Page K C, Mason P B, Lindstrom L, Swan J S, Nyquist S E
Department of Biology and Chemistry, Bucknell University, Lewisburg, PA 17837.
Biochem Cell Biol. 1992 Jun;70(6):496-503. doi: 10.1139/o92-077.
The relative contribution of the Sertoli cell and the pachytene spermatocyte to dolichol and N-linked oligosaccharide biosynthesis within the seminiferous tubule was investigated. Evidence is presented to show that the interaction between these two cell types affects dolichol and N-linked oligosaccharide biosynthesis. Analysis of the dolichol content of Sertoli cultures confirms earlier data suggesting that the Sertoli cell constitutes the major pool of dolichols within the seminiferous tubule. [14C]Acetate incorporation studies suggest that the Sertoli cell in culture synthesizes dolichol much more rapidly than does the isolated pachytene spermatocyte. This information, in addition to previous data in the literature, infers an interactive effect whereby the presence of the spermatogenic cell in the tubule stimulates dolichol synthesis in the Sertoli cell. The absence of normal Sertoli-spermatocyte interactions in in vitro incubations may also limit dolichol synthesis in the pachytene spermatocyte. The distribution of dolichol kinase between the Sertoli and the pachytene spermatocyte was also examined. The concentration of this enzyme in the Sertoli cell suggests the presence of an active salvage pathway within that cell. The correlation between the appearance of the pachytene spermatocyte and the previously described peak of dolichol kinase activity in the seminiferous tubules of the prepubertal animal implies cell-cell interactions. Radiolabelling studies of N-linked oligosaccharides were conducted using [3H]mannose and concanavalin A affinity chromatography to identify multiantennary, biantennary, and high-mannose oligosaccharide pools. An in vitro bicameral coculture system was used to demonstrate that pachytene spermatocytes stimulate incorporation of [3H]mannose into Sertoli cell oligosaccharides.(ABSTRACT TRUNCATED AT 250 WORDS)
研究了支持细胞和粗线期精母细胞对生精小管中多萜醇和N - 连接寡糖生物合成的相对贡献。有证据表明这两种细胞类型之间的相互作用会影响多萜醇和N - 连接寡糖的生物合成。对支持细胞培养物中多萜醇含量的分析证实了早期数据,表明支持细胞构成生精小管中多萜醇的主要储存库。[14C]乙酸掺入研究表明,培养中的支持细胞合成多萜醇的速度比分离的粗线期精母细胞快得多。这些信息以及文献中的先前数据推断出一种相互作用效应,即小管中生精细胞的存在刺激支持细胞中的多萜醇合成。体外孵育中缺乏正常的支持细胞 - 精母细胞相互作用也可能限制粗线期精母细胞中多萜醇的合成。还检测了多萜醇激酶在支持细胞和粗线期精母细胞之间的分布。该酶在支持细胞中的浓度表明该细胞内存在活跃的补救途径。粗线期精母细胞的出现与青春期前动物生精小管中先前描述的多萜醇激酶活性峰值之间的相关性暗示了细胞间相互作用。使用[3H]甘露糖和伴刀豆球蛋白A亲和色谱法进行N - 连接寡糖的放射性标记研究,以鉴定多天线、双天线和高甘露糖寡糖库。使用体外双室共培养系统证明粗线期精母细胞刺激[3H]甘露糖掺入支持细胞寡糖中。(摘要截短于250字)