Division of Surgery, Department of Urology and Uro-Oncology, Hannover Medical School, Hannover 30625, Germany.
J Sex Med. 2010 Feb;7(2 Pt 2):888-95. doi: 10.1111/j.1743-6109.2009.01539.x.
In contrast to research findings describing the localization of nitric oxide synthases (NOS), guanylyl cyclases, and cyclic adenosine monophosphate (cAMP)- and cyclic guanosine monophosphate (cGMP)-degrading phosphodiesterase isoenzymes in the human vagina, the distribution of proteins known as major targets for cyclic nucleotides has not yet been evaluated. cAMP- and cGMP-dependent protein kinases (cAK, cGKI) have been identified as important receptors for cyclic nucleotides downstream the signaling cascades.
To investigate, by means of immunohistochemistry, the expression of cAK and cGKI in relation to endothelial NOS (eNOS), vasoactive intestinal polypeptide (VIP), and protein gene product 9.5 (PGP 9.5) in the human vagina.
Expression and distribution of cAK and cGKI(alpha,beta) in relation to eNOS, VIP, and PGP 9.5 in human vaginal tissue.
Immunohistochemical techniques were applied to sections of human vaginal full wall specimens in order to evaluate the presence of cAK and cGKI(alpha,beta) in relation to VIP, PGP 9.5, and eNOS, respectively. Western blot analyses were conducted using cytosolic supernatants of homogenized specimens of the vaginal wall and epithelium.
Immunostaining specific for cGKIbeta was observed in vascular and nonvascular smooth muscle of the vagina. In the endothelial layer, cGKIbeta was found colocalized with eNOS. In contrast, no signals indicating cGKIalpha were registered. cAK-positive subepithelial vessels were found to be innervated by a dense meshwork of PGP-containing varicose nerve fibers, some of which presented expression of VIP. The expression of cAK and cGKIbeta was confirmed by Western blotting.
Our results demonstrate the expression of cAK and cGKIbeta in the human vagina. The colocalization with VIP and eNOS underlines the significance of both the cAMP and GMP pathway in the control of human vaginal vascular and nonvascular smooth muscle.
与描述一氧化氮合酶（NOS）、鸟苷酸环化酶、环磷酸腺苷（cAMP）和环鸟苷酸（cGMP）降解磷酸二酯酶同工酶在人阴道中的定位的研究结果相反，已知作为环核苷酸主要靶标的蛋白质的分布尚未得到评估。cAMP 和 cGMP 依赖性蛋白激酶（cAK、cGKI）已被确定为信号级联下游环核苷酸的重要受体。
通过免疫组织化学方法研究 cAK 和 cGKI 与内皮型一氧化氮合酶（eNOS）、血管活性肠肽（VIP）和蛋白基因产物 9.5（PGP 9.5）在人阴道中的表达关系。
cAK 和 cGKI（α、β）与阴道组织中 eNOS、VIP 和 PGP 9.5 的表达和分布关系。
应用免疫组织化学技术对人阴道全壁标本切片进行检测，以评估 cAK 和 cGKI（α、β）与 VIP、PGP 9.5 和 eNOS 的关系。使用阴道壁和上皮匀浆上清液进行 Western blot 分析。
在阴道的血管和非血管平滑肌中观察到特异性的 cGKIβ免疫染色。在内皮层中，cGKIβ与 eNOS 共定位。相比之下，未检测到 cGKIα的信号。在 subepithelial 血管中发现 cAK 阳性的神经纤维呈密集的 PGP 含曲张神经纤维网，其中一些表达 VIP。cAK 和 cGKIβ 的表达通过 Western blot 得到证实。
我们的研究结果表明 cAK 和 cGKIβ在人阴道中的表达。与 VIP 和 eNOS 的共定位强调了 cAMP 和 GMP 通路在人阴道血管和非血管平滑肌控制中的重要性。
