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1.
Androgens mediate a wide range of physiological responses and developmental processes in vertebrates, involving both reproductive and nonreproductive systems. The activity of androgens is mediated by the androgen receptor (AR), a member of the nuclear receptor superfamily. In this study, an AR gene was cloned from the large yellow croaker (Larimichthys crocea) for the first time. qRT-PCR revealed ubiquitous expression of AR in all adult tissues examined, with higher expression in the gonad and liver of both sexes and highest expression in the blastula stage of embryonic development. Using in situ hybridization, we detected positive signals of AR in the spermatogonium, spermatocyte, spermatid, and spermatozoon during spermatogenesis, in the cytoplasm of all oocytes during oogenesis and in the follicle cells of stage IV oocytes. Our findings support the important role that AR plays in gametogenesis, gonadal development, and the early stages of embryonic development.  相似文献   

2.
The transformer‐2 gene (tra‐2) plays a key role in the sexual differentiation regulatory hierarchy. In this study, tra‐2 gene homologs designated as Mntra‐2 was cloned and characterized from Macrobrachium nipponense. The full‐length cDNA of Mntra‐2 consists of 1724 bp with an open reading frame (ORF) encoding 192 amino acids, an 827 bp 5′‐untranslated region (UTR) and a 318 bp 3′‐UTR. The predicated molecular mass of Mntra‐2 was 20.805 kDa with an estimated theoretical isoelectric point of 10.36. The deduced amino acid sequence shares high homology with Penaeus monodon. Real‐time quantitative polymerase chain reaction (RT‐qPCR) analyses demonstrated that the expression levels of Mntra‐2 varied significantly during different developmental stages of embryogenesis, larvae, and post‐larvae and in various adult tissues. During embryogenesis, the expression level of Mntra‐2 was slightly higher at the cleavage stage than at the blastula stage, and reached the highest level at the nauplius stage. During the larvae, the Mntra‐2 expression gradually increased from 1 d larvae post hatch (L1) to L10 and decreased to a lowest level at the end of metamorphosis. During the post‐larvae, the Mntra‐2 expression was higher level at the 5 d after the metamorphosis (P5). RT‐qPCR showed the Mntra‐2 mRNA was expressed in ovary, testis, muscle, heart, abdominal ganglion, brain, and intestine with the highest level of expression in muscle and intestine. The results indicate that Mntra‐2 is an arthropods tra‐2 homolog and probably plays important roles in embryonic development and sex differentiation of M. nipponense .  相似文献   

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4.
为了解GnRH基因在中华鳖(Pelodiscus sinensis)性腺和胚胎发育过程中的表达特征,采用cDNA末端快速扩增(RACE)技术从中华鳖全脑中获得与生长生殖调控密切相关的GnRH1基因全长cDNA,并运用实时荧光定量PCR(qRT-PCR)技术检测GnRH1在成鳖不同组织和胚胎发育时期的表达水平。结果显示:中华鳖GnRH1基因cDNA全长546 bp,其中5′非编码区(5′UTR)99 bp,3′非编码区(3′UTR)168 bp,开放阅读框(ORF)279 bp,编码92个氨基酸,分子质量为10.23 ku,理论等电点pI为5.65,具有N端信号肽(1~23 aa)、核心十肽区域(24~33 aa)、断裂位点GKR(34~36 aa)及相关肽区域(37~92 aa),符合GnRH蛋白典型结构特征。系统进化树结果显示,中华鳖GnRH1基因和绿海龟(Chelonia mydas)、墨西哥箱龟(Terrapene carolina mexicana)及西部锦龟(Chrysemys picta bellii)GnRH1基因聚为一支。qRT-PCR结果表明,GnRH1基因在中华鳖雌雄个体的8个组织中均有表达,在脑和性腺组织中高表达,且具有性别差异,雄性中华鳖中的表达显著高于雌性(P<0.05);在10个胚胎发育时期均表达,且随发育时间的后移,表达量显著增加,在第16期达到峰值。GnRH1基因可能在中华鳖生长及性腺分化中具有重要作用。  相似文献   

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7.
采用RT-PCR和cDNA末端快速扩增技术(rapid amplification of cDNA ends,RACE),首次克隆了西伯利亚鲟(Acipenser baerii)的CXCR7a和CXCR7b基因的全长cDNA。CXCR7a的cDNA全长为2 325 bp(登录号为:JQ034508),包括207 bp的5'端非翻译区(UTR)、可编码362个氨基酸的1 086 bp的开放阅读框(ORF)及包括poly(A)尾巴在内的大小为1 032 bp的3'端非翻译区(UTR);CXCR7b的cDNA全长为2 423 bp(登录号为:JQ034509),包括209 bp的5'端非翻译区(UTR)、可编码370个氨基酸的1 110 bp的开放阅读框(ORF)及包括poly(A)尾巴在内的大小为1 104 bp的3'端非翻译区(UTR),并对它们编码的蛋白质序列的分子特征进行了分析。氨基酸序列相似性(similarity)分析表明,CXCR7a与人(Homo sapiens)、小鼠(Mus musculus)、非洲爪蟾(Xenopus laevis)、斑马鱼(Danio rerio)的CXCR7相似性分别为82.6%、81.4%、80.9%、81.0%;CXCR7b与人、小鼠、非洲爪蟾、斑马鱼的CXCR7相似性分别为82.9%、82.3%、80.8%、82.5%;西伯利亚鲟CXCR7a与其CXCR7b的相似性最高为96.5%。系统进化树分析显示,西伯利亚鲟与斑马鱼聚为一簇,两栖类聚为一簇,其它高等脊椎动物再进行聚类。对CXCR7a和CXCR7b在各发育时期胚胎及仔鱼的表达状况分别进行定性分析表明,CXCR7a在除卵裂期、囊胚期的胚胎外,其在原肠胚中期至出膜1 d的仔鱼的各时期中均具有表达,CXCR7b在卵裂期至出膜1 d的仔鱼的各时期中均具有表达。另外,从基因组DNA水平上表明CXCR7a和CXCR7b来自不同的基因拷贝。这些结果为进一步研究CXCR7在西伯利亚鲟侧线系统发育中的作用提供了新的基础资料。  相似文献   

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9.
Facilitative glucose transporter 1 (GLUT1) is a transporter protein for glucose transport via the plasma membrane of the cells to provide energy through carbohydrate metabolism. GLUT1 cDNA from Litopenaeus vannamei was obtained and analysed in this study. Full‐length GLUT1 cDNA is 2062 bp long and contained a 1506‐bp ORF encoding a 502 amino acid protein, a 270‐bp 5′UTR and a 284‐bp 3′UTR. When shrimp were under acute low salinity stress, the expression in hepatopancreas, muscle, gill and eyestalk was all up‐regulated at 12 h (P < 0.05) and 96 h (P < 0.05), while the expression in the four tissues was all down‐regulated at 6 h (P < 0.05) and 48 h (P < 0.05) . The expression in the muscle of shrimp at water salinity of 3 was lower than that at water salinity of 30 independent of dietary carbohydrate levels, while expression in hepatopancreas, gill and eyestalk was up‐regulated at 200 and 300 g kg?1 carbohydrate levels. The expression in all tissues fed glucose was up‐regulated when compared to the expression in shrimp held at a water salinity of 30. This study suggests that GLUT1 is a conserved protein in L. vannamei, and changes in expression due to environmental salinity and dietary carbohydrate level and source.  相似文献   

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11.
为探明鱼类消化液分泌的相关调控因子,采用RACE技术分别克隆了鳜(Siniperca chuatsi)两种肠道激素——胃泌素(gastrin,GAS)与胆囊收缩素(cholecystokinin,CCK)基因的cDNA序列。鳜GAS基因cDNA全长581 bp,5′非翻译区长100 bp,3′非翻译区长145 bp,开放阅读框长336 bp,编码111个氨基酸;鳜CCK具有2种类型:CCK1与CCK2,CCK1 cDNA全长为843 bp,5′非翻译区长60 bp,3′非翻译区长369 bp,开放阅读框414 bp,编码137个氨基酸;CCK2 cDNA全长846 bp,5′非翻译区长112 bp,3′非翻译区长332 bp,开放阅读框403 bp,编码134个氨基酸。鳜GAS与CCK成熟肽C末端具有相似的八肽结构(DYQGWVDF/DYLGWMDF),仅在C末端第3位和第6位氨基酸发生替换。荧光定量PCR分析表明,鳜GAS mRNA主要表达于肠和幽门垂,CCK1与CCK2 mRNA在脑中表达量最高,在肠和幽门垂也有较高表达,表明GAS与CCK同为消化调控因子,而CCK还是神经分泌因子。鳜GAS与CCK mRNA表达贯穿于整个幼体早期发育阶段(孵化后0～22 d),前期表达水平较高,后期表达水平较低,并趋于平稳,GAS与CCK mRNA发育表达水平变化可能与这一时期消化道生长发育旺盛有关。  相似文献   

12.
大黄鱼性别特异SNP标记的开发与验证   总被引：1，自引：1，他引：0  
大黄鱼是我国养殖量最大的海水经济鱼类,其雌鱼生长显著快于雄鱼,但两性的外部形态差异不明显,也没有异形性染色体,依靠传统方法无法对其活体准确进行生理性别和遗传性别的判别与鉴定,需要开发性别特异的分子标记。本研究从2尾雌鱼和2尾雄鱼、以及分别由50雌鱼与50尾雄鱼组成的2个混合样品的基因组重测序数据比较中筛选与性别显著关联的SNP位点,对其中11个位点分别设计引物在15尾雌鱼和15尾雄鱼中扩增出PCR产物进行Sanger测序验证,鉴定出1个与性别完全连锁的位点(SNP6,15尾雌鱼均为纯合、15尾雄鱼均为杂合)。然后,设计等位基因特异性PCR引物,其中包括2条雌性与雄性通用引物和1条雄性特异引物,在闽—粤东族与岱衢族大黄鱼合计近2 200个个体中进行扩增,结果在全部雌鱼中都只扩增出1个348 bp的条带,而在全部雄鱼中还扩增出1个194 bp的Y染色体特异条带,检出率达到100%。研究表明,大黄鱼属于XX♀-XY♂类型的性别决定。本研究鉴定出一个雄性特异SNP标记,并建立了一种新的大黄鱼遗传性别鉴定技术,为大黄鱼单性育种、基因组选择育种和性别决定分子机制研究提供了重要的技术手段。  相似文献   

13.
Moult‐inhibiting hormone (MIH), an important regulator of steroidogenesis, inhibits the synthesis of ecdysteroid in Y‐organ (YO) and plays a significant role in the regulation of moulting and post‐embryonic development of crustacean. Because unsuccessful moulting have been widely observed in precocious crabs, we investigated whether genetic variants in the 5′‐flanking region and 3′‐untranslated region (3′‐UTR) of the MIH gene are associated with precocity of the Chinese mitten crab. Thirty individual DNA samples were sequenced to search for SNPs in the 5′‐flanking region and 3′‐UTR of the MIH gene. Five SNPs (g.196 T>A, g.230 C>T, g.305 T>C, g.323 C>A and g.372 C>T) in the 5′‐flanking region and 6 SNPs (g.2677 C>T, g.2759 T>A, g.2807 T>C, g.3042 A>G, g.3088 T>G and g.3295 T>G) in the 3′‐UTR of the MIH gene were selected for the individual genotyping in a two‐stage association study. We found that a SNP g.3088 T>G in the 3′‐UTR of MIH gene was consistently associated with precocity of the Chinese mitten crab in stage 1 and stage 2, with a per‐allele OR (Odds Ratio) of 1.469 (95% CI: 1.169–1.844) after two stages combined (P = 0.001). However, no significant associations were observed between the other 10 SNPs and precocity of the Chinese mitten crab. To our best knowledge, this is the first association study between various SNP genotypes and phenotype attributes in Chinese mitten crab. Our findings suggest that the SNP g.3088 T>G may be a candidate marker for effective marker‐assisted selection to decrease the precocity of the Chinese mitten crab in future studies.  相似文献   

14.
李媛媛  蔡生力  刘红 《水产学报》2012,36(11):1667-1674
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。  相似文献   

15.
为建立大黄鱼肿大细胞病毒的培养方法,明确其分类地位,用肿大细胞病毒检测呈阳性的大黄鱼幼鱼病料 (FD201807和SA201808)肾组织匀浆液感染鳜仔鱼细胞系 (mandarin fish fry cell line-1,MFF-1)并连续传代,从病料组织匀浆液和细胞冻融液中提取病毒DNA,克隆病毒主要衣壳蛋白基因 (mcp),测序后与NCBI GenBank中的虹彩病毒科肿大细胞病毒属病毒mcp以及2018—2020年所检出的15株大黄鱼肿大细胞病毒mcp进行比对分析。结果显示,病毒传至第4代才可引起MFF-1细胞病变,细胞病变的主要特征为细胞脱壁、变圆、折光度增强;感染时间越长脱壁细胞越多,同时培养液中的颗粒增加;透射电镜下可见感染细胞的细胞质散在大小为130~150 nm的六边形病毒粒子和空壳。感染细胞的病变周期随传代代次的增加而缩短,第15代次的FD201807株感染细胞80%细胞病变的时间为3 d,第15代次的SA201808株感染细胞80%细胞病变的时间为7~8 d。mcp序列比对和聚类分析发现,SA201808株与FD201807株的mcp序列存在21个碱基差异,二者的mcp序列分别与大黄鱼虹彩病毒(large yellow croaker iridovirus, LYCIV) LYCIV-Zhoushan (GenBank: MW139932.1)和花鲈虹彩病毒 (Lateolabrax maculatus iridovirus, LMIV) (GenBank: MH577517.1)相近。15株从大黄鱼病料检出的肿大细胞病毒中,12株的mcp序列与SA201808株聚类;3株与FD201807聚类。本研究利用MFF-1细胞系分离培养了大黄鱼肿大细胞病毒,揭示了大黄鱼肿大细胞病毒存在差异,为更好地了解大黄鱼肿大细胞病毒提供了数据参考。  相似文献   

16.
O‐GlcNAc transferase gene (OGT) was considered as the sole rate‐limiting enzyme in the O‐GlcNAc modification. In the present study, the OGT gene of hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂) was cloned and characterized, and its expression in response to dietary carbohydrate level and acute glucose treatment was investigated. The full‐length of OGT (GenBank accession no. KY656469 ) was 4,063 bp, including a 302 bp 5′untranslated terminal region (UTR), a 3,165 bp coding region that encoded 1,054 amino acids residues and a 596 bp 3′ UTR. The highly conservation of OGT gene between fish and mammals was also observed through multiple sequences alignment and phylogenetic analysis. O‐GlcNAc transferase gene was ubiquitously expressed in all detected tissues with highest expressions in brain and liver, to a lesser degree, in eye, heart, kidney and intestine. The increasing dietary carbohydrate from 8.02% to 16.08% had no significant effect on the mRNA expression of OGT. However, the expression of OGT was slightly elevated at 6 hr post‐glucose injection, and the elevation became significant at 24 hr time‐point. These data may enhance our understanding on the nutritional regulation of OGT and O‐GlcNAc modification in fish species.  相似文献   

17.
Gonadotropin (GTH) hormones are glycoprotein which stimulates gonadal maturation in vertebrates. Follicle stimulating hormone is involved in initiation of gametogenesis and regulation of gonadal growth. FSHβ has been cloned and characterized from the brain of Catla catla. The FSHβ full‐length of cDNA sequence of 523 bp comprised 3, 394 and 128 bp of 5′‐UTR, open reading frame (ORF) 3′‐UTR respectively. The coding region of C. catla FSHβ encoded a peptide of 130 amino acids. Phylogenetic analysis of C. catla FSHβ deduced amino acid sequence showed high similarity with Gobiocypris rarus followed by goldfish, Carassius auratus. The qPCR result shows that FSHβ mRNA is mainly expressed in pituitary while moderate and low expression was observed in testis and ovary respectively. Chitosan‐nanoconjugated kisspeptin‐10 (CK‐10) of particle size 125 nm, polydispersity index of 0.335 to 0.65 and zeta potential of ?34.95 mV were synthesized and evaluated at against naked kisspeptin‐10 for their reproductive hormonal profile. Treatment of fish with CK‐10 showed controlled and sustained surge of the reproductive hormones (FSH & LH) with peak at 12 h. The hormone levels of naked kisspeptin‐10 treated fish decline after 6 h. The sustained release of this CK‐10 will help in reducing maturation age, synchronization of ovulation and spawning in fish. This is the first report on use of chitosan‐nanoconjugated kisspeptin‐10 (CK‐10) for reproduction in fish.  相似文献   

18.
The 26S proteasome, which includes 20S core and 19S regulatory particles, plays an important role in removing harmful and/or unnecessary proteins. In this study, we cloned a 2443‐bp full‐length cDNA sequence of the S7 subunit of 19S from the sea cucumber Apostichopus japonicus, including a 70‐bp 5′‐untranslated region (UTR) and 1070‐bp 3′‐UTR coding for a theoretical 435 amino acid protein. Sequence analysis showed that S7 belongs to the ATPases associated with a wide variety of cellular activities (AAA+) superfamily. S7 was widely expressed in different tissues as determined by real‐time quantitative polymerase chain reaction, and its expression was gradually upregulated in the regenerating body wall. Combined with histological observations, our results suggest that the S7 subunit or the 26S proteasome assembled based on S7 may be associated with the regulation of cell migration and cell cycle during body wall regeneration in A. japonicus.  相似文献   

19.
为比较日本鲭和大黄鱼肌肉中微生物和代谢功能的变化及其与鱼肉腐败特性之间的关系,本研究检测了2种鱼在冷藏过程中的理化指标和菌落总数的变化,利用Illumina Miseq测序技术分析细菌群落变化,并利用皮尔森相关性分析检验微生物与鱼肉腐败及组胺产生相关性,结合功能预测分析细菌群落组成与代谢功能之间的关系。结果显示,冷藏期间日本鲭和大黄鱼的pH、挥发性盐基氮、组胺、菌落总数等均呈上升趋势,且日本鲭上升较快;冷藏末期2种鱼TVB-N值和组胺含量分别达到76.34、59.98和59.92、3.11 mg/100 g;日本鲭肌肉中细菌丰富度和多样性先增加后减少,大黄鱼则整体呈下降趋势;2种鱼肌肉中的优势腐败菌均为希瓦氏菌属;日本鲭体内与TVB-N产生相关的菌共12种,其中10种与组胺产生具有显著相关性;大黄鱼体内与TVB-N产生相关的菌共7种,但未检测出与组胺产生具有相关性的细菌;冷藏过程中氨基酸代谢和碳水化合物代谢为最主要的代谢通路,日本鲭样品组氨酸、精氨酸、脯氨酸等氨基酸代谢相关基因和丁酸丁酯代谢、丙酸酯代谢及丙酮酸代谢丰度均显著高于同一时期的大黄鱼,本实验从微生物代谢水平解释了日本鲭比大黄鱼更易腐败的原因,为不同水产品腐败特性的研究提供新思路。  相似文献   

20.
为探究大黄鱼在高温胁迫条件下的蛋白表达情况，实验应用串联质谱标签tandem mass tag (TMT)标记结合液相色谱-串联质谱(LC-MS/MS)技术对大黄鱼耐高温组和不耐高温组的肝脏组织进行蛋白组学分析。共鉴定到3 369个蛋白，其中有687个差异表达蛋白，包括281个上调蛋白和406个下调蛋白。用平行反应监测(parallel reaction monitoring, PRM)对随机挑选的13个蛋白进行验证，其结果与TMT标记的蛋白组学分析结果一致。随后，通过生物信息学分析发现，高温胁迫对大黄鱼的蛋白质折叠、能量代谢有显著影响，其中热休克蛋白70 (heat shock proteins 70, HSP70)、钙网蛋白(calreticulin, CRT)和葡萄糖调节蛋白(glucose-regulated protein, GRP)参与调节蛋白质的正确折叠，丙酮酸激酶(pyruvate kinase, PK)参与高温条件下的能量供给。由此推测HSP70、CRT、GRP和PK在大黄鱼应对高温胁迫时发挥着重要的作用。  相似文献   

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