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.     

银鲳亲鱼不同组织的氨基酸及其随性腺发育的变化

测定了海捕银鲳亲鱼肌肉,肝脏和性腺组织的氨基酸,并分析了不同性腺发育阶段的氨基酸变化。结果表明,雄性V期亲鱼精巢中的牛磺酸含量最高（27.8mg/g）,显著高于肌肉和肝脏组织,而雄性V期亲鱼氨基酸总量在肌肉中最高（631.4 mg/g）,其次是精巢（544.8 mg/g）和肝脏（468.0 mg/g）组织。雌性亲鱼肝脏中的牛磺酸含量（13.7 mg/g）显著高于卵巢和肌肉组织。在卵巢从III期发育到V期过程中,雌性亲鱼肌肉和肝脏中的氨基酸总量在V期下降显著,而卵巢氨基酸总量维持稳定。在总氨基酸的组成上,肌肉中含量高的氨基酸为Glu,Lys, Asp,Leu和Arg;肝脏中含量高的为Glu,Lys, Asp,Val和Leu;精巢中含量高为Glu,Asp, Lys, Arg和Val;卵巢含量高中为Glu,Lys,Leu,Val和Asp。性别和性腺发育阶段对亲鱼肌肉总氨基酸组成无影响。游离氨基酸含量在III 、IV和 V期卵巢中的含量分别为17.7 mg/g ,41.5 mg/g和29.6 mg/g,且其组成随性腺发育阶段的不同而有显著差异。     

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。     

Direct actions of serotonin on gonadotropin-II and growth hormone release from goldfish pituitary cells: interactions with gonadotropin-releasing hormone and dopamine and further evaluation of serotonin receptor specificity

In this study, the direct actions of serotonin (5HT) on gonadotropin (GTH)-II and growth hormone (GH) release in the goldfish were tested at the pituitary cell level. 5HT (10 nM - 10 µM) stimulated GTH-II but inhibited GH release from perifused goldfish pituitary cells in a dose-dependent manner. The minimal effective dose of 5HT tested to suppress basal GH secretion (10 nM) was 10-fold lower than that to stimulate GTH-II release (100 nM). The GTH-II releasing effect of 5HT was abolished by repeated 5HT treatment (10 µM) whereas the corresponding inhibition on GH release was unaffected. These results suggest that 5HT receptors on goldfish gonadotrophs and somatotrophs exhibit intrinsic differences in terms of sensitivity to stimulation and resistance to desensitization. Salmon GTH-releasing hormone (sGnRH, 100 nM) stimulated GTH-II and GH release from goldfish pituitary cells. The GTH-II releasing action of sGnRH was unaffected by simultaneous treatment of 5HT (1 µM). However, the corresponding GH response to sGnRH (100 nM) was inhibited. In the goldfish, dopamine is known to stimulate GH release through activation of pituitary D1 receptors. In the present study, the GH-releasing action of dopamine (1 µM) and the D1 agonist SKF38393 (1 µM) was significantly reduced by 5HT (1 µM). To examine the receptor specificity of 5HT action, the effects of 5HT1 and 5HT2 analogs on GTH-II and GH release were tested in goldfish pituitary cells. The 5HT1 agonist 8OH DPAT (0.1 and 1µM) and 5HT2 agonist methyl 5HT (0.1 1µM) mimicked the GTH-II releasing effect of 5HT. The 5HT1 agonist 8OH DPAT (0.1 and 1µM) also stimulated GH release but the 5HT2 agonist methyl 5HT (0.1 and 1µM) was inhibitory to basal GH secretion. In addition, 5HT (1µM) -stimulated GTH-II release was abolished by the 5HT1 antagonist methiothepin (10µM) and 5HT2 antagonist mianserin (10µM). Similarly, the inhibitory action of 5HT (1µM) on basal GH release was blocked by the 5HT2 antagonist mianserin (10µM). The 5HT1 antagonist methiothepin (10µM) was not effective in this regard. These results, taken together, indicate that 5HT exerts its regulatory actions on GTH-II and GH release in the goldfish directly at the pituitary cell level, probably through interactions with other regulators including sGnRH and dopamine. The GTH-II releasing action of 5HT is mediated through 5HT2 and possibly 5HT1 receptors. The inhibition of 5HT on basal GH release is mediated through 5HT2 receptors only. Apparently, 5HT1 receptors are not involved in this inhibitory action. In this study, a paradoxical stimulatory component of 5HT on GH release by activating 5HT1 receptors is also implicated.     

Gonadal morphogenesis and sex differentiation in cultured chub mackerel,Scomber japonicus

The timing of primordial germ‐cell (PGC) migration with regard to the gonadal anlagen, gonad formation and sex differentiation was examined histologically in the chub mackerel (Scomber japonicus) at 5–190 days post hatching (dph). At 5 dph, PGCs appeared on the peritoneal epithelium surface or in the mesentery, on the dorsal side of the abdominal cavity. By 10 dph, stromal cells around the PGCs proliferated. The gonadal primordium was formed by 15 dph. The gonadosomatic index was 0.01% at 30 dph and increased thereafter (0.32% in females and 0.04% in males at 160 dph). Ovarian differentiation occurred at 30–40 dph, indicated by ovarian cavity formation (elongation and fusion of the upper and lower ovarian edges). Meiosis was subsequently initiated. A few meiotic oocytes surrounded the cavity at 50 dph; most were in the perinucleolus stage at 60 dph and attained a diameter of 60–70 μm at 190 dph. Testicular differentiation occurred at 30 dph, indicated by the formation of the sperm duct primordium. Spermatogonia gradually proliferated, developing into spermatocytes at the chromatin–nucleolus stage (after 90 dph) and subsequently into spermatids and spermatozoa (160 dph). These data could aid the development of seeding and cell‐engineering technologies for scombrid fish.     

生物絮团对罗氏沼虾体组成和消化酶活性的影响

通过向养殖水体中泼洒糖蜜构建生物絮团养殖模式,分析生物絮团营养组成,并探讨生物絮团对罗氏沼虾体组成和消化酶活性的影响。试验分对照组和试验组(生物絮团组),其中试验组在养殖过程中泼洒糖蜜。试验在室内水泥池内(2 m×2 m×0.6 m)进行,每个处理有3个重复,每个重复225尾虾(0.26 g±0.02 g),试验周期为90 d。养殖过程中不换水,糖蜜的泼洒量根据饲料投喂量进行计算(C/N为20)。结果显示:添加糖蜜能够显著促进生物絮团的形成,到第90天时,试验组的絮团体积达21.22 mL/L;而对照组为6.03 mL/L;试验组絮团粗蛋白含量为29.47%,粗脂肪含量为4.32%,二者均显著高于对照组,而粗灰分含量为11.36%,显著低于对照组;泼洒糖蜜对罗氏沼虾体组成的影响不显著,对照组和试验组肌肉粗蛋白含量分别为21.09%和21.20%,粗脂肪含量分别为2.91%和3.06%;另外,向水体中泼洒糖蜜对罗氏沼虾消化酶活性影响显著。试验组罗氏沼虾肠脂肪酶活性、胃脂肪酶活性和胰脂肪酶活性均显著高于对照组;试验组罗氏沼虾糜蛋白酶活性、胰蛋白酶活性也均显著高于对照组。但泼洒糖蜜对肠淀粉酶、胃蛋白酶、胃淀粉酶、胰淀粉酶和纤维素酶活性没有显著影响。试验表明,生物絮团营养组成丰富,能够有效提高消化酶活性。     

饥饿对斜带石斑鱼肝脏和肌肉脂肪酸组成的影响

在室外水泥池(4m×1.5m×1m)进行石斑鱼(62.3±0.6g)的饥饿实验,每池放养35尾,设3个重复,实验时间分别为0、1、2、3和4周。随着饥饿时间的延长,体重减轻,肥满度和肝体比显著降低;肌肉水分显著增加而蛋白质含量显著下降(P<0.05),脂肪含量在第1周下降后基本保持稳定。肝脏饱和脂肪酸在饥饿前3周基本保持不变,第4周显著下降。单不饱和脂肪酸在饥饿第1周保持不变,然后呈下降趋势。多不饱和脂肪酸和高不饱和脂肪酸的总和随着饥饿时间的延长,含量显著增加。与肝脏相比,肌肉中脂肪酸变化程度较小。随着饥饿时间的延长,肌肉中140减少而205增加。饥饿使肌肉中DHA/EPA比率呈下降趋势,但肝脏DHA/EPA比率显著上升;肌肉和肝脏中DHA EPA的百分比都随饥饿时间的延长而显著上升。     

Effect of Dietary Antarctic Krill Meal on Growth Performance,Muscle Proximate Composition,and Antioxidative Capacity of Juvenile Spotted Halibut,Verasper variegatus

A 7‐wk feeding trial was conducted to evaluate the effects of dietary Antarctic krill meal (AKM) on the growth performance, proximate composition of muscles, and antioxidative capacity of juvenile spotted halibut. Six diets were formulated to contain about 50% protein and 8% lipid. A control diet (R0) without AKM and the other five diets with 8.1, 16.2, 24.3, 32.4, and 42.5% AKM supplementation (R10–50) to replace 10, 20, 30, 40, and 50% fishmeal protein were used to feed to juvenile spotted halibut. The juveniles were fed with each diet using three replicates and cultivated in the indoor culture system. Results showed that the specific growth rate, feed intake, and protein efficiency ratio in the R30 and R40 groups were significantly higher than that in other groups (P < 0.05). Survival rate in the R50 group was significantly lower compared with the R0 group and the other four AKM supplementation groups. Moreover, the rising AKM levels in diets had significant effects on the chemical composition of juvenile spotted halibut, showing significantly decreased contents of crude protein, but increased lipid and ash contents (P < 0.05). The total superoxide dismutase activity and catalase activity of serum and liver in AKM groups were significantly higher than those in the control group (P < 0.05). In contrast, the contents of malondialdehyde in serum and liver were significantly lower compared with the control group. These findings illustrate that a moderate AKM level in diets can significantly improve the growth performance, feed utilization, and antioxidative capacity in juvenile spotted halibut, which support the finding that AKM may be used as a good protein source for halibut in the aquafeed industry.     

The changes in proximate compositions and glycogen contents in the dorsal ordinary muscles of the full-cycle cultured Pacific bluefin tuna Thunnus orientalis occurring with growth

ABSTRACT:   Using full-cycle cultured (FC) Pacific bluefin tuna (body length [BL], 42.6–66.4 cm; body weight [BW], 1.66–7.40 kg, n  = 15), the changes in chemical compositions and histological structure of the cephalal parts of the dorsal ordinary muscles (DOM) occurring with growth were investigated. A positive correlation ( r  = 0.9644, P  < 0.05) was observed between BL and BW with growth. The protein, lipid and ash contents of DOM and condition factor did not change with growth. However, the glycogen content of DOM increased ( P  < 0.05) from approximately 55 cm (BL) in this study. Using optical microscopic photographs, the various shapes of muscle fibers were observed and it was noted that the muscle fiber diameter increased ( P  < 0.05) with growth. Using transmission electron microscopic observation, many glycogen granules were observed in muscle fibers (especially, side of connective tissue) of DOM throughout the growth stage in this study. These results indicate that the glycogen content of DOM of FC Pacific bluefin tuna increases before the lipid with growth.     

刺参血系统的超微结构观察与功能探讨

在光学显微镜、扫描电镜和透射电镜下观察刺参血系统的组织细胞结构，探讨血系统的生理功能，结果显示：(1)刺参的血系统由复杂的管网构成，排列紧密的血管上皮细胞和上皮下连续成层的基膜，构成血系统中连续而封闭的管状结构，管腔中充满血液，血系统的管网承担了血管输送血液的功能，并通过与呼吸树的连接完成血液的气体交换。(2)血管上皮细胞游离面的细胞膜、微绒毛和吞饮小泡呈酸性磷酸酶阳性反应，表明细胞具有吸收功能。血管上皮细胞内的小泡在管壁内外的物质运输中起作用。由单层细胞和基膜构成的血管壁是血液与体腔液进行物质交换的场所。(3)血管上皮细胞富含内质网、脂滴、线粒体及分泌自噬体，具有类固醇激素分泌细胞的超微结构特点，可能与刺参体内的固醇合成有关。