GnRH及其受体在性成熟前后鲇、黄颡鱼脑、垂体和卵巢中的免疫细胞化学定位

用链霉亲和素 -生物素化过氧化物酶复合物 (StreptAvidinBiotin peroxidaseComplex ,SABC)免疫细胞化学方法 ,使用促性腺激素释放激素 (Gonadotropin releasinghormone ,GnRH)以及促性腺激素释放激素受体 (GnRHR) 2种抗血清对性成熟前后的黄颡鱼 (Pelteobagrusfulvidraco)和鲇鱼 (Silurusasotus)的脑、垂体、卵巢中的免疫活性内分泌细胞进行了免疫细胞化学定位。结果表明GnRH和GnRHR免疫活性在两种鱼的各脑区、垂体、卵巢中均有分布 ;两种鱼在性成熟时它们的下丘脑、垂体和卵巢中的GnRH和GnRHR免疫反应细胞数目和免疫反应强度明显高于性成熟前。本文讨论了GnRH、GnRHR直接或间接参与黄颡鱼和鲇鱼性腺发育成熟调节的可能性及形态学证据。可为下丘脑 垂体 性腺轴、神经 内分泌、GnRH功能的多样性等研究领域提供新的形态学依据。     

促性腺激素释放激素的结构及其生物学功能

促性腺激素释放激素(GnRH)是下丘脑分泌的十肽激素,是神经、免疫、内分泌三大调节系统互相联系的重要信号分子,对生殖调控具有重要意义.GnRH类似物是近年来应用最广的多肽类激素新药之一.就GnRH及其受体的结构及分布、GnRH在垂体和性腺水平调控生殖的一系列证据、影响GnRH释放的因素等进行了综述,并展望了GnRH研究的发展趋势及应用前景.     

下丘脑促性腺激素释放激素及其类似物对卵巢的作用

下丘脑促性腺激素释放激素(GnRH)及其类似物(GnRH-激动剂,GnRH-A)能直接作用于卵巢。对卵巢的各个成份、颗粒细胞、黄体细胞、内膜/间质细胞、卵子均有作用。本文着重介绍了 GnRH 和 GnRH-A 对卵巢的作用与其作用机制。     

卵巢内发现促性腺激素释放激素样蛋白质

近来许多报道表明,促性腺激素释放激素(GnRH)或GnRH样物质可直接影响卵巢功能,其中包括对离体颗粒细胞和黄体细胞功能的影响以及GnRH拮抗剂在体内加强FSH对卵巢内卵泡发育的作用。在未成年和成年大鼠离体颗粒细胞和黄体细胞均发现有 GnRH 受体。这提示,卵巢内可能合成 GnRH 或 GnRH样肽。美国学者 Aten 等用大鼠黄体化卵巢检测是否存在有 GnRH 或 GnRH 样肽。他们用50IU 孕马血清     

GnRH介导的GPCR信号转导系统在卵巢癌发生、发展中的作用

卵巢癌是最致命的妇科恶性肿瘤。虽然上皮性卵巢癌约占人类所有卵巢癌的90%,但该病的病因学了解甚少。促性腺激素释放激素(gonadotropin-releasing hormone,GnRH)是下丘脑-垂体-性腺轴关键的调节器,主要调节脊椎动物的繁殖。研究发现,GnRH已经显示出对卵巢癌细胞有直接的抗增殖作用。GnRH受体是G蛋白偶联受体(G protein–coupled receptor,GPCR)家族的成员。本文主要就GnRH介导的GPCR信号系统与卵巢癌细胞的抗增殖作用之间的关系展开综述。     

蒙药乌力吉-18对大鼠下丘脑-垂体-卵巢轴相关激素及受体表达的影响

目的:探讨蒙药乌力吉-18对大鼠下丘脑-垂体-卵巢轴相关激素及受体的影响。方法:选取40只健康雌性未孕SD大鼠,随机分为空白组、对照组、乌力吉-18高、低2个剂量组,每组10只。空白组灌胃等体积蒸馏水,对照组灌胃逍遥丸,高、低剂量组分别灌胃2.0 g·kg^-1·d^-1、1.0 g·kg^-1·d^-1乌力吉-18,连续给药31学艺术d。采用酶联免疫吸附法测定血清促性腺激素释放激素(GnRH)、促卵泡生成素(FSH)、黄体生成素(LH)、雌二醇(E₂)及孕酮(PROG)的含量;免疫组化法检测下丘脑组织促性腺激素释放激素(GnRH)、垂体组织促性腺激素释放激素受体(GnRHR)的表达;以蛋白免疫印迹技术检测卵巢组织促卵泡生成素受体(FSHR)、黄体生成素受体(LHR)蛋白表达量。以实时荧光定量PCR检测卵巢组织中FSHR、LHR基因表达量。结果:与空白组比较,乌力吉-18低剂量组可明显升高血清LH含量(P<0.05),上调下丘脑组织GnRH、垂体组织GnRHR表达及卵巢组织FSHR、LHR蛋白表达(P<0.05);乌力吉-18高剂量组可显著升高血清FSH、LH、E₂含量(P<0.05),上调下丘脑组织GnRH表达及卵巢组织FSHR表达量(P<0.05),并可显著升高卵巢组织中FSHR、LHR基因表达量(P<0.05);对照组可明显升高血清E₂含量(P<0.05)。结论:蒙药乌力吉-18可明显升高血清FSH、LH及E₂的含量,促进下丘脑组织GnRH、垂体组织GnRHR及卵巢组织中FSHR、LHR的表达,表明乌力吉-18能够对下丘脑-垂体-卵巢轴相关激素及受体表达产生影响。     

灵长类月经周期的调控

灵长类月经周期的调控与啮齿类不同,在下丘脑没有促性腺激素释放激素(GnRH)的周期性分泌中枢。排卵前促性腺激素(GTH)峰的出现无需下丘脑活动的增强和GnRH分泌的增加。GnRH对垂体GTH的周期性分泌不起控制作用,只起“允许作用”,起控制作用的是卵巢雌激素。雌二醇作用于垂体促性腺细胞的两个功能池,控制GnRH对它们的作用,完成调节GTH分泌的作用。     

卵巢中存在GnRH的mRNA

既往工作表明:促性腺激素释放激素(GnRH)具有多种垂体外作用。它可通过黄体和颗粒细胞上的特异性结合位点调节卵巢功能。由于血液循环中GnRH水平太低不足以与外周位点结合;而且,GnRH拮抗物对去垂体大鼠卵巢功能仍具有调节作用。因此推测卵巢能产生GnRH或GnRH样物质作为旁分泌激素。但是分离纯化这些肽类物质的工作未获得成功。作者在本工作中采用反向转录-聚合酶链锁反应(RT-PCR)     

首次发现血液循环中有天然抗激素

美国 Dahl 等将一种促性腺激素释放激素(GnRH)拈抗剂:A_(?)-△~3-_(?)~1,P-F-D-Phe~2,D-T_1P~(3,6)GnRH,注入性腺功能减退的妇女,注入速度为30μg/kg/h。注入后8h,测定血清卵泡刺激素(FSH)生物活性,发现其活性仅为处理前水平的62％,而 FSH 免疫活性减少约为30％;生物活性与免疫活性之比(B/I)从处理前的1.8降至1.0。将血清样本进行层析和分级分离,发现在 GnRH 拮抗剂处理后,血清中出现免疫活性 FSH     

5-HT和GnRH及其受体在长牡蛎卵巢的生理作用机制——双染和免疫细胞化学定位

用免疫细胞化学与双染技术对5-HT和GnRH及其受体在长牡蛎卵巢中的分布的研究结果提示,不同发育时期卵巢中卵母细胞均存在5-HT和GnRH受体,受体阳性物质沿胞膜分布,显深棕色,核显免疫阴性反应;5-HT和GnRH及其共存于不同时期卵母细胞胞质和胞膜上。以上结果表明长牡蛎的卵母细胞不仅能产生5-HT和GnRH,而且能表达5-HT和GnRH的受体,5-HT和GnRH可能以自分泌的方式对卵母细胞起调节作用。     

Distribution of lamprey gonadotropin-releasing hormone-III (GnRH-III) in brains of larval lampreys (Petromyzon marinus)

Two immunoreactive forms of gonadotropinreleasing hormone (GnRH), lamprey GnRH-I and lamprey GnRH-III, were found in neurons in larval sea lampreys (Petromyzon marinus). Using antisera preferentially directed against either lamprey GnRH-I or-III, dense reaction product was seen in cell bodies in the rostral hypothalamus and preoptic area. Reaction product was also dense in fibers to and within the neurohypophysis, in addition to numerous fibers which projected caudally, beyond the neurohypophysis through the mesencephalon. The majority of immunoreactive GnRH was lamprey GnRH-III, and when lamprey GnRH-I was seen, it was in cells that appeared to contain both forms of GnRH. A small number of cells found in the caudal hypothalamus contained only immunoreactive lamprey GnRH-III, and these may constitute a functional subgroup within the population of GnRH neurons. In animals undergoing metamorphosis there was a large increase in reaction product in all GnRH-containing cells and fibers. A striking change within the distribution of GnRH cells was localized to a distinct group of GnRH-immunoreactive cells (GnRH-I and-III) in the ventral anterior hypothalamic area. These cells were minimally detectable in larvae, but during metamorphosis became densely filled with immunoreactive product in perikarya and distal processes. The results are consistent with the hypothesis that lamprey GnRH-III is an important form of GnRH during the maturation of GnRH cells and fibers, and further indicates that these cells have attained their normal positions in the preoptic area and hypothalamus before metamorphosis.     

Insulin-, glucagonand somatostatin-like immunoreactivity in the endocrine pancreas of the lungfish,Neoceratodus forsteri

Summary The endocrine pancreas of the Australian lungfish,Neoceratodus forsteri, was investigated immunocytochemically for the presence of polypeptide hormone-producing cells. Three cell types were identified, namely insulin-, glucagon-, and somatostatin-immunoreactive elements. The insulin cells are confined solely to the center of the islets. Glucagon and somatostatin cells are distributed peripherally around the central mass of the insulin cells. Isolated cells or clusters of glucagon and somatostatin cells are also dispersed within the exocrine parenchyma. The immunoreactive cell types are compared with those staining with standard histological procedures. The spatial relationships of the different cell populations are examined.     

Immunocytochemistry of somatotrophs,gonadotrophs, prolactin and adrenocorticotropin cells in larval sea bream (Sparus auratus) pituitaries

Summary The chronological appearance of endocrine cells in the pituitary of sea-bream (Sparus auratus) larvae was studied using antisera against salmon prolactin, trout growth hormone, salmon gonadotropin and N-terminal human adrenocorticotropin. The larval pituitary (1–12 days after hatching) was oval in shape and was composed of a dense mass of cells with few neurohypophysial fibres. By 60 days after hatching it began to resemble the adult and was divisible into a distinct rostral pars distalis containing prolactin and adrenocorticotropin cells; a proximal pars distalis containing somatotrophs and gonadotrophs and a pars intermedia. Cells immunoreactive with antisera against growth hormone were observed immediately after hatching (2 days post-fertilization). Weakly staining prolactin cells were observed 2 days later in the region corresponding to the rostral pars distalis. Cells immunoreactive with anti-gonadotropin and anti-adrenocorticotropin sera were observed in the pituitary 6 and 8 days after hatching, respectively. All the cell-types studied were immunoreactive from the time they were first identified until the final samples 90 days after hatching.     

Localization of 5S rRNA Loci in Three Coregonid Species (Salmonidae)

In the present study the chromosome distribution of the 5S rDNA loci and its relation to the major rDNA genes were investigated in three Coregonid species (Salmonidae): Coregonus lavaretus, Coregonus peled and Coregonus albula, a family which has experienced large karyotype rearrangements along its evolution starting from a tetraploid ancestor. 5S PRINS/CMA₃ sequential staining together with previous data enabled us to locate 5S rRNA genes and nucleolar organizer regions (NORs) in the three species analyzed. PRINS revealed the 5S rDNA cluster at the distal part of the long arm of a similar submetacentric chromosome pair in the three species. Our data indicate that 5S rDNA clusters have probably conserved chromosomal location in the genus Coregonus, whereas 45S rDNA (NOR) sites are clearly differentiated, from a single locus in C. peled, to multiple loci in C. lavaretus and highly polymorphic multichromosomal location in C. albula.     

Nitric oxide (NO) synthase immunoreactivity in the starfish Marthasterias glacialis

The neuroendocrine system of the starfish Marthasterias glacialis was investigated immunocytochemically using antisera specific for rat neuronal, bovine aortic endothelial, and mouse macrophage, nitric oxide (NO) synthases. Immunoreactivity was detected only with the antibodies specific for the neural enzyme, in the ectoneural and hyponeural tissues of the radial nerve cords and in the basiepithelial plexus and endocrine cells of the digestive tract. The pyloric stomach showed more immunoreactive structures than the other digestive organs, with the rectal caeca showing the least activity. Immunoreactive endocrine cells were located in the cardiac and pyloric stomachs and in the pyloric caeca. Co-localization of the enzyme immunoreactivity, and the staining for NADPH-diaphorase, demonstrate the presence of NO synthase in echinoderms. These results provide further evidence that NO is a neuronal messenger of early phylogenetic origin which has been conserved throughout evolution.     

Gonadotropin-releasing hormone-immunoreactive neuronal structures in the brain and pituitary of the African catfish,Clarias gariepinus (Burchell)

Summary The distribution of gonadotropin-releasing hormone (GnRH) immunoreactivity was studied in the African catfish, Clarias gariepinus, by means of immunofluorescence and immunoperoxidase techniques. Immunoreactive neurons were found throughout the preoptic nucleus (NPO). However, only a portion of the secretory perikarya in the NPO showed a positive reaction by use of an anti-LHRH serum. Numerous immunoreactive fibres were found to enter the pituitary and to terminate in its proximal pars distalis, the site of concentration of the gonadotropic cells. Since GnRH is present in the brain and pituitary of the African catfish, the lack of spontaneous ovulation in captivity is apparently due to an insufficient release of GnRH.     

Co-localization of peptides in the Brockmann bodies of the cod (Gadus morhua) and the rainbow trout (Oncorhynchus mykiss)

Endocrine cells exhibiting immunoreactivity to FMRFamide-like, LPLRFamide-like, neuropeptide Y(NPY)-like and peptide YY(PYY)-like peptides were found in the periphery of the Brockmann bodies of the cod, Gadus morhua, and rainbow trout, Oncorhynchus mykiss. No immunoreactivity or very weak labelling was found with antisera to pancreatic polypeptide (PP). Vasoactive intestinal polypeptide (VIP)-like immunoreactivity was found in nerve fibres, whereas labelling with VIP antiserum in endocrine cells disappeared after preincubation with nonimmune serum. There were always more immunoreactive cells in the rainbow trout than in the cod. No immunoreactivity could be seen with antisera to gastrin/cholecystokinin (CCK) or enkephalin. Double-labelling studies were performed to study the colocalization of the peptides in peripheral endocrine cells. Cells immunoreactive to NPY were also labelled with antisera to FMRFamide, LPLRFamide and PYY. The co-localization pattern of NPY varied; in some Brockmann bodies, a population of the immunoreactive cells showed co-localization and others contained NPY-like immunoreactivity only, whereas in other Brockmann bodies, all NPY-labelled cells also contained FMRFamide-like, LPLRFamide-like and PYY-like immunoreactivity. Cells immunoreactive to PYY similarly contained FMRFamide-like, LPLRFamide-like and NPY-like immunoreactivity, comparable to the patterns observed with NPY. Glucagon-like immunoreactivity was found at the periphery of the Brockmann bodies. A subpopulation of the glucagon-containing cells contained NPY-like immunoreactivity. PYY-like immunoreactivity was also found co-localized with glucagon-like immunoreactivity, as were FMRFamide-like and LPLRFamide-like immunoreactivity. Therefore, either NPY-like and PYY-like immunoreactivity together with FMRFamide-like and LPLRFamide-like immunoreactivity occur in the same endocrine cells of the Brockmann body of the cod and rainbow trout, or a hybrid NPY/PYY-like peptide recognized by both NPY and PYY antisera is present in the Brockmann body.     

Distribution and ontogeny of VIP-like immunoreactivity in the gastro-entero-pancreatic system of a cartilaginous fish Scyliorhinus stellaris

Summary The presence, distribution and development of vasoactive intestinal polypeptide (VIP)-like immunoreactivity in the gastro-entero-pancreatic system of a cartilaginous fish Scyliorhinus stellaris (L.) was investigated by immunohistochemical methods utilizing mammalian VIP antisera. In the gut VIP-like immunoreactivity was observed in both nerves and endocrine cells. Endocrine cells with VIP-like material were only detected in the intestinal epithelium while nerve fibres containing VIP-like material were noted along the whole gastro-entero-pancreatic system, being more numerous in the pyloric sphincter and in the intestinal portion. Immunoreactive nerve cell bodies were encountered in the stomach and intestinal portions localized in the submucosa and in the myenteric plexus. Intestinal immunoreactive endocrine cells were already present in the first developmental stage considered (embryos aged 4 months). They grow in number and before birth reach a frequency higher than in adults. Nerves and cell bodies showing VIP-like immunoreactivity, appear later, before birth, as a few elements in the smooth muscular layer, but only after birth their distribution and frequency are similar to those found in adults. The faint immunofluorescence shown by the immunoreactive endocrine cells and their developmental pattern, which is always different from that observed in nervous elements, suggest the presence of at least two VIP-like substances in the gastro-entero-pancreatic system of S. stellaris.     

水鼩鼱消化道5-羟色胺阳性细胞的形态与分布

应用免疫酶标技术ABC( avidin-biotin-peroxidase complex)法,对水鼩鼱(Neomys fodiens)消化道内5-羟色胺(5-HT)阳性细胞的分布及形态进行研究.结果显示,5-羟色胺阳性细胞仅在肠内分布,食管和胃处没有分布.细胞分布密度曲线呈抛物线型,空肠部分布密度最高,十二指肠部次之...