones of interconnected cells and also can differentiate into endopolyploid pseudonurse cells. Raising the temperature speeds the rate at which tumors form; lowering it increases the probability of pseudonurse cell differentiation. Df(1)C159 includes fs(1)1621. The pattern of ovarian chamber production is more temperature sensitive in hemizygous females than in homozygous ones. The morphology of hemizygous tumors and the number of dividing cells within them also differ from homozygotes. These observations support the hypothesis that fs(1)1621 is producing a product, that less is produced by one gene than by two, and that the product plays a role in the mitosis and cytokinesis of ovarian cystocytes.
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  • 171
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    Developmental Genetics 2 (1981), S. 237-252 
    ISSN: 0192-253X
    Keywords: Drosophila ; embryonic cultures ; protein synthesis ; stage specific ; differentiation ; two-dimensional gel electrophoresis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The patterns of proteins synthesized during embryonic development in Drosophila melanogaster have been examined by two-dimensional gel electrophoresis. Primary cell cultures prepared from donor embryos synchronized to ± 1 hr were labeled with [35S]methionine at 5, 11.5, 14.5, and 26 hr after oviposition. Of approximately 400 to 500 proteins detected, the synthesis of about 50 is developmentally modulated. The greatest number of changes in the synthesis of stage-specific proteins occurs at 11.5 and 14.5 hr after oviposition, periods just prior to and during the times of the greatest overt morphological and biochemical changes. At 11.5 hr, 35 stage-specific proteins are synthesized, including 19 that are not present at the previous stage examined. At 14.5 hr, 34 stage-specific proteins can be detected, including 11 newly synthesized proteins. However, 12 proteins from the previous stage are no longer synthesized. At the completion of embryonic differentiation, at 26 hr, no new proteins are synthesized and the synthesis of many present in earlier stages has decreased or stopped. Comparison of patterns of embryonic proteins to those synthesized by two Drosophila continuous cell lines reveals that the majority of proteins are common to all. However, only about 40% of the embryonic stage-specific proteins are present in either cell line. In addition, there are several proteins unique to each cell line that are not observed in any of the embryonic stages.
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  • 172
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    Developmental Genetics 2 (1981), S. 1-12 
    ISSN: 0192-253X
    Keywords: β-galactosidase ; preimplantation ; mouse ; Bgl ; paternal effect ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Lysosomal acid hydrolase expression during preimplantation mouse embryogenesis has proved useful in estimating when mRNA transcription commences during this period. Previous work from this laboratory has shown that α-galactosidase and β-glucuronidase undergo 50- to 100-fold increases in activity between the two-cell stage and the blastocyst stage [1, 2]. Here we show that β-galactosidase activity levels undergo a similar change. We also demonstrate that mouse strains with the Bgl-sh allele produce cleavage stage embryos with 2-4-fold higher activity levels than strains with the Bgl-sd allele. Bgl has been shown to control β-galactosidase levels in adult mouse tissues [3]. Unfertilized egg β-galactosidase levels are also regulated by Bgl, but loci distant from Bgl modify egg expression. The distant sites are not observed to act during cleavage. Hybrid embryos (Bgl-sd/h) show intermediate activity levels to the parental types. The timing of the deviation of hybrid embryo β-galactosidase activity levels from maternal-type activity levels is used to estimate when transcription of genes governing β-galactosidase expression occurs.
    Additional Material: 5 Ill.
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  • 173
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    Developmental Genetics 2 (1981), S. 23-34 
    ISSN: 0192-253X
    Keywords: Paramecium tetraurelia ; trichocysts, nuclear differentiation ; cytoplasmic inheritance ; determination ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The mtFE mutation isolated in Paramecium tetraurelia affects mating type differentiation, trichocyst excretion, and viability. Its effect on mating type has already been shown to correspond to a restriction to the E mating type interpreted by an inefficiency of nuclear O-determining factors. In this paper we study the other two phenotypic characteristics whose hereditary transmission displays two unusual features. (1) In crosses between a wild-type strain and the mutant strain, the mutant characteristics do not reappear in F2 in the wild-type cytoplasmic lineage but only in F3 after the homozygous clones have undergone an additional nuclear reorganization. (2) Some F2 wild-type clones, in the mutant cytoplasmic lineage, retain some of the phenotypic characteristics of the mutant. We propose that the mtF gene product plays a role in the control of several macronuclearly differentiated functions.
    Additional Material: 3 Ill.
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  • 174
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    Developmental Genetics 2 (1981), S. 75-87 
    ISSN: 0192-253X
    Keywords: Drosophila melanogaster ; homoeosis ; imaginal discs ; aldehyde oxidase ; pattern formation ; compartments ; selector genes ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In a study of the regulation of enzyme patterns in imaginal discs the aldehyde oxidase pattern was determined for some homoeotic mutations of D. melanogaster. Earlier indications that suggested that this pattern follows the determinitive state of compartments within imaginal discs were confirmed by the aldehyde oxidase (AO) pattern of both the wing and haltere discs from en1; bx3, en1; pbx, and en1; bx3 pbx larvae and the antennal discs from Antp73b and ssa larvae.We additionally analyzed whether AO activity depended on the determinative state of an entire compartment or was expressed autonomously in clones. Homozygous engrailed clones were induced by mitotic recombination. From the AO clones found in normally negative areas of the posterior compartment it was concluded that enzyme activity depended upon the determinative state of the cells and was not a function of the compartment as a whole.The results are described with reference to a scheme in which compartmental and subcompartmental selector genes are thought to determine a binary code on which AO patterns depend.
    Additional Material: 6 Ill.
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  • 175
    ISSN: 0192-253X
    Keywords: Drosophila ; dopa decarboxylase ; female fertility ; temperature sensitive lethal mutant ; differential tissue-specific responses to temperature ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The mutation Ddcts1 effects female sterility when homozygous, hemizygous, or heterozygous over a series of Ddc null alleles (Ddcx) indicating that some aspect of Ddc gene function is necessary for female fertility. Ovary transplant experiments demonstrate that the female sterility phenotype is ovary autonomous. Two to 3% of the total DDC activity measurable in newly hatched females is localized in their previtellogenic ovaries. The degree to which females heterozygous for Ddcts1 over different Ddc null alleles are fertile at 22°C reflects a continuous spectrum of allelic complementation similar to that observed for the effects of these genotypes on viability at 30°C. Fertility of all the Ddcts1/Ddcx females tested is significantly depressed at 30 vis-a-vis 22°C providing evidence that it is the DDC enzyme activity itself which is required for female fertility. Ddcts1/Ddcts1 homozygous and Ddcts1/Df hemizygous females are nonconditionally, completely sterile at 18, 20, 22, 25, and 30°C. Although all homo- and hemizygous females do lay some eggs, no evidence of embryogenesis or fertilization has ever been detected. The absolute, nonconditional sterility of Ddcts1 homo- and hemizygous females stands in stark contrast to the conventional temperature dependent effects of these same genotypes on viability and to the temperature sensitive effects of Ddcts1/Ddcx heterozygous females on both fertility and viability. Reasons for these tissue-specific and genotypic differences are discussed.
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  • 176
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    Developmental Genetics 2 (1981), S. 253-267 
    ISSN: 0192-253X
    Keywords: myxomycete ; Physarum polycephalum ; mating type ; sexual compatibility ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The conversion of the uninucleate amoebal form of Physarum polycephalum to the multi-nucleate plasmodial form is under the control of a genetic region which contains matA (or mt), a determinant of mating specificity. The region is the site of most gad mutations, which give amoebae the ability to produce plasmodia in clones without mating (ie, to self). In the present study, nonselfing revertants were isolated from two matA2-derived gad mutants and two matA3-derived gad mutants. Some revertants were found to have regained exactly, or nearly, the same phenotype as the original matA2 or matA3 strain. Others expressed new mating types, having gained the ability to mate with strains of the parental matA type. The results are compatible with a model in which new mating types arise from forward mutations (gad) and back mutations (npf or no plasmodium formation) occurring successively in a single gene, matA.
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  • 177
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    Developmental Genetics 2 (1981), S. 269-277 
    ISSN: 0192-253X
    Keywords: β-glucuronidase ; androgen ; receptor ; development ; mouse ; kidney ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: During postnatal development of mouse kidney the androgen responsiveness of epithelial cells for β-glucuronidase induction, cellular hypertrophy, and other enzyme inductions appears coincidentally with a rise in androgen receptor protein. Initially, a low level of receptor is present but no response is seen. Beginning at about 12 days of age responsiveness begins to increase, reaches a half-maximal level at 18-20 days, and full responsiveness by 28-30 days. The limiting factor appears to be levels of androgen receptor protein.Our experiments shed no light on the question of why each androgen responsive cell type in the organism differentiates the capacity to induce a different array of proteins. However, they do suggest that responsiveness of the β-glucuronidase gene does not appear until a minimum threshold level of receptor is exceeded, and that the response of the gene may not be saturated even at the highest levels of receptor reached.
    Additional Material: 5 Ill.
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  • 178
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    Developmental Genetics 2 (1981), S. 305-315 
    ISSN: 0192-253X
    Keywords: X-chromosome inactivation ; mouse ; PGK-1 ; embryonic cell lineage ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have used a sensitive electrophoretic technique for estimating the activity, or ratio, of two allozymes of the X-chromosome-linked enzyme phosphoglycerate kinase (PGK-1), in order to investigate the randomness of X-chromosome expression in the derivatives of the three primary cell lineages of the early mouse conceptus. The maternally derived Pgk-1 allele is preferentially expressed in the derivatives of the primitive endoderm and trophectoderm lineages at 6 1/2 days post coitum in Pgk-1a/Pgk-1b heterozygous conceptuses, and in the one informative 5 1/2-day heterozygous conceptus analysed. This evidence for preferential expression of the maternally derived X chromosome (Xm), so soon after the time of X-chromosome inactivation, favors the possibility that the preferential expression of Xm is a consequence of primary non-random X-chromosome inactivation, rather than a secondary selection phenomenon. The majority of embryos analysed at 4 1/2 and 5 1/2 days pc produced only a single PGK-1 band, corresponding to the allozyme produced by the Pgk-1 allele on Xm, although 50% of these embryos should have been heterozygous females. Possible explanations are discussed.
    Additional Material: 2 Ill.
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  • 179
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    Developmental Genetics 2 (1981), S. 319-336 
    ISSN: 0192-253X
    Keywords: maize ; mitochondrial DNA ; recombinant DNA ; cms-T ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Recombinant DNA and hybridization techniques have been used to compare the organization of mitochondrial DNA (mtDNA) from normal (N) and Texas male sterile (T) cytoplasms of maize. Bam H1 restriction fragments of normal mtDNA were cloned and used in molecular hybridizations against Southern blots of Bam H1 digested N and T mtDNA. Fifteen of the 35 fragments were conserved in both N and T as indicated by hybridization to comigrating bands in their restriction patterns. Only three fragments produced autoradiographs whose differences could reasonably be attributed to single changes in the cleavage site of the enzyme while approximately half (17/35) of the clones resulted in more complicated differences between N and T. The autoradiographs produced by these 17 clones indicated multiple cleavage site changes and/or sequence rearrangements of the mtDNA. Patterns of six of these 17 clones indicated partial duplication of the sequence and two showed variation in the intensity of hybridization between N and T, which may be related to the molecular heterogeneity phenomenon found in maize mitochondrial genomes. The large proportion of changes observed between N and T mtDNA indicates that rearrangements may have played an important role in the evolution of the maize mitochondrial genome.
    Additional Material: 7 Ill.
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  • 180
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    Developmental Genetics 2 (1981), S. 185-202 
    ISSN: 0192-253X
    Keywords: somatic DNA alteration ; nuclear differentiation ; mating types ; ciliate genetics ; immunoglobulin genes ; Tetrahymena thermophila ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Experimental data on mating type determination in T. thermophila, collected by Nanney, Allen, and their collaborators over a period of 25 years, are reinterpreted in the light of our current understanding of macronuclear genetics. A strong case is developed supporting the idea that mating type determination involves the developmental alteration of somatic DNA that occurs regularly in developing macronuclei in conjugating pairs. A. testable DNA deletion/splicing model is developed that although based on a few simple, plausible assumptions, explains the observations remarkably well. The model is in (at least) superficial analogy to the mechanism that must be involved to explain the somatic differentiation and alteration of DNA sequences that ultimately constitute an expressed vertebrate immunoglobulin gene. Because of the genetic, biochemical, and micromanipulative versatility of Tetrahymena, it may well turn out to be a uniquely suitable microbial eukaryotic experimental system for the study of developmental alterations of somatic DNA.
    Additional Material: 7 Ill.
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  • 181
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    Developmental Genetics 2 (1981), S. 317-317 
    ISSN: 0192-253X
    Keywords: Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
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  • 182
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    Developmental Genetics 2 (1981), S. 291-303 
    ISSN: 0192-253X
    Keywords: mouse ; trisomy ; gene dosage ; enzyme activity pattern ; phosphoglycerate mutase (PGAM) ; glutamate oxaloacetate transaminase (GOT) ; isozyme ; developmental pattern ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Activity patterns of cytosolic and mitochondrial enzymes of carbohydrate and amino acid metabolism have been measured in murine trisomy 19. In spite of marked hypoplasia, no significant alterations of the patterns (per gram of organ weight) were observed, with the exception of glutamate oxaloacetate transaminase (GOT-1), and phosphoglycerate mutase (PGAM). Clear-cut gene dosage effects in liver, brain, heart, skeletal muscle, and erythrocytes of fetal and newborn mice, confirm the assignment of GOT-1 to chromosome 19. Data obtained for PGAM demonstrate that one of the two different subunits leading to organ-specific isozyme patterns of the dimer enzyme protein is coded on chromosome 19 (gene Pgam-1). Dosage effects are fully expressed in liver, brain, and erythrocytes (AA-type isozyme), but not in skeletal muscle (BB-type isozyme). Dosage effects on the hybrid AA-AB-BB-isozyme pattern in the course of development of the heart muscle, were demonstrated by means of quantitative activity measurement after electrophoretic separation. The comparison of enzyme patterns of eusomic and trisomic erythrocytes, produced after injection of fetal stem cells into irradiated adult carriers (transplantation chimaeras), revealed enzyme activity ratios that were similar to those produced by erythrocytes of adult euploid and trisomic mice. This is in agreement with the chromosome assignments and dosage effects mentioned above.
    Additional Material: 5 Ill.
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  • 183
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    Developmental Genetics 2 (1981), S. 349-356 
    ISSN: 0192-253X
    Keywords: agouti locus ; lethal yellow gene ; MSH ; dibutyryl cyclic AMP ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Alleles at the agouti locus determines the type of pigment produced in hair-bulb melanocytes. In order to analyze the mechanism of agouti gene function, an attempt was made to induce the shift in melanin synthesis in vitro. Skin explants from newborn yellow mice with genotype Ay/a were cultured with the method using membrane-filter and roller tube. Production of black pigment in the hair bulbs was observed when the explants were cultured in the presence of α-melanocyte stimulating hormone (α-MSH). Electron-microscopic observation indicates that the induced black pigments are eumelanin that is normally found in hair-bulb melanocytes of genotypically black mice. The eumelanin synthesis was also induced by cAMP, DbcAMP, or theophylline. This α-MSH-induced eumelanin synthesis was suppressed by actinomycin D or cycloheximide, suggesting that the α-MSH-induced eumelanogenesis requires de novo transcription and/or translation.
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  • 184
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    Developmental Genetics 2 (1981), S. 385-405 
    ISSN: 0192-253X
    Keywords: allelic isozyme variants ; mammalian development ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The past decade has seen an explosion of interest in mammalian embryos. Techniques of molecular and genetic analysis coupled with advances in in vitro culture and experimental manipulation of mammalian embryos have provided important insights into mechanisms of embryogenesis. Many of these recent advances have been facilitated by the use of allelic isozyme variants as autonomous cell markers or representative gene products. Investigations aimed at exploring cell lineages and cell commitment, the timing and regulation of gene expression, X chromosome inactivation, and cell interactions have depended on the availability of appropriate isozyme variants. Results from such experiments are summarized here in order to demonstrate the usefulness of this approach and to stimulate its wider application in developmental biology.
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