Fillion C, Malassine A, Tahri-Joutei A, Allevard A M, Bedin M, Gharib C, Hugues J N, Pointis G
Laboratoire d'Endocrinologie et de Biologie de la Reproduction, UFR Santé Médecine Biologie Humaine, Bobigny, France.
Biol Reprod. 1993 Apr;48(4):786-92. doi: 10.1095/biolreprod48.4.786.
The presence of arginine vasopressin (AVP)-like peptide(s) and AVP receptors has been previously demonstrated in the testis of several species. In this study we examined the immunocytochemical localization of AVP and of its associated neurophysin (NPII) within the mouse testis, and the testicular immunoreactive (IR)-AVP content in normal pubertal and adult mice and in unilaterally cryptorchid animals. Immunostaining was conducted on fresh frozen adult testicular sections and on cultured testicular cells using the avidin-biotin immunoperoxidase technique. Immunoreactivities for AVP and NPII were localized at the periphery of the seminiferous tubules. Staining for AVP and NPII was reduced in testes showing impaired spermatogenesis. AVP and NPII immunoreactivities were present in cultured Sertoli cells, but staining was undetectable in cultured Leydig cells. Negative controls were obtained by applying antiserum that had been preabsorbed with excess peptides in place of primary antiserum. Mouse testes were found to contain IR-AVP, which co-eluted with synthetic AVP on HPLC and diluted in parallel in a specific RIA for AVP. Levels of IR-AVP expressed as pg/mg wet-weight testis were significantly higher (p < 0.05) in pubertal (36.6 +/- 1.5) than in adult animals (27.4 +/- 1.5). Experimental unilateral cryptorchidism in adult animals resulted 2 wk later in a marked reduction (p < 0.01) in IR-AVP levels in the abdominal testis (18.1 +/- 1.0 pg/mg testis) as compared to the contralateral scrotal testis (29.8 +/- 1.1 pg/mg testis).(ABSTRACT TRUNCATED AT 250 WORDS)
先前已证实在多个物种的睾丸中存在精氨酸加压素(AVP)样肽和AVP受体。在本研究中,我们检测了AVP及其相关神经垂体素(NPII)在小鼠睾丸内的免疫细胞化学定位,以及正常青春期和成年小鼠及单侧隐睾动物睾丸中的免疫反应性(IR)-AVP含量。使用抗生物素蛋白-生物素免疫过氧化物酶技术,对新鲜冷冻的成年睾丸切片和培养的睾丸细胞进行免疫染色。AVP和NPII的免疫反应定位于生精小管的周边。在精子发生受损的睾丸中,AVP和NPII的染色减少。在培养的支持细胞中存在AVP和NPII免疫反应性,但在培养的间质细胞中未检测到染色。通过使用用过量肽预吸收的抗血清代替一抗获得阴性对照。发现小鼠睾丸含有IR-AVP,其在高效液相色谱上与合成AVP共洗脱,并在AVP的特异性放射免疫分析中平行稀释。以pg/mg湿重睾丸表示的IR-AVP水平在青春期小鼠(36.6±1.5)中显著高于成年动物(27.4±1.5)(p<0.05)。成年动物实验性单侧隐睾在2周后导致腹部睾丸中的IR-AVP水平(18.1±1.0 pg/mg睾丸)与对侧阴囊睾丸(29.8±1.1 pg/mg睾丸)相比显著降低(p<0.01)。(摘要截断于250字)