Faculty of Allied Health Sciences, Burapha University, Chonburi 20131, Thailand.
Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Road, Bangkok 10400, Thailand.
Acta Histochem. 2020 Jan;122(1):151454. doi: 10.1016/j.acthis.2019.151454. Epub 2019 Oct 9.
Gonadotropin releasing hormone (GnRH) is a peptide brain hormone that is involved in the regulation of reproduction in vertebrates via stimulation of the secretion of the pituitary hormones, luteinizing hormone (LH) and follicle-stimulating hormone (FSH), which in their turn stimulate sexual development and sex steroid hormone secretion by the gonads. The tropical abalone, Haliotis asinina, in common with many other invertebrates contains a peptide with a similar structure to GnRH. This study looks at its possible involvement in reproduction by injecting groups of one-year-old female abalone at the mature phase by injecting them with synthetic H. asinina (Has) GnRH at doses of 0, 250 and 500 ng/g and then measuring the amount of material in nerve ganglia, ovary and hemolymph that cross-reacted with enzyme-linked immunosorbent assays (ELISA) for vertebrate LH and steroid, estradiol. Immunohistochemistry, using antibodies for the same two compounds, was also carried out to examine the location of immunoactivity in the tissues of the animals. There were slight (in some cases statistically significant) increases in LH-immunoactivity and estradiol in the hemolymph and tissues. However, this applied to the lower dose only (i.e the dose-response relationship was non-monotonic). Using immunohistochemistry, LH-immunoreactive cells were observed in types 1 and 2 neurosecretory (NS1 and NS2) cells within the cerebral and pleuropedal ganglia of H. asinina. In addition, LH-immunoreactive nerve fiber bundles were strongly detected in both ganglia. The immunoactivity against the estrogen appeared to be localized in the granulated cells within the connective tissue and trabeculae of the mature ovary. There was no positive staining in the cytoplasm of any stage of the germ cells. The interpretation of these findings is presently hindered by the fact that the homologous gene for vertebrate LH has not yet been identified in the genomes of any mollusks (so the cause of the immunostaining is as yet unknown) and also by the fact that mollusks are known to readily absorb steroids from the environment and store them long-term in the form of fatty acid esters. More work, involving identification of the protein that cross-reacts with the LH antiserum and also exclusion of the possibility that the estradiol is of exogenous origin, will have to be carried out before these findings can be used to manipulate reproduction in this species.
促性腺激素释放激素(GnRH)是一种肽类脑激素,通过刺激垂体激素黄体生成素(LH)和卵泡刺激素(FSH)的分泌,参与脊椎动物的生殖调节,而这两种激素反过来又刺激性腺的性发育和性激素分泌。热带鲍鱼 Haliotis asinina 与许多其他无脊椎动物一样,含有一种与 GnRH 结构相似的肽。本研究通过向处于成熟阶段的 1 岁龄雌性鲍鱼注射合成 H. asinina (Has) GnRH,剂量分别为 0、250 和 500ng/g,然后测量神经节、卵巢和血淋巴中与酶联免疫吸附测定(ELISA)检测脊椎动物 LH 和甾体激素雌二醇交叉反应的物质含量,来研究其在生殖中的可能作用。还使用针对这两种化合物的抗体进行了免疫组织化学,以检查动物组织中免疫活性的位置。在血淋巴和组织中,LH 免疫活性和雌二醇略有增加(在某些情况下具有统计学意义)。然而,这只适用于较低的剂量(即剂量反应关系是非单调的)。使用免疫组织化学,在 H. asinina 的脑和侧神经节中观察到 1 型和 2 型神经分泌(NS1 和 NS2)细胞中的 LH 免疫反应细胞。此外,在两个神经节中均强烈检测到 LH 免疫反应神经纤维束。针对雌激素的免疫活性似乎定位于成熟卵巢结缔组织和小梁中的颗粒细胞中。在任何阶段的生殖细胞细胞质中均未发现阳性染色。目前,这些发现的解释受到以下事实的阻碍:脊椎动物 LH 的同源基因尚未在任何软体动物的基因组中被识别(因此免疫染色的原因尚不清楚),并且已知软体动物很容易从环境中吸收类固醇,并以脂肪酸酯的形式长期储存它们。在这些发现可用于操纵该物种的繁殖之前,还需要进行更多的工作,包括鉴定与 LH 抗血清发生交叉反应的蛋白质,并排除雌二醇来源于外源性的可能性。