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Journal of Oceanology and Limnology 2018年第6期36卷 2288-2296页

作者：孙虎山王宜艳刘小莉刘东武 School of Life Science Ludong University Analysis and Testing Center Shandong University of Technology

Little is known about the roles of dynorphin and Kappa opioid receptor(KOR) in mollusks. In this study, we aim to determine the distribution of dynorphin A and KOR-1 in the digestive system of the scallop Chlamys farreri. Using immunohistochemical staining, we confirmed the expression of dynorphin A and KOR-1 in the digestive system of C. farreri. Dynorphin A immunopositive cells were identified in intestine and hepatopancreas. In intestine, small and spherical dynorphin A immunopositive cells(4–9 μm in diameter) were scattered among the long columnar epithelial cells(ECs). In hepatopancreas, cells containing masses(5–14 μm in diameter) of dynorphin A immunopositive products were observed in epithelium of acinis. These immunopositive cells may be synthetic and/or secretory cells of dynorphin A. Dynorphin A immunoreactive products were commonly observed in epithelium and connective tissue(CT) of labial palps, mouth labia and stomach, which presented in forms of grains, fibers or flakes. KOR-1 immunoreactive material was observed in ECs and CTs of labial palps, mouth labia and stomach, intestine and hepatopancreas. The distribution of both dynorphin A and KOR-1 in the digestive organs suggests an involvement of dynorphin via KOR-1 in the functional regulation of the digestive system of C. farreri.

关键词： Chlamys farreri dynorphin digestive system kappa opioid receptors immunohistochemistry

A Population Genetic Analysis of Continuously Selected Chlamys farreri Populations

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Journal of Ocean University of China 2018年第4期17卷 913-919页

作者： ZHANG Lu LI Yuli LI Yangping YANG Zhihui LI Yuqiang WANG Yangfan WANG Shi BAO Zhenmin Key Laboratory of Marine Genetics and Breeding (MGB) of Chinese Ministry of Education College of Marine Life Sciences Ocean University of China

This study applied an optimized one-step 2b-RAD library construction strategy and performed simplified genome sequencing of 539 individuals from three continuously selected Chlamys farreri populations. SNP screening was performed using RAD typing software and population genetic parameters for the continuously selected populations from three generations(G1, G2, G3) were determined. The results showed that the optimized one-step 2b-RAD library construction strategy greatly simplified the experimental process, making it suitable for efficiently constructing a large number of libraries. A total of 18450 SNP markers were identified, which evenly distributed throughout the genome. Population genetic analysis of these three generations showed that the mean value of observed heterozygosity was 0.275 ± 0.177, 0.272 ± 0.181 and 0.275 ± 0.166, respectively. Meanwhile, the mean value of expected heterozygosity was 0.275 ± 0.141, 0.274 ± 0.145 and 0.280 ± 0.133, respectively. The Wright's fixation index(F) was 0.04291, 0.04976 and 0.06685, respectively. Markers deviated from Hardy–Weinberg equilibrium accounted for 10.34%, 12.64%, and 23.11%, and the Shannon diversity index was 0.0999 ± 0.0404, 0.0921 ± 0.0388 and 0.0733 ± 0.0308. FIS(also known as the inbreeding coefficient) of the three populations was 0.0256, 0.0323 and 0.0468, respectively. We suggested that the 2b-RAD method is well suited to population genetic studies of aquacultured organisms. Moreover, our results indicated that the continuous selection affected the population genetic structure of the cultured Penglai-Red scallop, but the change was not significant; therefore, population selection should continue.

关键词： 2b-RAD population genetic Chlamys farreri SNP

Expression Characteristics of Odf3 in Gonad Are Different Between Chlamys farreri and Vertebrates

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Journal of Ocean University of China 2017年第1期16卷 121-128页

作者： MA Xiaoshi LIANG Shaoshuai YANG Dandan JI Aichang MA Xiaoru ZHANG Zhifeng Ministry of Education Key Laboratory of Marine Genetics and Breeding College of Marine Life Sciences Ocean University of ChinaQingdao 266003P.R.China

Outer dense fibers are specific subcellular components of mammal sperm flagella, apart from the axoneme and its associated proteins. As one of the outer dense fibers components, ODF3 is essential for the formation of mammal sperm flagella. In the present study, we isolated the Odf3 gene, 1033 bp in length, from Zhikong scallop Chlamys farreri, which was named as Cf-Odf3. It was predicted that Cf-Odf3 encodes a protein of 240 amino acids which contains 5 discontinuous Pro-Gly-Pro repeats. The Cf-Odf3 transcripts were detected in both testis and ovary of C. farreri at various stages through q RT-PCR. The Cf-ODF3 protein synthesized by prokaryotes was purified using Ni^(2+)-NTA affinity chromatography and used to produce its polyclonal antibody. The antibody product was effective and specific as was examined by Western blotting. The Cf-Odf3 transcript and Cf-ODF3 protein were located in intragonadal somatic cells(ISCs), spermatogonia and spermatocytes of C. farreri testes, while no transcription of Cf-Odf3 and synthesis of Cf-ODF3 were detected in spermatids and spermatozoa. The location of Odf3 expression is different from that of vertebrates where ODF3 is synthesized specifically in spermatids and sperm flagella. Moreover, the Cf-Odf3 expression was also detected in ISCs, oogonia and oocytes of the scallop ovaries. Our finding revealed a different characteristic of Odf3 expression between scallop and vertebrates, which implied that Cf-Odf3 played a role in the gametogenesis of C. farreri.

关键词： Odf3 germ cell gametogenesis gonad Chlamys farreri

Mapping Toll-Like Receptor Signaling Pathway Genes of Zhikong Scallop(Chlamys farreri) with FISH

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中国海洋大学学报：英文版 2015年第6期14卷 1075-1081页

作者： ZHAO Bosong ZHAO Liang LIAO Huan CHENG Jie LIAN Shanshan LI Xuan HUANG Xiaoting BAO Zhenmin Key Laboratory of Marine Genetics and Breeding of Ministry of Education College of Marine Life SciencesOcean University of China

Toll-like receptor(TLR) signaling pathway plays a pivotal role in the innate immune system. Studies on TLR signaling pathway genes in Zhikong scallop(Chlamys farreri) have mainly focused on sequence analysis and expression profiling, no research has been carried out on their localization. The chromosomal position of TLR signaling pathway genes can be valuable for assemblying scallop genome and analysizing gene regulatory networks. In the present study, five key TLR signaling pathway genes(Cf TLR, Cf Myd88, Cf TRAF6, Cf NFκB, and Cf IκB) containing bacterial artificial chromosomes(BACs) were isolated and physically mapped through fluorescence in situ hybridization on five non-homologous chromosome pairs, showing a similar distribution to another five model species. The isolation and mapping of these key immune genes of C. farreri will aid to the research on innate immunity, assignment of interested genes to chromosomes, and integration of physical, linkage and cytogenetic maps of this species.

关键词： immunogenetics Chlamys farreri TLR signaling pathway FISH

Expression Pattern of phb2 and Its Potential Function in Spermatogenesis of Scallop(Chlamys farreri)

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Journal of Ocean University of China 2015年第6期14卷 1034-1040页

作者： HAN Tiantian MA Xiaoshi LIANG Shaoshuai GAO Beibei ZHANG Zhifeng Ministry of Education Key Laboratory of Marine Genetics and Breeding College of Marine Life SciencesOcean University of China

Prohibitin(PHB) participates in several biological processes including apoptosis, transcription regulation and suppression of cell proliferation in mammals. In this study, we cloned the full-length c DNA of prohibitin 2(Cf-phb2) from the testis of scallop(Chlamys farreri). The deduced amino acid sequence presented a characteristic of PHB family with the PHB domain, and clustered with PHB2 of other species. Temporal and spatial expression of Cf-phb2 in testis during the reproductive cycle was detected by quantitative real-time PCR(q RT-PCR) and in situ hybridization. The expression of Cf-phb2 in the testis increased when testis developed from the resting stage to mature stage. The m RNA abundance of Cf-phb2 was the highest at mature stage, which was about 15-fold higher than that at proliferative stage. The expression of Cf-phb2 could be detected by in situ hybridization in all types of germ cells in testis, including spermatogonia, spermatocytes, spermatids and spermatozoa. The intensity of the signal increased with the spermatogenesis and was the highest in spermatids, which suggested that CF-PHB2 might affect the spermatogenesis of C. farreri.

关键词： phb2 spermatogenesis testis Chlamys farreri

EFFECTS OF DIFFERENT COMBINATIONS OF CRYOPROTECTANTS ON SPERMATOZOON VITALITY OF CHINESE SCALLOP,CHLAMYS farreri,DURING LOW TEMPERATURE EQUILIBRIUM

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Chinese Journal of Oceanology and Limnology 1995年第4期13卷 354-361页

作者：薛钦昭相建海 Experimental Marine Biology Laboratory Institute of Oceanology Chinese Academy of Sciences Qingdao

The effects of membrane penetrating anti-freezing agents(MPAAs), DMSO(dimethyl sulfoxide),glycerol,EG(ethylene glycol) and methanol in combination with different cryoprotective additives suchas carbohydrates,macromolecules and inorganic compounds on the spermatozoon vitality of Chinesescallop,Chlamys farreri,during 1 h 0℃ equilibrium were *** only MPAAs existed,the detrimental effects of different MPAAs ranked in the following order:DMSO

关键词： spermatozoon vitality Chlamys farreri cryoprotectants equilibrium

TETRAPLOID INDUCTION BY INHIBITING MITOSIS I IN SCALLOP CHLAMYS farreri

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Chinese Journal of Oceanology and Limnology 1999年第4期17卷 350-358,344页

作者：杨蕙萍郭希明陈再忠王永平

Tetraploid induction was carried out by inhibiting mitosis I in fertilized eggs of Chlamys farreri . Mitosis I was blocked with cold shock (5 7℃), Cytochalasin B (0.75 mg/L) and 6 dimethylaminopurine (6 DMAP) (60 75 mg/L) when 60% fertilized eggs released polar body II at 20℃. At 4 cells embryo stage, the ploidy was determined by counting chromosome number. In control groups, most embryos were diploids (72.22%) and aneuploids (24.78%). In Cytochalasin B, cold shock and 6 DMAP treated groups, tetraploids were respectively 10.51%, 4.08%, and 13.34%; aneuploids were 43.10%, 35.93% and 29.16%,and triploids were 7.84%, 8.52% and 18.33%. At D larva stage, ploidy was determined by flow cytometry (FCM). The ploidy analysis of day 2 larvae showed diploids in control group and also in three treated groups. Juvenile scallops (0.2 0.3cm) which were harvested in two control groups and two CB treated groups were all diploids through checking ploidy individually by FCM.

关键词： scallop Chlamys farreri tetraploid Cytochalasin B mitosis I

STUDIES ON THE ANTITUMOR ACTIVITY AND ISOLATION OF THE GLYCOPROTEIN FROM THE GONAD OF CHLAMYS (AZUMAPECTEN) farreri

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Chinese Journal of Oceanology and Limnology 1998年第4期16卷 359-363页

作者：顾谦群方玉春王长云肖航秦玉青

After being degreased with acetone, the feInal gonad of Chlamys (Azumapecten)farreri was extracted with watr, and then subjected to DEAE - cellulose column chromatogrphyy and pu-rified by Sephadex G - 100 chromatography to obain a pure female gonad glycoprotein (FGGP). The IRspectra, electrophoresis and chemical analysis of FGGP indicated tha it was a kind of acid glycoproteinand that the content of total protein and sugar were 54. 8% and 40%, respectively. Pharmacological testsshawd that FGGP could inhibit (inhibition rate of 40. 87% ) the growth of Sarorna 180 of mice.

关键词： Chlamys (Azumapecten) farreri gonad glycoprotein antitumor activity

Isozyme expression of Chinese and Japanese populations of Chlamys farreri and their reciprocal hybrids

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Chinese Journal of Oceanology and Limnology 2005年第2期23卷 158-162页

作者：李太武刘艳宋林生孙修勤 Faculty of Life Science and Technology Ningbo University Ningbo 315211 China Ningbo Polytechnic College Ningbo 315800 China Institute of Oceanology The Chinese Academy of Sciences Qingdao 266071 China First Institute of Oceanography State Oceanic Administration Qingdao 266003 China

Chinese and Japanese population of Chlamys farreri and their reciprocal hybrids were surveyed in isozyme variability at 13 loci by polyacrytamide gel electrophoresis (PAGE). Isozyme banding patterns indicated these hybrids were diploid. Loci that were observed as being monomorphic in inbred populations of C. farreri were also found to be monomorphic in filial progeny; loci that observed to be polymorphic in parental type gen- erations were also polymorphic in hybrid generations. Differences existed among allelic frequency of the four types of cross. Within the reciprocal hybrids the expression of malic enzyme (ME) isozyme was sufficient to distinguishing individual hybrids because of the band, Rf=0.38. However, there were no noticeable variations among all the samples to differentiate one from another. Inbreeding was likely to be the main problem in aqua- culture. The introduction of new broodstock can improve the genetic diversity. Hybrid vigor has manifested to a certain extent in the present study.

关键词：同功酶表达式互相混合作用同系繁殖人种后裔 Chlamys farreri 基因变化

因子胁迫对栉孔扇贝（Chlamys farreri）血细胞免疫响应的影响

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因子胁迫对栉孔扇贝（Chlamys farreri）血细胞免疫响应的影响

作者：张冬冬中国海洋大学

学位级别：硕士

本论文研究了水温突变、不同温度干露、饥饿及繁殖后，栉孔扇贝全血细胞数量、颗粒血细胞数量及比例的变化;以及不同温度下两种浓度鳗弧菌感染后，栉孔扇贝血细胞数量、颗粒血细胞数量及血细胞6种免疫相关基因，脂多糖β-1,3葡聚糖结合... 详细信息

本论文研究了水温突变、不同温度干露、饥饿及繁殖后，栉孔扇贝全血细胞数量、颗粒血细胞数量及比例的变化;以及不同温度下两种浓度鳗弧菌感染后，栉孔扇贝血细胞数量、颗粒血细胞数量及血细胞6种免疫相关基因，脂多糖β-1,3葡聚糖结合蛋白(LGBP)、C-型凝集素(Lec-2)、Toll样受体(TLR)、溶菌酶(LYZ)、超氧化物歧化酶(SOD)和酚氧化酶(PO)表达量的变化。 1．四种因子变化对栉孔扇贝血细胞数量变化血细胞及其亚群的数量是栉孔扇贝(Chlamys farreri)重要的免疫指标之一。本章研究了栉孔扇贝在17℃下，分别经水温突变(11℃、23℃、28℃)，不同温度(5℃、11℃、17℃、23℃)干露、饥饿和雌雄扇贝繁殖刺激，应用以抗栉孔扇贝全血细胞单抗、颗粒血细胞单抗分别为第一抗体的间接酶联免疫吸附实验和流式细胞术测定全血细胞数量、颗粒血细胞数量及比例的变化。结果表明：水温突变后全血细胞数量均先增加然后逐渐下降，水温变化幅度大全血细胞数量变化较大，颗粒血细胞数量及比例在11℃组始终显著高于对照组，23℃和28℃组表现出先下降后上升的趋势。不同温度干露后全血细胞和颗粒血细胞数量均显著降低，海水恢复后5℃组较11℃、17℃组更接近对照组，23℃组干露12h扇贝全部死亡。饥饿处理组、产卵或者排精后过夜全血细胞数量、颗粒血细胞数量及比例均显著降低。结论认为：全血细胞及颗粒血细胞数量对温度变化的反应敏感;较低水温有利于细胞数量的提升，较高水温下细胞数量下降显著;低温干露后细胞数量恢复较好，高温干露扇贝存活率差。饥饿和繁殖压力会导致全血细胞数量、颗粒血细胞数量及比例及减少。 2.弧菌感染后栉孔扇贝血细胞数量及免疫相关基因表达的变化本章研究了11℃、17℃、23℃、28℃下，栉孔扇贝(Chlamys farreri)分别注射10、10CFU/ml的鳗弧菌(Vibrio anguillarum)和PBS后，应用以抗栉孔扇贝全血细胞单抗和颗粒血细胞单抗分别为第一抗体的间接酶联免疫吸附实验(ELISA)测定全血细胞和颗粒血细胞数量的变化，同时应用实时定量荧光PCR技术检测免疫相关基因(LGBP、Lec-2、TLR、SOD、LYZ和PO)表达量的变化。结果表明：感染10CFU/ml鳗弧菌全血细胞及颗粒血细胞数量无显著变化;10CFU/ml组全血细胞数量除28℃均显著增加后下降，且23℃增加更明显;28℃36h下降至最低值然后增加;10CFU/ml组颗粒血细胞数量均降低，温度越高降低幅度越大;17℃、23℃基因的表达水平较高，28℃下基因的表达量最终几乎均显著的低于对照组。综上研究认为鳗弧菌感染栉孔扇贝，血细胞的数量及免疫相关基因表达均具有显著的温度相关性，较高温度下变化更加显著;血细胞数量及免疫相关基因表达量变化幅度10CFU/ml组均高于10CFU/ml组。

关键词：栉孔扇贝Chlamys farreri 血细胞数量免疫相关基因因子胁迫