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大鼠精子发生中核基质 |
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劳萍 徐炎 章静波 陈延文 刘世广 孙英丽
【摘要】 目的 研究精子发生中的核基质-核纤层-中间丝体系(nuclear matrix-lamina-intermediate filament,NM-L-IF)的变化趋向。 方法 利用整装电镜技术、SDS-PAGE和Southwestern blot等技术观察与分析经单位重力沉降法分离的大鼠精母细胞、早期精子细胞和晚期精子细胞的NM-L-IF的形态结构、蛋白组成及其与A-T重复序列的相互关系。 结果 精母细胞核基质由核纤层、核仁残余物及内部纤维网络组成。精子细胞中，中间丝丰富，核基质结构被致密的染色质团簇所掩盖。精子形成Ⅷ～Ⅸ期时，核基质分化出两部分：疏松的头端(Ap)和致密的尾端(Ca)，中间丝连接细胞核与细胞膜；精子形成Ⅹ～Ⅺ期时，头端和尾端间形成弯曲；ⅩⅡ～ⅩⅣ期，头端进一步弯曲，一部分中间丝连接头端的顶部和头尾端的连接部。SDS-PAGE显示分子量小于40kD的蛋白在不同细胞中组成有差异，而分子量大于40kD的蛋白没有明显差异。Southwestern blot显示精母细胞的低分子量蛋白对A-T重复序列的吸附能力强，而只有早期精子细胞的部分高分子量蛋白对该序列具有很强的亲和力。 结论 精子发生中NM-L-IF的形态结构、蛋白组成及与DNA的相互作用发生显著变化，这可能与精子细胞核的浓缩变形以及形态发生密切相关。
【关键词】 精子发生 核基质-核纤层-中间丝体系 整装电镜技术
STUDY ON THE NUCLEAR MATRIX-LAMINA-INTERMEDIATE
FILAMENT SYSTEM IN THE RAT SPERMATOGENESIS
Lao Ping, Xu Yan*△，Zhang Jingbo*，Chen Yanwen*，
Liu Shiguang*，Sun Yingli**
(Centre Laboratory of Shandong Provincial Hospital, Ji nan;*Department of Cell Biology, Institute of Basic medical Sciences,
Chinese Academy of Medical Sciences, Beijing; **Department of Biology, Peking University, Beijing)
【Abstract】 Objective To study the changes of the nuclear matrix-lamina-intermediate filament(NM-L-IF) during the rat spermatogenesis. Methods Spermatocytes, early stage spermatids and the later stage spermatids were separated from the adult rat testis by the velocity gravitative sedimentation technique. Using the whole mount electron microscopy, SDS-PAGE and Southwestern blot, the morphology, components of NM-L-IF and the relationship between an A-T repetitive DNA fragment and the proteins of NM-L-IF in those cells were analyzed. Results: In spermatocytes, the NM is composed of a lamina, a large mass of residual nucleolar material and fibers. In spermatids, there are many IFs and the NM is concealed beneath the clumps of chromatin. NM becomes to form two portions, a less condensed apical(Ap) and a more condensed caudal(Ca) in spermatogenic stages Ⅷ-Ⅸ.IFs of these cells [1] [2] [3] 下一页
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