Bentley George E, Ubuka Takayoshi, McGuire Nicolette L, Chowdhury Vishwajit S, Morita Yoshihiro, Yano Tetsu, Hasunuma Itaru, Binns Molly, Wingfield John C, Tsutsui Kazuyoshi
Laboratory of Reproductive Neuroendocrinology, Department of Integrative Biology and Helen Wills Neuroscience Institute, University of California at Berkeley, Berkeley, CA 94720-3140, USA.
Gen Comp Endocrinol. 2008 Mar 1;156(1):34-43. doi: 10.1016/j.ygcen.2007.10.003. Epub 2007 Oct 22.
Many hormones that are classified as neuropeptides are synthesized in vertebrate gonads in addition to the brain. Receptors for these hormones are also expressed in gonadal tissue; thus there is potential for a highly localized autocrine or paracrine effect of these hormones on a variety of gonadal functions. In the present study we focused on gonadotropin-inhibitory hormone (GnIH), a neuropeptide that was first discovered in the hypothalamus of birds. We present different lines of evidence for the synthesis of GnIH and its receptor in the avian reproductive system including gonads and accessory reproductive organs by studies on two orders of birds: Passeriformes and Galliformes. Binding sites for GnIH were initially identified via in vivo and in vitro receptor fluorography, and were localized in ovarian granulosa cells along with the interstitial layer and seminiferous tubules of the testis. Furthermore, species-specific primers produced clear PCR products of GnIH and GnIH receptor (GnIH-R) in songbird and quail gonadal and other reproductive tissues, such as oviduct, epididymis and vas deferens. Sequencing of the PCR products confirmed their identities. Immunocytochemistry detected GnIH peptide in ovarian thecal and granulosa cells, testicular interstitial cells and germ cells and pseudostratified columnar epithelial cells in the epididymis. In situ hybridization of GnIH-R mRNA in testes produced a strong reaction product which was localized to the germ cells and interstitium. In the epididymis, the product was also localized in the pseudostratified columnar epithelial cells. In sum, these results indicate that the avian reproductive system has the capability to synthesize and bind GnIH in several tissues. The distribution of GnIH and its receptor suggest a potential for autocrine/paracrine regulation of gonadal steroid production and germ cell differentiation and maturation.
许多被归类为神经肽的激素除了在大脑中合成外,还在脊椎动物的性腺中合成。这些激素的受体也在性腺组织中表达;因此,这些激素对多种性腺功能具有高度局部化的自分泌或旁分泌作用的潜力。在本研究中,我们聚焦于促性腺激素抑制激素(GnIH),这是一种最初在鸟类下丘脑发现的神经肽。通过对雀形目和鸡形目两个鸟类目进行研究,我们提供了不同的证据,证明GnIH及其受体在包括性腺和附属生殖器官在内的鸟类生殖系统中合成。GnIH的结合位点最初通过体内和体外受体荧光成像鉴定,并定位在卵巢颗粒细胞以及睾丸的间质层和生精小管中。此外,物种特异性引物在鸣禽和鹌鹑的性腺及其他生殖组织(如输卵管、附睾和输精管)中产生了清晰的GnIH和GnIH受体(GnIH-R)的PCR产物。PCR产物的测序证实了它们的身份。免疫细胞化学检测到GnIH肽存在于卵巢的卵泡膜细胞和颗粒细胞、睾丸间质细胞和生殖细胞以及附睾的假复层柱状上皮细胞中。GnIH-R mRNA在睾丸中的原位杂交产生了强烈的反应产物,该产物定位于生殖细胞和间质。在附睾中,产物也定位于假复层柱状上皮细胞。总之,这些结果表明鸟类生殖系统有能力在多个组织中合成并结合GnIH。GnIH及其受体的分布表明其对性腺类固醇生成以及生殖细胞分化和成熟具有自分泌/旁分泌调节的潜力。