Localization of specific glycoproteins secreted by the rabbit and hamster epididymis.
Localization of specific glycoproteins secreted by the rabbit and hamster epididymis.
复制标题
兔子和仓鼠附睾分泌的特定糖蛋白的定位。
DOI：
10.1093/biolreprod/22.3.705
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发表时间：
1980
影响因子：
3.6
通讯作者：
H. Moore
中科院分区：
文献类型：
--
作者：
H. Moore
为了研究附睾中一些特异性蛋白质的起源和去向，采用逆行灌注法收集了性成熟新西兰白色兔和叙利亚火腿的附睾尾管腔内容物。用10%Ficolh溶液离心分离附睾液与精子，然后在聚丙烯酰胺平板凝胶上进行等电聚焦。根据等电点和对刀豆蛋白A的亲和力，鉴定了3种兔附睾特异性蛋白（p13.45、4.15和4.65）和2种仓鼠附睾特异性蛋白（pH3.40和4.15）为糖蛋白。将这些级分分离并皮下植入。用免疫扩散法和免疫电泳法分别在豚鼠和家兔体内制备了单特异性抗血清。用未标记抗体酶法在速冻附睾纵切片上进行糖蛋白抗原的定位。家兔和仓鼠的糖蛋白主要由附睾头和近体区的上皮细胞分泌。然而，最初的节段和睾丸缺乏抗原。个别糖蛋白的染色模式也存在明显差异。在兔中，糖蛋白首先与精子的远端头和近端附睾体，而仓鼠附睾中的糖蛋白首先与精子的近端头区域。在这两个物种的附睾尾部的糖蛋白的反应产物存在于精子和上皮细胞的adhuminal边界。这些结果的意义方面的变化，精子质膜在成熟过程中的表面组成进行了讨论。
To investigate the origin and fate of a number of specific epididymal proteins, the luminal con tents from the cauda epididymides of sexually mature New Zealand White rabbits and Syrian ham sters were collected by retrograde perfusion. Epididymal fluid, separated from spermatozoa by centrifugation over a 10% Ficolh solution, was then subjected to isoelectric focussing on pohyacrylamide flat bed gels. Three epididymal specific proteins from the rabbit, p1 3.45, 4.1 5 and 4.65 and two from the hamster, ph 3.40 and 4.1 5 were identified as glycoproteins by their how isoehectric point and their affinity for con canavalin A. These fractions were isolated and implanted s.c. in guinea pigs and rabbits, respec tively, to produce monospecific antisera as verified by immunodiffusion and immunoehectropho resis. Localization of glycoprotein antigen within the epididymis was performed on longitudinal sections of snap -frozen epididymis using the unlabeled antibody enzyme method. Both rabbit and hamster glycoproteins were secreted mainly by epididymal epithehial cells in the caput and proxi mal corpus region. However, the initial segments and testis were devoid of antigen. Differences were also apparent in the staining pattern of individual glycoproteins. In the rabbit, glycoproteins were first associated with spermatozoa in the distal caput and proximal corpus epididymidis while glycoproteins in the hamster epididymis were first associated with spermatozoa in the proximal caput region. Reaction product to glycoproteins in the cauda epididymidis of both species was present on spermatozoa and at the adhuminal border of epithehial cells. The significance of these re suhts with regard to changes in the surface composition of the sperm plasmalemma during matura tion is discussed.