Djakiew D, Pflug B, Dionne C, Onoda M
Department of Cell Biology, Georgetown University Medical Center, Washington, District of Columbia 20007.
Biol Reprod. 1994 Aug;51(2):214-21. doi: 10.1095/biolreprod51.2.214.
Because nerve growth factor beta (NGF beta) and its corresponding receptors have been implicated in the paracrine regulation of spermatogenesis, we examined the postnatal developmental expression of the low- and high-affinity NGF receptors in the rat testis, and localized their expression to specific testicular cell types. The neurotropin receptors consist of a low-affinity p75 nerve growth factor receptor (LNGFR) and a family of high-affinity tyrosine receptor kinases (trk). Both the p75 LNGFR gene product and the trk receptor gene product were detected in immature rat testes, with maximal expression in 10- and 20-day-old rats. Expression of the testicular p75 LNGFR and the trk receptor progressively declined in older animals so that they were barely detectable in 90-day-old adult rats. The 75-kDa LNGFR was detected in membrane fractions of Sertoli cells, whereas the p75 LNGFR was not detected by Western blot in membrane fractions of round spermatids and primary spermatocytes. Interestingly, microsomal fractions of peritubular myoid cells were immunoreactive for a 65-kDa band on Western blots with the p75 LNGFR monoclonal antibody. Immunoblot analysis of the trk receptor in cell lysates of isolated cell types was inconclusive. Excess NGF beta and round spermatid protein, which is known to contain a NGF-like protein, were both capable of displacing the binding of 125I-NGF beta from the surface of Sertoli cells.(ABSTRACT TRUNCATED AT 250 WORDS)
由于神经生长因子β(NGFβ)及其相应受体参与了精子发生的旁分泌调节,我们研究了大鼠睾丸中低亲和力和高亲和力NGF受体的出生后发育表达,并将它们的表达定位到特定的睾丸细胞类型。神经营养素受体由低亲和力的p75神经生长因子受体(LNGFR)和高亲和力酪氨酸受体激酶家族(trk)组成。在未成熟大鼠睾丸中检测到了p75 LNGFR基因产物和trk受体基因产物,在10日龄和20日龄大鼠中表达最高。在年龄较大的动物中,睾丸p75 LNGFR和trk受体的表达逐渐下降,以至于在90日龄成年大鼠中几乎检测不到。在支持细胞的膜组分中检测到了75 kDa的LNGFR,而在圆形精子细胞和初级精母细胞的膜组分中通过蛋白质印迹未检测到p75 LNGFR。有趣的是,用p75 LNGFR单克隆抗体进行蛋白质印迹时,睾丸肌样细胞的微粒体组分对一条65 kDa的条带有免疫反应性。对分离细胞类型的细胞裂解物中trk受体的免疫印迹分析尚无定论。过量的NGFβ和已知含有NGF样蛋白的圆形精子细胞蛋白都能够从支持细胞表面取代125I-NGFβ的结合。(摘要截短于250词)