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...     

石鲽脑垂体组织生理学研究

石鲽(Kareius bicoloratus)脑垂体呈鸡心形,由神经垂体和腺垂体两部分组成。腺垂体又包括前外侧部(RPD)、中外侧部(PPD)和垂体中间部(PI)。神经纤维束延伸至腺垂体的各个部分。采用HE、Jafri、HA、Mallory和PAS染色法,结合免疫组织化学方法,对石鲽脑垂体的6种内分泌细胞进行鉴别和定位。RPD部位主要含有PRL和ACTH细胞,PI部位主要含有MSH细胞。PPD部位有4类细胞,分别为GH、Gt H、TSH和嫌色细胞。GH、Gt H和TSH细胞数量随鱼体生长发育而变化,Gt H在性腺成熟期达到高峰,GH细胞和TSH细胞在生长早期数量多。     

大黄鱼脑垂体组织学与免疫组织化学

用组织学和免疫组织化学方法研究不同发育时期大黄鱼（Pseudosciaena crocea）脑垂体的形态结构和各种促激素分泌细胞的分布。结果表明：（1）大黄鱼脑垂体呈“鸡心“形,由神经垂体和腺垂体组成,腺垂体可分为前外侧部（RPD）、中外侧部（PPD）和垂体中间部（PI）。（2）RPD部有3种细胞,分别为催乳激素（PRL）细胞,促肾上腺皮质激素（ACTH）细胞,促性腺激素（GtH）细胞。（3）PPD部位于腺垂体腹面,也有3种细胞,其中1种为促生长激素（GH）分泌细胞,呈嗜酸性,另2种呈嗜碱性,分别为GtH细胞与促甲状腺激素（TSH）分泌细胞。（4）垂体中间部有2种细胞,1种为促黑激素（MSH）细胞,呈嗜酸性,另1种为GtH细胞,呈嗜碱性。随性腺发育成熟,大黄鱼GtH细胞的分布从中外侧部扩展至前外侧部和中间部背面;GtH细胞胞质空泡可作为其分泌活动的标志。     

长吻鮠脑垂体的组织学观察

应用Azan、PAS-MB、AF以及H．E染色方法，时长吻鮠脑垂体的形态组织结构和注射催产激素前后的GTH分泌细胞分别进行了观察研究。结果表明。长吻鮠脑垂体由腺垂体和神经垂体构成。神经垂体较简单,腺垂体结构较复杂，明显分为前腺垂体（RPD）、中腺垂体（PPD）和后腺垂体（PI）。腺垂体内可鉴别出7种激素分泌细胞，其中RPD内2种：为促肾上腺皮质激素（ACTH）分泌细胞和催乳激素（PROL）分泌细胞；PPD内3种：为生长激素（GH）分泌细胞、促甲状腺激素（TSH）分泌细胞、促性腺激素（GTH）分泌细胞；PI内有2种：为促黑色素激素（MSH）分泌细胞和1种嫌色细胞。     

中华鳖脑垂体的显微与超微结构观察

为了探讨中华鳖脑垂体的组织结构及其季节性变化,采用H.E染色、三色染色和免疫组织化学染色法在光镜下观察中华鳖脑垂体的显微结构,结合电镜对鳖脑垂体进行超微显微观察。结果发现中华鳖脑垂体同样分为神经垂体和腺垂体,但是缺乏中间部。腺垂体包括结节部和远侧部,远侧部可分头叶和尾叶两部分,头叶大于尾叶,神经部很小。腺垂体中的细胞可分为嗜酸性细胞、嗜碱性细胞和嫌色细胞,各类细胞的分布和形态有明显区别。免疫组化染色证明促肾上腺皮质激素(ACTH)细胞密集分布于结节部,在远侧部也有零星分布;促性腺激素(GTH)细胞和生长激素(GH)细胞数量在繁殖季节明显多于冬季。电镜下根据细胞大小、细胞核的形状和胞质中分泌颗粒特征可将腺垂体中的细胞分为GH细胞、ACTH细胞、GTH细胞、催乳激素(LTH)细胞、促甲状腺激素(TSH)细胞和嫌色细胞。在这些细胞间有明显的细胞连接。细胞形态和数量在繁殖季节和冬季有明显变化。     

牙鲆生长抑素基因的表达及细胞定位

采用RACE和荧光定量PCR技术克隆牙鲆生长抑素(SS)基因cDNA序列并且分析了其在仔鱼期的表达规律,同时通过免疫细胞化学染色,对牙鲆胃肠道和脑垂体等器官的生长抑素分泌细胞进行定位。最终获取包含完整编码区的牙鲆SS基因cDNA序列,其编码区长为381 bp,推测编码的蛋白质为126个氨基酸,其前23个氨基酸为信号肽,分子量为14.3 ku,理论等电点为8.33。牙鲆仔鱼不同发育时期的SS基因表达量比较结果表明,出膜后仔鱼体内SS基因表达不断增加,到变态发育期变化不大,变态后期又开始显著增加。免疫组织化学染色结果表明,SS细胞存在于胃、胰腺和脑垂体中。在胃中,生长抑素分泌细胞主要分布于胃上皮层;在胰腺中,生长抑素分泌细胞零散地分布,血管周围较多;在脑垂体中细胞密度高,集中在腺垂体的周边区域。     

硬骨鱼下丘脑对腺垂体的控制

<正> 鱼类的性腺发育和生殖活动是由脑下垂体控制和调节的,而脑下垂体又在下丘脑控制之下。脑受到刺激和冲动又影响下丘脑分泌激素释放因子的活动。神经冲动通过单胺类神经递质调节激素的分泌。故而研究下丘脑和脑垂体之间的关系对鱼类的性腺发育和繁殖是极其重要的。一下丘脑和脑垂体解剖学上的联系从形态解剖来看,鱼类的下丘脑通过垂体柄与脑垂体相连。神经垂体的分泌细胞位于下丘脑内,神经纤维广泛分布于腺垂体内,从而把脑和腺垂体联系起来。硬骨鱼有由正中隆起和门脉系统合成的神经垂体咀部(Holmes,Ball,1974)。正中隆起的毛细血管丛是供给神经垂体的纵行毛细管丛。     

大西洋牙鲆(Paralichthys dentatus)仔、稚、幼鱼形态观察

文中对大西洋牙鲆仔、稚、幼鱼各期发育时间及特征进行观察记录,并对各发育阶段进行显微摄像。大西洋牙鲆室内人工育苗水温18℃～22℃,盐度30,pH8.2,光控500Lx左右。饵料为强化的轮虫、卤虫无节幼体和配合饲料。初孵仔鱼肠道细而直,孵化后3d～4d肛门开通并开口摄食,6d～7d卵黄囊吸收完毕,仔鱼前期阶段结束。仔鱼后期阶段,消化道弯曲,冠状幼鳍完全形成,右眼开始上升。稚鱼期右眼完全迁移至左侧,变态完成,营底栖生活。幼鱼期基本具有成鱼的外部形态和习性。     

漠斑牙鲆胚胎及仔稚鱼发育的形态学观察

对漠斑牙鲆(Paralichthys lethostigma)胚胎及胚后发育进行观察,描述各发育时期的发育时序和形态特征.在水温18.5±0.5℃,盐度32～33的条件下,受精卵历时49 h脱膜而出,孵化后仔鱼在水温20.5±0.5℃时,4日龄开口摄食,26日龄进入变态期,45日龄左右完成变态成为幼鱼.漠斑牙鲆的卵子为浮性卵,油球一个.刚孵化仔鱼的消化道细直,3日龄肛门开通,4日龄卵黄囊消耗殆尽,开口摄食.在原肠期胚体下包约1/2时,在视囊和胚盘上出现点状星状的黑色素细胞.孵化后体两侧菊花状黑色素均匀分布;进入变态期后,体两侧的幼体色素逐渐收缩消失,同时在有眼侧出现个体较小的圆形成体色素细胞.在早期发育过程中,伴随着冠状鳍条的发生、伸长和缩短吸收.无鳔器官的发生.     

褐牙鲆(♀)×犬齿牙鲆( ♂)杂交子一代胚胎发育及仔稚鱼形态学观察

杂交是目前水产育种中应用最为广泛、最有成效的育种途径,为了选育一个能够促进中国海水鱼类养殖业发展,改善目前养殖褐牙鲆发病严重的现状的牙鲆类养殖新品种,对褐牙鲆(Paralichthys olivaceus)(♀)×犬齿牙鲆(P.dentatus)(♂)的杂交进行了研究。在Nikon MS800解剖镜下对褐牙鲆(♀)×犬齿牙鲆(♂)人工杂交子一代的胚胎及胚后发育进行观察,并使用Nikon数码相机拍摄其发育特征,描述了各发育时期的发育时序和形态特征。杂交子一代胚胎发育可分为卵裂前期、卵裂期、囊胚期、原肠期、神经胚期、器官发生期、尾芽期、肌肉效应期、脱膜孵化期共9个时期,除克氏泡出现时期以及胚体开始扭动时期具有相对特异性外,胚胎发育过程与褐牙鲆相比无显著差异。在水温(16.5±0.5)℃,盐度32~33的条件下,受精卵历时67 h完成孵化。(19±0.5)℃水温下,仔鱼3 d以前营内源性营养(卵黄囊仔鱼),3 d开口摄食营外源性营养(前弯曲期仔鱼),20 d左右进入变态期(弯曲期仔鱼),40 d后基本完成变态进入稚鱼阶段,生活方式由浮游转为底栖生活,50 d左右全身被鳞。同时观察了自胚胎发育时期至幼鱼期体表色素的分布及形态变化过程,描述了仔鱼变态期冠状鳍条、眼位置和尾部的发育、变化过程。早期生活史无鳔器官发生。     

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.     

Pituitary gland morphogenesis and ontogeny of adenohypophyseal cells of Salminus brasiliensis (Teleostei,Characiformes)

In this study, we describe for the first time the details of the pituitary gland morphogenesis and the ontogeny of adenohypophyseal cells of a South American Characiform species with great importance for Brazilian Aquaculture, Salminus brasiliensis (Characiformes, Characidae), from hatching to 25 days after hatching (dah), by histochemical and immunocytochemical methods. The pituitary placode was first detected at hatching (0 dah), and the pituitary anlage became more defined at 0.5 dah. The neurohypophysis (NH) development started at 3 dah, and the early formation of its stalk at 12.5 dah. An increase in adenohypophyseal and NH tissues was also observed, and in juveniles at 25 dah, the pituitary displayed similar morphology to that found in adults of this species, displaying the main features of the teleost pituitary. PRL cells were detected at 0.5 dah, together with ACTH and α-MSH cells, followed by GH and SL cells at 1.5 dah. β-FSH cells were detected at 25 dah, while β-LH cells at 5 dah. The pituitary development in this species comprises a dynamic process similar to other teleosts. Our findings in S. brasiliensis corroborate the heterogeneity in the ontogeny of adenohypophyseal cells in teleosts and suggest a role for adenohypophyseal hormones in the early development of this species.     

Positive feedback control by the gonads on gonadotropin (GTH) and gonadoliberin (GnRH) levels in experimentally matured female silver eels,Anguilla anguilla

Treatment of sham-operated female silver eels with carp pituitary extract stimulated ovarian development and induced increases in pituitary gonadotropin (GTH) and gonadoliberin (GnRH) contents. Both effects of carp pituitary extract were abolished in ovariectomized eels, indicating the involvement of the gonads. Endogenous sexual steroids, the secretion of which was increased during sexual maturation, should be responsible for the stimulation of GTH and GnRH levels. Ovariectomy itself had no significant effect on pituitary GTH and GnRH contents, reflecting the fact that, at the silver stage, sexual steroid levels are too low to exert any significant effect on pituitary GTH and GnRH. The positive feedback control exerted by the gonads on GTH and GnRH levels during sexual maturation, in the eel as well as in some other teleosts, would produce an amplification of the pubertal stimulation of the hypothalamo-pituitary-gonadal axis.     

Effects of LED light spectra on the growth of the yellowtail clownfish <Emphasis Type="Italic">Amphiprion clarkii</Emphasis>

Growth hormone (GH) is an essential polypeptide required for the normal growth and development of vertebrates. We have studied the effects of light-emitting diodes (LEDs) emitting different spectra (red, green, and blue) on the GH of yellowtail clownfish Amphiprion clarkii. Full-length GH cDNA from the pituitary of the yellowtail clownfish was first cloned and then the expression of GH mRNA under different light spectra was measured. GH mRNA expression was significantly higher under green and blue light than under red light spectra. These results indicate that in yellowtail clownfish, short-wavelength LED enhances growth more than long-wavelength LED, and that LED lights are more effective for enhancing growth than white fluorescent bulbs. Injection of melatonin resulted in significantly higher expression levels of GH mRNA compared to the control. We therefore conclude that green and blue light enhance GH levels and that melatonin plays a role in modulating growth of the yellowtail clownfish.     

Effect of photoperiod manipulation on daily rhythms of melatonin and reproductive hormones in caged European sea bass (Dicentrarchus labrax)

There are few studies regarding the daily changes in reproductive hormone profiles in teleosts. The aim of the present research was to investigate the effects of photoperiod manipulation on daily rhythms of melatonin and reproductive hormones (pituitary and plasma LH, testosterone [T] and 11-ketotestosterone [11KT]) in juvenile sea bass, kept in net cages under farming conditions in winter. Our results provide the first evidence for the presence of daily variations in pituitary and plasma LH and plasma T in sea bass. Artificial lights suppressed circulating plasma melatonin levels and significantly affected the daily rhythm of LH storage and release.     

Immunocytochemical localization of gonadotropins during the development of XX and XY Nile tilapia

Gonadal development and steroidogenesis in teleosts is regulated by two gonadotropic hormones; luteinizing hormone (LH) and follicle-stimulating hormone (FSH). Earlier studies in tilapia have shown that FSH-β and LH-β appear by 14 days after hatching (dah), results from the current study corroborate with these previous reports in tilapia. Here we demonstrate the appearance of LH in pituitary between 14 and 20 dah. In addition to this the present study primarily focuses on any possible differences in appearance of LH-β and FSH-β immunoreactivity between XX and XY population of Nile tilapia. LH immunoreactivity was found to be lower in pituitary of XX fish when compared to XY fish. The development of FSH-β immunoreactivity in pituitary of the Nile tilapia is also presented. Overall, it remains to be established what significance these findings on the appearance of gonadotropins hold for sex differentiation in tilapia. F. Sakai and I. Swapna contributed equally.     

半滑舌鳎(Cynoglossus semilaevis)新型膜孕激素受体基因(mPRL)在卵母细胞成熟过程中的表达特征

为进一步提高半滑舌鳎(Cynoglossus semilaevis)人工育苗技术的水平,对半滑舌鳎新型膜孕激素受体(mPR-like,mPRL)基因的表达特征和作用机制进行了研究.应用实时定量PCR方法分析mPRL mRNA在卵子形成过程中的时序表达,发现半滑舌鳎mPRL mRNA相对表达量的最高值出现在性成熟阶段卵巢的Ⅴ时相卵母细胞.原位杂交分析mPRL mRNA在繁殖相关组织的细胞学定位,发现mPRL mRNA分布在半滑舌鳎性成熟阶段卵巢的卵母细胞膜上;在脑的神经元和垂体内分散的细胞中,mPRL mRNA阳性信号较强.制备半滑舌鳎mPRL的多克隆抗体,采用Westem blotting方法检测半滑舌鳎mPRL蛋白在不同组织的表达特征,发现mPRL蛋白的表达量在卵巢、脑、垂体中相对较高,在肝脏、头肾、肾脏中表达量相对较少.免疫组化结果显示,半滑舌鳎mPRL蛋白在卵巢、脑和垂体的细胞学定位与mPRL mRNA定位一致.应用实时定量PCR和Westem blotting方法检测促性腺激素调控下半滑舌鳎不同时相卵母细胞mPRL基因mRNA和蛋白的表达变化,结果显示,促性腺激素对半滑舌鳎卵母细胞中mPRL基因mRNA和蛋白表达都有一定的上调作用,特别是对Ⅴ时相卵母细胞中mPRL mRNA和蛋白的表达量提升明显;发现表达量与促性腺激素调控作用具有剂量依存关系.半滑舌鳎mPRL在繁殖相关组织的表达特征表明其通过脑-垂体-卵巢轴参与繁殖调控,同时也揭示了mPRL介导卵母细胞成熟机制.     

Stimulation of gonadal and germ cell development in larval and juvenile carp (Cyprinus carpio L.) by homologous pituitary extract

The data presented show that in larval carp, gonadal size has increased distinctly after treatment with homologous pituitary extract (PE). Moreover, the precocious onset of primordial germ cell proliferation and of sex differentiation into male and female gonads was induced. Body weight of treated and control specimens did not differ significantly from each other throughout the experiment. Treatment of juvenile carp with homologous PE led also to an increase in gonadal size without concomitant increase in body weight. Induction of precocious spermatogenesis was observed too. GTH-levels in control and PE-treated carp were monitored by means of a homologous radioimmunoassay, showing that in PE-treated carp the GTH-level was distinctly elevated. The possible role of pituitary hormones in larval and juvenile gonadal development is discussed.