卵黄蛋白原研究及其进展

甲壳动物和昆虫由于进化上亲缘关系近,被统称泛甲壳动物,大多数泛甲壳动物卵黄蛋白原为雌性特异性蛋白,是卵母细胞中储存的卵黄蛋白的前体,可为胚胎发育提供营养和能量,是决定繁殖性能的关键因素,在生殖发育中起着十分重要的作用。已有研究证明,调控泛甲壳动物卵黄蛋白原的主要激素有保幼激素、蜕皮激素、神经肽和胰岛素样肽等。昆虫保幼激素和蜕皮激素发挥促进作用,而神经肽和胰岛素样肽对卵黄蛋白原合成的调控作用因种类不同而存在不同。除了以上激素,甲壳动物特有的眼柄高血糖激素家族以及促雄腺激素(IAG)对卵黄蛋白原的合成起负调控作用。本文总结了近年来泛甲壳动物卵黄蛋白原合成调控机制的相关研究,概括并比较了调控昆虫与甲壳动物的卵黄蛋白原合成的主要激素及调控机制,为养殖虾蟹甲壳动物生殖调控相关研究提供参考。

     

EH和MIH调控甲壳动物蜕皮和生殖的分子机制进展

甲壳动物性早熟是抑制甲壳养殖业发展的主要原因之一,而影响甲壳动物发育主要激素:蜕皮激素(Ecdysteroid hormone,EH)和蜕皮抑制激素(Molt-inhibiting hormone,MIH)是决定甲壳性早熟的主要原因,本文对甲壳动物Y器官和蜕皮激素、X器官窦腺复合体和蜕皮抑制激素研究上的应用进展,并着重综述蜕皮抑制激素对蜕皮激素的调控机制,以期为水生甲壳类的相关研究提供参考。     

甲壳动物眼柄神经内分泌系统研究概述

眼柄X器官-窦腺复合体(XO-SG)被称为甲壳动物的内分泌调控中心,如同开关一样控制着虾蟹等甲壳动物的生长代谢和生殖发育等重要的生理活动。文中综述了眼柄神经内分泌系统的结构和功能、神经肽类激素的种类和作用机制,重点阐述了甲壳动物眼柄神经肽类激素分泌和调控研究进展,以期为研究和实现在生产上控制虾蟹的繁殖和生长提供参考。     

甲壳动物CHH家族神经激素结构和功能研究进展

甲壳动物主要利用温度、光照周期等外界因子调节其生理状态 ,使它们的生殖活动处于最适条件下 ,来自外界因子的这些信息作用于甲壳动物的中枢神经系统 ,后者将其传递到神经内分泌系统和内分泌系统 ,神经内分泌系统和内分泌系统能够分泌一些促进因子和抑制因子以实施对性腺活动的调控。由于甲壳动物成体的生殖和蜕皮常常交替出现 ,因此 ,神经内分泌系统和内分泌系统的精确调控非常重要。甲壳动物高血糖激素 (ＣＨＨ)家族神经激素是甲壳动物特有的多肽激素 ,它们主要由眼柄的Ｘ -器官窦腺复合体合成 ,它们包括 :甲壳动物高血糖激素 (ＣＨＨ)、…     

脊尾白虾翻译控制肿瘤蛋白基因TCTP克隆及自噬调控相关基因在卵巢发育期的表达

为深入了解脊尾白虾(Exopalaemon carinicauda)卵巢发育和卵黄蛋白原发生的分子机制,本研究克隆得到脊尾白虾翻译控制肿瘤蛋白基因TCTP,并结合自噬调控基因TCTP、Hif-1α、Beclin1和Bcl-2在卵巢发育期的表达水平,阐释脊尾白虾卵黄蛋白原合成过程中的分子调控特征。研究显示,脊尾白虾TCTP基因c DNA全长为732 bp,编码168个氨基酸,具有典型的TCTP1和TCTP2功能域以及PKC和TKⅡ等mTOR信号通路相关的磷酸化位点。同时发现,甲壳动物TCTP普遍缺乏在其他动植物中高度保守的C末端的cys残基。进化分析显示,脊尾白虾TCTP与中华绒螯蟹(Eriocheir sinensis)亲缘关系最近。4种自噬调控基因在脊尾白虾卵巢发育期的表达结果显示,肝胰腺TCTP基因从增殖期到产后恢复期呈递减趋势;肝胰腺Hif-1α、Beclin1和Bcl-2基因表达趋势相似,即从增殖期到小生长期高表达,大生长期极显著下降,表达量最低(P0.01),这与外源性卵黄蛋白原的合成趋势大致相反。这些自噬调控基因可能通过自噬作用共同调节外源性卵黄蛋白原的合成。卵巢TCTP基因在小生长期表达量最高;卵巢Hif-1α基因从增殖期到产后恢复期持续升高,这与内源性卵黄蛋白原表达趋势相似;卵巢Beclin1基因在大生长期表达量最高,与脊尾白虾卵巢Ec R表达趋势相似,与卵巢Bcl-2基因表达趋势相反,这些自噬调控基因可能通过自噬作用共同促进内源性卵黄蛋白的合成。本研究表明,自噬调控基因TCTP、Hif-1α、Beclin1和Bcl-2在脊尾白虾卵巢发育时期相互协调共同作用,可能通过自噬作用调节脊尾白虾卵黄蛋白原的合成和卵巢发育。     

甲壳动物蜕皮的调节机制研究进展

综述了甲壳动物蜕皮现象调节机制的研究进展,包括Y-器官、X-器官及其窦腺复合体、蜕皮激素、蜕皮抑制激素及其起始表达和作用机理。     

十足目甲壳动物卵黄蛋白原的生化特性及生物合成

十足目在甲壳动物中种类最多,经济价值最大,分为枝鳃亚目(对虾类)和腹胚亚目(真虾、螯龙虾、蟹类)两大类。十足目种类卵黄蛋白原基因c DNA全长基本都在8 kb左右,编码2500~2600个氨基酸组成的多肽,具有高度保守性。然而这两类生物的卵黄发生机制却存在显著差异。人工育苗过程中,对虾类卵巢不易发育,通常需要切除眼柄促熟,而腹胚亚目种类容易成熟甚至需要控制早熟。对虾类的卵黄蛋白原由卵巢和肝胰腺共同合成,一般卵巢贡献更大。尤其在卵巢发育前期,卵巢合成卵黄蛋白原较肝胰腺显著旺盛,具有"卵巢内源基因表达型"的特征。在真虾等腹胚亚目种类中,卵巢虽然也参与合成卵黄蛋白原,但以肝胰腺为主,绝大多数种类肝胰腺的贡献率超过90%。本文探讨了两类甲壳动物在卵黄发生机制中的显著差异与其在人工控制条件下性腺发育的不同表现之间的相互关系。     

实时荧光定量PCR检测凡纳滨对虾和罗氏沼虾卵黄蛋白原mRNA在卵巢和肝胰腺中的表达

卵黄磷蛋白作为卵黄蛋白的主要成分, 可为甲壳动物胚胎和早期幼体发育提供能量, 为研究其来源及合成规律, 实验应用SYBR GreenⅠ荧光定量PCR法检测了凡纳滨对虾和罗氏沼虾性腺不同发育时期卵巢和肝胰腺两种组织中卵黄蛋白原mRNA的表达水平。结果发现,凡纳滨对虾和罗氏沼虾的卵巢和肝胰腺中都有卵黄蛋白原mRNA的表达。随着卵巢的发育, 凡纳滨对虾卵巢中卵黄蛋白原mRNA的相对表达量在前5个阶段不断增加, 分别为1.1, 5.9, 10.4, 26.9, 85.0, 恢复期急剧减少, 为1.6。肝胰腺中的相对表达量也不断增加, 分别为1.3, 3.3, 7.1, 37.3, 51.6, 恢复期急剧减少, 为1.0。罗氏沼虾肝胰腺中卵黄蛋白原mRNA的相对表达量在前四个阶段不断增加, 分别为3.4, 12.6, 15.2, 38.9, 抱卵期急剧减少, 为2.9;而卵巢在整个发育过程中对卵黄蛋白原合成的贡献比较小, 分别为1.0, 1.3, 1.7, 4.8, 1.5。研究表明, 两种虾类的肝胰腺和卵巢均具合成卵黄蛋白的功能, 而且在不同的卵巢发育阶段呈现明显的规律性。     

GnIH多肽对半滑舌鳎(Cynoglossus semilaevis)下丘脑生殖相关基因表达的影响

本研究首次通过下丘脑离体孵育的方法研究促性腺激素抑制激素(Gonadotropin-inhibitory hormone,GnIH)多肽对半滑舌鳎(Cynoglossus semilaevis)下丘脑中生殖相关基因的表达调控。研究结果显示,tsGnIH-1促进了gnrh2和gnih的表达,对gnrh3和kiss2的表达无影响;tsGnIH-2抑制了gnrh3的表达,对gnrh2、kiss2和gnih的表达无影响。GnIH多肽对生殖相关基因的不同调控表明同一前体蛋白编码的不同GnIH多肽在生殖调控中的作用不尽相同。本研究结果增加了对GnIH参与鱼类生殖调控机制的认识,为深入研究奠定了基础。     

虾蟹类眼柄激素的生理生化及基因克隆研究进展

甲壳动物内分泌学研究主要集中在虾蟹类、虾蟹类内分泌学研究对于揭示甲南动物的生长与繁殖以及指导经济虾蟹类养殖生产实践具有重要意义，尤其是眼柄内分泌激素在调控甲壳运物的蜕皮、变态发育、生殖、血糖平衡及体内的渗透压等方面起着重要的作用，一直是神经内分泌学的研究热点^[1,2]。本主要对虾蟹类的眼柄神经内分泌系统的结构、激素和种类及其生理生化特性，特别是基因结构及克隆等方面作一综述，为进一步开展虾蟹类内分泌学研究提供参考资料。     

Thyroid hormone modulation of ovarian recrudescence of air-breathing catfish Clarias gariepinus

In the present study, thiourea-induced thyroid hormone depletion and thyroxine (T₄) ‘overdose’ were used as a strategy to understand the influence of thyroid hormones on ovarian recrudescence of juvenile (3-months-old), immature (8-months-old) and adult (1-year-old) air-breathing catfish, Clarias gariepinus. Thiourea-induced thyroid hormone depletion in juvenile catfish impaired ovarian development, but no significant effect was observed in immature catfish and during late stage of ovarian recrudescence of mature catfish. T₄ treatment in females undergoing late stages of ovarian recrudescence induced rapid oocyte growth by promoting its early entry into maturational phase as evident from the presence of more number of vitellogenic and post-vitellogenic follicles, decrease in aromatse immunoreactivity and reduced estradiol–17β levels. Hence, thyroid hormones have an important role to play during early stages of ovarian development and vitellogenesis of catfish and also indicating that thyroid has a stage dependent effect on ovary.     

In vitro methyl farnesoate secretion by mandibular organs isolated from different molt and reproductive stages of the crab Oziotelphusa senex senex

ABSTRACT:   Considerable evidences indicate that mandibular organ (MO) of crustaceans secrete terpenoid hormone (methyl farnesoate [MF]), which regulates molting and reproduction. In the freshwater south Indian rice field crab Oziotelphusa senex senex , MO isolated during premolt and vitellogenesis, secreted greater amounts of MF, which indicates that the regulation of the crustacean molt and reproduction is complex and also involves MF, besides steroid (ecdysteroid) and peptide (sinus gland peptide) hormones.     

中国对虾肝胰腺_卵巢及血淋巴中的_省略_雌二醇含量的生殖周期变化

首次证实了孕酮和雌二醇这两种类固醇激素在中国对虾体内的存在.在性腺未发育阶段,肝胰腺、卵巢和血液中两种激素的含量均很低氐,难以检测到.而在卵黄发生前期(核仁周边期),三种组织中孕酮和雌二醇含量迅速上升,卵巢的雌二醇含量达到高峰(450.1±86.7).进入初级卵黄发生阶段,三种组织中,两种激素均具较高含量,卵巢和肝胰腺的孕酮含量(分别为1975 1±175.2和902.6±130.5pg/g)以及血淋巴中的雌二醇含量(451.3±73.7)达到高峰.到了次级卵黄发生阶段,孕酮和雌二醇的含量迅速下降,肝胰腺等组织中儿乎检测不到.对虾性腺指数(GSl)的增长既显著且有规律性,每一期增长幅度都达到或超过l00%.肝胰腺指数(HSI)从性腺未发育期(3.4±04)到卵黄发生前期(4.9±0 7)以及从卵黄发生前期到初级卵H黄发生期(6.3±1.0)有显著的增长,而从初级卵黄发生期到次级卵黄发生期HSI增长不显著(6.7±1.2).肝胰腺指数的增长与两种类固醇激素含量的变化具相似的趋势.上述结果显示,孕酮和雌二醇可能具有刺激和凋控中国对虾性腺发育的作用,肝胰腺可能是卵黄蛋白原的合成场所.     

Oocyte development and serum profiles of vitellogenin and steroid hormone levels in captive female Pacific herring Clupea pallasii during their first maturational cycle

ABSTRACT:   The present study investigates the relationship between oocyte development and serum steroid hormone levels in captive Pacific herring, Clupea pallasii , during the first reproductive cycle. The process of oocyte development in Pacific herring belongs to the group-synchronous type. Maturity of the ovary was divided into six periods based on histological observation (i.e. immature (April to September), onset of vitellogenesis (August to October), progress of vitellogenesis (October to December), completion of vitellogenesis (December to March), maturation and spawning (March to April) and spent (late April)). The pattern of seasonal change in the gonadosomatic index (GSI) well reflected the ovarian maturity. Serum vitellogenin levels showed good correlation with change in GSI, which increased from September to a peak (4.2 ± 0.3 mg/mL) in March. Serum estradiol-17β (E2) levels elevated from September and reached a peak (15.8 ± 4.2 ng/mL) in December, and remained comparatively high until March, suggesting that the active vitellogenin synthesis during vitellogenesis is controlled by the high E2 level. 17,20β-Dihydroxy-4-pregnen-3-one showed a single sharp peak (2.4 ± 0.28 ng/mL) in early April of the second year, suggesting it was a maturation-inducing steroid in this species.     

中国对虾卵黄蛋白原合成部位的研究

十足目甲壳类卵巢中卵黄的来源,在过去的几十年研究中一直存在争议,内源性合成和外源性合成都有报道。本文以雌性中国对虾为实验材料,根据组织学观察确定其发育阶段,试验虾可以分为:卵巢未发育期;卵黄发生前期;初级卵黄发生期和次级卵黄发生期4个时期。从处于不同卵巢发育期对虾的卵巢和肝胰腺中提取总RNA,用RT-PCR的方法初步探讨不同发育期的中国对虾卵巢和肝胰腺中卵黄蛋白原mRNA的表达,确定是否存在卵黄蛋白原的合成功能。结果发现,从卵巢未发育期到次级卵黄发生期的卵巢和肝胰腺中都有卵黄蛋白原mRNA的表达,说明二者为卵黄蛋白的合成部位。     

克氏原螯虾大颚器对卵巢发育的影响

采用埋植和离体方法进行克氏原螯虾的大颚器（ＭＯ）对卵巢发育作用的研究,埋植大颚器７次能显著提高成熟系数和促进卵径增大,处于不同卵巢发育时期的克氏原螯虾大颚器提取物（ＭＯＥ）的离体研究发现,卵黄发生期的ＭＯＥ对初级和次级卵黄发生期卵径增大均有极显著的作用,而对卵黄发生前期的卵径增大无作用,处于卵巢发育早期的ＭＯＥ对卵黄发４生期卵巢小块总ＲＮＡ含量升高无显著作用,处于卵黄发生前期,卵黄发生期和恢复期卵     

Stimulation of ovarian growth by methyl farnesoate and eyestalk ablation in penaeoidean model shrimp,Sicyonia ingentis Burkenroad, 1938

The effect of methyl farnesoate (MF) administration on the vitellogenesis of the penaeoidean shrimp, Sicyonia ingentis, was studied. The short‐ and long‐term treatment effects as well as the effect of two MF injection regimens (0.1 and 1.0 μg MF/injection) were evaluated. The studies were also carried out to understand the pattern of vitellogenesis in eyestalk ablated adult and juvenile shrimps. A combination of endpoints, haemolymph vitellogenin (Vg) levels, gonadosomatic index (GSI) and histology, was used to study the effect of these treatments. The GSI increased in all the MF‐treated shrimp compared with the control shrimps. Although haemolymph Vg levels declined over the experimental period in all the treatments, the Vg levels decreased significantly only in the short‐term treatment with 1.0 μg MF. Similarly, haemolymph protein level also declined over the experimental period in all the treatment groups. However, except in the long‐term treatment with 0.1 μg MF, all treatments showed a significant decrease in haemolymph protein level. Conversely, in all eyestalk ablated adults and juveniles, haemolymph Vg, total protein and GSI increased over the experimental period, all of which were higher than the concurrent control. The discrepancy in the vitellogenic pattern between MF‐treated and eyestalk ablated shrimp was possibly due to the difference in the ovarian phase of the initial control. Although unilateral eyestalk ablation failed to induce vitellogenesis in juveniles, bilateral ablation induced vitellogenesis, which indicates that juveniles are competent to undergo vitellogenesis.     

Endocrine changes during the onset of vitellogenesis in spring in the mosquitofish

We examined short-term changes in neuroendocrine function during the onset of vitellogenesis by introducing female mosquitofish into a warm environment. Activation of FSH cells occurred prior to vitellogenesis, which was characterized by the production of estradiol-17β (E2) by follicles and following vitellogenin (Vg) synthesis in hepatocytes. Incorporation of Vg into the oocytes was detected less than three days after fish were transferred into a warm aquarium.     

In vitro effect of pituitary,interrenal and gonadal hormones on vitellogenin synthesis in primary hepatocyte cultures of Chalcalburnus tarichi

Vitellogenin (Vtg) is an important precursor yolk protein in egg‐laying vertebrates, including fish. The 17β‐oestradiol (E2) plays a crucial role in the Vtg synthesis; moreover, certain hormones can stimulate Vtg synthesis. We investigated the possible role of E2, carp recombinant growth hormone (crGH), insulin (Ins), progesterone (P4) and 11‐deoxycortisol (11‐DOC) hormones in Vtg synthesis on primary juvenile Chalcalburnus tarichi hepatocyte culture. The amount of Vtg in the medium was measured at 2‐day intervals using enzyme‐linked immunosorbent assay (ELISA). The hepatocytes were maintained in culture for more than 2 weeks without the addition of serum components. Vitellogenin localization was visualized with the immunofluorescence method in E2‐supplemented hepatocytes. Among hormones applied to the culture, only E2 had an influence on Vtg synthesis in a time‐dependent manner, while crGH, Ins, P4 and 11‐DOC had no effect. However, in hepatocytes stimulated with E2 in combination with P4, a lower Vtg production was seen compared with Vtg produced when hepatocytes were stimulated with E2 alone. P4 proved to have potentiating effects on co‐treatment with E2‐induced Vtg production. As a result, E2 and P4 are the most important hormones for Vtg synthesis in juvenile C. tarichi hepatocyte culture.