鲤鱼肥胖基因的分子克隆及在大肠杆菌中的表达

为了研究鲤鱼肥胖基因的结构特点和体外表达产物的生物学活性 ,利用RT PCR技术从鲤鱼肠系膜脂肪组织中扩增出鲤鱼肥胖基因的cDNA编码序列 ,分析表明该cDNA序列由 4 38个核苷酸组成 ,编码 14 6个氨基酸组成的多肽 ,鲤鱼肥胖基因与人、猪、鼠的相比 ,核苷酸同源性分别为 :84 %、 86 %、 95 % ;氨基酸的同源性分别为 84 %、 82 %、 96 %。构建了原核表达载体 pET 2 8a li,利用IPTG在大肠杆菌中进行了诱导表达 ,并对表达产物进行了初步纯化和生物活性检测 ,结果表明 ,鲤鱼肥胖基因在大肠杆菌中进行了高效特异性融合表达 ,融合蛋白质分子量约为 2 0kD ,经薄层扫描分析 ,目的蛋白占菌体总蛋白的 2 0 3%。表达产物经过纯化和复性能够明显抑制小鼠的摄食和生长 ,说明表达产物Leptin具有明显的生物学活性     

Neural Regulation of Dark-Induced Abundance of Arylalkylamine N-Acetyltransferase (AANAT) and Melatonin in the Carp (Catla catla) Pineal: An In Vitro Study

In all the vertebrates, synthesis of melatonin and its rhythm-generating enzyme arylalkylamine N-acetyltransferase (AANAT) reaches its peak in the pineal during the night in a daily light-dark cycle, but the role of different neuronal signals in their regulation were unknown for any fish. Hence, the authors used specific agonist and antagonists of receptors for different neuronal signals and regulators of intracellular calcium (Ca²⁺) and adenosine 3',5'-cyclic monophosphate (cAMP) in vitro to study their effects on the abundance of AANAT and titer of melatonin in the carp (Catla catla) pineal. Western blot analysis followed by quantitative analysis of respective immunoblot data for AANAT protein, radioimmunoassay of melatonin, and spectrophotometric analysis of Ca²⁺ in the pineal revealed stimulatory effects of both adrenergic (α₁ and β₁) and dopaminergic (D₁) agonists and cholinergic (both nicotinic and muscarinic) antagonists, inhibition by both adrenergic and dopaminergic antagonists and cholinergic agonists, but independent of the influence of any agonists or antagonists of α₂-adrenergic receptors. Band intensity of AANAT and concentration of melatonin in the pineal were also enhanced by the intracellular calcium-releasing agent, activators of both calcium channel and adenylate cyclase, and phophodiesterase inhibitor, but suppressed by inhibitor of calcium channel and adenylate cyclase as well as activator of phophodiesterase. Moreover, an inhibitory effect of light on the pineal AANAT and melatonin was blocked by both cAMP and proteasomal proteolysis inhibitor MG132. Collectively, these data suggest that dark-induced abundance of AANAT and melatonin synthesis in the carp pineal are a multineuronal function, in which both adrenergic (α₁ and β₁, but not α₂) and dopaminergic signals are stimulatory, whereas cholinergic signals are inhibitory. This study also provides indications, though arguably not conclusive evidence, that in either case the neuronal mechanisms follow a signal-transduction pathway in which Ca²⁺ and cAMP may act as the intracellular messengers. It also appears that proteasomal proteolysis is a conserved event in the regulation of AANAT activity in vertebrates. (Author correspondence: dgp_skmaitra@yahoo.co.in)     

Verdaulichkeit,wachstum und körperzusammensetzung von karpfen (Cyprinus carpio L.) nach verfütterung verschiedener stärkearten

Maisstärke, Maisquellstärke, Weizenstärke, Weizenquellstärke, Kartoffelstärke oder Maniokmehl werden in Anteilen von 41% bzw. 46,7% (Maniokmehl) einer Basalration zugemischt. Jede Versuchsration wurde an insgesamt 33 Karpfen (Cyprinus carpio L.) (drei Becken mit je 11 Karpfen) in einer Menge von 1,8‐ 2,0 % der Lebendmasse über eine einheitliche Versuchsdauer von 118 Tagen ab einer mittleren Lebendmasse von 146 g verfüttert. Die 4 Rationen auf Basis von Mais‐ bzw. Weizenstärke weisen insgesamt die gleiche, hohe Energieverdaulichkeit von etwa 90% auf. Die Zumischung von Kartoffelstärke bzw. von Maniokmehl verringert die Energieverdaulichkeit um 5 % bzw. 3% signifikant. Das Wachstum und der Futteraufwand werden durch die unterschiedlichen Stärkearten nicht beeinflußt. Zu Versuchsende weisen die Karpfen eine mittlere Masse von 520 g auf. Die Ganzkörper haben insgesamt mittlere Gehalte an Rohasche, Rohprotein und Gesamtfett bzw. Energie von 2,8, 15,7,9,4% bzw. 7,4 kJ pro g Frischmasse. Allerdings führt die Zumischung von Kartoffelstärke bzw. Maniokmehl zu einer deutlich geringeren Energieretention gegenüber Mais‐ und Weizenstärke, da die Trockensübstanz‐, Fett‐ und Energiegehalte dieser Karpfen signifikant niedriger sind.     

Temperature acclimation of gross cardiovascular morphology in common carp (Cyprinus carpio)

Temperature is of fundamental importance in the functioning of the cardiovascular system of ectothermic fish, with cold-induced ventricular hypertrophy and increased red muscle mass being reported in a number of fish species upon cold acclimation. This study demonstrates a non-linear cold-induced ventricular hypertrophy in common carp (relative ventricular mass (RVM)=0.086±0.003%, 0.074±0.005% and 0.074±0.004% at 5, 15 and 25 °C, respectively), but a cold-induced atrophy of the lateral red muscle mass (RMM) with respect to total muscle mass (2.504±0.554%, 3.982±0.818% and 4.490±0.256% at 5, 15 and 25 °C, respectively). The ventricular hypertrophy in carp acclimated to 5 °C reflects a compensation against the increased workload of pumping viscous blood at low temperatures and the negative inotropic effects of the cold, whereas atrophy of the red muscle is likely due to disuse resulting from reduced locomotory activity in the cold.     

热休克法抑制第一次卵裂实现草鱼雌核发育的细胞学观察

用组织切片方法系统地观察了草鱼卵被经辐射处理的鲤精子激活后进行第一次卵裂的发育过程。实验表明：在24℃孵化水温下草鱼卵在被激活后24min进入第一次卵裂胶期,27－30min处于中期,33min进入后期。由此可知被激活的草鱼卵子在第24min时已经完成染色体的复制,使草鱼卵子雌核染色体人工加倍的最佳时期是在被激活后的27－30min这一时间区段内。此外,用不同热休克温度和不同的热休克强度处理已完成染色体复制的被激活草鱼卵,表明草鱼卵经41℃处理2min可得到较高比例的基因纯合型雌核发育二倍体鱼。     

四株草鱼呼肠孤病毒毒株的细胞感染特性比较研究

本文首次对低温保存的三株草鱼呼肠孤病毒GCRV873 、GCRV875、GCRV876与新分离的GCRV991毒株进行了细胞培养与病毒感染特性等比较研究。结果表明 ,GCRV873 、GCRV875、GCRV876在 - 30℃保存 10年后仍然具有一定的感染性 ,其滴度均在 10 2 TCID50 /mL以上 ,略低于从病鱼组织分离的GCRV991毒株的滴价。经传代培养后 ,四株GCRV的毒力逐渐升高 ,并趋于稳定 ;当感染复数 (MOI)为 0 .0 5PFU/cell时 ,测定四株GCRV的滴度均高于 10 8TCID50 /mL ,但略有差异。GCRV873 的滴度最高 ,可达到 6 .4× 10 11TCID50 /mL。连续传代的GCRV毒株在不同温度 (2 8℃、31℃、34℃、37℃、41℃ )条件下 ,均可感染CIK细胞 ;在 2 8℃时 ,感染效价最高 ,随着温度的升高 ,其感染效价逐渐降低     

The carp homeobox gene Ovx1 shows early expression during gastrulation and subsequently in the vagal lobe, the facial lobe and the ventral telencephalon

 The homeobox gene Carp-Ovx1 shows similarity to vertebrate and invertebrate Ovx genes and to Drosophila unplugged. Its expression pattern was studied by in situ hybridization in carp embryos and juveniles. During segmentation, expression becomes gradually limited to the neural tube. In juveniles up to 9 weeks old, cells in the ventral telencephalon, the facial lobe and the vagal lobe show Ovx1 expression, confining expression to parts with chemosensory projections. Received: 29 October 1997 / Accepted: 12 November 1997