Breed W G, Leigh C M, Washington J M, Soon L L
Department of Anatomy and Histology, University of Adelaide, South Australia.
Mol Reprod Dev. 1994 Jan;37(1):78-86. doi: 10.1002/mrd.1080370111.
The organization of sperm chromatin in the dasyurid marsupial, Sminthopsis crassicaudata, was investigated using various morphological techniques. Transmission electron microscopy indicates two quite distinct chromatin regions became evident late in spermiogenesis with an outer globular region containing blocks of very electron-dense chromatin. Fluorescent light microscopical studies after staining with DNA dyes and 7-amino actinomycin D of testicular, caput, and cauda epididymal spermatozoa showed that this region fluoresced less brightly than the rest of the nucleus, indicating the presence of fewer DNA binding sites. Freeze fracture showed that the chromatin in most of the nucleus had randomly arranged particles of various sizes, but that of the outer region was composed entirely of small particles. This outer region was more resistant to low concentrations of the ionic detergent, SDS, whereas both guanidine hydrochloride and urea together with sodium chloride generally dispersed all the chromatin except that in the outer globular region and in a localized area of the nucleus beneath the acrosome. This study has thus revealed that the outer globular chromatin of these spermatozoa responds differently to ionic detergents and protein denaturing agents and has a different chromatin organization than most of the rest of the nucleus. The significance of these differences remains, however, to be determined.
利用各种形态学技术,对袋鼬科有袋动物肥尾袋鼩(Sminthopsis crassicaudata)精子染色质的组织情况进行了研究。透射电子显微镜观察表明,在精子发生后期出现了两个截然不同的染色质区域,外部球状区域含有电子密度极高的染色质块。对睾丸、附睾头和附睾尾精子用DNA染料和7-氨基放线菌素D染色后进行荧光显微镜研究发现,该区域的荧光亮度低于细胞核的其他部分,表明其DNA结合位点较少。冷冻断裂显示,细胞核大部分区域的染色质具有大小各异的随机排列颗粒,而外部区域则完全由小颗粒组成。该外部区域对低浓度离子去污剂十二烷基硫酸钠(SDS)具有更强的抗性,而盐酸胍、尿素与氯化钠共同作用时,通常会使除外部球状区域和顶体下方细胞核局部区域之外的所有染色质分散开来。因此,这项研究表明,这些精子的外部球状染色质对离子去污剂和蛋白质变性剂的反应不同,其染色质组织与细胞核的其他大部分区域也不同。然而,这些差异的意义仍有待确定。