翘嘴鲌鱼种对磷的需求量

以磷酸二氢钠为磷源,制成含磷水平为0.53%—1.61%的7种半精制试验饲料,饲养7组三重复的翘嘴隔冬鱼种。试验鱼每尾平均体重3.79±0.20g。饲养期为8周。试验结果表明,饲料磷含量不足会导致鱼体生长不良,饲料效率低下。饲料磷为0.88%或以上时,鱼体增重和饲料效率显著提高。全鱼脂肪含量随饲料磷水平上升而呈下降趋势,但各试验组间无显著差异(p>0.05)。全鱼的粗灰分含量与饲料磷水平呈正相关。饲料磷水平为0.88%或更高时,全鱼磷含量和脊椎骨的灰分含量稳定在同一水平上。饲料磷水平对全鱼水分、全鱼粗蛋白、鳞片灰分、鳞片磷和脊椎骨磷含量无显著影响(p>0.05)。因此,满足翘嘴鱼种生长所需的饲料磷为0.88%。     

武昌东湖蒙古红鲌和翘嘴红鲌的食性及其种群控制问题的研究

本文主要报道蒙古红鲌和翘嘴红鲌对放养鳙、鲢鱼种的危害性及控制其种群发展的途径。通过肠道内食物的检查,发现不同大小的蒙古红鲌和翘嘴红鲌与所吞食“家鱼”鱼种规格的大小有一定的相关,根据所得的数据分别求出这两种凶猛鱼的全长与被吃鱼种全长的迥归方程式及95％的可信限,确定不同大小的蒙古红鲌和翘嘴红鲌危害不同规格鱼种的范围。并通过对东湖这两种凶猛鱼种群长度的调查,提出鳙、鲢鱼种放养的合理规格。蒙古红鲌和翘嘴红鲌全年各月的摄食强度随着水温的不同和生殖季节的来临而有所改变,根据这种情况,对于什么时间放养鱼种较为有利也进行了探讨。从两种鱼的食物组成和摄食强度来看,它们对鳙、鲢的危害性是很明显的。为了遏制其种群发展,进行了围捕产卵群体、药物杀卵和投放棕榈皮鱼巢诱其产卵等试验。试验证明,在生殖季节当它们大量集群时进行围捕,效果较为显著。近年来这两种凶猛鱼的产量和种群的长度组成都有所下降。文中附有不同长度的蒙古红鲌和翘嘴红鲌吞食不同鳙鱼种的范围表,供有关单位参考。     

饲料碳水化合物水平对南方鲇幼鱼餐后糖酵解酶活性及血糖浓度的影响

配制含0%、15%和30%糊化玉米淀粉的等能饲料分别作为对照、中水平和高水平碳水化合物(CHO)饲料,以体重为50.5±0.6g的南方鲇幼鱼为实验对象,在水温27.5±0.5℃的条件下以对照饲料驯化15d后,分别测定了以三种饲料投喂的南方鲇在餐后3、61、2、24和48h的己糖激酶(HK)、葡萄糖激酶(GK)和磷酸果糖激酶(PFK-1)的活性和血糖水平。结果发现,中、高水平CHO组HK活性在餐后24h时显著高于对照组(P<0.05),其余时间点各组间无显著差异;GK活性随CHO水平增加而增加,但其活性的绝对值远低于HK;PFK-1活性在各组间及同组内各时间点均无显著差异。通过分析认为,南方鲇体内葡萄糖磷酸化主要由HK催化,但其活性受饲料CHO水平的影响不明显;GK活性受饲料CHO的诱导而增高,使葡萄糖磷酸化加速,但最大增幅不超过30%;此外,催化果糖-6-磷酸进一步转化为果糖-1,6-二磷酸的PFK-1活性不受饲料CHO水平的影响,因此糖酵解过程的这两个代谢环节均不能在鱼体吸收高糖营养后有效地加速,这应当是该肉食性鱼类在高水平CHO营养条件下产生高血糖积累的重要原因。实验结果表明,血糖变化呈现先升高后降低的趋势,中水平和高水平CHO组的血糖峰值均出现在餐后12h,这作为在该类研究中设定血液取样时间的实验依据。     

饲料蛋白对翘嘴鲔生长和内分泌激素的影响

选择健康的翘嘴鲌（Culter alburnus）为试验鱼,以红鱼粉为蛋白源,配制5个蛋白水平的等能、等必需氨基酸（EAA）平衡关联度的半精制饲料,又以豆粕替代鱼粉,大豆蛋白分别替代不同水平的鱼粉蛋白,配制5个EAA关联度的等蛋白、等能的半精制饲料,探讨饲料蛋白对鱼类生长和内分泌激素等的影响。结果表明：饲料蛋白水平对翘嘴鲌的特定增重率（SGR）具有显著影响（P〈0．05）,40．89％饲料蛋白组的SGR显著高于31．04％、35．51％饲料蛋白组（P〈0．05）,但与46．62％和50．33％饲料蛋白组没有显著性差异（P〉0．05）。血清T3水平与饲料蛋白水平正相关（P〈0．05）。血清GH水平与饲料蛋白水平和生长负相关（P〈0．05）,血清IGF—I水平与饲料蛋白水平和生长正相关（P〈0．05）。适宜蛋白水平通过提高翘嘴鲌血清T,和IGF—I的水平来调控生长。当大豆蛋白分别替代54．0％的鱼粉蛋白时,翘嘴鲌的SGR与对照组差异显著（P〈0.05）。翘嘴鲌血清T1与饲料大豆蛋白对鱼粉蛋白的替代量负相关（P〈0．05）。40．5％和54．0％替代组的血清GH显著高于对照组（P〈0．05）,且与饲料中豆粕对鱼粉替代水平正相关（P〈0．01）,与生长负相关（P〈0．05）。血清IGF—I与饲料中大豆蛋白对鱼粉蛋白替代量负相关（P〈0．05）,与生长正相关（P〈0．05）。大豆蛋白的替代亦通过对内分泌激素的影响来调控生长。     

草鱼、鳡和翘嘴鲌消化道组织的早期发育

研究仔稚鱼消化机能的发育变化对于掌握鱼类早期发育阶段的消化特点、营养需要及提高仔稚鱼成活与生长等均有重要意义。采用HE、PAS等染色方法,对草鱼(Ctenopharyngodon idellus)、鳡(Elopichthys bambusa)和翘嘴鲌(Culter alburnus)消化道组织的早期发育进行了研究,结果表明:(1)初孵仔鱼卵黄囊的相对体积以鳡的最大;(2)均在孵后2d和3d分别出现肠管和口裂,在孵后3d、4d和2d分别出现肠腔;(3)在孵后4d、7—9d和4d其肠腔内分别出现食物团,表明此时草鱼、鳡和翘嘴鲌已分别开始外源性摄食;(4)在孵后5d、6d和6d其肠道内表面分别出现黏膜褶,随后在稚鱼中其黏膜褶的高度和数量不同程度的发育;(5)草鱼和鳡的肠道分别在孵后14d和30d出现盘曲,而在翘嘴鲌的切片图中未发现其肠道的盘曲;(6)草鱼、鳡和翘嘴鲌的肠道分别于孵后17—23d、30d和24—29d出现数量较多的黏液细胞,此时标志着食性的转换和分化过程基本完善。     

翘嘴红鲌(♀)×黑尾近红鲌(♂)杂种F_1的AFLP分析

采用AFLP分子标记技术对翘嘴红鲌(♀)、黑尾近红鲌(♂)及杂种F1三个群体各12个个体进行遗传差异分析。结果表明:9对引物共扩增出257条带,多态性比例为51.36%;三个群体间特异条带共24条,其中黑尾近红鲌群体和杂种F1群体共有特异带15条、翘嘴红鲌群体和杂种F1群体共有特异带2条、翘嘴红鲌群体特异带7条;黑尾近红鲌群体、翘嘴红鲌群体及杂种F1群体的Shannon’s信息指数分别为0.2326、0.0794和0.1774,Nei’s基因多样性指数分别为0.1602、0.0537和0.1229,期望杂合度分别为0.1582、0.0464和0.1062;杂种F1群体和黑尾近红鲌群体及翘嘴红鲌群体遗传距离分别为0.0523、0.2165,遗传相似系数分别为0.9490、0.8053,遗传数据表明杂种F1更接近于父本黑尾近红鲌。     

锦江翘嘴鲌的繁殖生物学特征

对2015年1月至12月采集于锦江的翘嘴鲌(Culter alburnus)进行了繁殖生物学特征的研究。锦江的翘嘴鲌繁殖期集中在6~7月,属分批产卵类型。繁殖群体年龄在3+龄至6+龄之间,体长250~537 mm,体重184.9~2 587.5 g。雌雄性比1.14︰1,体表差异表现在泄殖孔和腹部膨胀程度,雌雄群体间体长-体重关系存在显著性差异(0.01P0.05)。性成熟雌鱼绝对繁殖力25 067~54 274粒,相对繁殖力24.2~36.9粒/g,平均卵径(1.1±0.3)mm。性成熟系数在1~5月份逐渐增大,6月明显上升,7月达到最高峰,8月显著下降,9~12月逐渐趋于平缓。采用Logistic方程推算了初次性成熟个体特征,雄性体长273 mm,体重192.0 g,年龄3.4龄;雌性体长311 mm,体重249.4 g,年龄4.2龄。锦江种群繁殖期与其他地理种群相近,但绝对繁殖力和相对繁殖力明显偏小。     

淀山湖翘嘴鲌的年龄结构与生长特性

研究翘嘴鲌的年龄结构与生长特征可为探其繁殖、性成熟年龄、存活率等习性积累有效数据, 并可为优化鱼类种群结构、科学利用其种质资源提供参考依据。以2016年5月至2017年4月逐月于淀山湖采集到的452尾翘嘴鲌(Culter alburnus)为研究材料, 研究其年龄结构与生长特征间的紧密联系。结果表明: 翘嘴鲌体长15.32—77.91 cm; 体重范围43—5567 g。雌雄群体间的体长和体重的差异不显著(P>0.05), 体长和体重拟合关系式为W=0.00002L2.9211 (R2=0.9143, n=452), 符合匀速生长特性; 选用鳞片鉴定年龄、测量鳞径, 并建立von Berta-lanffy生长方程, L_t=99.65[1–e–0.1357(t+0.6287)]; W_t=11874.27[1–e–0.1357(t+0.6287)]2.9211。采集的翘嘴鲌样本由1—6龄组成, 优势年龄组3龄, 占样本总数的55.71%, 表明生长趋于低龄化、小型化; 生长拐点年龄为7.2711龄时对应的体长和体重分别为65.54 cm和3471.79 g。     

不同养殖密度对翘嘴鳜生长、血液生化和抗氧化能力指标的影响

研究旨在探讨不同养殖密度对翘嘴鳜(Siniperca chuatsi)生长、血液生化和抗氧化能力指标的影响。在循环水系统的水箱中(60 cm×60 cm×40 cm)于5种密度下[超低养殖密度组(ELD; 3.34 kg/m³)、低养殖密度组(LD; 9.51 kg/m³)、中等养殖密度组(MD; 15.82 kg/m³)、高养殖密度组(HD; 21.05 kg/m³)和超高养殖密度组(EHD; 25.99 kg/m³)]养殖翘嘴鳜[平均体重(175.76±15.85) g]40d。结果表明, ELD组和LD组在增重率(WGR)和饲料转化率(FCR)方面表现出更好的生产性能(P<0.05),并增加血浆超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)的活性,同时减少丙二醛(MDA)的形成(P<0.05)。此外, EHD组的血浆葡萄糖水平、总胆固醇水平和甘油三酯水平均较高(P<0.05)。LD组刺鼠基因相关蛋白(agrp)的mRN...     

翘嘴鲌胰岛素样生长因子-Ⅰ的克隆及序列分析

为探讨胰岛素样生长因子-Ⅰ(Insulin like growth factor-Ⅰ, IGF-Ⅰ)对于翘嘴鲌(Culter alburnus)生长性状的影响, 对其DNA序列进行了克隆。翘嘴鲌IGF-Ⅰ全长14567 bp, 由5个外显子和4个内含子组成。其5个外显子长度分别为298、160、182、36 和1360 bp。推测的阅读框为486 bp, 编码由161个氨基酸组成的IGF-Ⅰ前体蛋白。前体肽由信号肽、成熟肽、E肽三部分组成, 其中信号肽44个氨基酸, 成熟肽70个氨基酸, E肽47个氨基酸。成熟肽由B、C、A、D四个区域组成, 其中B结构域和A结构域的保守性最高, 在这2个区域包含由6个半胱氨酸残基形成的3个二硫键。翘嘴鲌B区域还包含保守的IGF-Ⅰ受体识别序列(PheB23-TyrB24-PheB25)。E肽的长度表明翘嘴鲌IGF-Ⅰ属Ea-2型。同源性分析表明翘嘴鲌与鲤科鱼类的IGF-Ⅰ编码氨基酸同源性较高, 为94%—100%, 但在聚类分析中翘嘴鲌并不是首先和鲌亚科的鱼类聚集在一起。Real-time qPCR组织特异性表达结果显示IGF-Ⅰ mRNA在肝脏组织中的表达量最高, 脾、心脏、精巢、脑次之, 肾、鳃、胃和卵巢中表达量较低。翘嘴鲌IGF-Ⅰ基因4个内含子长度分别为1170、9364、251 和1746 bp。相对外显子来说, 种间内含子变异较大, 其中第三内含子变异最大。翘嘴鲌IGF-Ⅰ基因中包含6个微卫星, (GATG)₅AATAT (ATAG)₁₁位于第一内含子中, (CT)₈、(TTA)₅、(AC)₁₃、(TG)₁₂和(ATT)₅位于第二内含子中。其中4个微卫星位点具有多态性, 将它们在120尾同塘养殖的翘嘴鲌中进行基因型与生长性状的关联性分析, 均未达到显著水平(P>0.05)。结果为进一步研究该基因的表达、功能及其转录调控特征奠定了分子基础。     

抗菌肽GK1在大肠杆菌中的融合表达

为高效表达抗菌肽GK1并避免GK1的高抗菌活性对大肠杆菌宿主菌的致命影响, 将经改造后的人胰岛素原(mhPI)与GK1的融合基因(mhPI-GK1)克隆到表达载体pET28a中, 构建出表达质粒pET28a-mhPI-GK1, 转化至大肠杆菌BL21(DE3)中进行表达。融合蛋白在大肠杆菌中以包涵体形式表达, 表达量占菌体总蛋白的20%。经CNBr裂解、阳离子交换层析和RP-HPLC纯化后, 每升发酵液可获得5.7 mg纯度大于97%的重组GK1。质谱检测显示重组GK1的分子量为2794.0 D, 抑菌活性实验表明纯化后的重组GK1和化学合成GK1具有相同的抗菌活性。为利用基因工程方法大规模生产GK1奠定了基础。     

Discovery of novel 3,6-disubstituted 2-pyridinecarboxamide derivatives as GK activators

A novel class of 3,6-disubstituted 2-pyridinecarboxamide derivatives was designed based on X-ray analysis of the 2-aminobenzamide lead class. Subsequent chemical modification led to the discovery of potent GK activators which eliminate potential toxicity concerns associated with an aniline group of the lead structure. Compound 7 demonstrated glucose lowering effect in a rat OGTT model.     

Comparison of GK1.5 and chimeric rat/mouse GK1.5 anti-CD4 antibodies for prolongation of skin allograft survival and suppression of alloantibody production in mice.

GK1.5 is a rat mAb that recognizes the mouse CD4 Ag. It has been shown to deplete CD4+ cells in vivo and to be immunosuppressive. To evaluate the effect of the C region of this antibody in achieving cell depletion, chimeric antibodies, each having the rat GK1.5 V regions and one of the four mouse IgG C region isotypes, were compared with the native rat antibody. The chimeric antibodies and the native antibody were tested for their ability to mediate in vitro C-dependent cytotoxicity, in vivo cell depletion, and prolongation of allogeneic skin graft survival and suppression of alloantibody production. In vitro C-dependent cytotoxicity assays revealed that rat IgG2b and the chimeric antibodies containing mouse IgG2a, mouse IgG2b, and mouse IgG3 were effective in lysing CD4+ lymphocytes whereas mouse IgG1 was ineffective. In vivo studies of CD4+ cell depletion showed that mouse IgG2a, rat IgG2b, and mouse IgG2b were effective isotypes, mouse IgG1 was less effective, and mouse IgG3 did not deplete CD4+ cells. A correlation was found between the ability of an isotype to deplete CD4+ cells in vivo and its ability to prolong the survival of skin allografts and to suppress alloantibody production. The nondepleting mouse IgG3 was ineffective in these assays. Overall the most effective mouse isotype was IgG2a which was as effective as rat IgG2b. These results indicate 1) that syngeneic isotypes of mAb can cause cell depletion and consequently the prolongation of allograft rejection and suppression of alloantibody production; 2) that not all isotypes are equally effective; and 3) that the ability of a given isotype to deplete cells in vivo does not correlate with its ability to mediate C-dependent lysis in vitro. Our results are consistent with the hypothesis that in vivo depletion of cells is mediated by opsonization and binding through the FcR.     

Down-regulated expression of exocytotic proteins in pancreatic islets of diabetic GK rats

Exocytosis is regulated by exocytotic proteins, which are present in insulin-secreting beta-cells and play regulatory roles in insulin secretion. Non-insulin dependent diabetes mellitus (type 2 diabetes) is a disease characterized by impaired insulin secretion and insulin resistance. Exocytotic protein immunoreactivities were studied in pancreatic islets of type 2 diabetic Goto-Kakizaki (GK) rats using immunofluorescence histochemistry. The immunoreactivities for vesicle-associated membrane protein-2 (VAMP-2), synaptotagmin III, cysteine string protein (CSP), mammalian homologue of the unc-18 gene (Munc-18), alpha-soluble N-ethylmaleimide-sensitive attachment protein (alpha-SNAP), N-ethylmaleimide-sensitive factor (NSF) and synaptosomal-associated protein of 25 kDa (SNAP-25) exhibited weaker immunofluorescence intensity in islets of GK rats as compared to control Wistar rats. Insulin immunoreactivity was also decreased in GK rat beta-cells, whereas no detectable alterations in the expression of actin immunoreactivity could be detected. The data suggest that reduced expression of exocytotic proteins and decreased insulin content may contribute to the diabetic syndrome in the GK rat.     

Comparison of the pharmacological properties of GK11 and MK801, two NMDA receptor antagonists: towards an explanation for the lack of intrinsic neurotoxicity of GK11

Over-stimulation of NMDA receptors (NMDARs) is involved in many neurodegenerative disorders. Thus, developing safe NMDAR antagonists is of high therapeutic interest. GK11 is a high affinity uncompetitive NMDAR antagonist with low intrinsic neurotoxicity, shown to be promising for treating CNS trauma. In the present study, we investigated the molecular basis of its interaction with NMDARs and compared this with the reference molecule MK801. We show, on primary cultures of hippocampal neurons, that GK11 exhibits neuroprotection properties similar to those of MK801, but in contrast with MK801, GK11 is not toxic to neurons. Using patch-clamp techniques, we also show that on NR1a/NR2B receptors, GK11 totally blocks the NMDA-mediated currents but has a six-fold lower IC₅₀ than MK801. On NR1a/NR2A receptors, it displays similar affinity but fails to totally prevent the currents. As NR2A is preferentially localized at synapses and NR2B at extrasynaptic sites, we investigated, using calcium imaging and patch-clamp approaches, the effects of GK11 on either synaptic or extrasynaptic NMDA-mediated responses. Here we demonstrate that in contrast with MK801, GK11 better preserve the synaptic NMDA-mediated currents. Our study supports that the selectivity of GK11 for NR2B containing receptors accounts contributes, at least partially, for its safer pharmacological profile.     

Significant Improvement of Insulin Resistance of GK Rats by Treatment with Sodium Selenate

We studied the effect of sodium selenate on insulin resistance of Goto-Kakizaki (GK) rats. Rats were kept on standard laboratory chow with and without i.p. injections of sodium selenate (0.173 mg/kg body weight) for 14 days, and then subjected to the glucose clamp. The glucose clamp studies confirmed an improvement in insulin-stimulated glucose disposal in GK rats treated with sodium selenate, with respect to both insulin sensitivity and responsiveness. This amelioration of insulin resistance may be partly due to a direct effect of the sodium selenate on peripheral tissues. 2-Deoxyglucose uptake in sodium selenate-treated adipocytes was increased and the insulin findings suggest that sodium selenate increases not only insulin sensitivity but also insulin responsiveness. Sodium selenate also accelerated glucose incorporation into adipocytes of rats, suggesting that the action of sodium selenate is peripheral. Interestingly, sodium selenate at a high concentration (about 1 mmol/L) was more effective than insulin in enhancing glucose uptake. The present study suggested that sodium selenate treatment led to substantial improvement in peripheral insulin resistance.     

Adenylyl cyclase isoform expression in non-diabetic and diabetic Goto-Kakizaki (GK) rat pancreas. Evidence for distinct overexpression of type-8 adenylyl cyclase in diabetic GK rat islets

Glucose-induced insulin release is markedly decreased in the spontaneously diabetic Goto-Kakizaki (GK) rat pancreas. This defect was recently shown to be reversed by forskolin which markedly enhances cAMP generation in GK islets. These effects of forskolin were associated with overexpression of type-3 adenylyl cyclase (AC) mRNA due to the presence of two functional point mutations in the promoter region of AC3 gene in GK rat. Nine AC isoforms have been described, but their expression pattern in relation to the main pancreatic islet cell types, as well as their involvement in the diabetic state, is still unknown. Using antibodies raised against AC1–8, we have studied by double immunofluorescence the localisation of these AC isoforms in different endocrine cell types in both normal and diabetic GK rat pancreas. Our results demonstrated a clear immunoreaction (IR) to AC1–4 and 6 in normal and GK islet β-cells, while a smaller number of ACs were expressed in α- and δ-cells. No AC-IR was observed in pancreatic polypeptide cells. Moreover, we have found an increased IR of the Ca²⁺-stimulated AC1, AC3 and AC8 in diabetic β- and α-cells, compared with the corresponding IR in control pancreas. Most noticeable was the eliciting of a markedly enhanced AC8-IR in GK rat β- and α-cells, in contrast to a barely discernible AC8-IR in corresponding normal cells. In conclusion, AC expression exhibits a complex pattern in the endocrine pancreas, with specific differences between the normal and diabetic state. Accepted: 25 November 1999     

Differential activations of PKC/PKA related to microvasculopathy in diabetic GK rats

Microvasculopathy is the most serious and predictable threat to the health of diabetic patients, which often results in end-stage renal disease, blindness, and limb amputations. Up to the present, the underlying mechanisms have remained elusive. Here, it was found that the differential activations of PKC/PKA were involved in diabetic microvasculopathy in diabetic GK rats. By real-time PCR, Western blot, immunohistochemistry, and enzyme activity assay, upregulation of PKC was prominent in kidney but was not significant in liver and brain. The expression and activity of PKA were lowered in kidney but comparable in brain and liver during diabetic nephropathy. Furthermore, the generation of reactive oxygen species, production of nitric oxide, and expression of inducible nitric oxide synthase induced by advanced glycation end products were inhibited by PKCβ inhibitor LY-333531 or a PKA agonist in rat glomerular microvascular endothelial cells. Finally, albuminuria was significantly lowered by a PKA agonist and boosted by a PKA antagonist. It suggested that the differential activations of PKC/PKA related to microvasculopathy in diabetes and that activation of PKA may protect the diabetic microvasculature.