企鹅珍珠贝生殖细胞的发生

对不同发育阶段的企鹅珍珠贝[Ptedapenguin(Rding)]的性腺结构、生殖细胞结构与分布进行组织学观察,描述精/卵子发生过程中各级精/卵细胞的特征。结果表明:根据雌性生殖细胞形态和卵黄的变化特点,卵子发生经历卵原细胞、无卵黄初级卵母细胞、卵黄形成期卵母细胞及成熟卵子阶段;依据雄性生殖细胞大小和细胞核形态变化,精子发生经历精原细胞、初级精母细胞、次级精母细胞、精子细胞和精子阶段,其中精原细胞存在形态上有差异的A型和B型两种类型。此外还观察到企鹅珍珠贝的性别转化现象。     

CYTOLOGICAL COMPARISON OF GAMETOGENESIS OF SCALLOPS,ARGOPECTEN IRRADIANS AND CHLAMYS FARRERI

ＩＮＴＲＯＤＵＣＴＩＯＮＫｎｏｗｌｅｄｇｅｏｆｇａｍｅｔｏｇｅｎｅｓｉｓｉｎｖａｌｕａｂｌｅｍａｒｉｎｅａｎｉｍａｌｓｉｓｖｅｒｙｉｍｐｏｒｔａｎｔｔｏｃｏｍｍｅｒｃｉａｌｆｉｓｈ ｅｒｙ.Ｇａｍｅｔｏｇｅｎｅｓｉｓｉｎｂｉｖａｌｖｅｓｗａｓｅｘｔｅｎｓｉｖｅｌｙｓｔｕｄｉｅｄａｔｄｉｆｆｅｒｅｎｔｌｅｖｅｌｓ (Ｄｏｈｍｅｎ ,1 983 ;ＤｏｒａｎｇｅａｎｄＬｅＰｅｎｎｅｃ ,1 989ａ,ｂ) .Ｓｃａｌｌｏｐｓａｎｄｏｔｈｅｒｍａｒｉｎｅｂｉｖａｌｖｅｓｈａｖｅｓｉｍｉｌａｒｒｅｐｒｏｄｕｃｔｉｖｅｃｙｃｌｅ…     

Three-dimensional fluorescence characteristics of dissolved organic matter produced by <Emphasis Type="Italic">Prorocentrum donghaiense</Emphasis> Lu

Filtration and cross-flow ultrafiltration techniques were used to separate culture media of Prorocentrum donghaiense at the exponential growth,stationary and decline stages into <0.45 μm filtrate,100 kDa-0.45 μm,10-100 kDa and 1-10 kDa retentate and <1 kDa ultrafiltrate fractions.The fluorescence properties of different molecular weights of dissolved organic matter(DOM) were measured by excitation-emission matrix spectra.Protein-like and humic-like fluorophores were observed in the DOM produced by P.donghai...     

CYTOLOGICAL COMPARISON OF GAMETOGENESIS OF SCALLOPS, ARGOPECTEN IRRADIANS AND CHLAMYS FARRERI

Histological characteristics of gametogenesis of two kinds of scallops, gonochoric Chinese scallop, Chlamys farreri and hermaphroditic bay scallop, Argopecten irradians were investigated in this study. Spermatogenesis in C. farreri has different developmental stages: spermatogonia, primary spermatocyte, second spermatocyte, spermatid and spermatozoon. A large number of same developmental stage spermatic cells converge at a definite area of the testis. Premeiotic, previtellogenic and vitellogenic oocytes can be found during oogenesis in C. farreri, where oocyte distribution is obviously irregular. The A. irradians gonad consists of two different parts in one individual: one part functions as testis, the other as ovary. Between these two parts is a special appearance area, where a large number of spermatic cells are bound with two layers of acellular substance with many oocytes in it.     

Seasonal Changes of Reproductive Activity and Biochemical Composition of Pen Shell Atrina pectinata Linnaeus,1767 in Bohai Sea,China

Seasonal variations in the biochemical composition and the reproductive cycle of pen shell Atrina pectinata in Bohai Sea were investigated from May 2013 to April 2014. Histological analysis indicated that the reproductive cycle of A. pectinata can be divided into two phases, inactive stage and gametogenesis, which were equally and significantly influenced by seawater temperature and food availability. Gametogenesis began in late autumn(October), and completed in June and July. Spawning took place in August, coinciding with the highest water temperature and the richest phytoplankton. The significantly high glycogen content in adductor muscles sustained throughout the late active and ripeness stages, but plummeted during spawning and inactive stages. The protein content in female gonads exhibited a synchronous increase along with oocyte diameter and lipid content, suggesting that the female gonads could accumulate protein and lipid for vitelline in A. pectinata. Furthermore, the RNA/DNA ratio was found to be a useful index to indicate the level of gonad maturation in both males and females. The findings of the present study provided a foundation for the fishery resource administration and the aquaculture development of this species.     

Changes during late-stage embryonic development from egg-juvenile to free-living hatchling in Chinese freshwater crab Sinopotamon yangtsekiense (Decapoda, Brachyura, Potamidae)

This study expands on recent reports that direct development in the Chinese potamid freshwater crab Sinopotamon yangtsekiense involves the completion of all brachyuran larval stages (nauplius, zoea, and megalopa) inside the egg case during embryonic development. Detailed studies of embryonic development in this species revealed the presence of an additional larval stage (the egg-juvenile) between the megalopa and the free-living hatchling crab. We described and compared the appendages of the head, thorax, and abdomen of the egg-juvenile with those of the hatchling crab in S. yangtsekiense. Significant differences were found between most of the appendages of these two stages with a soft exoskeleton in the egg-juvenile, no joint articulation, a slimmer appearance, and a lack of setae when compared with the newly emerged free-living hatchling crab. These modifications of the appendages are related to the confinement within the egg case of the egg-megalopa and egg-juvenile during direct development, and the need for the free-living hatchling freshwater crab to move, feed, and respire. In marine crabs, the megalopa gives rise to the first crab stage whereas in freshwater crabs the egg-juvenile follows the megalopa and immediately precedes the free-living first crab stage.     

Characterization,expression and function analysis of DAX1 gene of scallop (Chlamys farreri jones and preston 1904) during its gametogenesis

DAX1, a member of nuclear receptor superfamily, has a function in the sex determination and gonadal differentiation of several vertebrate species. However, little information about DAX1 of invertebrates is available. Here we cloned a homolog of scallop (Chlamys farreri Jones and Preston 1904) dax1, Cf-dax1, and determined its expression characteristics at mRNA and protein levels. The cDNA sequence of Cf-dax1 was 2093 bp in length, including 1404 bp open reading frame (ORF) encoding 467 amino acids. Unlike those of vertebrates, no conserved LXXLL-related motif was found in the putative DNA binding region of Cf-DAX1. Fluorescence in situ hybridization showed that Cf-dax1 located on the short arm of a pair of subtelocentric chromosomes. Tissue distribution analysis using semi-quantitative RT-PCR revealed that Cf-dax1 expressed widely in adult scallop tissues, with the highest expression level found in adductor muscle, moderate level in mantle, gill and testis, and low level in kidney, ovary and hepatopancreas. The result of quantitative real-time PCR indicated that the expression of Cf-dax1 was significantly higher (P<0.05) in testis than in ovary at the same stage, showing a sex-dimorphic expression pattern. Furthermore, immunohistochemical detection found that Cf-DAX1 mainly located in spermatogonia and spermatocytes of testis and in oogonia and oocytes of ovary, implying that DAX1 may involve in gametogenesis of bivalves.     

Effects of mercuric chloride on antioxidant system and DNA integrity of the crab Charybdis japonica

Mercury (Hg) is one of the commonly encountered heavy metals, which is widespread in inshore sediments of China. In order to investigate the toxicity of Hg on marine invertebrates, we studied the effects of the divalent mercuricion (Hg²⁺) (at two final concentrations of 0.0025 and 0.0050 mg L⁻¹, prepared with HgCl₂) on metallothionein (MT) content, DNA integrity (DNA strand breaks) and catalase (CAT) in the gills and hepatopancreas, antioxidant enzyme activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx), in the hemolymph, gills and hepatopancreas of the portunid crab Charybdis japonica for an experiment period up to 15 d. The results indicated that MT was significantly induced after 3 d, with a positive correlation with Hg²⁺ dose and time in the hepatopancreas and a negative correlation with Hg²⁺ dose and time in the gills. While CAT in the hemolymph was not detected, it increased in the hepatopancreas during the entire experiment; SOD and GPx in the three tissues were stimulated after 12 h, both attained peak value and then reduced during the experimental period. Meanwhile, DNA strand breaks were all induced significantly after 12 h. These results suggested the detoxification strategies against Hg²⁺ in three tissues of C. japonica.     

Seasonal changes of reproductive activity and biochemical composition of pen shell <Emphasis Type="Italic">Atrina pectinata</Emphasis> Linnaeus, 1767 in Bohai Sea,China

Seasonal variations in the biochemical composition and the reproductive cycle of pen shell Atrina pectinata in Bohai Sea were investigated from May 2013 to April 2014. Histological analysis indicated that the reproductive cycle of A. pectinata can be divided into two phases, inactive stage and gametogenesis, which were equally and significantly influenced by seawater temperature and food availability. Gametogenesis began in late autumn (October), and completed in June and July. Spawning took place in August, coinciding with the highest water temperature and the richest phytoplankton. The significantly high glycogen content in adductor muscles sustained throughout the late active and ripeness stages, but plummeted during spawning and inactive stages. The protein content in female gonads exhibited a synchronous increase along with oocyte diameter and lipid content, suggesting that the female gonads could accumulate protein and lipid for vitelline in A. pectinata. Furthermore, the RNA/DNA ratio was found to be a useful index to indicate the level of gonad maturation in both males and females. The findings of the present study provided a foundation for the fishery resource administration and the aquaculture development of this species.     

Levels of Metabolic Enzymes and Nitrogenous Compounds in the Swimming Crab Portunus trituberculatus Exposed to Elevated Ambient Ammonia-N

This study investigated the effects of ammonia-N(control and 1 and 5 mg L~(-1)) on the changes of metabolic enzymes activities and nitrogenous compounds levels in the gills, hepatopancreas, muscles, and hemolymph of the swimming crab Portunus trituberculatus. Results showed that the levels of hemolymph ammonia increased significantly at 6 h and then decreased to control level at 12 h. The activities of key enzymes involved in biosynthesis pathways of glutamine(Gln) and urea, such as glutamate dehydrogenase(GDH), glutamine synthetase(GS), and arginase(ARG), increased remarkably in different tissues of P. trituberculatus. Gln level was the highest in muscles, while urea content was the largest in hepatopancreas. The m RNA expression level of xanthine dehydrogenase(XDH) was downregulated sharply in gills but upregulated in the hepatopancreas and muscles. A small amount of uric acid was detected in the hemolymph. This result suggests that Gln in muscles may be used as an intermediary nitrogen repository, while urea may be primarily synthesized in the hepatopancreas and then transferred to other tissues via the hemolymph. Uric acid could be synthesized by the anabolism of purine nucleotides, in additon to the inhibitory activities of gills. In addition, the levels of Gln, urea and uric acid exhibited dose-and time-dependent effects with ammonia content. Therefore, P. trituberculatus could possibly convert ammonia into Gln, urea, or uric acid to detoxify ammonia during high ambient ammonia-N stress.     

Expression and localization of a novel phosducin-like protein from amphioxus Branchiostoma belcheri

A full length amphioxus cDNA, encoding a novel phosducin-like protein (Amphi-PhLP),was identified for the first time from the gut cDNA library of Branchiostoma belched. It is comprised of 1 550 bp and an open reading frame (ORF) of 241 amino acids, with a predicted molecular mass of approximately 28 kDa. In situ hybridization histocbemistry revealed a tissue-specific expression pattern of Amphi-PhLP with the high levels in the ovary, and at a lower level in the hind gut and testis, hepatic caecum, gill, endostyle, and epipharyngeal groove, while it was absent in the muscle, neural tube and notochord. In the Chinese Hamster Ovary (CHO) cells transfected with the expression plasmid pEGFP-NIIAmphi-PhLP, the fusion protein was targeted in the cytoplasm of CHO cells, suggesting that Amphi-PhLP is a cytosolic protein. This work may provide a framework for further understanding of the physiological function of Amphi-PhLP in B. belcheri.     

Growth,reproduction & population structure of the freshwater crabSinopotamon yangtsekiense bott, 1967, from Zhejiang,China

Monthly investigations were mae on the population of Chinese freshwater crab,Sinopotamon yangtsekiense Bott, 1967 from April, 1984 to March, 1985. The data on 4413 specimens show that the growth was affected mainly by temperature. During the April to November growth period, the crabs' major development occurred from June through October. One year was required for a fine white oocyte to develop into a mature egg. The reproduction period was June–October. Females bearing eggs were taken from June–August, and crabs with young were found from July–October. The females reproduced once a year but could for more than one year. The number of eggs carried by a female varied greatly according to the size of the crab, ranging from 30 to 100 eggs. New-born crabs become mature after 1–2 years. The sex ratio was approximately 1∶1 in the overall population. However, the larger crabs are predominantly male. The age distribution ofS. yangtsekinese was estimated from size frequency histograms. There were more adult crabs (over 70%) from June to October and more immature crabs (over 50%) from November to May.     

以江蓠为栖息环境养殖的双齿围沙蚕性腺发育

研究以江蓠为养殖基质的双齿围沙蚕的配子发生及性腺发育。结果表明:1)根据双齿围沙蚕卵细胞形态特征、内部结构、卵径大小,卵母细胞的发生可划分为第Ⅰ时相(卵原细胞)、第Ⅱ时相(小生长期初级卵母细胞)、第Ⅲ时相(大生长期初级卵母细胞,卵黄形成前期、卵黄形成中期、卵黄形成后期)和第Ⅳ时相(成熟卵母细胞,胶膜形成前期、胶膜形成后期)等4时相;根据各时相卵子的数量变化及体腔中其他结构的形态变化,将雌性性腺分为Ⅰ期卵巢、Ⅱ卵巢、Ⅲ卵巢、Ⅳ卵巢和Ⅴ期卵巢等5个阶段。2)根据双齿围沙蚕体腔内精母细胞的大小、形态和所处的位置,将双齿围沙蚕的精子发生划分为精原细胞、初级精母细胞、次级精母细胞、精细胞和精子;根据各时相精子的形态和数量变化以及体腔中其他组织器官的形态变化,将双齿围沙蚕的雄性性腺划分为Ⅰ期精巢、Ⅱ期精巢、Ⅲ期精巢、Ⅳ期精巢和V期精巢等5个阶段。3)Ⅰ-Ⅳ期的性腺主要包含对应时相的生殖细胞,同时,也存在有少量其他时相的生殖细胞,生殖细胞的发育速度并不完全一致。     

Amino acid composition and functional properties of giant red sea cucumber (Parastichopus californicus) collagen hydrolysates

Giant red sea cucumber (Parastichopus californicus) is an under-utilized species due to its high tendency to autolysis.The aim of this study was to evaluate the functional properties of collagen hydrolysates from this species.The degree of hydrolysis (DH),amino acid composition,SDS-PAGE,emulsion activity index (EAI),emulsion stability index (ESI),foam expansion (FE),and foam stability (FS) of hydrolysates were investigated.The effects of pH on the EAI,ESI FE and FS of hydrolysates were also inves-tigated.The results indicated that the β and α1 chains of the collagen were effectively hydrolyzed by trypsin at 50℃ with an En-zyme/Substrate (E/S) ration of 1:20 (w:w).The DH of collagen was up to 17.3% after 3 h hydrolysis with trypsin.The hydrolysates had a molecular weight distribution of 1.1 17 kDa,and were abundant in glycine (Gly),proline (Pro),glutamic acid (Glu),alanine (Ala) and hydroxyproline (Hyp) residues.The hydrolysates were fractionated into three fractions (< 3 kDa,3 10 kDa,and > 10 kDa),and the fraction of 3 10 kDa exhibited a higher EAI value than the fraction of > 10 kDa (P<0.05).The fraction of > 10 kDa had higher FE and FS values than other fractions (P<0.05).The pH had an important effect on the EAI,ESI,FE and FS.All the fractions showed undesirable emulsion and forming properties at pH 4.0.Under pH 7.0 and pH 10.0,the 3 10 kDa fraction showed higher EAI value and the fraction of > 10 kDa showed higher FE value,respectively.They are hoped to be utilized as functional ingredients in food and nutraceutical industries.     

GROWTH, REPRODUCTION & POPULATION STRUCTURE OF THE FRESHWATER CRAB SINOPOTAMON YANG TSEKIENSE BOTT, 1967, FROM ZHEJIANG, CHINA

Monthly investigatiom were made on the population of Chinese freshwater crab, Sinopotamon yangteskiense Bott, 1967 from April, 1984 to Match, 1985. The data on 4413 specimens show that the growth was affected mainly by temperature. During the April to November growth period, the crabs' major development occured from June through October. One year was required for a fine white oocyte to develop into a mature egg. The reproduction period was June-October. Females bearing eggs were taken from June-August, and crabs with young were found from July-October. The females reproduced once a year but could for more than one year. The number of eggs carried by a female varied greatly according to the size of the crab, ranging from 30 to 100 eggs. New-bom crabs become mature after 1-2 yeats. The sex ratio was approximately 1:1 in the overall population. However, the larger crabs are predominantly male. The age distribution of S. yangtsekinese was estimated from size frequency histograms. There were more adult crabs     

The role of phosphatidylinositol signaling pathway in regulating serotonin-induced oocyte maturation in Mercenaria mercenaria

Serotonin (5-HT) has been found to stimulate meiotic maturation of oocytes in many molluscs. During maturation, several signaling pathways are involved, especially the phosphatidylinositol and cAMP pathways. In order to examine the possible role of the phosphatidylinositol signaling pathway in regulating oocyte maturation in Mercenaria mercenaria, the effects of the activator/inhibitor of phospholipase (PLC) and protein kinase C (PKC) on serotonin-induced maturation were studied. Results show that high-concentrations of neomycin (inhibitor of PLC) blocked oocyte maturation, while 9, 10-dimethyl-1, 2-benzanthracene (DMBA, activator of PLC) promoted oocyte maturation in the presence of serotonin. It was also found that in the presence of serotonin, phorbol 12-myristate 13-acetate (PMA, activator of PKC) inhibited oocyte maturation, while sphingosine (inhibitor of PKC) stimulated oocyte maturation. These results indicate that serotonin-induced oocyte maturation requires the activation of the phosphatidylinositol pathway. Decrease of PLC inhibited serotonin-induced oocyte maturation, whereas a decrease of PKC stimulated the maturation. Thus, our study indicates that serotonin promotes maturation of M. mercenaria oocytes through PLC stimulated increase in calcium ion concentration via inositol-1, 4, 5-trisphosphate (IP3) but not PKC.     

Seasonal variation of biochemical components in clam (<Emphasis Type="Italic">Saxidomus purpuratus</Emphasis> Sowerby 1852) in relation to its reproductive cycle and the environmental condition of Sanggou Bay,China

Seasonal variation of biochemical components in clam (Saxidomus purpuratus Sowerby 1852) was investigated from March 2012 to February 2013 in relation to environmental condition of Sanggou Bay and the reproductive cycle of clam. According to the histological analysis, the reproductive cycle of S. purpuratus includes two distinctive phases: a total spent and inactive stage from November to January, and a gametogenesis stage, including ripeness and spawning, during the rest of the year. Gametes were generated at a low temperature (2.1°C) in February. Spawning took place once a year from June to October. The massive spawning occurred in August when the highest water temperature and chlorophyll a level could be observed. The key biochemical components (glycogen, protein and lipid) in five tissues (gonad, foot, mantle, siphon and adductor muscle) were analyzed. The glycogen content was high before gametogenesis, and decreased significantly during the gonad development in the gonad, mantle and foot of both females and males, suggesting that glycogen was an important energy source for gonad development. The protein and lipid contents increased in the ovary during the gonad development, demonstrating that they are the major organic components of oocytes. The lipid and protein contents decreased in the testis, implying that they can provide energy and material for spermatogenesis. The results also showed that protein stored in the mantle and foot could support the reproduction after the glycogen was depleted.