Role of kisspeptin/GPR54 signaling pathways in prompting formation of precocious puberty

AIM: To explore the role of kisspeptin/GPR54 signaling pathway in the pathogenesis of precocious puberty based on female precocious puberty rat model induced by the single dose of danazol. METHODS: Female SD rats aged 3 d were randomly divided into normal group, vehicle group and model group. On the 5th day, the model rats were given a single subcutaneous injection of danazol with ethanol and ethylene glycol mixture. All rats were executed on 15 d, 25 d, 30 d, 35 d and 40 d, and the samples were collected to observe the sexual organ development. The levels of E₂, FSH and LH in peripheral blood were measured by ELISA. The Kiss-1 mRNA expression of Kiss-1, GPR54 and GnRH in hypothalamus was detected by real-time PCR. The kisspeptin expression in rat hypothalamus was observed by immunofluorescence. RESULTS: The time for puberty onset and sexual maturation in the model rats was significantly earlier than that in normal group and vehicle group. On days 25 and 30, the levels of peripheral sex hormones and uterine coefficients in model group were significantly higher than those in normal group and vehicle group. On days 25 and 30, the ovarian morphological development in the model rats was significantly earlier than that in normal group. On day 25, the mRNA expression of hypothalamic Kiss-1 and GnRH, and kisspeptin expression of hypothalamic arcuate nucleus in the model rats significantly increased compared with normal group and vehicle group. On day 30, kisspeptin expression of hypothalamic arcuate nucleus in the model rats decreased compared with normal group and vehicle group. On day 35, the mRNA expression of Kiss-1 and GnRH in the model rats decreased compared with normal group and vehicle group. The mRNA expression of GPR54 had no obvious difference among all groups. CONCLUSION: The Kiss-1 mRNA and kisspeptin expression in the model rats with precocious puberty is significantly increased in the hypothalamus during onset of puberty, suggesting that kisspeptin is an initiating factor for precocious puberty. Kisspeptin/GPR54 signaling pathway may play an important role in the occurrence of precocious puberty.     

Kisspeptin-10对蛋鹌鹑卵巢发育与内分泌的影响

为研究Kisspeptin-10(Kp-10)对禽类生殖内分泌的影响,将75羽22日龄雌鹌鹑随机分为3组,分别腹腔注射300μL生理盐水(对照组),注射0.1 nmol Kp-10(低剂量组)和1 nmol Kp-10(高剂量组),每日注射1次,连续注射3周。记录鹌鹑开产情况,并于60日龄时采集血液和卵泡组织,测定生殖激素与受体的mRNA表达。结果:与对照组相比,注射Kp-10组鹌鹑产蛋率显著提高,且大卵泡数显著增加,血液中雌激素水平显著升高;低剂量组输卵管重显著增加,但体重、肝脏重和小卵泡重无显著变化。Real-time PCR结果显示,Kp-10处理显著下调小卵泡及F1级卵泡上促黄体激素受体(LHR)、促卵泡激素受体(FSHR)和促性腺激素释放激素Ⅰ(GnRH-Ⅰ)的基因表达。     

光照强度对Kisspeptin及其受体GPR54在鸡脑表达的影响

随机选用100日龄京红Ⅰ号蛋鸡,分别接受光照强度30,20,10,1 lx处理,运用免疫组化技术测定Kisspeptin和GPR54免疫反应阳性神经元在鸡中脑和间脑的分布和表达。结果显示:Kisspeptin免疫反应阳性神经元主要分布于中脑SGC、Imc、Ipc、顶盖前核(Pt)、外侧螺旋核(SPL)。在SGC、Imc、Ipc,平均灰度值和面积百分比,30,20 lx组显著高于10,1 lx组(P<0.05),在Pt和SPL,30 lx平均灰度值和面积百分比最高。GPR54免疫反应阳性神经元主要分布于中脑的lmc、Ipc、SP/IPS、nbor、SGC,平均灰度值和面积百分比,光照强度30,20 lx组显著大于l0,1 lx组(P<0.05)。Kisspeptin免疫反应阳性神经元主要分布于间脑的下丘脑后内侧核(MPH)、丘脑室旁核前部(PVa)、PHN、PVN、Rot。在Rot、PHN、PVN和PVa,Kisspeptin免疫反应阳性神经元平均灰度值和面积百分比,30,20 lx组显著大于l0,1 lx组(P>0.05);在MPH,30 lx高于20 lx,20 lx高于10 lx,10 lx高于1 lx组。GPR54免疫阳性神经元主要分布于鸡间脑PVN、GLv、Rot、PHN和CPa。平均灰度值和面积百分比在Rot,30 lx组最大,但各组间差异不显著(P>0.05),在PVN、PHN、GLv和CPa,30,20 lx组显著大于10,1 lx组(P<0.05)。Kisspeptin免疫反应阳性细胞在腺垂体的平均灰度值,光照强度20 lx组最大,面积百分比30 lx最大,但差异不显著(P>0.05)。结果表明,光照强度能促进Kisspeptin和GRP54在蛋鸡中脑和间脑表达,随光照强度增加,表达量增加。Kisspeptin接受光信息调控,通过调节GnRH分泌,参与调节鸡的生殖机能。     

Kisspeptin对鱼类生殖轴的调控机制研究

Kisspeptin(简称Kiss或者Kp)是由KISS1/Kiss1基因编码的一种下丘脑神经肽,通过其受体KissR(也称作GPR54)的介导参与了多种生理过程,如抑制肿瘤转移和参与生殖调控。目前,尽管在鲤形目(Cypriniformes)、鲈形目(Perciforms)、鲽形目(Pleuronectiforms)、鲀形目(Tetraodontiforms)、颌针目(Beloniforms)、鲉形目(Scorpaeniformes)、鲑形目(Salmoniformes)及鳕形目(Gadiformes)等多种鱼类中均鉴定出了kiss/kissr基因,但Kiss/KissR系统在鱼类生殖调控中的精确作用及其分子机制尚未完全阐明。尤其是在鱼类中存在2种kiss及3种kissr基因,Kiss/KissR系统对鱼类生殖调控的作用方式更加复杂。本文简要总结鱼类Kiss及其受体的研究进展,并对Kiss的生理学功能、信号转导机制以及kiss/kissr表达调控研究进行概括讨论,旨在加深对鱼类Kiss/KissR系统的认识和了解,为后续研究指明方向。     

Kiss-1基因功能的研究进展

Kiss-1基因编码的Kisspeptins可通过其受体GPR54的介导来调控哺乳动物生殖内分泌。近年来的研究表明,Kiss-1基因具有抑制肿瘤转移、影响滋养细胞浸润、启动动物青春发育、调控能量平衡以及生物节律的功能。因此,本文就Kiss-1基因功能的研究进展做一综述。     

Dynorphin-Kappa Opioid Receptor Signaling Partly Mediates Estrogen Negative Feedback Effect on LH Pulses in Female Rats

Accumulating evidence suggests that the arcuate nucleus (ARC) kisspeptin/neurokinin B (NKB)/dynorphin (KNDy) neurons play a role in estrogen negative feedback action on pulsatile gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) release. The present study aimed to determine if dynorphin (Dyn) is involved in estrogen negative feedback on pulsatile GnRH/LH release. The effect of the injection of nor-binaltorphimine (nor-BNI), a kappa-opioid receptor (KOR) antagonist, into the third cerebroventricle (3V) on LH pulses was determined in ovariectomized (OVX) adult female rats with/without replacement of negative feedback levels of estradiol (low E₂). The mean LH concentrations and baseline levels of LH secretion in nor-BNI-injected, low E₂-treated rats were significantly higher compared with vehicle-treated controls. On the other hand, the nor-BNI treatment failed to affect any LH pulse parameters in OVX rats without low E₂ treatment. These results suggest that Dyn is involved in the estrogen negative feedback regulation of pulsatile GnRH/LH release. The low E₂ treatment had no significant effect on the numbers of ARC Pdyn (Dyn gene)-,Kiss1- and Tac2 (NKB gene)-expressing cells. The treatment also did not affect mRNA levels of Pdyn and Oprk1 (KOR gene) in the ARC-median eminence region, but significantly increased the ARC kisspeptin immunoreactivity. These findings suggest that the negative feedback level of estrogen suppresses kisspeptin release from the ARC KNDy neurons through an unknown mechanism without affecting the Dyn and KOR expressions in the ARC. Taken together, the present result suggests that Dyn-KOR signaling is a part of estrogen negative feedback action on GnRH/LH pulses by reducing the kisspeptin release in female rats.     

Differential Effects of Continuous Exposure to the Investigational Metastin/Kisspeptin Analog TAK-683 on Pulsatile and Surge Mode Secretion of Luteinizing Hormone in Ovariectomized Goats

The aim of the present study was to determine if the estradiol-induced luteinizing hormone (LH) surge is influenced by the constant exposure to TAK-683, an investigational metastin/kisspeptin analog, that had been established to depress the pulsatile gonadotropin-releasing hormone (GnRH) and LH secretion in goats. Ovariectomized goats subcutaneously received TAK-683 (TAK-683 group, n=6) or vehicle (control group, n=6) constantly via subcutaneous implantation of an osmotic pump. Five days after the start of the treatment, estradiol was infused intravenously in both groups to evaluate the effects on the LH surge. Blood samples were collected at 6-min intervals for 4 h prior to the initiation of either the TAK-683 treatment or the estradiol infusion, to determine the profiles of pulsatile LH secretion. They were also collected at 2-h intervals from –4 h to 32 h after the start of estradiol infusion for analysis of LH surges. The frequency and mean concentrations of LH pulses in the TAK-683 group were remarkably suppressed 5 days after the start of TAK-683 treatment compared with those of the control group (P<0.05). On the other hand, a clear LH surge was observed in all animals of both groups. There were no significant differences in the LH concentrations for surge peak and the peak time of the LH surge between the TAK-683 and control groups. These findings suggest that the effects of continuous exposure to kisspeptin or its analog on the mechanism(s) that regulates the pulsatile and surge mode secretion of GnRH/LH are different in goats.     

母猪情期启动营养调控分子机制的探讨

养猪生产中后备母猪年更新率高达40％～50％,哺乳母猪断奶后情期启动失败被淘汰比例占30％～40％,50％的母猪终身仅提供30～40头断奶仔猪就被淘汰,给养猪生产带来巨大经济损失.大量研究表明,下丘脑Kisspeptin信号系统在调控雌性动物情期启动和卵泡发育方面发挥重要作用.本课题组前期研究证实,猪的情期启动只有在营养充足、机体处于合成代谢时才能发生和维持.营养是推动后备母猪由生长向繁殖转换,或断奶母猪由泌乳向繁殖转换的物质基础.本文提出了营养调控母猪情期启动的几种理论假设,综述了能量、蛋白质(氨基酸)、维生素D3和部分矿物元素对机体代谢和下丘脑Kisspeptin信号的影响及其在调控母猪情期启动方面的分子机制.     

Sociosexual stimuli and gonadotropin-releasing hormone/luteinizing hormone secretion in sheep and goats

Sociosexual stimuli have a profound effect on the physiology of all species. Sheep and goats provide an ideal model to study the impact of sociosexual stimuli on the hypothalamic-pituitary-gonadal axis because we can use the robust changes in the pulsatile secretion of luteinizing hormone as a bioassay of gonadotropin-releasing hormone secretion. We can also correlate these changes with neural activity using the immediate early gene c-fos and in real time using changes in electrical activity in the mediobasal hypothalamus of female goats. In this review, we will update our current understanding of the proven and potential mechanisms and mode of action of the male effect in sheep and goats and then briefly compare our understanding of sociosexual stimuli in ungulate species with the "traditional" definition of a pheromone.