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  • 21
    ISSN: 1573-4927
    Keywords: carbonic anhydrase ; isozymes ; immunological cross-reactions ; monoclonal antibodies ; protein evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Cross-reactions among carbonic anhydrases (CAs) I, II, and III were studied using a variety of antisera: (1) a rabbit antiserum to bovine CA III, (2) mouse antisera to human CA I, CA II, and CA III; and (3) five monoclonal antibodies prepared by the hybridoma technique using splenocytes from a mouse immunized with human CAs I and II and bovine CA III. Cross-reactions between CAs were readily found by binding assays using these antisera. Human CA I, but not human CA II, inhibited the reaction of the rabbit anti-CA III with its homologous antigen. Mouse antisera to CA I or CA II bound the homologous I or II with nearly as great efficiency as the autologous isozyme and sometimes weakly bound CA III. Mouse antisera to CA III frequently bound CA I or II. These cross-reactions were confirmed by the first use of hybridoma-prepared, monoclonal antibodies to CAs. The mouse monoclonal antibodies to CA isozymes varied in the amount of cross-reactivity among I, II, and III: at one extreme, one monoclonal was highly specific for the autologous CA III; at the other extreme, one monoclonal weakly reacted with some examples of CAs I, II, and III.
    Type of Medium: Electronic Resource
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  • 22
    ISSN: 1573-4935
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract cDNA clones to poly(A)+ mRNA from spermatids have been obtained to study gene transcription in post-meiotic germ cells. Four cDNA clones detect mRNAs that increase in abundance in post-meiotic germ cells. One clone, pPM459, was shown to correspond to an mRNA that is transcribed after meiosis. Pulse-labelling experiments demonstrate transcription o5 the message in spermatids. These data constitute further evidence for post-meiotic gene transcription in spermatids.
    Type of Medium: Electronic Resource
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  • 23
    ISSN: 1573-4927
    Keywords: mouse mutant microsomes ; biochemical analysis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Investigations of the lethal effects of a series of radiation-induced deletion alleles in the mouse have identified severe ultrastructural abnormalities of endoplasmic reticulum liver membranes in lethal albino homozygous newborns. The ultra-structural defects were accompanied by deficiencies of several enzymes, some of them microsomal membrane bound. Chemical and immunological studies were therefore undertaken in order to identify a possible biochemical alteration of mutant endoplasmic reticulum membranes. Patterns of gel electrophoresis produced by several different methods showed no differences between the microsomal proteins of mutant and normal mice. Immunological methods also failed to detect any changes in the major proteins. Thus in spite of the severe ultrastructural defects no major differences between microsomal proteins of mutant and normal membranes could be identified. Since several of the enzymes affected are inducible by cAMP in newborn mice, microsomal and supernatant cAMP-binding activities were also measured in mutants and normals but showed no differences. As yet, the primary cause of the severe effects of the X-ray-induced deletions on membrane structure and enzyme activities remains unknown.
    Type of Medium: Electronic Resource
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  • 24
    ISSN: 1573-4927
    Keywords: β-glucuronidase ; lysosomal enzymes ; mucopolysaccharidosis ; glycosaminoglycans ; mice
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Young (60–80 days) mice of the low β-glucuronidase strain, C3H/HeJ, showed no differences in hepatic levels of glycosaminoglycans (GAGs) when compared to the randombred, “normal” Swiss-Webster mice of the same age. However, by 12 months of age hepatic GAG is nearly twice as high in C3H/HeJ mice as in Swiss-Webster mice. Studies of β-glucuronidase, β-galactosidase, and N-acetyl-β-glucosaminidase in four tissues of the two types of mice at the two ages revealed that glucuronidase was the only enzyme with lower activity in the C3H/HeJ strain.
    Type of Medium: Electronic Resource
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  • 25
    ISSN: 1573-4927
    Keywords: transgenic mice ; carbonic anhydrase ; promoter analysis ; transcription ; DNA control regions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract Although the proximal, 5′ 115 bp of the human carbonic anhydrase II (CA II) gene was sufficient for expression of a reporter gene in some transfected cell lines, we found previously that 1100 bp of this promoter (or 500 bp of the mouse CA II promoter) was not sufficient for expression in transgenic mice. We have now studied the expression of linked reporter genes in mice transgenic for either (1) 11 kb of the human 5′ promoter or (2) 8 kb of the human 5′ promoter with mouse sequences from the first exon, part of the first intron (since a CpG island spans this region), and the 3′ sequences of the gene. Expression was found in both cases, but the tissue specificity was not appropriate for CA II. Although there was a difference in the sensitivity of the assays used, the first construct led to expression in many tissues, while the second construct was expressed only in spleen. These findings indicate considerable complexity of DNA control regions for in vivo CA II expression.
    Type of Medium: Electronic Resource
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  • 26
    ISSN: 1573-4935
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract We have used a human phosphoglycerate kinase-1 (PGK-1) cDNA clone to study expression of PGK-2 during mouse spermatogenesis. Hybrid selection, in vitro translation with product identification by 2-D gel electrophoresis demon-strated that the PGK-1 cDNA clone hybridized to PGK-2 mRNA in mouse testes. Northern analyses of RNA purified from separated spermatogenic cells demonstrated a large increase in abundance of PGK-2 mRNA in post-meiotic cells. Thus, post-meiotic transcription of PGK-2 mRNA is demonstrable with cloned DNA probes.
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  • 27
    Electronic Resource
    Electronic Resource
    Springer
    Mammalian genome 11 (2000), S. 69-71 
    ISSN: 1432-1777
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
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  • 28
    ISSN: 1432-1777
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Inter-α-inhibitor (IαI) and related molecules in human are comprised of three evolutionarily related, heavy (H) chains and one light (L) chain, also termed bikunin. The latter originates from a precursor molecule that is cleaved to yield the bikunin and another protein designated α-1-microglobulin (A1m). The four H and L chains are encoded by four distinct genes designated H1, H2, H3, and L. The L and H2 genes are localized onto human chromosomes (chr) 9 and 10, respectively, whereas the H1 and H3 genes are tandemly arranged on chr 3. Mouse poly(A)+ RNAs or endonuclease-restricted mouse DNA were analyzed by standard and pulsedfield gel electrophoresis (PFGE) techniques in agarose gels and blot-hybridized with human H1, H2, H3 or L cDNA probes. The variable sized transcripts and unique restriction fragment patterns detected with each probe indicate that four genes, including one common L gene for Alm and bikunin also exist in mouse. The co-migration of H1- and H3-hybridizing fragments on PFGE suggest that the mouse H1 and H3 genes are also tandemly arranged. An Msp I restriction fragment length polymorphism (RFLP) in the mouse L gene (proposed symbol, Intin-4) links this gene to other genes already mapped at mouse Chr 4 near the brown (b) locus, a homologous region to the human chr 9q32-34 band where the human IαI L gene is located. Therefore, a similar number and arrangement of IαI genes is found in mouse and human, including the triplication of an H gene ancestor. These results point to an ancient origin of this complex set of genes.
    Type of Medium: Electronic Resource
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  • 29
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 37 (1994), S. 370-381 
    ISSN: 1040-452X
    Keywords: Sex determination ; Sex determining region Y ; Postmeiotic expression ; HMG box containing proteins ; Interstitial cells ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Although its expression in adult testis was immediately apparent, the role for Sry (sex determining region, Y) in testicular function remains elusive. We have performed transcriptional studies in an effort to elucidate potential roles of Sry by studying the time and location of its transcription in mouse testes. Northern analyses and more sensitive nuclease protection assays detected transcripts in 28-day-old testes and beyond. The highly sensitive technique of reverse transcription polymerase chain reaction (RTPCR) could not detect Sry expression in 14-day testes when primers for the most conserved portion of the gene, the high mobility group (HMG) box, were used, but primers for the circular form detected Sry transcription at all postnatal stages studied. The same HMG box primers were able to detect expression of Sry in XX, Sxra or Sxrb testes. This suggested that Sry is expressed in cells other than germ cells, which was confirmed with studies on fractionated cells - RTPCR detected transcription of Sry in the highly pure interstitial cell fraction. However, Leydig cells and a Leydig cell tumor were negative for Sry expression. We performed in situ studies in an attempt to localize the expression of Sry in the testes. Abundant expression of an Sry cross-hybridizing transcript was found in spermatogonia, in early spermatocytes, and in some interstitial cells with antisense probes to the HMG box or a more specific, 3′ region, whereas the sense probe gave little or no hybridization. It is probable that the circular transcripts, which are seen in reverse transcriptase positive (RT+) and RT- reactions by PCR because of the RT activity of Taq polymerase, are responsible for the hybridization seen in spermatogonia and spermatocytes, whereas linear and circular forms are detected later. Thus Sry is expressed in pre- and postmeiotic germ cells and in somatic cells of the testes. © 1994 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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  • 30
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Molecular Reproduction and Development 1 (1988), S. 27-34 
    ISSN: 1040-452X
    Keywords: LDH-X ; Poly(A) ; Translational regulation ; cDNA probe ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The expression of the mRNA for mouse testicular lactate dehydrogenase (LDH-X) was examined by Northern analyses of meiotic and postmeiotic spermatogenic cell populations. Silver grains accumulated in cells inside the second layer from the periphery of the seminiferous tubule, confirming previous findings that LDH-X mRNA first appears in the spermatocyte and continues to accumulate until the late spermatid stage. Northern analyses showed that meiotic and postmeiotic cells contained 1.2 and 1.3 kb classes of hybridizing mRNA, respectively. RNase H digestion of oligo(dT)-hybridized RNA and poly(U)-Sepharose column chromatography with differential elution by formamide revealed that the difference in size of the two classes of mRNAs was due to the poly(A) tail length of the LDH-X mRNA. When the distribution of the LDH-X mRNA was examined across polysome gradients, both mRNAs were partially associated with polysomes. These results suggest that the changes in the polyadenylation of LDH-X mRNA were associated with the meiotic division during spermatogenesis in the mouse. They raise the possibility that the stable accumulation of the LDH-X mRNAs in the postmeiotic cells is enhanced by poly(A) tails of increased length.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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