Please use this identifier to cite or link to this item: https://hdl.handle.net/2440/5387
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Type: Journal article
Title: Isolation of histones and related chromatin structures from spermatozoa nuclei of a dasyurid marsupial, Sminthopsis crassicaudata
Author: Soon, L.
Ausio, J.
Breed, W.
Power, J.
Muller, S.
Citation: Journal of Experimental Zoology Part A: Comparative Experimental Biology, 1997; 278(5):322-332
Publisher: WILEY-LISS
Issue Date: 1997
ISSN: 1552-499X
1097-010X
Statement of
Responsibility: 
L. L. L. Soon, J. Ausio, W. G. Breed, J. H. T. Power and S. Muller
Abstract: The spermatozoa of a dasyurid marsupial, Sminthopsis crassicaudata, have two distinct nuclear regions: uniformly electron-dense chromatin (C1) in the interior and fissured chromatin (C2) at the periphery. To investigate whether the differences in morphology are due to incorporation of different packaging proteins, spermatozoa nuclear proteins were characterised by acetic acid-urea polyacrylamide gel electrophoresis (PAGE) and fractionated by reverse-phase high-pressure liquid chromatography (HPLC). The main protein component was protamine I, but a complete histone complement (H1, H2A, H2B, H3, and H4) was also detected. Immunocytochemistry showed localisation of H4, H2B, and H2A histones to the periphery of the nuclei, a region that corresponded to the C2 chromatin. The fissures in the chromatin of this region disappeared following incubation with fish protamines, indicating that the nucleohistone C2 region may be incompletely condensed relative to nucleoprotamines. This observation is consistent with the view that 60% of phosphodiester charges remain negative in nucleohistone DNA, whereas all DNA charges are neutralised in highly compact nucleoprotamines. Treatment of spermatozoa with micrococcal nuclease showed that the C1 chromatin was resistant to digestion, whereas the C2 region was cleaved into 30- to 38-nm agglomerates and 11-nm nucleosomal-size structures. Thus, this study demonstrates that spermatozoa nuclei of this marsupial species contain peripherally localised histones, and the nucleohistone chromatin accounts for the different morphology of the C2 region compared with the rest of the nucleus.
Keywords: Spermatozoa
Cell Nucleus
Chromatin
Animals
Marsupialia
Histones
Protamines
Electrophoresis, Polyacrylamide Gel
Amino Acid Sequence
Molecular Sequence Data
Male
Rights: © 1997 Wiley-Liss, Inc.
DOI: 10.1002/(SICI)1097-010X(19970801)278:5<322::AID-JEZ6>3.0.CO;2-R
Published version: http://dx.doi.org/10.1002/(sici)1097-010x(19970801)278:5%3C322::aid-jez6%3E3.0.co;2-r
Appears in Collections:Anatomical Sciences publications
Aurora harvest
Environment Institute publications

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