Power R F, Bishop A E, Wharton J, Inyama C O, Jackson R H, Bloom S R, Polak J M
Department of Histochemistry, Royal Postgraduate Medical School, London, United Kingdom.
Ann N Y Acad Sci. 1988;527:314-25. doi: 10.1111/j.1749-6632.1988.tb26989.x.
Vasoactive intestinal polypeptide has a widespread distribution in the body, occurring in both the central and peripheral nervous systems and considerable information is available on its distribution, physiology, and pharmacological actions. Receptors for VIP have been demonstrated previously in peripheral tissues by conventional binding techniques using isolated membrane preparations. However, information on their precise localization is limited. We therefore localized binding sites in a variety of guinea pig and rat tissues by in vitro autoradiography and made a parallel study of the distribution of VIP nerves in these tissues using immunocytochemistry. [125I]VIP was prepared by the chloramine T method and shown to be pharmacologically active. After a preincubation procedure to remove endogenously bound VIP, unfixed cryostat sections were incubated with 1 nM [125I]VIP. To determine specific binding, sections were incubated in the presence or absence of 1 microM unlabeled VIP. Autoradiograms were generated by exposing the sections to LKB-Ultrofilm or emulsion-coated coverslips. Dense binding occurred in discrete locations within the gastrointestinal, respiratory, and genital tracts, correlating with known actions of VIP and, to various extents, with the distribution of VIP nerves. For example, there was precise localization to respiratory epithelium, smooth muscle of airways and blood vessels, and alveolar walls, in keeping with the effects of VIP on vascular and airway smooth muscle and mucus secretion.
血管活性肠肽在体内分布广泛,存在于中枢和外周神经系统,关于其分布、生理学及药理作用已有大量信息。此前通过使用分离膜制剂的传统结合技术已在外周组织中证实了血管活性肠肽的受体。然而,关于其精确定位的信息有限。因此,我们通过体外放射自显影术在多种豚鼠和大鼠组织中定位结合位点,并使用免疫细胞化学方法对这些组织中血管活性肠肽神经的分布进行了平行研究。[125I]血管活性肠肽通过氯胺T法制备,并显示具有药理活性。在经过预孵育程序以去除内源性结合的血管活性肠肽后,将未固定的低温恒温切片与1 nM [125I]血管活性肠肽一起孵育。为了确定特异性结合,切片在存在或不存在1 microM未标记血管活性肠肽的情况下孵育。通过将切片暴露于LKB-Ultrofilm或涂有乳剂的盖玻片上来生成放射自显影片。密集结合出现在胃肠道、呼吸道和生殖道内的离散位置,这与血管活性肠肽的已知作用相关,并在不同程度上与血管活性肠肽神经的分布相关。例如,在呼吸道上皮、气道和血管平滑肌以及肺泡壁有精确定位,这与血管活性肠肽对血管和气道平滑肌以及黏液分泌的作用一致。