Please use this identifier to cite or link to this item: http://hdl.handle.net/2440/5430
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Type: Journal article
Title: Atomic force microscopy and cytochemistry of chromatin from marsupial spermatozoa with special reference to Sminthopsis crassicaudata
Author: Soon, L.
Bottema, C.
Breed, W.
Citation: Molecular Reproduction and Development, 1997; 48(3):367-374
Publisher: WILEY-LISS
Issue Date: 1997
ISSN: 1040-452X
1098-2795
Statement of
Responsibility: 
L.L.L. Soon, C. Bottema, W.G. Breed
Abstract: Atomic force microscopy (AFM) of the nuclear topology of spermatozoa from two marsupial species, Sminthopsis crassicaudata and Trichosurus vulpecula was investigated to determine the structural organisation of the chromatin subunits. That of the former species is of special interest as it has a peripheral nucleohistone region (C2) as well as a nuclease-resistant, nucleoprotamine core region (C1). Atomic force microscopy showed that the C2 region contained clusters of 120-160 nm nodules, whereas the C1 region exhibited smaller 50-80 nm nodules. The spermatozoa nuclei of Trichosurus, which has mainly nucleoprotamines, contained higher order chromatin structures of similar size to those from the C1 region of Sminthopsis. This study shows that nucleohistones and nucleoprotamines of marsupial sperm form distinct higher order conformations. For the second part of this work, the chromatin density and affinity for cationic stains of Sminthopsis spermatozoa were determined. Spermatozoa were observed with the transmission electron microscopy (TEM) either unstained or stained with metal salts. In the unstained specimens, the C2 nucleohistone region appeared more electron-lucent than the C1 region. When large cations such as uranyl were used, the reverse situation was observed. Therefore, the electron-dense appearance of the C2 chromatin in conventionally stained material may be due to the presence of net negative DNA charges that attract the cations used for EM staining, whereas the C1 chromatin may lack excess DNA negative charges that attract these stains and thus appears less electron-dense.
Keywords: Spermatozoa; Chromatin; Animals; Marsupialia; Osmium Tetroxide; Citric Acid; Organometallic Compounds; Microscopy, Atomic Force; Histocytochemistry; Coloring Agents; Male
Rights: © 1997 Wiley-Liss, Inc.
RMID: 0030006244
DOI: 10.1002/(SICI)1098-2795(199711)48:3<367::AID-MRD10>3.0.CO;2-T
Appears in Collections:Anatomical Sciences publications
Environment Institute publications

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