Localization and synthesis of entactin in seminiferous tubules of the mouse.
Localization and synthesis of entactin in seminiferous tubules of the mouse.
复制标题
小鼠生精小管中巢蛋白的定位和合成。
DOI：
10.1095/biolreprod47.3.316
复制
发表时间：
1992
影响因子：
3.6
通讯作者：
Enders,GC
中科院分区：
文献类型：
--
作者：
Lian,G;Miller,KA;Enders,GC
用免疫细胞化学方法研究了巢蛋白在小鼠睾丸曲细精管中的定位和合成。对成年小鼠睾丸的冷冻切片进行抗巢蛋白和抗层粘连蛋白免疫荧光检测。巢蛋白和层粘连蛋白均定位于生精小管基底膜和睾丸管间区。加入过量的巢蛋白（但不是纤连蛋白），与抗巢蛋白抗血清预混，取消免疫染色。免疫荧光显微镜检测显示，原代Sertoli细胞和管周肌样细胞培养6 d后，富集的部分能够合成并分泌entactin。免疫荧光和Western blotting检测到，在原代培养的细胞系和TM 3、TM 4细胞系中，巢蛋白和层粘连蛋白的分布是不同的。Englyn主要出现在细胞外。上述结果提示：1）巢蛋白存在于成年小鼠睾丸生精小管基底膜和管间区，与层粘连蛋白共定位，2）巢蛋白由原代培养的支持细胞、管周肌样细胞和TM 3、TM 4细胞合成; 3）与层粘连蛋白的细胞内积累相反，巢蛋白被胞外化，细胞内几乎没有积累。
Localization and synthesis of entactin in seminiferous tubules of mouse testis was studied by immunocytochemistry. Frozen sections from adult mice testes were subjected to anti-entactin and anti-laminin immunofluorescence. Both entactin and laminin were localized within the seminiferous tubule basement membrane and intertubular region of the testis. The addition of excess amount of entactin (but not fibronectin), premixed with anti-entactin antiserum, abolished the immunostain. Western blotting showed that a protein extract from a seminiferous tubule basement membrane preparation was recognized by anti-entactin anti-serum and comigrated with recombinant entactin.Enriched fractions of isolated primary Sertoli cells and peritubular myoid cells cultured for 6 days on a glass coverslip were able to synthesize and secrete entactin as detected by immunofluorescence microscopy. Entactin was also produced by TM3 (Leydig-like) and TM4 (Sertoli-like) cell lines as detected by both immunofluorescence and Western blotting.The distribution of entactin vs. laminin within both the cultured primary cells and the TM3 and TM4 cell lines differed. Entactin appeared mainly localized extracellularly. In contrast, laminin was mainly localized intracellularly.The above findings suggested that 1) entactin existed in the seminiferous tubule basement membrane and intertubular region of adult mice testis, co-localized with laminin; 2) entactin was synthesized by the cultured primary Sertoli cells and peritubular myoid cells and the TM3 and TM4 cell lines; 3) entactin was exocytosed with little intracellular accumulation, in contrast to an intracellular accumulation of laminin.