拟穴青蟹受精膜形成过程中环形颗粒的发生与胞吐

为了揭示拟穴青蟹受精膜形成机制,采用透射电子显微镜研究了其受精膜形成过程中环形颗粒的形成与胞吐。结果显示,拟穴青蟹精子入卵后,发生皮层反应而形成受精膜。皮层反应包括致密颗粒首先胞吐以及环形颗粒的相继多轮胞吐。每轮环形颗粒的胞吐参与到受精膜的平缓期、增厚期和举起期3个阶段。环形颗粒的形成主要发生在受精膜的增厚期,受精卵的皮层和靠近皮层的内质中都在形成大量的环形颗粒,并且迅速移到皮层质膜附近发生爆发性胞吐,参与到受精膜的快速形成。环形颗粒的形成与2种卵黄颗粒以及脂滴密切相关,在皮层中发现了线粒体参与其中,推测在拟穴青蟹受精卵的皮层和内质可能存在环形颗粒合成的两套不同机制。本文证实了卵黄颗粒和脂滴及环形颗粒在拟穴青蟹受精膜形成过程中发挥重要作用。国家“八六三”高技术研究发展计划(2002AA603013);;福建省教育厅科技项目(JAT160256);;集美大学学科建设基金(C517064

甲壳动物精子学研究概况Ⅰ精子的形态与结构

甲壳动物精子学研究概况Ⅰ精子的形态与结构王艺磊1，2）张子平3）李少菁2）（集美大学水产学院养殖系1）厦门361021）（厦门大学海洋学系2）厦门361005）（香港城市大学生物与化学系3）九龙香港）关键词甲壳动物精子形态结构甲壳纲是整个动物界中...福建省自然科学基金;中国科学院昆明动物研究所细胞与分子进化开放实验室资

十足目受精生物学研究概述

由于十足目精子的特殊结构———无鞭毛、不运动、核呈凝絮状、顶体结构复杂，且受精过程迅速，特别是精卵识别、顶体反应，常常在瞬间发生，加上卵子富含卵黄而不易制片，使其受精生物学研究与其它精子具鞭毛的动物相比报道较少。本文就十足目受精生物学的研究做一简要的...福建省自然科学基

甲壳动物精子学研究概况Ⅱ.精子发生与精子的生化组成

甲壳动物精子学研究概况Ⅱ.精子发生与精子的生化组成王艺磊1，2）张子平3）李少菁2）（集美大学水产学院养殖系1）厦门361021）（厦门大学海洋学系2）厦门361005）（香港城市大学生物与化学系3）九龙香港）关键词甲壳动物精子发生生化组成在前文阐...福建省自然科学基金;中国科学院昆明动物研究所细胞与分子进化开放实验室资

Construction of Normalized cDNA Library from Hepatopancreas of Abalone Haliotis diversicolor supertexta Exposure to Tributyltin

用rdP试剂提取三丁基锡暴露诱导的杂色鲍肝胰腺总rnA,经OlIgOTEX纯化得到MrnA;应用SMArT技术合成双链CdnA,双链特异核酸酶(dSn)进行双链CdnA的均一化,构建了杂色鲍三丁基锡暴露诱导下的均一化CdnA文库。原始文库的库容为4.3x106Cfu/Ml,重组率为97.9%。从文库中随机挑选了3288个克隆进行测序,得到3048个高质量EST序列,其中有370条COnTIgS,2103条SInglETS,unIgEnES共2473条,冗余率为18.86%。以上结果说明该文库质量较好,为进一步筛选相关功能基因打下基础;较低的冗余率说明该文库值得继续使用大规模ESTS测序的方法寻找相关功能基因,并为进一步使用基因芯片技术研究相关功能基因的表达谱提供便利。A normalized cDNA library from hepatopancreas of abalone Haliotis diversicolor supertexta exposure to Tributyltin was constructed.Total RNA were prepared using RDP reagent.Oligotex(QIAGEN)was used to separate the mRNA from total RNA.The first strand cDNA was synthesized by transcription of mRNA with the SMART technique.The LD-PCR was performed using a modified SMART primer as the primer set,and the first-strand cDNA as the template to synthesize double strand cDNA.Double strand cDNA was normalized using Duplex-Specific Nuclease(DSN).The containing capacity of the unamplified cDNA library was 4.3×106 CFU/ml with a recombinant rate of 97.9%.A total of 3 288 clones were random selected to be sequenced,and 3 048 high quality ESTs were generated.After processing,a total of 2 473 unigenes comprising 370 contigs and 2 103 singlets were obtained.The redundancy was 18.86%.These results indicate that the normalized cDNA library is suitable to be used for further cloning and analysis of genes related to tributyltin exposure.厦门市科技计划项目(3502Z20055024);国家自然科学基金(20877034);集美大学创新团队基金(2008A001

中草药淫羊藿和菟丝子对日本鳗鲡卵巢发育的影响

在水温(21±1)℃下,给体重(592.5±52.5) g的日本鳗鲡(Anguilla japonica)雌亲鱼投喂每千克体重含0.034 g淫羊藿和0.034 g菟丝子浸膏的饲料90 d,对照组投喂人工配合饲料,研究淫羊藿和菟丝子对卵巢发育的影响。结果显示,饲料中添加淫羊藿和菟丝子的雌鱼卵巢大部分卵母细胞属第Ⅱ时相,性腺指数和肝体比均显著高于对照组(P<0.05);卵母细胞油滴明显增多,部分卵母细胞胞质已充满油滴,核仁变小增多;血清雌二醇(E2)和11-酮基睾酮(11-KT)水平显著升高(P<0.05),实验组11-KT含量约为对照组的4倍;肝脏卵黄蛋白原基因vtg表达量升高,实验组和对照组卵巢cyp19a1仅微量表达,肝脏未检测到erα和erβ的表达;实验组肌肉总脂肪酸、饱和脂肪酸(SFA)、单不饱和脂肪酸(MUFA)和高度不饱和脂肪酸(HUFA)含量均显著高于对照组(P<0.05),最主要的高度不饱和脂肪酸花生四烯酸(AA)、二十碳五烯酸(EPA)和二十二碳六烯酸(DHA)含量均显著高于对照组(P<0.05)。本研究表明,在亲鱼培育饲料中添加中草药淫羊藿和菟丝子促进了日本鳗鲡卵母细胞油滴和肝脏卵黄蛋白原增加,提高了肌肉高不饱和脂肪酸的积累,为卵黄生成和卵母细胞进一步发育做好更充分的准备。国家自然科学基金项目(41806193);;厦门市科技局项目(3502Z20133015);;鳗鲡现代产业技术教育部工程研究中心开放基金项目(RE201604,RE201503

Acrosome Reaction of Sperm in the Mud Crab Scylla serrata as a Sensitive Toxicity Test for Metal Exposures

【英文摘要】In order to test the sensitivity of the sperm cell of the mud crab Scylla serrata to heavy metals, the toxic effects of Ag+, Cd2+, Cu2+, and Zn2+ on the acrosome reaction (AR) were studied by artificially inducing the AR of sperm exposed to heavy metals, counting the AR rates by light microscopy, and observing structural changes in sperm by transmission electron microscopy. The AR in S. serrata occurs at two stages. The first stage (ARI) is the eversion of the subacrosomal material. The second stage (ARII) is the ejection of the acrosomal filament. The results showed the EC50 values of the AR based on (ARI + ARII)% for Ag+, Cd2+, Cu2+, and Zn2+ were 10.02, 2.14, 13.69, and 2.21 μg/L, and the EC50 values based on ARII % of Ag+, Cd2+, Cu2+, and Zn2+ were 1.96, 0.20, 1.46, and 0.34 μg/L. The order of toxicity is Cd2+ > Zn2+ > Cu2+ > Ag+ based on the percentage of reacted sperm at the second stage. Sperm cells exposed to heavy metals showed an increased rate of swelling, shape irregularities, and the acrosomal filament of some sperm cells was, crooked, ruptured, and even dissolved. The AR of the sperm cell from S. serrata is more sensitive to the tested heavy metals compared to sea urchin sperm cell toxicity tests

Studies on the Vision of Red of Sea Bream Chrysophrys major Temmick et Schlegel

本文采用组织学与电生理学相结合的方法,综合报导了浅海 底层经济鱼类-真鲷（Chrysophrysophry5maioir TemmincketSchlegel）的视网膜、视顶盖的结构，超微 结构和视觉特性，并探讨了与其栖息环境的相互关系以及结构本 身的功能意义。结果如下： 1、真鲷视网膜存在大量视杆、单锥和孪生双锥三种光感受 细胞。视网膜颞侧区为视锥高密度区，该区单位面积（0.01ｍｍ2） 有双锥128－134个，单锥64—70个，双锥与单锥之 比约为2：1。视网膜中央区和鼻侧区为视杆高密度区，该区单 位面积（0.01mm2)有视杆细胞5236—5089个。视 杆与视锥...By means of this histological and electrophysiological method, these studies deals with the morphological structure, ultrastructure and visual characteristics in the retina and optic tectum of the shallow sea benthic economic fish—red sea bream ( Chrysophrys major Temminck et Schlegel) comprehensively, and probe into the their relationship with its dwelling environment and the functional significa...学位：理学硕士院系专业：海洋与环境学院海洋学系_海洋生物学学号：xy00022

Reference gene validation for quantification of gene expression during final oocyte maturation induced by 17α, 20β-dihydroxy-4-pregnen-3-one in common carp(Cyprinus carpio)

本研究检测了18S rrnA(18S)、28S rrnA(28S)、组织蛋白酶z(CATHEPSIn z,CTSz)、延伸因子1-α(ElOngATIOn fACTOr 1-α,Ef-1α)、3-磷酸甘油脱氢酶(glyCErAldEHydE-3-PHOSPHATE dEHydrOgEnASE,gAPdH)、β肌动蛋白(β-ACTIn)6个候选内参基因在17α,20β双羟孕酮(dHP)诱导鲤(CyPrInuS CArPIO)卵母细胞最终成熟过程的表达情况,并应用不同的内参分析软件对数据进行了分析。bESTkEEPEr软件分析表明Ef-1α的变异系数和标准差是6个候选内参基因中最低的,且bESTkEEPEr指数在6个候选内参基因中最高,说明其表达稳定性最高,适合做为内参基因;gEnOrM软件分析认为需要同时使用Ef-1α和CTSz两个内参基因来校正目标基因的表达;nOrMfIndEr软件分析同样表明Ef-1α是6个候选内参基因中表达最稳定的;最后通过rEffIndEr软件综合比较分析,确定Ef-1α相对于其他5个候选内参基因,在17α,20β双羟孕酮诱导卵母细胞最终成熟阶段表达最为稳定,可作为内参校正17α,20β双羟孕酮诱导鲤卵母细胞最终成熟阶段各基因的表达情况。Final oocyte maturation is a key step for successful spawning and fertilization.Like most other vertebrates, teleosts have full-grown postvitellogenic oocytes in the ovary that are physiologically arrested at the G2/M border in the first meiotic prophase and cannot be fertilized.Shortly before ovulation, the oocytes must become fully mature, which involves breakdown of the germinal vesicle(GVBD), chromosome condensation, assembly of the meiotic spindle, and exclusion of the first polar body.Meiosis is again arrested at the second metaphase.The time period from the resumption of meiosis to the second meiotic metaphase has been referred to as final oocyte maturation.Knowledge of the molecular events and the role of various factors involved in final oocyte maturation is a focus of research in the field of reproductive biology.Quantitative real-time PCR(qPCR) is often used to quantify transcript abundance in such studies; however the technique is subject to considerable experimental error and variation.A stably expressed housekeeping gene is typically used as a reference gene to normalize this variation.However an ideal universal reference gene that is stable under all experimental circumstances has not been described.Researchers should validate their reference genes before performing qPCR analysis.We documented the expression of six candidate reference genes: 18 S rRNA(18S), 28 S rRNA(28S), cathepsin Z(CTSZ), elongation factor 1-α(EF-1α), glyceraldehyde-3-phosphate dehydrogenase(gapdh), and β-actin during 17α, 20β-dihydroxy-4-pregnen-3-one(DHP) induced final oocyte maturation in common carp(Cyprinus carpio).We used a range of software to evaluate the expression stability of these reference genes.Analysis with Bestkeeper revealed that EF-1α had the lowest CV%(2.65) and STDEV(0.71) values, but had the highest Bestkeeper index value(0.956) among the six candidate reference genes, suggesting that EF-1α was the best reference gene in the present study.Analysis with geNorm revealed that EF-1α and CTSZ had the lowest M(gene expression stability) values which indicated that they were the most stable genes among the six.Additionally, the value of pairwise variation of these two genes(V2/3) was much lower than the default(0.15), suggesting that EF-1α and CTSZ should be used concurrently as a reference gene pair to normalize the expression of the target genes.Consistently, analysis with NormFinder and RefFinder demonstrated that EF-1α was the most stable gene among the six candidates.Hence, EF-1α can serve as a reference gene to normalize the expression of target genes during final oocyte maturation induced by DHP in common carp.The mRNA expression of luteinizing hormone receptor gene(LHR) during final oocyte maturation induced by DHP was normalized by 6 candidate reference genes respectively, and the relative expression was varied significantly depending on the reference gene that was used.Selection of a stably expressed reference gene is critical for all qPCR analysis to ensure accurate target gene mRNA expression information.集美大学创新团队基金项目(2010A001

STUDIES ON STRUCTURE of THE RETINA AND VISUAL CHARACTERISTICS of PAGROSOMUS MAJOR -Ⅲ. VISUAL CHARACTERISTICS of PAGROSOMUS MAJOR

视网膜电图（Erg）结果显示，真鲷PAgrOSOMuSMAJOr视网膜具混合型特征。强光明适应后，b波阈值在80MIn回复至暗视水平。明适应过程中，b波振幅存在d型和r型。其光谱敏感曲线在强背景光下（lOgIb＝1.8）发生浦氏位移，峰值从484nM移至525nM，且从颜色光适应对Sλ曲线的影响等都表明，真鲷存在两种以上的光感受系统，且具辨色能力。浦氏位移的光强水平较高，说明辨色能力不会很强。The waveForms of ERG reveal the Features of mixed retina.AFter preexposure to high intensity light, the threshold For eliciting a contant b-wave return to the dark adapted level in a period of 80 mintltes.According to the level of light adaptation, the D-type and the R-type of the time course of change in b-wave sensitivity are observed in the retina.When the intensity of white background light is higher (logIB=-1.8) the spectral sensitivity curve occurs Purkinje shiFt, the peak shiFts From 484nm to 524nm.The change of the spectral sensitivity curve under chromatic light adaptation.All these show that the retina has more than two types of receptor system and has the ability of color discrimination, but is not strong