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Impact of Selected Environmental Pollutants on the Ultrastructure of the Gills in pinctada radiatafrom Coastal Zones, Egypt

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Open Journal of Ecology 2014年第14期4卷 907-917页

作者： Sherifa Shaker Hamed Eman Hashem Radwan Gaber Ahmed Saad Department of Zoology College of Science King Saud University Saudi Arabia KSA Department of Zoology Faculty of Science Alexandria University Alexandria Egypt Department of Zoology Faculty of Science Damnhour University El Behara Egypt Department of Biology College of Medicine Dammam University Saudi Arabia KSA

There has been an increasing interest in marine oysters (order: Petridae) in recent years because their numbers are declining in many parts of the world and also because they are used as monitors of pollution. The present study describes the microscopic structure of gills as viewed by light and electron microscopy in two locations selected in Alexandria coast, Eastern Harbor (E.H.) and El Asafra. The specimens in the E.H. represent the presence of extracellular mineralized granules.

关键词： Gill Bivalve Transmission Electron Microscope pinctada radiata

Gill Histopathological Effects of PAHs on Adult Pearl Oyster, pinctada radiataat Al-Khiran Coast in Kuwait

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Journal of Environmental Protection 2017年第2期8卷 109-119页

作者： Mona A. Al-Hashem Department of Biological Sciences Faculty of Science Kuwait University Safat State of Kuwait

The hazardous effect of water pollution on the histopathology of gill organ of the pearl oyster, pinctada radiata was studied with respect to polycyclic aromatic hydrocarbon contamination in Al-Khiran coast at Kuwait. Thirty oyster samples from each site, site one where dead oysters were located, site two which is two kilometers away from site one and the control site which is ten kilometers away from site one were collected from Al-Khiran area where a massive number of dead oysters were reported. The gills of oysters were immediately removed and transferred to Bouin’s solution for fixation and then processed for sectioning, staining and mounting and gill tissues were ready for examination. Histopathological changes in gills of oysters exposed to polycyclic aromatic hydrocarbons in site one and site two included necrosis and edemas of branchial lamellae, complete degeneration of gill filaments, loss of regular shape and haemolysis, and inflammation. Gill tissues of oysters from the control site had normal appearance. The study showed a clear evidence that PAHs caused severe histopathological changes in gills of pearl oyster pinctada radiata.

关键词： Gills Histopathology PAHs Oysters pinctada radiata Kuwait

Molecular Cloning and Tissue Distribution of Troponin I from the Japanese Pearl Oyster, pinctada fucata

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American Journal of Molecular Biology 2019年第2期9卷 29-40页

作者： Daisuke Funabara Yoshinori Urakawa Satoshi Kanoh Graduate School of Bioresources Mie University Tsu Japan

Troponin is a complex of three proteins (troponin I, troponin C, and troponin T) that binds Ca2+ and is a thin filament-associated regulator of vertebrate striated muscle contraction. The function of troponin I (TnI) in vertebrates has been extensively characterized, but its role in molluscan muscles has not yet been elucidated. Our previous work suggested that the troponin C subunit has a role in adductor phasic muscle but not in catch muscle. Here, we investigated the molecular characteristics of TnI from the bivalve Japanese pearl oyster, pinctada fucata to aid the elucidation of the function of molluscan muscle troponin. We determined the primary structure of the full-length TnI protein from the P. fucata adductor muscle (Pifuc-TnI) and found that it is composed of 286 amino acid residues with a predicted molecular weight of 33,737. Motif structure predictions and multiple sequence alignments revealed that Pifuc-TnI has a 138 residue extension at its N-terminus compared with rabbit TnI. This is analogous to characterized TnIs from other mollusks. However, unlike scallop TnI, Pifuc-TnI is predicted to contain two cAMP-dependent protein kinase phosphorylation sites, at residues 39 - 45 (RRGTEDD) and 145 - 151 (KKKSKRK). Phylogenetic analysis indicated that Pifuc-TnI and molluscan TnIs were grouped into the same clade. Pifuc-TnI gene structure predictions using Splign alignment of our obtained cDNA and genome sequences indicated that Pifuc-TnI consists of fifteen exons, with the start and stop codons located in exon 2 and exon 11, respectively. Using quantitative real-time PCR, we determined that the Pifuc-TnI gene is predominantly expressed in adductor phasic muscle, weakly in adductor catch muscle, and is not expressed in the gill, mantle or foot. These findings suggest that TnI, as a component of the troponin complex, plays a regulatory role in adductor phasic muscle contraction, but not in catch contraction.

关键词： Adductor Muscle Catch Contraction pinctada fucata Troponin Troponin I

Troponin T from the Japanese Pearl Oyster pinctada fucata: Molecular Cloning, Tissue Distribution, Gene Structure, and Interaction Analysis with Tropomyosin

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American Journal of Molecular Biology 2020年第1期10卷 61-73页

作者： Daisuke Funabara Yoshinori Urakawa Daisuke Ishikawa Satoshi Kanoh Graduate School of Bioresources Mie University Tsu Japan

Troponin (Tn) is composed of three subunits (TnI, TnC and TnT) that bind Ca2+ and regulate striated muscle contraction in vertebrates. TnT’s function has been extensively described in vertebrates, but its role has been obscure in molluscan muscles. Our previous work indicated that the TnC and TnI subunits work in adductor phasic muscle, but not in catch muscle. Here, we have characterized TnT from the Japanese bivalve pearl oyster pinctada fucata to start to explain the function of Tn in molluscan muscle contraction. We determined the primary structure of the full-length TnT protein from the P. fucata adductor muscle (Pifuc-TnT), and found that it is composed of 316 amino acid residues with a predicted molecular mass of 37.4 kDa. Multiple sequence alignment showed that Pifuc-TnT has an extension of >60 residues at the C-terminus that are not present in vertebrate TnTs, including known TnTs from other mollusks. Pifuc-TnT gene structure predictions using Splign alignment of the cDNA generated in this study and genome sequences indicated that Pifuc-TnT consists of 13 exons. Start and stop codons are located in exons 2 and 12, respectively. Quantitative real-time PCR revealed that the Pifuc-TnT gene was predominantly expressed in adductor phasic muscle, weakly in adductor catch muscle, slightly in gill, and not at all in mantle and foot. These findings suggest that TnT plays a regulatory role in adductor phasic muscle contraction, but not in catch contraction. Isothermal titration calorimetry revealed that unlike vertebrate TnTs, Pifuc-TnT does not interact with P. fucata tropomyosin-1 nor with tropomyosin-2. These findings in P. fucata imply that Tn functions differently in molluscan muscle than it does in vertebrates.

关键词： Adductor Muscle Catch Contraction pinctada fucata Troponin T Tropomyosin

Production of Aneuploid pinctada martensii Dunker in Tetraploid Induction

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Marine Science Bulletin 2001年第1期3卷 63-68页

作者：何毛贤林岳光沈琪胡建兴姜卫国 South China Sea Institute of Oceanology Chinese Academy of SciencesGuangzhou 510301 South China Sea Institute of Oceanology Chinese Academy of SciencesGuangzhou 510301neuploidy embryos of Pinctada martensii Dunker are produced during tetraploid induction by inhibiting the first polar body in eggs from triploid fertilized with haploid sperms with cytochalasin B treatment. Chromosome analysis reveals that there are 88.18±6.79% aneuploidy embryos and 28.70% aneuploids in pearl oysters of one-year age. These aneuploids have five chromosomal conditions such as 2n + 1(29) 2n + 2 (30) 3n-2 (40) 3n -1(41) and 3n + 1 (43). Results of growth measurement show that there is no significant difference between aneuploids (as a group) and diploids in body size and weight (p > 0.10) but the aneuploide is obviously different from triploid (p < 0.01). The mean body size and weight of aneuploids in diploid condition (2n ± 1 and 2n ± 2) are significantly smaller than those of diploids (p < 0.01) but aneuploids within triploid condition (3n ± 1 and 3n ± 2) are not smaller than diploids in body size and weight (p > 0.1).This study indicates Pinctada martensii Dunker could tolerate aneuploidy by 7 - 14% of the haploid genome and that aneuploids of this species are viable under certain conditions.

Aneuploidy embryos of pinctada martensii Dunker are produced during tetraploid induction by inhibiting the first polar body in eggs from triploid fertilized with haploid sperms with cytochalasin B treatment. Chromosome analysis reveals that there are 88.18 ±6.79% aneuploidy embryos, and 28.70% aneuploids in pearl oysters of one-year age These aneuploids have five chromosomal conditions, such as 2n + 1(29), 2n + 2 (30), 3n-2 (40), 3n-1(41) and 3n + 1 (43). Results of growth measurement show that there is no significant difference between aneuploids (as a group) and diploids in body size and weight (p ＞ 0.10), but the aneuploide is obviously different from triploid (p ＜ 0.01). The mean body size and weight of aneuploids in diploid condition (2n ± 1 and 2n ± 2) are significantly smaller than those of diploids (p ＜ 0.01),but aneuploids within triploid condition (3n ± 1 and 3n ± 2) are not smaller than diploids in body size and weight (p ＞ 0.1).This study indicates pinctada martensii Dunker could tolerate aneuploidy by 7 ～ 14% of the haploid genome, and that aneuploids of this species are viable under certain conditions.

关键词： aneuploid, triploid, pinctada martensii Dunker

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马氏珠母贝家系的生长比较

热带海洋学报 2007年第1期26卷 39-43页

作者：何毛贤管云雁林岳光黄良民中国科学院南海海洋研究所广东省/中国科学院应用海洋生物学重点实验室广东广州510301

采用单对配对方法建立马氏珠母贝pinctada martensii第一代家系7个,对家系间的生长和家系内大小变异进行了分析比较。7个家系的壳高大小顺序为F15>F20>F18>F19>F12>F17>F21,总重大小顺序为F15>F18>F19>F20>F12>F17>F21。在同批家系中,F1... 详细信息

采用单对配对方法建立马氏珠母贝pinctada martensii第一代家系7个,对家系间的生长和家系内大小变异进行了分析比较。7个家系的壳高大小顺序为F15>F20>F18>F19>F12>F17>F21,总重大小顺序为F15>F18>F19>F20>F12>F17>F21。在同批家系中,F15的壳高、总重显著大于其它2个家系(F12、F17)(p<0.05),其壳高比F12大11.64%,比F17大16.01%;其总重比F12重27.40%,比F17重38.98%,而且其个体大小变异较小,表现出明显的生长优势,是一个生长性状优良的家系。F18在壳高和总体重上比F19大2.88%和19.49%。F20显著大于F21(p<0.05),壳高比F21大19.49%,总重大53.72%,该家系生长较快,可通过进一步的选择培育成一个生长快、个体大的家系。

关键词：马氏珠母贝 pinctada martensii 家系生长

马氏珠母贝IKKε同源基因的克隆及表达分析

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基因组学与应用生物学 2014年第2期33卷 266-272页

作者：焦钰杜晓东王庆恒黄荣莲邓岳文广东海洋大学水产学院湛江524025

IKKε(inhibitor of NF-κB kinasesε)是NF-κB(nucleur factorκB)信号通路中的关键成员,参与调控细胞的分化、发育、凋亡及免疫反应。本研究根据马氏珠母贝转录组数据库中注释为IKKε的unigene序列设计引物,应用cDNA末端快速扩增(RA... 详细信息

IKKε(inhibitor of NF-κB kinasesε)是NF-κB(nucleur factorκB)信号通路中的关键成员,参与调控细胞的分化、发育、凋亡及免疫反应。本研究根据马氏珠母贝转录组数据库中注释为IKKε的unigene序列设计引物,应用cDNA末端快速扩增(RACE)技术克隆获得了马氏珠母贝(pinctada martensii)IKKε基因cDNA的全长序列(PmIKKε)并对其序列特征进行分析;同时利用荧光定量PCR(Real-time PCR)技术检测了马氏珠母贝六个组织中PmIKKε基因mRNA的表达。结果表明:PmIKKε基因cDNA全长2 843 bp,其中5'UTR为116 bp,3'UTR为516 bp,含有27 bp polyA,开放阅读框为2 211 bp,编码737个氨基酸残基。预测其分子量为85.07 kD,等电点为6.17。多序列比对显示PmIKKε与其它物种IKKε有较高的同源性,与牡蛎的序列相似性有45%。软件分析结果显示PmIKKε的N端含有一个蛋白激酶功能结构域和一个MAPKK(mitogen-activated protein kinase kinase)的激活位点,C端含有一个亮氨酸拉链(leucin zipper,LZ)和一个螺旋-环-螺旋结构(helix-loop-helix,HLH)。荧光定量PCR分析表明PmIKKε在马氏珠母贝肝胰脏、性腺、血淋巴、鳃、外套膜、闭壳肌六个组织中均有表达,肝胰脏中表达量最高。本研究为进一步阐述PmIKKε在马氏珠母贝生长、发育及先天性免疫中的作用提供理论基础。

关键词： IKKε pinctada martensii 基因克隆荧光定量PCR