Miteva K, Valkov N, Goncharova-Peinoval J, Kovachev K, Zlatarev S, Pironcheva G, Russev G
Institute of Biology and Immunology of Reproductional Development of Organisms, Bulgarian Academy of Sciences, Sofia, Bulgaria.
Cytobios. 1995;83(333):85-90.
A whole mount electron microscopic technique facilitated a direct visualization of transcripts in ram spermatozoa in run-on experiments. The localization of transcripts in sperm chromatin by spreading enabled identification of regions where transcription complexes, presumably pre-mRNA species, were seen related to chromatin. In another series of experiments the localization of polymerase II was demonstrated using a specific antibody against the conservative tail domain of the polymerase II molecule, followed by protein A-gold visualization on spread chromatin and on thin sections from mature spermatozoa. Incorporation of bio-UTP in transcripts was visualized by streptavidin-gold on spreading and on thin sections. The data suggest that transcription occurs at the periphery of the mature spermatozoa.
在连续标记实验中，整装电子显微镜技术有助于直接观察公羊精子中的转录本。通过铺展精子染色质来定位转录本，能够识别出与染色质相关的区域，在这些区域可以看到转录复合物（推测为前体mRNA种类）。在另一系列实验中，使用针对聚合酶II分子保守尾部结构域的特异性抗体来证明聚合酶II的定位，随后通过蛋白A-金标法在铺展的染色质和成熟精子的薄切片上进行可视化观察。通过链霉亲和素-金标法在铺展和薄切片上观察生物UTP掺入转录本的情况。数据表明转录发生在成熟精子的周边。
