Characterization,cerebral distribution and gonadotropin release activity of neuropeptide Y (NPY) in the goldfish

The presence of a peptide closely related to porcine NPY has been demonstrated in the goldfish brain and pituitary by means of radioimmunoassay (RIA) and high performance liquid chromatography (HPLC). The RIA data demonstrate that displacement curves of brain extracts are parallel to a porcine NPY standard and that in HPLC a compound present in brain extracts is co-eluted with porcine NPY. The distribution of this NPY-like factor within the central nervous system was studied by radioimmunoassay and immunohistochemistry. The results indicated that NPY has a widespread distribution with the highest concentrations being found in the telencephalon and diencephalon. In the pituitary gland, NPY immunoreactive terminals characterized at the electron microscope level were found in the different lobes and, in particular, in close association with the gonadotrophin (GTH) secreting cells. Using anin vitro perifusion system, it was shown that NPY causes a dose dependent increase of GTH release from anterior lobe fragments. These data indicate for the first time in teleosts that NPY is present and widely distributed in the brain and pituitary, and that among other putative functions, could be implicated in the multihormonal release of GTH from the pituitary.     

The regulatory effects of thyrotropin-releasing hormone on growth hormone secretion from the pituitary of common carp in vitro

The effects of thyrotropin-releasing hormone (TRH) on growth hormone (GH) and gonadotropin (GtH) release, and the influences of somatostatin (SRIF), the dopamine agonist apomorphine (APO) and extracellular calcium on basal and TRH-induced GH release were examined using an in vitro perifusion system for pituitary fragments of common carp (Cyprinus carpio). Five minute pulses of different dosages of TRH stimulated a rapid and dose-dependent increase in GH release from the perifused pituitary fragments with an ED₅₀ of 9.7 ± 2.3 nM. TRH was ineffective on GtH release. SRIF significantly inhibited basal and TRH-induced GH release from the perifused pituitary fragments, and the effects of SRIF were dose-dependent. APO induced a dose-dependent increase in basal and TRH-stimulated GH release from the perifused pituitary fragments. Increasing the concentrations of extracellular calcium from 0 mM to 1.25 mM resulted in an increase in basal and TRH-induced GH release. The high dose of calcium (6.25 mM) caused a slight decrease in basal and TRH-induced GH release compared with those at a concentration of 1.25 mM.

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Résumé Les effets de la thyrotropine (TRH) sur la sécrétion d'hormone de croissance (GH) et de gonadotropine (GTH), et de la somatostatine (SRIF), de l'apomorphine (APO), antagoniste dopaminergique, et du calcium extracellulaire sur les sécrétions basale et stimulée de GH ont été étudiées in vitro par périfusion, de fragments d'hypophyses de carpe (Cyprinus carpio). Des applications de 5 minutes de TRH à différentes concentrations induisent une stimulation rapide et dose dépendante de la sécrétion de GH (ED₅₀ = 9.7 ± 2.3 nM). Le TRH est sans effet sur la sécrétion de GTH. Le SRIF inhibe la sécrétion basale de GH ainsi que la résponse hypophysaire à l'action du TRH. Son action est dose dépendante. L'apomorphine induit une augmentation dose dépendante de la sécrétion basale de GH et potentialise l'action du TRH sur la stimulation de la sécrétion de GH. Des effets équivalents sont induits par des concentrations croissantes de calcium extra cellulaire de 0 à 1.2 mM, alors qu'à une concentration de 6.25 mM des effets opposés sont obtenus.

     

鸡胚垂体生长激素细胞的发生及其变化

为研究鸡胚垂体中生长激素细胞的发生及其在发育过程中的变化 ,分别在鸡胚发育的第 3 5～ 2 0 5天采集鸡胚垂体 ,用免疫组织化学方法研究鸡胚垂体中生长激素细胞的发生和细胞形态、数量、在垂体中的分布特点和在发育过程中的变化。结果表明 ,鸡胚发育中期第 9 5天可观察到少量的生长激素阳性细胞散在分布于垂体后叶 ,细胞多排列成索状或团状 ,细胞之间边界不清 ,细胞核较大。在鸡胚发育的第 12 5天以后 ,生长激素细胞的细胞浆与细胞核比值逐渐变小 ,且均匀分布于腺垂体的后叶。生长激素细胞数在鸡胚发育的第 12 5天之后显著升高 (P <0 0 1) ,在鸡胚发育的第 16 5天 ,生长激素细胞占垂体细胞总数的 9 4 %。以上结果证明鸡胚垂体中生长激素细胞增殖和分化过程主要发生在发育的中期至出壳之前 ,而生长激素细胞的分泌功能在鸡胚发育的后期最为活跃。     

The effects of gonadal development and sex steroids on growth hormone secretion in the male tilapia hybrid (Oreochromis niloticus × O. aureus)

Profiles of plasma growth hormone (GH) in male tilapia hybrid (Oreochromis niloticus x O. aureus) were measured and compared at different times of the year. The profiles did not appear to be repetitive, however, differences in their nature were observed at the different seasons; the most erratic profiles were seen in the height of the reproductive season (July), while the peaks were more subdued in the spring and disappeared in the autumn. Peaks in male fish were more prominent than in the females when measured in July. Perifused pituitary fragments from fish with a high GSI responded to salmon gonadotropin-releasing hormone (sGnRH) analog (10 nM-1 M), while those from fish with a low GSI barely responded to even the highest dose. Exposure of perifused pituitary fragments from sexually-regressed fish to carp growth hormone-releasing hormone (cGHRH; 0.1 M) or sGnRH (I M) stimulated GH release only after injection of the fish with methyl testosterone (MT; 3 injections of 0.4 mg kg ¹). The same MT pretreatment did not alter the response to dopamine (DA; 1 or 10 M). GH pituitary content in MT-treated fish was lower than in control fish, which may be explained by the higher circulating GH levels in these fish, but does not account for the increased response to the releasing hormones. Castration abolished the response of cultured pituitary cells to sGnRH (I fM-100 nM) without altering either their basal rate of secretion or circulating GH levels. Addition of steroids to the culture medium (MT or estradiol at 10 nM for 2 days) enabled a GH response to sGnRH stimulation in cells from sexually regressed fish. Pituitary cells which had not been exposed to steroids failed to respond to sGnRH, although their response to forskolin or TPA was similar to that of steroid-exposed cells. It would appear, therefore, that at least one of the effects of the sex steroids on the response to GnRH is exerted proximally to the formation of cAMP, or PKC, presumably at the level of the receptor. An increase in the number of receptors to the GH-releasing hormones, following steroid exposure, would explain also the changing nature of the GH secretory profile in different stages of the reproductive season.     

Distinct alpha (α)-subunits of salmon glycoprotein hormones: production sites in the pituitary with sexual maturity

Salmon pituitary glands contain two structurally distinct -subunit proteins (1 and 2) of glycoprotein hormones: the 2-subunit is common to all salmon gonadotropins (GTH I and GTH II), whereas the 1-subunit is present in only some GTH I molecules. GTH I is predominant in the pituitary gland and plasma during gametogenesis of salmon, but the roles of the 2 GTHs in gametogenesis remain unclear. To understand the roles of GTH I, it is important to clarify patterns of 1- and 2-subunit production with sexual maturity. Thus, we produced antisera that recognized the 1- or 2-subunit, and then immunohistochemically examined the production sites of these subunits in the trout pituitary gland during ovarian development. In all pituitary glands examined, the immunoreactivity of both the 1- and 2-subunits was strong in the GTH II-producing cells, although salmon GTH II, both 1- and II-subunits, has not been detected. However, GTH I-producing cells showed a less dense immunoreactivity for 1- and 2-subunits, whereas the I-subunit was abundant. On the other hand, TSH cells, reacted with 2 but not with 1.     

Effects of salinity, dietary level of protein and 17α-methyltestosterone on growth hormone (GH) and prolactin (tPRL177 and tPRL188) levels in the tilapia, Oreochromis mossambicus

In the present study, we examined the long-term effects of environmental salinity, diet (35% and 25% crude protein) and 17-methyltestosterone (MT) on corresponding levels of pituitary and serum growth hormone (GH) and prolactins (tPRL₁₇₇ and tPRL₁₈₈) in the tilapia (Oreochromis mossambicus). We observed no discernible patterns in serum GH that would suggest an effect of salinity, diet or MT. However, serum GH levels in all treatments declined at 1 and 3h after first feeding. Serum tPRL₁₇₇ and tPRL₁₈₈ were significantly higher in freshwater (FW) than in seawater (SW) and levels were significantly affected by dietary protein. tPRL₁₇₇ levels were higher in all groups fed a 35% protein diet, but tPRL₁₈₈ levels were higher only in the groups fed the MT-treated 35% protein diet; only serum tPRL₁₈₈ levels were affected by MT. Moreover, serum tPRL₁₇₇ and tPRL₁₈₈ increased throughout the sampling time-course. Subsequent work using fasted tilapia suggests that first feeding is likely to initiate the post-prandial suppression of serum GH levels. In contrast with the picture observed in blood, pituitary glands of SW animals showed higher levels of GH than FW fish. Pituitary GH was elevated by MT in both FW and SW. We also observed that pituitary tPRL₁₇₇ and tPRL₁₈₈ levels were higher in FW fish than in SW fish; tPRL₁₇₇ and tPRL₁₈₈ levels were elevated by MT only in FW animals. To assess the somatomedin activity of plasma from FW- and SW-reared tilapia, we measured [35S]-sulfate incorporation into ceratobranchial cartilage explants in vitro. Plasma from SW-adapted tilapia showed greater activity in this assay than plasma from FW-reared tilapia, suggesting that the GH-dependent IGF bioactivity of plasma is higher in SW-reared tilapia. Collectively, these studies suggest that the growth-promoting actions of SW rearing and of MT administration in tilapia may be linked to elevations in GH and/or prolactin (tPRL₁₇₇ and tPRL₁₈₈)levels.     

藏鸡与白羽肉鸡垂体转录组差异表达研究

试验旨在通过对藏鸡和白羽肉鸡垂体转录组测序和生物信息学分析,筛选出与鸡生长发育相关的差异表达基因（differentially expression genes,DEGs）及信号通路。本研究选用42日龄健康藏鸡和白羽肉鸡各3只,分别采集垂体组织利用Illumina HiSeq 2000平台进行转录组mRNA测序,对筛选到的DEGs筛选DEGs,GO和KEGG数据库功能注释和富集分析,实时荧光定量PCR验证随机挑选的DEGs表达水平。结果显示,藏鸡和白羽肉鸡分别获得126 094 302和125 666 442条clean reads。与白羽肉鸡相比,藏鸡垂体组织中共有DEGs 392个,其中196个为上调基因,196个为下调基因（P<0.05）,其中包括生长激素（GH）基因、生长激素释放激素受体（GHRHR）基因和胰岛素样生长因子2 mRNA结合蛋白1（IGF2BP1）基因。GO和KEGG富集分析发现,DEGs显著富集于细胞黏附分子信号通路和神经活性配体-受体相互作用信号通路等细胞通讯相关的信号通路,GH、GHRHR和IGF2BP1基因也显著富集于神经活性配体-受体相互作用信号通路。实时荧光定量PCR结果显示,转录组测序结果准确可靠。综上研究,本研究初步揭示了影响藏鸡和白羽肉鸡生长发育速度差异的关键候选基因和信号通路,为了解鸡垂体组织调节生长发育的分子机制提供了理论依据。     

大豆黄酮对体外灌流垂体前叶组织促黄体素分泌的影响

对德国格丁根微型猪垂体前叶组织进行体外灌流培养，研究大豆黄酮对垂体促黄体素（ＬＨ）分泌的影响。结果表明，灌流２４ｈ后，大豆黄酮灌流组（Ⅰ组１００ｎｇ／ｍｌ；Ⅱ组２μｇ／ｍｌ）和雌二醇灌流组（Ⅲ组１００ｐｍｏｌ／Ｌ；Ⅳ组２００ｐｍｏｌ／Ｌ）ＬＨ分泌水平低于对照组（Ⅴ组空白对照；Ⅵ组助溶剂对照），雌激素抑制效应明显较强。灌流４８ｈ，Ⅰ、Ⅱ组ＬＨ峰值和平均水平均显著高于对照组，Ⅲ、Ⅳ组则明显低于对照组。促性腺激素释放激素（ＧｎＲＨ）刺激后１～２ｈ，Ⅰ、Ⅱ组ＬＨ平均水平、峰值和脉冲频率均明显高于刺激前，而且显著高于对照组；Ⅲ、Ⅳ组却明显低于对照组和Ⅰ、Ⅱ组。大豆黄酮、雌激素对垂体体外ＬＨ分泌没有明显的剂量差异。无ＧｎＲＨ刺激时，大豆黄酮对离体垂体ＬＨ分泌表现为抑制效应，但抑制强度较弱；长期以大豆黄酮灌流培养垂体，或加以ＧｎＲＨ刺激，大豆黄酮对ＬＨ分泌具促进作用。     

驼背鲈脑垂体显微和超微结构的观察

采用PAS、Mallory三色法和腺垂体不同细胞改良染色法3种组织学染色方法,以及透射电镜技术,对驼背鲈(Cromileptes altivelis)脑垂体的形态、组织结构和超微结构进行了研究。结果表明,驼背鲈脑垂体为圆球形实心腺体,位于间脑腹面,由神经垂体和腺垂体2部分组成。根据神经纤维中所含分泌颗粒形态和大小把神经分泌纤维分为A1、A2和B型。神经纤维中存在2种垂体细胞,即呈椭圆形的Ⅰ型垂体细胞和呈梭形的Ⅱ型垂体细胞。腺垂体由3部分组成:前外侧部(RPD)、中外侧部(PPD)和中间部(PI)。腺垂体部可以鉴别出6种内分泌细胞,其中RPD有2种内分泌细胞:促肾上腺皮质激素(ACTH)分泌细胞和催乳激素(PRL)分泌细胞;PPD有3种内分泌细胞:生长激素(GH)分泌细胞、促性腺激素(GTH)分泌细胞和促甲状腺激素(TSH)分泌细胞;PI只有1种内分泌细胞,即促黑激素(MSH)分泌细胞。此外,腺垂体还有1种非分泌细胞星状细胞(SC)。驼背鲈腺垂体中6种内分泌细胞的分布情况为:前外侧部分布有2种细胞,即嗜酸性的PRL分泌细胞和嗜碱性的ACTH分泌细胞;中外侧部有GH、GTH、TSH 3种分泌细胞;MSH分泌细胞分...