三疣梭子蟹卵巢发育期间孕激素受体的免疫识别与分布

为探讨三疣梭子蟹卵巢发育过程中孕激素受体（Progestin receptor, PR）的调控作用,采用免疫印迹和免疫组化方法,首次对三疣梭子蟹卵巢、肝胰腺、大颚器、Y器官、视神经节和胸神经团中的PR进行免疫识别和定位, 并系统研究了卵巢发育期间PR在这些器官中的分布和变化。结果表明：(1) 三疣梭子蟹视神经节中存在PR阳性条带,分子量约100kDa;(2) PR仅在早期(Ⅰ-Ⅲ期)生殖细胞胞质和后期(Ⅲ-Ⅴ期)细胞核呈强阳性, 但在各期滤泡细胞中始终呈强阳性; (3) PR阳性主要存在于Ⅰ-Ⅴ期肝胰腺F细胞和Ⅲ期R细胞核中, 其余肝胰腺细胞中均无PR阳性; (4) 大颚器腺细胞中的PR阳性仅发现于Ⅳ期腺细胞胞质中, 各期Y器官腺细胞中均呈PR阴性; (5) 就视神经节而言, PR主要存在于Ⅲ-Ⅴ期的神经细胞。就胸神经团而言, Ⅰ-Ⅴ期神经细胞中的PR阳性呈增强趋势, 而Ⅰ-Ⅴ期神经髓质中PR始终呈PR强阳性。以上结果暗示, 孕酮不仅可以通过与卵巢和肝胰腺中PR结合直接调控三疣梭子蟹的卵黄发生和卵巢发育, 还可能通过影响其它内分泌激素的合成和分泌来间接调控其卵巢发育。     

三疣梭子蟹卵巢发育过程中雌二醇的免疫定位和变化

雌二醇(estradiol,E_2)是甲壳动物体内重要的性类固醇激素,对其卵巢发育和卵黄发生起着十分重要的调控作用。本实验采用免疫组织化学方法系统研究了三疣梭子蟹(Portunus trituberculatus)卵巢发育过程中E_2在卵巢、肝胰腺、胸神经节、脑神经节和大颚器中的分布及变化。结果表明:(1)蟹卵巢中E_2的免疫阳性主要分布于滤泡细胞和卵巢发育中后期的卵母细胞细胞质(II-V期),卵巢发育过程中卵母细胞细胞质和细胞核中的E_2免疫阳性均为先上升后下降的趋势,最大免疫阳性分别出现在IV期和II-III期,滤泡细胞中始终存在较强的E_2免疫阳性;(2)肝胰腺中的E_2免疫阳性主要分布于F细胞及R细胞细胞核中,R细胞细胞核中的E_2免疫阳性从III期开始显著下降,F细胞中始终存在较强的E_2免疫阳性;(3)卵巢发育(I-V期)的神经组织,E_2主要分布于胸神经团的神经细胞和神经髓质以及脑神经节神经细胞细胞核内,其中脑神经节神经细胞细胞核中始终保持着强免疫阳性,卵巢发育早期(I-II期)胸神经团中的神经髓质中为E_2中等免疫阳性,神经细胞中一直为弱免疫阳性;(4)就大颚器而言,E_2强免疫阳性始终存在于大颚器腺细胞的核仁以及细胞核周围的细胞质内。以上结果表明,E_2在三疣梭子蟹卵巢、肝胰腺、神经组织和大颚器中广泛分布,且免疫阳性与卵巢发育具有一定的相关性,E_2可能通过作用于多个靶器官来调控三疣梭子蟹的卵巢发育。     

三疣梭子蟹B型清道夫受体蛋白的原核表达及细胞、组织分布研究

B型清道夫受体是清道夫受体超家族成员,可识别多种配体,在机体脂类转运及免疫防御过程中发挥重要作用。为深入研究三疣梭子蟹B型清道夫受体(Pt-SRB)的功能,本实验在前期工作基础上构建了Pt-SRB胞外结构域原核表达载体,并成功获得了重组蛋白rPt-SRB。利用亲和层析方法获得纯化的rPt-SRB,并将纯化rPt-SRB蛋白免疫大鼠获得抗rPt-SRB重组蛋白免疫抗血清以用于后续研究。SDS-PAGE检测发现,体外诱导表达重组rPt-SRB以包涵体的形式出现在大肠杆菌BL21裂解液的沉淀中,分子量大小约为49.18 ku。Western-Blot分析表明,大鼠抗rPt-SRB血清能与rPt-SRB特异性结合。本实验还利用免疫荧光技术,对Pt-SRB在三疣梭子蟹血淋巴细胞的定位及不同组织中的分布进行研究。结果显示,Pt-SRB在三疣梭子蟹消化道、肝胰腺、鳃、心脏、肌肉组织中均有分布,但不同组织中Pt-SRB的分布不同。免疫荧光结果显示,绿色荧光信号在消化道及腺体结缔组织中较强,另外在肝小管上皮、鳃丝上皮细胞中亦有较强的阳性信号,表明PtSRB蛋白在上述组织结构中分布较广;在血淋巴细胞中,绿色荧光信号主要分布于细胞质和细胞膜,在细胞核中没有明显荧光信号,提示Pt-SRB在三疣梭子蟹血淋巴细胞的细胞质和细胞膜上表达。本结果将为三疣梭子蟹B型清道夫受体蛋白的生理及免疫学功能的研究奠定基础。     

池塘养殖三疣梭子蟹的生长规律

为探讨池塘养殖三疣梭子蟹的生长规律和雌雄差异，本研究对1～6月龄池塘养殖三疣梭子蟹各生长指标进行了比较分析，并采用Logistic模型分别拟合三疣梭子蟹的体重(BW)、体长(BL)、体高(BH)、全甲宽(FCW)、甲宽(CW)、大螯长节长(MLC)、大螯不动指长(FFLC)、第一步足长节长(MLFP)等8个形态性状的生长特征。结果显示，三疣梭子蟹雌性和雄性的生长存在差异，早期雄性生长较快，后期雌性生长较快，而雌雄混合分析介于二者之间。三疣梭子蟹各生长性状的Logistic模型拟合结果显示，除雌性MLC和MLFP以及雌雄混合分析的MLFP之外，其余性状R²均达到0.990以上；雌性和雄性体质量的极限生长值分别为290.27和195.91g，快速生长区间分别为2.74～5.10月龄和2.33～4.14月龄，拐点分别为3.92和3.24月龄。研究表明，三疣梭子蟹的生长过程均符合“慢-快-慢”的特征，雄性比雌性更早进入快速生长期，但是快速生长期的持续时间不及雌性。本研究对三疣梭子蟹不同生长指标的规律特征，以及各指标在混合养殖条件下雌性和雄性的优势阶段进行了研究，为实现三疣...     

[4]东海三疣梭子蟹精子发生及精荚形成

为探究东海三疣梭子蟹(Portunus trituberculatus Miers,1876)雄性群体的性腺发育规律,通过定期采样,运用形态学、组织学方法对东海三疣梭子蟹精子发生及精荚形成过程进行了研究,并结合精小叶生殖节段法对处于不同发育阶段的精巢进行了定性分析.结果表明,三疣梭子蟹精巢属于叶型,由精小叶和精小管组成的叶状管交织而成.精子发生过程与其他高等短尾派蟹类相似,精子成熟后即进入精小管中.输精管可能起源于精小管.三疣梭子蟹精巢和输精管的发育存在一定的年际变化.4-5月份,精巢小叶内精原细胞占优势,输精管内精荚零星分布;6-7月份,精母细胞与精细胞比例增加;8月份精母细胞为主.到了9月份,整个精巢完全被精细胞等发育程度较高的精小叶所占据,输精管内充满精液和精荚,为即将到来的交配季节做好准备.三疣梭子蟹交配活动主要集中在10月份.12月份精巢精小叶中出现游离精子.此后,精巢逐渐退化(1-3月),但输精管中一直储存着精液和精荚.三疣梭子蟹精荚形成过程始于前输精细管,精荚膜两层,分别由来自前输精细管分泌的第一种不定物质和其中、后端分泌的第二种不定物质.精荚内的基质可能直接源于精巢的精小叶部分.输精管中的强嗜酸性颗粒与第二种不定物质有融合作用,使输精管中贮存的精荚膜嗜酸性.     

长江口及毗邻海域三疣梭子蟹种群生物学特征及与环境的关系

根据2006年6月、8月和10月在长江口及其毗邻海域三疣梭子蟹底拖网资源调查资料,分析此区域三疣梭子蟹生物量和资源密度的分布特征及其与环境因子的关系。分析结果表明,三疣梭子蟹主要分布在N 31°~33°之间的海域,其生物量和资源密度在调查中有明显的变化,10月最高,8月次之,6月最少;种群组成中的雌雄比季节性变化明显,6月,雌性个体占优,之后雌雄比趋于相等,至10月,两者比为1∶1;头胸甲宽和体质量呈现幂函数的关系,7—9月是三疣梭子蟹体质量增长最快的时期;在所得环境因子中,溶解氧和NO2是影响三疣梭子蟹分布的最为重要的环境因子。与20世纪90年代数据相比,三疣梭子蟹的资源量有了一定的恢复,这可能与增殖放流和生态环境的改善有关。     

海州湾秋季三疣梭子蟹肥满度时空变化及其影响因素

根据2011年及2013―2021年秋季于海州湾及其邻近海域进行的10个航次的渔业资源底拖网调查数据,采用Le cren状态指数作为肥满度指标,分析了三疣梭子蟹(Portunus trituberculatus)肥满度的时间变化及空间分布规律,并应用广义可加模型(generalized additive model, GAM)评估不同性别三疣梭子蟹肥满度的影响因素。研究表明,海州湾秋季三疣梭子蟹肥满度的雌雄差异不明显且变化趋势相似,从2013年到2020年平均肥满度呈现递减的趋势,但在2021年出现高值。在空间分布上,肥满度较高的个体主要分布于20 m等深线以深区域。在肥满度的影响因素方面,头胸甲宽、水深、种群密度以及底层盐度对雌性个体肥满度影响明显,而叶绿素a浓度、头胸甲宽、底层水温以及底层盐度对雄性三疣梭子蟹的肥满度具有显著影响。本研究初步阐明了不同性别三疣梭子蟹肥满度的影响因素,为资源的开发和保护提供了科学依据。     

基于图像识别技术研究不同海区三疣梭子蟹甲壳白色斑纹特征及蜕壳前后斑纹特征的变化

为进行三疣梭子蟹外观特征的研究,本研究通过计算机视觉技术建立三疣梭子蟹背部白色斑纹评估方法,经人工对比验证其准确性达100%。在此基础上,对来自海域的30只三疣梭子蟹[平均体质量(2.8±0.56)g]进行跟踪观察,发现蜕壳前后白色斑纹数量和位置保持不变,而白色斑纹面积和斑纹区域面积则会随着蜕壳后蟹壳面积的增加而扩大;蜕壳后,白色斑纹面积增加89.33%±8.61%,斑纹区域面积增加90.51%±7.95%,蟹壳面积增加94.66%±8.26%,均显著正相关。为进一步研究不同海区梭子蟹白色斑纹分布特征,对来自朝鲜、中国辽宁和浙江等不同海区的三疣梭子蟹进行白色斑纹个数(N)、白色斑纹面积(S1)、斑纹区域面积(S2)、白色斑纹面积百分比(R1)、斑纹区域面积百分比(R2)和密度(D)等6项参数分析,结果发现,不同海区三疣梭子蟹斑纹性状存在显著特点,朝鲜海区呈现斑纹小、分布面积少、占蟹壳总面积比例低的特点;辽宁海区呈现斑纹小、分布面积多、占蟹壳总面积比例高的特点;浙江海区呈现斑纹大、分布面积少、占蟹壳总面积比例低的特点。本研究将计算机视觉技术应用于水产养殖研究,通过建立评估参数阐明三疣梭子蟹白色斑纹的生长规律,揭示了不同群体的三疣梭子蟹白色斑纹具有地理多样性的特征。     

放流三疣梭子蟹遗传多样性和贡献率初步研究

近年来我国沿海三疣梭子蟹自然资源明显下降,为恢复现有资源,辽宁省自2012年开始,连续实施了4年大规模人工放流工作。在目前的增殖放流中,为检测参与繁殖的三疣梭子蟹亲本和即将放流的子代间的遗传差异以及亲本对其子代的繁殖贡献率水平,利用10对具有丰富遗传多态性的微卫星分子标记,分别对9只三疣梭子蟹雌性亲本和179只即将放流的子代进行了遗传多样性分析及亲子鉴定。结果发现,子代的平均观测杂合度、平均期望杂合度和平均多态性信息含量数值均低于参与繁殖的雌性亲本,亲本和放流子代之间在观测杂合度、多态性信息含量参数方面并无显著差异(P0.05),但期望杂合度遗传参数存在显著差异(P0.05),子代的遗传多样性较雌性亲本呈下降的趋势。使用4个微卫星分子标记时,累积排除率≥0.998,亲子鉴定的准确率为56.98%;微卫星分子标记为6个时,累积排除率≥0.9999,准确率达到97.21%;当使用8个微卫星分子标记鉴定时,准确率达100%。同时发现,9只雌性亲本对子代均有贡献,最高为29.61%,最低为3.35%,不同亲本之间的贡献率存在显著差异(P0.05)。研究表明,微卫星可以作为有效的标记手段用于三疣梭子蟹增殖放流遗传评估中。上述试验结果将为我国三疣梭子蟹增殖放流的科学发展提供基础资料。     

盐度对三疣梭子蟹成熟前后呼吸代谢的影响

为了探讨三疣梭子蟹对不同盐度水平的生理响应以及成熟前后的生理变化,实验采用水化学和酶学方法,研究了盐度(15、20、25、30和35)对成熟前后三疣梭子蟹耗氧率、排氨率、氧氮比(O∶N)以及己糖激酶(HK)、丙酮酸激酶(PK)、琥珀酸脱氢酶(SDH)和乳酸脱氢酶(LDH)4种呼吸代谢酶活力的影响。结果发现:1.盐度与规格的交互作用对三疣梭子蟹呼吸代谢无显著影响(P>0.05);2.盐度对三疣梭子蟹耗氧率的影响不显著(P>0.05),但显著影响其排氨率(P<0.05);成熟前后的三疣梭子蟹耗氧率和排氨率差异显著(P<0.05);3.盐度对三疣梭子蟹O∶N未产生显著影响(P>0.05),三疣梭子蟹成熟前O∶N显著高于成熟后(P<0.05);4.除LDH外,盐度对三疣梭子蟹呼吸代谢酶活力影响不显著(P>0.05);除鳃中SDH和LDH外,三疣梭子蟹呼吸代谢酶活力随生长发育发生显著变化(P<0.05)。研究表明,规格对三疣梭子蟹呼吸代谢的影响高于盐度,长时间盐度驯化可能减弱甚至消除三疣梭子蟹在不同盐度水平下代谢的差异。     

Gonadotropin-releasing hormone (GnRH) neuronal systems in the pejerrey, Odontesthes bonariensis (Atheriniformes)

The brain of the pejerrey (Odontesthes bonariensis) has recently been shown to contain three forms of gonadotropin-releasing hormone (GnRH): salmon GnRH (sGnRH), chicken GnRH-II (cGnRH-II) and pejerrey GnRH (pjGnRH), nevertheless neuroanatomical studies on the distribution of these peptides are lacking. In this study we investigated the distribution of immunoreactive GnRH in the brain of adult pejerrey. Four different policlonal antisera and a monoclonal antibody against different GnRH variants were applied on cryosections and visualized using the ABC method. Three antisera (PBL#49, sGnRH#2 and cII741) revealed three different immunoreactive areas: the terminal nerve ganglion (at the junction between the olfactory bulbs and the anterior telencephalon), the preoptic area just anterior to the hypothalamus and the midbrain tegmentum. Fibers immunoreactive to GnRH were detected in different brain areas: the olfactory bulbs, the ventral thelencephalon, the hypothalamus, the mesencephalic area and an important innervation entering into the pituitary gland. Two other antibodies (LRH13 and s1668) labeled the two nuclei corresponding to the forebrain but not the midbrain tegmentum. As both antibodies have low crossreactivity to cGnRH-II, the data suggest that this group of cells express cGnRH-II. In summary, three different areas with immunoreactivity to GnRH were detected in the pejerrey brain. The distribution of sGnRH, pjGnRH and cGnRH-II expressing neurons, is discussed.     

Pituitary gonadotropin-releasing hormone (GnRH) receptor activity in goldfish and catfish: seasonal and gonadal effects

The goldfish pituitary contains two classes of gonadotropin-releasing hormone (GnRH) binding sites, a high affinity/low capacity site and a low affinity/high capacity site (Habibiet al. 1987a), whereas the catfish pituitary contains a single class of high affinity GnRH binding sites (De Leeuwet al. 1988a). Seasonal variations in pituitary GnRH receptor binding parameters, and the effect of castration on pituitary GnRH receptor binding were investigated in goldfish and catfish, respectively. In goldfish, GnRH receptors undergo seasonal variation with the highest pituitary content of both high and low affinity sites occurring during the late stages of gonadal recrudescence. The observed changes in pituitary GnRH receptor content correlate closely with responsiveness to a GnRH agonistin vivo in terms of serum gonadotropin (GTH) levels. In catfish, castration results in a two-fold increase in pituitary GnRH receptor content, which can be reversed by concomitant treatment with androstenedione, but not by the non-aromatizable androgen 11β-hydroxyandrostenedione; changes observed in GnRH receptor content correlate with variations in serum GTH levels and responsiveness to a GnRH agonist. In summary, the present study provides a clear evidence for seasonal variation in pituitary GnRH receptor activity in goldfish, and demonstrates a gonadal feedback mechanism regulating GnRH receptor activity in the catfish pituitary.     

Role of neuropeptide Y (NPY) in the regulation of reproduction: study based on catfish model

Significance of NPY in the regulation of GnRH–LH axis was evaluated. Considerable NPY immunoreactivity was seen in the components like olfactory system, basal telencephalon, preoptic and tuberal areas, and the pituitary gland that serve as neuroanatomical substrates for processing reproductive information. Close anatomical association as well as colocalizations of NPY and GnRH were seen in the olfactory receptor neurons, olfactory nerve fibers and their terminals in the glomeruli, ganglion cells of nervus terminalis, medial olfactory tracts, fibers in the ventral telencephalon and pituitary. In the pituitary, NPY fibers seem to innervate the GnRH as well as LH cells. Intracranial administration of NPY resulted in significant increase in the GnRH immunoreactivity in all the components of the olfactory system. In the pituitary, NPY augmented the population of GnRH fibers and LH cells. HPLC analysis showed that salmon GnRH content in the olfactory organ, bulb, preoptic area+telencephalon and pituitary was also significantly increased following NPY treatment. NPY may play a role in positive regulation of GnRH throughout the neuraxis and also up-regulate the LH cells in the pituitary.     

The existence of gonadotropin-releasing hormone (GnRH) immunoreactivity in the ovary and the effects of GnRHs on the ovarian maturation in the black tiger shrimp Penaeus monodon

Immunocytochemical localization using antibodies against five isoforms of gonadotropin-releasing hormone (GnRH), namely, luteinizing hormone-releasing hormone (LHRH), salmon (s)GnRH, octopus (oct)GnRH, lamprey (l)GnRH-I, and lGnRH-III, showed that only lGnRH-I immunoreactivity (ir-lGnRH-I) was localized in follicular cells of proliferative, vitellogenic, and mature ovaries. The effects of exogenous GnRHs on the ovarian maturation cycle of Penaeus monodon were compared by treating female broodstocks with LHRH, sGnRH, and lGnRH-I. The cycle of ovarian maturation in both eyestalk-ablated and eyestalk-intact shrimp administered with the three isoforms of GnRH was significantly shorter than that of the control animals. Moreover, administrations of all GnRH isoforms showed similar numbers of spawned eggs and the percentage of successful fertilization as in the control animals. These findings suggest that GnRHs may be highly conserved peptides that play an important role in inducing the ovarian maturation in the shrimp.     

Distinct expression of GnRH genes in the red seabream brain

This paper reports the molecular cloning of a cDNA encoding the precursor of seabream gonadotropin-releasing hormone (prepro-sbGnRH) and the localization of salmon GnRH (sGnRH) and seabream GnRH (sbGnRH) expressing neurons in the brain of the red seabream (Pagrus major). The cloned prepro-sbGnRH cDNA has a 285 bps open reading frame encoding a 23 amino acid signal peptide, a 10 amino acid sbGnRH, the cleavage site (Gly-Lys-Arg), and a 59 amino acid GnRH-associated peptide. The expression of sGnRH and sbGnRH peptides, and prepro-sGnRH and prepro-sbGnRH mRNA were studied using immunocytochemistry and non-radioactive in situ hybridization, respectively. We found cell bodies that reacted positively with both the sGnRH cRNA probe and anti-sGnRH serum, but not with the sbGnRH cRNA probe or anti-sbGnRH serum in the ganglion of the terminal nerve. Cell bodies that reacted positively with the sbGnRH cRNA probe, anti-sbGnRH serum, and anti-sGnRH serum, but negatively with the sGnRH cRNA probe were found in the preoptic area (POA). Immunocytochemistry showed that a distinct bundle of axons arises in the POA which projected to the pituitary gland. These results suggest that sbGnRH is the most relevant hypophysiotropic form of GnRH.     

Extrapituitary gonadotropin-releasing hormone (GnRH) binding sites in goldfish

In teleosts, as in other vertebrates, the secretion of pituitary gonadotropin (GTH) is mediated by the hypothalamic decapeptide, gonadotropin-releasing hormone (GnRH). Recent findings in teleosts indicate that GnRH receptors are not restricted to the pituitary gonadotropes and are also associated with somatotropes as well as being present in a number of other tissues. In the present study, we provide novel information on GnRH binding in a number of extrapituitary tissues in goldfish. However, we do not intend to provide full characterization of GnRH binding sites in various extrapituitary tissues in goldfish as this would clearly be outside the scope of this paper. In this study we examined GnRH binding in a number of extrapituitary tissues in goldfish and observed specific binding in ovary, testis, brain, liver and kidney. No specific GnRH binding was observed in muscle, skin, gut, gill and heart. In general, the present findings together with the results of other studies carried out in our laboratory demonstrate that mature goldfish ovary and testis contain two classes of GnRH binding sites, high affinity/low capacity and low affinity/high capacity sites with binding characteristics similar to those of the pituitary GnRH receptors. The brain of goldfish was also found to contain two classes of GnRH binding sites, a super-high affinity/low capacity and a low affinity/high capacity sites. Furthermore, study of goldfish liver and kidney demonstrated the presence of a single class of GnRH binding sites with characteristics different from those of pituitary, ovary, testis and brain. Overall, it is evident that goldfish contains a family of GnRH binding sites which can be classified into four groups based on binding affinities: 1) A class of high affinity binding sites present in the pituitary, ovary and testis, 2) a class of super high affinity sites so far only detected in the brain, 3) a class of intermediate-affinity GnRH binding sites in the liver and kidney, and 4) a class of low affinity binding sites present in all the tissues containing specific GnRH binding sites except for liver and kidney.     

In vivo activity of native GnRHs and their analogues on ovulation in the African catfish, Clarias gariepinus (Burchell)

Four distinct forms of native gonadotropin‐releasing hormone (GnRH) and two newly designed analogues were tested for their in vivo activity to induce ovulation in African catfish. The effects of these peptides on ovulatory parameters were compared with those of carp pituitary and [d ‐Ala⁶, Pro⁹‐NEt]‐mammalian GnRH analogue (mGnRHa), two tested ovulation‐inducing agents in African catfish. Assessment of ovulation was carried out by determining the ovulation ratio and the relative quantity of egg produced. From the results of the experiments, the order of potency of the native GnRH peptides is summarized as chicken GnRH‐II (cGnRH‐II) >salmon GnRH (sGnRH) >mammalian GnRH >chicken GnRH‐I (cGnRH‐I). Chicken GnRH‐II was as potent as mGnRHa while cGnRH‐I was totally ineffective. The new d ‐Orn⁶‐cGnRH‐II and d ‐Orn⁶‐sGnRH with a substitution at position 6 with d ‐isomer residue were as potent as the most extensively used mGnRHa, indicating that the position 6 modification might be more crucial than the substitution at the C‐terminal. On the basis of our results, the potential use and incorporation of cGnRH‐II and sGnRH for the development of more generic spawning induction therapies are suggested.     

Characterization of the receptor for gonadotropin-releasing hormone in the pituitary of the African catfish,Clarias gariepinus

Receptors for gonadotropin-releasing hormone (GnRH) were characterized using a radioligand prepared from a superactive analog of salmon GnRH (sGnRH), D-Arg⁶-Pro⁹-sGnRH-NEt (sGnRHa). Binding of¹²⁵I-sGnRHa to catfish pituitary membrane fractions reached equilibrium after 2 h incubation at 4°C. Displacement experiments with several GnRH analogs as well as other peptides, demonstrated the specificity of¹²⁵I-sGnRHa binding. Specific binding was enhanced in the presence of the cation chelator ethylene bis (oxyethylenenitrilo) tetra-acetic acid (EGTA), indicating an inhibitory effect of cations on GnRH-receptor binding. The binding of¹²⁵I-sGnRHa to pituitary membranes was found to be saturable at radioligand concentrations of 5 nM and above. A Scatchard analysis of the saturation data suggested the presence of a single class of high-affinity binding sites (Ka=0.901±0.06×10⁹M^–1, Bmax=1678±150 fmol/mg protein). A comparative study on¹²⁵I-sGnRHa binding to pituitary membrane fractions of male and female catfish, indicated that there were no differences in binding affinity and binding capacity between both sexes. The results demonstrate the presence of specific, saturable GnRH receptors in the African catfish pituitary.