Culture of mantle epithelial cells expressing shell matrix proteins from scallop Patinopecten yessoensis

ABSTRACT:   In molluscs, mantle epithelial cells secrete organic matrix proteins to form shells. In this study, we established a culture of mantle epithelial cells by using the mantle pallial layer of scallops. We aimed to identify the mantle epithelial cells expressing scallop shell matrix proteins and establish a culture system of epithelial cells. After the mantle pallial layer was carefully isolated from the mantle tissue, explant culture was performed at 4°C. Most cells that migrated from the explant tissue were round cells. Most of the adhered cells retained round morphology, while some of the cells adhered to the dish and showed morphology similar to that of epithelial-like and fibroblast-like cells. When the cultured cells were immunostained with a polyclonal antibody against the shell matrix protein, the antibody recognized many of the adhered cells. An estimation of the number of epithelial cells revealed that approximately 70% of the adhered cells were epithelial cells. This is the first report to describe epithelial cells in cultured mantle cells, which express shell matrix proteins. This culture system may be a useful method for characterization of the mantle epithelial cells.     

牙鲆体质量与形态性状的异速生长分析

为分析牙鲆(Paralichthys olivaceus)体质量与形态性状之间异速生长的遗传规律,通过人工受精的方法建立牙鲆19个全同胞家系,测量了牙鲆不同日龄的体质量和形态性状。采用逐步回归方法建立最优联合异速生长模型,将此模型镶嵌到体质量动物模型的每个遗传和环境效应中,进一步分析对多个形态性状异速生长遗传规律。静态异速生长分析表明:体质量与全长之间存在最大异速生长指数(1.415 5),表现为正异速生长,剩余形态性状与体质量之间的异速生长指数为0.061 5~0.718 0,皆表现为负异速生长。全长与尾柄高之间异速生长指数的正遗传相关最大(0.907 8);全长与尾柄长之间的负遗传相关最大(-0.946 8)。不同模型通过统计标准比较,确定模型Ⅰ为进行牙鲆动态异速生长分析的最优随机回归模型。     

牙鲆养殖群体遗传变异的微卫星标记研究

应用10对微卫星引物对中国牙鲆一个养殖群体的30个个体进行了群体遗传结构分析。结果显示，微卫星标记比其他标记具有更高的多态性，10个微卫星座位的等位基因数在4～10之间，有效等位基因数在2．23～5．82之间，平均等位基因数为7．6，群体平均杂合度为0．6960，Hardy-weinberg遗传偏离指数的平均值为0．1774。     

Fine structure and differentiation of the alimentary canal in captive-bred Japanese eel Anguilla japonica preleptocephali

ABSTRACT:   The fine structure of the alimentary canal in preleptocephali produced by artificially matured Japanese eel was examined. At 1 day posthatch (dph), the alimentary canal was found only above the dorsal side of the yolk mass, and the epithelium was composed of a single layer of epithelial cells. By 5 dph, the alimentary canal was divided into three segments based on the structure of the epithelial cells: foregut, midgut and hindgut, corresponding to the future esophagus, intestine and rectum, respectively. After 7 dph, the epithelium in the foregut was surrounded by a circular muscle layer, suggesting a role in the transportation of food materials. The epithelial cells of the midgut exhibited well-developed membranous structures, which are deduced to be invaginations of the cytoplasmic membrane. Pinocytotic invaginations and vacuoles were observed in the epithelial cells of the hindgut; this observation suggests that this region is involved in the uptake of food. Significant changes in morphological features of the epithelial cells in each segment were observed until 7 dph; however, these were not evident between 7 dph and 13 dph. Consequently, the differentiation of the alimentary canal was completed by 7 dph, and preleptocephalus had developed the ability to absorb food by 7 dph.     

Epidermal proteases of the Japanese eel

A fluorescent-sensitive assay was used to demonstrate the protease activity in the dorsal skin of Japanese eel (Anguilla japonica). Two distinct extracts were separately prepared from skin mucus and epidermal cell layers, with no mutual contamination. The epidermal extract was sensitive to various substrates, whereas there was no, or only marginal, susceptibility to the same substrates for the mucous extract. Optimum hydrolysis pHs of the epidermal extract was variable and below pH 7.0, and the optimum hydrolysis temperatures were between 40 and 50 °C. In addition, Tos-Phe-Ch₂Cl, chymostatin, CdCl₂, CuCl₂, HgCl₂ and ZnCl₂ inhibited protease activities to different extents. Several other reagents specifically affected the protease activities, and their induced effects were useful for the identification of epidermal proteases. The findings indicate that a proteolytic factor, exhibiting various enzymological specificities, is retained within epidermal cell layers of Japanese eel. This factor is composed of 4 distinct proteases, such as cathepsins L and B-like proteases, a serine protease and an aminopeptidase.     

虾夷扇贝横纹肌和平滑肌的蛋白分布及理化性质

为探讨虾夷扇贝闭壳肌中横纹肌(ST)和平滑肌(SM)蛋白分布特征及理化性质差异。对肌肉组织进行切片、苏木精—伊红染色后,在光学显微镜下观察肌肉组织形态,SDS-PAGE分析蛋白组成,通过盐溶性和Ca~(2+)-ATPase对理化性质进行了比较研究。研究表明,横纹闭壳肌的肌纤维更粗,肌细胞间更致密;Myorod蛋白仅存在于平滑闭壳肌中,在横纹闭壳肌中不存在;副肌球蛋白在平滑闭壳肌中含量为31%,横纹肌中仅为11%。2种肌肉盐溶曲线也完全不同,在0.05~1.0 mol/L盐浓度范围内平滑肌的溶解性呈S型曲线上升,而横纹肌呈直线型上升,这种差异可能与副肌球蛋白含量有关。此外,平滑肌和横纹肌肌原纤维蛋白Ca~(2+)-ATPase最适温度分别为35和30°C,活化能分别为23.4和11.3 k J/mo L。     

Efficacy of Netminder® silicone release coating for biofouling reduction in bay scallop grow‐out and comparative effects on scallop survival,growth and reproduction

Netminder^®, a water‐based silicone barrier release coating, was highly effective in reducing biofouling of lantern nets during grow‐out/overwintering of bay scallops, Argopecten irradians irradians, in a large‐scale restoration programme in New York, USA. Compared with untreated nets, scallops held in Netminder^®‐coated nets experienced reduced survival after 8 months, probably due to higher loads of epibionts on their shells. However, this was countered by higher reproductive and overall condition of scallops in treated vs. untreated nets; there were no consistent differences in shell growth in different net treatments. Although further testing is recommended under a wider range of conditions, we conclude that for the purposes of our scallop restoration work, coating nets with Netminder^® would eliminate the need for a gear change in spring, thus reducing labour costs; the coating process should be refined so that Velcro closures of nets do not become sealed shut.     

Comparison of early life history stages of the bay scallop, Argopecten irradians: Effects of microalgal diets on growth and biochemical composition

The culture of bay scallops, Argopecten irradians, is limited by a reliable and affordable supply of spat and the ability to ensure that animals attain market size within a single growing season. The main goals of our study were thus: (1) to develop growth-optimizing algal diets for implementation in hatcheries, and (2) to identify and compare bay scallop postlarval and juvenile dietary requirements, especially of lipids and fatty acids, which if met may enhance production. Nutritional needs of postlarval bay scallops (present study) are compared with those of sea scallops, Placopecten magellanicus, offered the same diets in a previous companion study. To this end, postlarval (initial shell height, SH = 240 μm) and juvenile (initial SH = 10 mm) bay scallops were offered 6–7 microalgal diet combinations at 20 °C, for 3 weeks. A similar growth ranking among diets was observed between the two developmental stages. A combination diet of Pavlova sp. (CCMP 459) and Chaetoceros muelleri was far superior to any other diet tested, yielding growth rates of 58 and 357 μm day^− 1 which were 65% and 25% higher than the next highest performing diet of Tetraselmis striata/C. muelleri in postlarvae and juveniles, respectively. The T. striata/C. muelleri diet, which is limited in the n-3 fatty acid docosahexaenoic acid (DHA), yielded very poor growth of sea scallop postlarvae in a prior study, indicating that bay scallops may have less stringent requirements for DHA than sea scallops. The Pav 459/C. muelleri diet, which also supported the highest growth of sea scallop postlarvae, is characterized by elevated levels of the n-6 fatty acids, arachidonic (AA) in C. muelleri and 4,7,10,13,16-docosapentaenoic (DPA) in Pav 459. The two diets deficient in AA and n-6 DPA, Pavlova lutheri/Thalassiosira weissflogii and P. lutheri/Fragilaria familica, yielded the lowest growth rates in both bay scallop postlarvae and juveniles. Tissue enrichment of these two fatty acids relative to the diet, as well as overall enrichment in ∑n-6 fatty acids was observed across developmental stages and dietary treatments. A similar pattern has previously been observed in sea scallop postlarvae, suggesting a dietary requirement for n-6 fatty acids in pectinids that has often been overlooked in the past.     

黄海北部大连沿岸虾夷扇贝天然苗采集技术研究

在调查了2005、2006年黄海北部大连海域虾夷扇贝的繁殖期、浮游幼虫的时空分布的基础上,对虾夷扇贝海区天然采苗进行了研究。结果表明,2005、2006年该区域的獐子岛海域、大长山岛海域、广鹿岛海域养殖的虾夷扇贝的繁殖期在4月初至5月上旬,盛期在4月15～25日。4月上旬至6月中旬,在此区域均可发现大量的虾夷扇贝浮游面盘幼虫,不同区域浮游幼虫密度变化很大。5月底至6月初是面盘幼虫集中的附着变态时期,在海区投放附着袋可进行虾夷扇贝天然苗的采集。从各海区虾夷扇贝浮游幼虫的拖网调查和采苗结果看,在虾夷扇贝主要养殖区獐子岛、大长山岛海域和广鹿岛海域,浮游幼虫在浮游的初始阶段能够形成较高的密度,但在附着阶段密度极低,不能采集到大量苗种,但在远离虾夷扇贝养殖区的大李家湾和凌水湾,浮游幼虫的出现比养殖区晚,在幼虫附着期能达到较高密度。能够采集到大量天然苗种。2005年在凌水湾的采苗数量达526±131个/袋(壳长0.6～1 mm),2006年在大李家湾的采苗数量达673±200个/袋(壳长0.6～1mm)。     

饥饿和再投喂对日本黄姑鱼代谢率和消化器官组织学的影响

采用封闭式呼吸仪测定日本黄姑鱼(8.14±1.07 g)在饥饿和恢复投喂过程中的耗氧率;用石蜡切片法观察和分析饥饿和再投喂对日本黄姑鱼消化器官形态结构和组织学的影响。结果表明:随着饥饿时间的延长,其耗氧率逐渐降低;恢复投喂之后,其耗氧率开始逐步上升。从组织结构看,食道无明显变化,而胃、肠和肝胰脏则随着饥饿时间的延长出现不同程度的损伤;饥饿5 d组无明显变化,而饥饿15 d组变化明显,表现为胃腺细胞收缩,皱壁和上皮细胞高度减少,分泌颗粒减少,肠胃绒毛退化,肝组织致密,肝细胞内脂滴减少,胰腺泡缩小,排列不规则;饥饿10 d组介于饥饿5 d组和饥饿15 d组之间,再投喂各主要结构均有恢复。