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催产素和加压素刺激人和猪输精管上皮细胞的阴离子分泌。

Oxytocin and vasopressin stimulate anion secretion by human and porcine vas deferens epithelia.

作者信息

Hagedorn Travis M, Carlin Ryan W, Schultz Bruce D

机构信息

Department of Anatomy and Physiology, Kansas State University, Manhattan, Kansas 66506, USA.

出版信息

Biol Reprod. 2007 Sep;77(3):416-24. doi: 10.1095/biolreprod.106.056762. Epub 2007 Apr 18.

Abstract

Experiments were conducted to characterize the effects of oxytocin (OT) and vasopressin (VP) on epithelial cells isolated from human (1 degree HVD) and porcine (1 degree PVD) vas deferens and an immortalized epithelial cell line derived from porcine vas deferens (PVD9902 cells). Cultured monolayers were assessed in modified Ussing flux chambers and the OT- or VP-induced change in short circuit current (I(SC)) was recorded. All cell types responded to basolateral OT or VP with a transient increase in I(SC) that reached a peak of 3-5 microA cm(-2). Concentration-response curves constructed with 1 degree PVD and PVD9902 cells revealed that the apparent K(D) (k(app)) for OT was approximately 100-fold less than the k(app) for VP. Amplicons for the OT receptor (OXTR) and vasopressin type 2 and type 1a receptors (AVPR2 and AVPR1A) were generated with RT-PCR and the identification of each amplicon confirmed by sequence analysis. A selective antagonist for OXTR and AVPR1A fully blocked the effects of OT and partially blocked the effects of VP when assessed in both 1 degree PVD and PVD9902 monolayers. APVR2 antagonists blocked the effects of low (< or =30 nM) but not high concentrations of VP, indicating that VP was affecting both AVPR2 and a second receptor subtype, likely OXTR or AVPR1A. Experiments employing chelerythrine demonstrated that OT stimulation of vas deferens monolayers requires PKC activity. Alternatively, VP (but not OT) increased the accumulation of cytosolic cAMP in vas deferens epithelial cells. Results from this study demonstrate that OT and VP can modulate ion transport across vas deferens epithelia by independent mechanisms. OT and VP have the potential to acutely change the environment to which sperm are exposed and thus, have the potential to affect male fertility.

摘要

进行实验以表征催产素(OT)和加压素(VP)对从人(1度HVD)和猪(1度PVD)输精管分离的上皮细胞以及源自猪输精管的永生化上皮细胞系(PVD9902细胞)的影响。在改良的尤斯灌流室中评估培养的单层细胞,并记录OT或VP诱导的短路电流(I(SC))变化。所有细胞类型对基底外侧OT或VP的反应都是I(SC)短暂增加,达到3 - 5微安/平方厘米的峰值。用1度PVD和PVD9902细胞构建的浓度 - 反应曲线显示,OT的表观解离常数(k(app))比VP的k(app)小约100倍。通过逆转录聚合酶链反应(RT-PCR)产生OT受体(OXTR)、加压素2型和1a型受体(AVPR2和AVPR1A)的扩增子,并通过序列分析确认每个扩增子的鉴定。在1度PVD和PVD9902单层细胞中评估时,OXTR和AVPR1A的选择性拮抗剂完全阻断了OT的作用,并部分阻断了VP的作用。APVR2拮抗剂阻断了低浓度(≤30 nM)但不阻断高浓度VP的作用,表明VP同时影响AVPR2和第二种受体亚型,可能是OXTR或AVPR1A。使用白屈菜红碱的实验表明,OT对输精管单层细胞的刺激需要蛋白激酶C(PKC)活性。另外,VP(但不是OT)增加了输精管上皮细胞中细胞溶质环磷酸腺苷(cAMP)的积累。本研究结果表明,OT和VP可通过独立机制调节离子跨输精管上皮的转运。OT和VP有可能急性改变精子所处的环境,因此有可能影响男性生育能力。

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