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  • 1
    Electronic Resource
    Electronic Resource
    Dynamic changes in autosomal spindle fibers during prometaphase in crane fly spermatocytes (1982)
    Springer
    Chromosoma 87 (1982), S. 363-371 
    Details
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Different prometaphase stages of Pales ferruginea spermatocytes were serially sectioned and the regions between kinetochores and poles analysed by counting and measuring spindle microtubules. These regions are characterized by an intermingling of kinetochoric (kMTs) and non-kinetochoric microtubules (nkMTs). A considerable proportion of nkMTs is skewed with respect to kMTs, thus being responsible for microtubule disorder in these spindle areas. The degree of disorder expressed by the percentage of skew microtubules was found to decrease from early prometaphase to metaphase, parallel with an increase in kMT number. A possible causal relation between pulling forces and morphological changes in the spindle is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00327637
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Three-dimensional architecture of chromosome fibres in the crane fly: co-oriented autosomal bivalents and amphitelic sex univalents during prometaphase (1986)
    Springer
    Chromosoma 94 (1986), S. 107-114 
    Details
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A technique for the fixation of cells during live observation (Nicklas et al. 1979) was used to investigate chromosomes which were moving at the time of fixation. Chromosome fibres were reconstructed by tracking their microtubules in longitudinal serial sections. A considerable proportion of non-kinetochoric microtubules (free microtubules, fMTs) is skewed with respect to the fibre axis. These skew fMTs contribute to the degree of disorder. It was found that the difference in the relative proportion of skew fMTs between “active” fibres (oriented in the direction of movement) and “passive” fibres (oriented backwards) is significantly correlated with the chromosome velocity (correlation coefficient r=0.796, P=0.01). It can be concluded that the pulling force generated in the chromosome fibre is a function of skew fMTs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00286988
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    Relationship between the arrangement of microtubules and chromosome behaviour of syntelic autosomal univalents during prometaphase in crane fly spermatocytes (1986)
    Springer
    Chromosoma 94 (1986), S. 412-418 
    Details
    ISSN: 1432-0886
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract During meiotic prometaphase of crane fly spermatocytes, syntelic autosomal univalents are able to move between the spindle poles several times. The relationship between the behaviour of chromosomes and the arrangement of microtubules during this stage was studied using a fixation technique (Nicklas et al. 1979) which makes it possible to examine a certain cell in an electron microscope after live observation. After reorientation, when a syntelic univalent had started moving towards the opposite spindle pole, a short chromosome fibre was found. When a univalent had reached the equator, a chromosome fibre could be traced up to the spindle apex. During the movement towards the opposite spindle pole the degree of disorder in the chromosome fibre was high, whereas it was low in the fibre of a motionless univalent. The degree of disorder was determined by the relative proportion of skew fibre microtubules. At the beginning of reorientation a chromosome fibre was still present, but later, it was no longer possible to recognize such a fibre. Instead of a chromosome fibre, a bundle of microtubules laterally associated with the kinetochore was observed. Some microtubules of this bundle had a direct contact with the kinetochore. These observations strongly hint that the laterally associated microtubules have an important function in the reorientation of syntelic autosomal univalents.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00328642
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  • 4
    Electronic Resource
    Electronic Resource
    Relationship between tektins and intermediate filament proteins: An immunological study (1989)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 14 (1989), S. 359-371 
    Details
    ISSN: 0886-1544
    Keywords: chymotrypsin digest ; multiple immunoblot ; keratin ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Affinity-purified antibodies raised against three flagellar tektins (tektin A, B, and C) from each of two sea urchin species (Lytechinus pictus and Strongylocentrotus purpuratus) were used to study the immunological relationship between tektins and intermediate filament proteins. By immunofluorescence microscopy, several antitektins revealed a staining of intermediate filament-like arrays in three vertebrate cell lines tested. Immunoelectron microscopy substantiated the cross reaction of antitektins with intermediate filaments. When the cells were treated with cytochalasin B, the arrangement of the filaments recognized by anti-(Lp)-tektin B was altered; the alteration observed is typical for keratin filaments. By immunoblot, it was found that anti-(Lp)-tektin B cross reacted with two isoforms or different proteins of ∼54 kD with pIs of 6.1 and 6.2 in human carcinoma epithelia (HeLa) cells and with two isoforms or different proteins of ∼55 kD with pIs of 6.1 and 6.3 in pig kidney epithelia (LLC-PK1) cells. Furthermore, when antitektin antibodies were affinity purified with the 54 kD HeLa keratin, these keratin-specific antibodies again restained the original tektins on immunoblots. From these observations, it can be concluded that tektins and keratins are to a certain extent immunologically related. To determine the degree of the immunological relationship, tektin filaments and purified intermediate filaments from HeLa cells were cleaved with α-chymotrypsin and examined by quantitative immunoblot analysis. On immunoblots of digested tektins from L. pictus, anti-(Lp)-tektin B recognized several cleavage products in the range of 20 kD to 46 kD. However, when immunoblots of digested intermediate filaments from HeLa cells were probed, the cross reaction of anti-(Lp)-tektin B with HeLa keratins was eliminated by more than 98% within 2 min, suggesting that tektins have epitopes in common with the end domains of certain keratins.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970140306
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    Paper (German National Licenses)
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  • 5
    Electronic Resource
    Electronic Resource
    Multiple immunoblot: A sensitive technique to stain proteins and detect multiple antigens on a single two-dimensional replica (1989)
    Weinheim : Wiley-Blackwell
    Electrophoresis 10 (1989), S. 714-718 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A multiple immunoblotting technique was developed to positively identify up to three different antigens on a single nitrocellulose replica of a two-dimensional isoelectric focusing/sodium dodecyl sulfate-polyacrylamide gel. Three highly sensitive immunoblot assays were selected, including: horseradish peroxidase/luminescence, alkaline phosphatase, and silver-enhanced immunogold. As a major advantage, the method permits a simultaneous detection of up to three different antigens without eluting the antibody-dye complex between staining of single polypeptides, thus providing a highly accurate identification of closely migrating components. The staining procedure is summarized in a flow chart. In addition to the multiple immunoblot staining, some suggestions are provided for a sensitive protein staining.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150101010
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  • 6
    Electronic Resource
    Electronic Resource
    The effect of sodium tetradecyl sulfate on mobility and antigen detectability of microtubule proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (1997)
    Weinheim : Wiley-Blackwell
    Electrophoresis 18 (1997), S. 1955-1959 
    Details
    ISSN: 0173-0835
    Keywords: Immunoblot ; Microtubules ; Sodium tetradecyl sulfate ; Sodium dodecyl sulfate-polyacrylamide gel electrophoresis ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Several factors been reported to influence the mobility of polypeptide in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) including the brand of SDS. Using microtubule proteins from axonemes of Lytechinus pictus and Spisula solidissima sperm and meiotic spindles of Spisula solidissima we demonstrate that the change in mobility was caused by sodium tetradecyl sulfate (STS), a major contaminant of many commercial SDS brands. We also examined the use of sodium tetradecyl sulfate and different SDS brands as a tool in extracting more information from immunoblot studies. Commercial SDS containing contaminants other than sodium tetradecyl sulfate reduced or eliminated the immunosignal from certain polypeptides and the loss of antigenicity could not even be recovered by immunoblot under “renaturing” conditions. It can thus be concluded that STS can be useful in separating and identifying comigrating polypeptides and in detecting additional immunobands in immunoblots.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150181113
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