刀鲚肌间骨新类型的发现

文章对刀鲚(Coilia nasus)的肌间骨数目、形态、分布特征及组织学结构进行了系统研究。结果表明刀鲚肌间骨的形态种类与普通鲤科鱼类没有显著差别,但其分布类别却有很大不同,刀鲚不仅有髓弓小骨、椎体小骨和脉弓小骨,且在其背部上方和腹部下方仍存在大量肌间骨,这两类肌间骨的形态为简单的"1"形和"("形,分别位于背部上方和腹部下方肌膈的皱褶部;在短颌鲚(C. brachygnathus)背部上方和腹部下方也发现这两类肌间骨的存在,统称为肌骨杆(Myorhabdoi)。刀鲚个体肌间骨数目范围为492—543,其中髓弓小骨为114—142 (■=133),椎体小骨为28—51 (■=42),脉弓小骨为138—153 (■=142),背上肌骨杆为92—135 (=■114),腹下肌骨杆为66—98 (■=89)。采用茜素红与阿利新蓝组织染色观察不同类型肌间骨的组织结构特征,结果显示肌间骨由肌膈结缔组织包被,在髓弓小骨和脉弓小骨两种类型中,肌间骨之间通过结缔组织相互连接。研究在鱼类背上肌骨杆和腹下肌骨杆的发现,是对我国鱼类肌间骨系统进化研究的进一步补充。     

鲂属鱼类杂交子代肌间骨的形态学比较

本研究对团头鲂(Megalobrama amblycephala)与鲂属其他3种鱼类三角鲂(M.skolkovii)、广东鲂(M.terminalis)和厚颌鲂(M.pellegrini)正反交及其亲本自交共10种组合子代的肌间骨数目、形态、分布和长度进行了比较分析。10种不同繁育组合子代个体的肌间骨总数范围为96~134,平均值为119,各组合肌间骨的数目无显著性差异(P0.05),其中,厚颌鲂自交组合子代个体的肌间骨数目最少(平均值为114)。不同杂交组合子代体重和体长与肌间骨的长度均呈线性相关。不同部位肌间骨长度存在差异,其中躯干轴上肌的肌间骨长度显著大于躯干轴下肌、尾部轴上肌和尾部轴下肌三个部位的肌间骨长度(P0.05)。躯干轴下肌的肌间骨数目显著少于其他部位(P0.05),尾部轴上肌和尾部轴下肌的肌间骨数目无显著性差异(P0.05)。杂交鲂肌间骨形态复杂多样,分别为"1"形、"卜"形、"y"形、一端多叉形、两端多叉形和"("形6种类型,肌间骨越靠近鱼体前端形态越复杂,且与存在部位有一定的关系,躯干轴上肌的肌间骨形态最复杂。研究结果表明,鲂属鱼类近缘杂交子一代的肌间骨相关性状特征无显著改变。     

团头鲂肌间骨发育的形态学观察

研究利用整体骨骼染色、形态学解剖和X光透射的方法, 对团头鲂(Megalobrama amblycephala)仔稚鱼肌间骨的出现时期、形态以及成鱼肌间骨数目、形态、分布和长度变化进行了观察与分析。结果表明: 团头鲂的肌间骨在孵出后20d (体长为1.33 cm)骨化出现, 首先出现在尾部, 然后向头部方向依次出现, 到第40天(体长为2.36 cm)基本全部出现; 肌间骨出现与分化的时间受生长发育的影响大于日龄的影响。团头鲂肌间骨数目在108129, 平均为119 根, 其中躯干部轴上肌中的肌间骨数目最多(4045根), 尾部轴上肌与轴下肌中的肌间骨数目相近(3239根)。肌间骨形态包括1形、卜形、y形、一端多叉形、两端多叉形和(形6种类型, 各种形态的肌间骨均是从1形发展而来; 肌间骨越靠鱼体前端形态越复杂。团头鲂躯干轴上肌中的肌间骨显著长于尾部肌肉中的肌间骨(P 0.05), 躯干轴下肌中的肌间骨最短, 并且肌间骨长度与个体体重与体长呈正相关。研究结果为今后揭示团头鲂肌间骨发生与发育的分子机制, 抑制团头鲂肌间骨骨化, 培育无肌间骨的团头鲂提供了形态学基础。       

不同倍性鱼肌间骨的比较分析

采用常规测量法和解剖法对野生鲫(Carassius auratus,2n=100)、彭泽鲫(Carassius auratus variety pengze,3n=162)、改良三倍体鲫鱼(Triploid crucian carp,即湘云鲫2号,3n=150)及其亲本改良二倍体红鲫(Carassius auratus red var,改良红鲫,♀,2n=100)和改良异源四倍体鲫鲤(Allotetreploid hybrid,四倍体鲫鲤,♂,4n=200)5种不同倍性鱼肌间骨的数目、形态和分布进行研究.结果显示,野生鲫肌间骨数目在78~83之间,平均值为81根,彭泽鲫肌间骨数目在80~86之间,平均值为84,四倍体鲫鲤肌间骨数目在77~84之间,平均值为82,但是湘云鲫2号和改良红鲫的肌间骨数目比较少,湘云鲫2号在77~82之间,平均值为79,改良红鲫在58~77之间,平均值为71.考虑到不同倍性的鱼体大小和肌节数目的不同,进一步统计了每一肌节的平均肌间骨数目,野生鲫最多(0.721),彭泽鲫次之(0.673),改良红鲫最少(0.608),湘云鲫2号次之(0.633),四倍体鲫鲤位于中间(0.653),除了两组湘云鲫2号与四倍体鲫鲤、四倍体鲫鲤和彭泽鲫外,两两间都存在显著差异.5种鱼的各种肌间骨有"I"形、"卜"形、"Y"形、一端多叉形、两端两分叉形、两端多叉形和树枝形7种类型.肌间小骨越靠前端,形态越复杂.每条鱼左右两侧的肌间骨数目不完全相等,但总体上两侧肌间骨的数目接近.在保持鱼的营养、形态和活动等基本生理功能的前提下,湘云鲫2号的肌间刺数目比野生鲫和彭泽鲫都少,所以它的食用价值较高.本研究结果说明,改良二倍体红鲫和改良三倍体鲫鱼等人工培育的杂交鱼比野生鲫的肌间刺少,为鱼类骨骼发育生物学和鱼类遗传育种提供了形态学基础.     

鲢肌间小骨发育的形态学观察

分别利用形态解剖和整体骨骼染色的方法,对鲢(Hypophthalmichthys molitrix)成鱼肌间小骨的形态、分布,以及仔、稚鱼肌间小骨的形态发生和出现进行观察.结果表明,鲢肌间小骨有I形、卜形、y形、一端多叉形、两端多叉形和树枝形6种类型,肌间小骨越靠前端形态越复杂.肌间小骨在43 dpf首先出现在尾部,然后往前依次出现,到55 dpf全部出现.出现一枚髓弓小骨的仔鱼最小体长是10.50 mm,出现一枚脉弓小骨的仔鱼最小体长是13.66 mm.各种复杂形态的肌问小骨均是从I形发展而来.本文结果为今后研究肌间小骨发生的分子机制提供了形态学基础.     

异育银鲫A+系和F系肌间骨的比较分析

研究采用常规测量法和解剖法, 并结合CT透视, 比较分析了异育银鲫主养品种“中科3号”(A+系)和候选新品系F系的肌间骨的数目、形态和分布。结果表明, 6月龄和18月龄异育银鲫F系平均总肌间骨数分别为71.8±2.9和83.3±1.4, 均极显著少于相应月龄银鲫A+系总肌间骨数目(78.6±3.9和87.0±1.5)(P<0.01)。并统计了2个品系的每一肌节的平均肌间骨数目, 6月龄和18月龄异育银鲫F系的每一肌节的平均肌间骨数目极显著少于相应月龄银鲫A+系(P<0.01)。银鲫A+系和F系均有髓弓小骨和脉弓小骨, 没有椎体小骨。6月龄和18月龄银鲫F系髓弓小骨的平均数目为48.2±1.1和55.8±0.52, 均极显著少于相应月龄银鲫A+系的髓弓小骨(53.7±1.6和58.7±0.5)(P<0.01)。异育银鲫两个品系的脉弓小骨数目差异不显著; 2个品系均具有“I”形、“卜”形、“Y”形、一端多叉形、两端两分叉形、两端多叉形和树枝形7种类型的肌间骨, 髓弓小骨比脉弓小骨数量多且形状复杂。随着月龄的增长, 2个品系具有的总肌间骨数目以及复杂肌间骨数目均增加。6月龄和18月龄银鲫F系躯轴上肌节间的复杂“Y”形髓弓小骨数目均比相应月龄的银鲫A+系少, 而简单的“I”形髓弓小骨数目比相应月龄的A+系多, 表现出一种有利于食用的优势。研究结果为异育银鲫候选新品系F系提供了一个品质评价指标, 同时也为后续进行异育银鲫肌间骨遗传改良提供了形态学基础资料。     

不同鱼类肌间骨的骨化模式研究

为研究鱼类肌间骨的骨化模式, 采用整体骨骼染色方法, 对黄鳝(Monopterus albus)和泥鳅(Misgurnus anguillicaudatus) 2种处于不同进化地位且游动模式不同的鱼类肌间骨发生发育进行了系统观察。结果显示合鳃鱼目合鳃鱼科且游动模式属于鳗鲡模式的黄鳝在孵出后30d (体长40 mm)时, 其头部开始出现椎体小骨; 随后依次向尾部骨化, 在孵出后55d (体长约100 mm)时, 肌间骨相继出现完毕, 所有椎体小骨均为“I”形。鲤形目鳅科且游动模式为鲹科模式的泥鳅在孵出后27d (体长17 mm)时, 肌间骨开始在尾部出现, 包括髓弓小骨和脉弓小骨; 随后依次向头部骨化, 在孵出后40d (体长35 mm)时, 肌间骨全部出现, 形态包括“I”、“Y”和“卜”形。此外, 研究利用成骨细胞特异性转录因子绿色转基因荧光蛋白(Osterix GFP)斑马鱼品系, 通过观察此osterix GFP活体斑马鱼, 可发现斑马鱼肌间骨从尾部向头部依次骨化的过程。研究结果揭示鱼类肌间骨的骨化规律与其游动模式密切相关, 且肌间骨的形态种类的多态性与其游动模式和体型有一定的关系。     

禾花鲤与建鲤肌间骨miRNAs测序与分析比较

为了研究禾花鲤(Rice flower carp)肌间骨柔软的分子调控机制,采集禾花鲤和建鲤(Jian carp)的肌间骨进行micro RNAs (miRNAs)的高通量测序,并进行了生物信息学分析。从禾花鲤肌间骨的小RNA文库测序获得18 32 nt的高质量miRNA序列25474895条、鉴定出成熟miRNAs 595种,而从建鲤文库获得miRNA序列24625715条、成熟miRNAs 570种。表达差异分析结果显示,与建鲤相比,禾花鲤肌间骨中84个miRNAs呈上调表达、267个呈下调表达。禾花鲤下调表达的miRNAs中有7条为已报道的促进人类成骨的miRNAs,上调表达的miRNAs中有6条为已报道的抑制人类成骨的miRNAs。因此推测禾花鲤可能是通过下调表达促进成骨的miRNAs及上调表达抑制成骨的miRNAs来抑制成骨过程,从而维持其肌间骨细小及柔软的特性。研究为国内首次开展禾花鲤肌间骨发育的分子调控机制研究,为水产动物肌间骨发育的研究奠定了基础。     

鲌鲂F1、F2及其亲本肌间骨的比较分析

为研究新型鲌鲂杂交鱼的肌间骨,采用常规测量和解剖法对鲌鲂F1 (female Culter alburnus Basilewskymale Megalobrama amblycephala Yih)、鲌鲂F2 (self-crossing of F1 hybrid of female C. alburnusmale M. amblycephala)及其母本翘嘴鲌(C. alburnus)、父本团头鲂(M. amblycephala)肌间骨的数目、形态和分布进行统计分析,结果表明,翘嘴鲌肌间骨数目为134-139,平均为136.8根; 团头鲂的肌间骨数目为121-129,平均为124.2根; 鲌鲂F1肌间骨数目为129-134,平均为131.6根; 鲌鲂F2肌间骨数目为127-134,平均为130.1根; 鲌鲂F1、F2与翘嘴鲌、团头鲂之间肌间骨数目差异显著(P0.05); 鲌鲂F1每一肌节所含肌间骨数目最多,为0.8024; 鲌鲂F2最少,为0.7744; 翘嘴鲌和团头鲂介于鲌鲂F1和F2之间,分别为0.7953和0.7763。4种鱼均含有I形、卜形、Y形、一端多叉形、两端两分叉形、两端多叉形和树枝形7种类型肌间骨,髓弓小骨比脉弓小骨多且复杂; 鱼体左右两侧肌间骨的数目不完全相等,形态也不完全对称,但较为接近,且肌间骨越靠前端,形态越复杂; 研究获得的鲌鲂F1和F2在肌间骨总数、复杂型肌间骨数目和躯体轴下肌肌间骨数目均较母本有所减少,鲌鲂F2较F1还呈下降的趋势,且每一肌节所含肌间骨最少,表现出一种有利于提高食用品质和精深加工的优势。研究结果为鲌鲂属间远缘杂交培育少肌间骨新品种提供了基础资料。     

鲔鲂F_1、F_2及其亲本肌间骨的比较分析

为研究新型鲐鲂杂交鱼的肌间骨,采用常规测量和解剖法对鲌鲂F_1(female Culter alburnus Basilewsky×male Megalobrama amblycephala Yih)、鲌鲂F_2(self-crossing of F_1 hybrid offemale C.alburnus×male M.amblycephala)及其母本翘嘴鲌(C.alburnus)、父本团头鲂(M.amblycephala)肌间骨的数目、形态和分布进行统计分析,结果表明,翘嘴鲌肌间骨数目为134—139,平均为136.8根;团头鲂的肌间骨数目为121—129,平均为124.2根;鲌鲂F_1肌间骨数目为129—134,平均为131.6根;鲌鲂F_2肌间骨数目为127—134,平均为130.1根;鲐鲂F_1、F_2与翘嘴鲌、团头鲂之间肌间骨数目差异显著(P0.05);鲌鲂F_1每一肌节所含肌间骨数目最多,为0.8024;鲌鲂F_2最少,为0.7744;翘嘴鲐和团头鲂介于鲐鲂F_1和F_2之间,分别为0.7953和0.7763。4种鱼均含有"I"形、"卜"形、"Y"形、一端多叉形、两端两分叉形、两端多叉形和树枝形7种类型肌间骨,髓弓小骨比脉弓小骨多且复杂;鱼体左右两侧肌问骨的数目不完全相等,形态也不完全对称,但较为接近,且肌间骨越靠前端,形态越复杂;研究获得的鲌鲂F_1和F_2在肌间骨总数、复杂型肌间骨数目和躯体轴下肌肌间骨数目均较母本有所减少,鲌鲂F_2较F_1还呈下降的趋势,且每一肌节所含肌间骨最少,表现出一种有利于提高食用品质和精深加工的优势。研究结果为鲐鲂属间远缘杂交培育少肌间骨新品种提供了基础资料。     

草鱼雌核发育后代不同群体的微卫星遗传分析及指纹识别

以紫外线灭活的团头鲂精子激活草鱼卵子,冷休克抑制第二极体排出的方法诱导出长江水系优良F2代草鱼减数雌核发育子代。在后代中不仅存在雌核发育后代,还存在草鲂杂交后代,雌核发育后代的体型与草鱼一致,而草鲂杂交后代的体型介于草鱼与团头鲂之间。Partec CyFlow倍性分析仪测定结果显示:普通草鱼与雌核发育草鱼的相对DNA含量分别为23.01和22.72,二者的DNA含量接近;而高体型子代的相对DNA含量为25.38,介于草鱼与团头鲂（DNA含量28.21）之间,属于草鲂杂交后代。选取17个微卫星标记对草鱼群体、雌核发育草鱼群体和草鲂杂交后代的遗传多样性进行了检测,共检测出59个等位基因,其中43.18个有效等位基因。草鱼对照群体、草鲂杂交后代和雌核发育草鱼群体的平均等位基因依次为3.57、2.86和2.79,平均有效等位基因依次为2.93、2.37和1.96,平均期望杂合度在依次为0.6502、0.5573和0.3775,多态信息含量（PIC）平均值依次为0.5738、0.4649和0.3791。与草鱼对照群体相比,雌核发育草鱼群体的遗传多样性显著下降,表明通过减数雌核发育方法可获得纯合性较高的草鱼个体。构建了草鱼后代不同群体的DNA指纹模式图,筛选到不同群体的9个特异微卫星标记,为草鱼优良群体的选育提供了基础资料。     

The formation of the polyploid hybrids from different subfamily fish crossings and its evolutionary significance

This study provides genetic evidences at the chromosome, DNA content, DNA fragment and sequence, and morphological levels to support the successful establishment of the polyploid hybrids of red crucian carp x blunt snout bream, which belonged to a different subfamily of fish (Cyprininae subfamily and Cultrinae subfamily) in the catalog. We successfully obtained the sterile triploid hybrids and bisexual fertile tetraploid hybrids of red crucian carp (RCC) (female symbol) x blunt snout bream (BSB) (male symbol) as well as their pentaploid hybrids. The triploid hybrids possessed 124 chromosomes with two sets from RCC and one set from BSB; the tetraploid hybrids had 148 chromosomes with two sets from RCC and two sets from BSB. The females of tetraploid hybrids produced unreduced tetraploid eggs that were fertilized with the haploid sperm of BSB to generate pentaploid hybrids with 172 chromosomes with three sets from BSB and two sets from RCC. The ploidy levels of triploid, tetraploid, and pentaploid hybrids were confirmed by counting chromosomal number, forming chromosomal karyotype, and measuring DNA content and erythrocyte nuclear volume. The similar and different DNA fragments were PCR amplified and sequenced in triploid, tetraploid hybrids, and their parents, indicating their molecular genetic relationship and genetic markers. In addition, this study also presents results about the phenotypes and feeding habits of polyploid hybrids and discusses the formation mechanism of the polyploid hybrids. It is the first report on the formation of the triploid, tetraploid, and pentaploid hybrids by crossing parents with a different chromosome number in vertebrates. The formation of the polyploid hybrids is potentially interesting in both evolution and fish genetic breeding.     

Genetic Diversity of and Differentiation among Five Populations of Blunt Snout Bream (Megalobrama amblycephala) Revealed by SRAP Markers: Implications for Conservation and Management

The blunt snout bream (Megalobrama amblycephala) is an important freshwater aquaculture fish throughout China. Because of widespread introductions of this species to many regions, the genetic diversity of wild and natural populations is now threatened. In the present study, SRAP (sequence-related amplified polymorphism) markers were used to assess genetic diversity of blunt snout bream. Three natural populations (Liangzi Lake, Poyang Lake and Yuni Lake, one cultured population (Nanxian) and one genetic strain (‘Pujiang No. 1’) of blunt snout bream were screened with 88 SRAP primer combinations, of which 13 primer pairs produced stable and reproducible amplification patterns. In total, 172 bands were produced, of which 132 bands were polymorphic. Nei''s gene diversity (h) and Shannon''s information index (I) values provided evidence of differences in genetic diversity among the five populations (Poyang Lake>Liangzi Lake>Nanxian>‘Pujiang No. 1’>Yuni Lake). Based on cluster analysis conducted on genetic distance values, the five blunt snout bream populations were divided into three groups, Poyang Lake and Liangzi Lake (natural populations), Nanxian and ‘Pujiang No. 1’ (cultured population and genetically selected strain), and Yuni Lake (natural population). Significant genetic differentiation was found among the five populations using analysis of molecular variance (AMOVA), with more genetic divergence existing among populations (55.49%), than within populations (44.51%). This molecular marker technique is a simple and efficient method to quantify genetic diversity within and among fish populations, and is employed here to help manage and conserve germplasm variability of blunt snout bream and to support the ongoing selective breeding programme for this fish.     

Host species as a strong determinant of the intestinal microbiota of fish larvae

We investigated the influence of host species on intestinal microbiota by comparing the gut bacterial community structure of four cohabitating freshwater fish larvae, silver carp, grass carp, bighead carp, and blunt snout bream, using denaturing gradient gel electrophoresis (DGGE) of the amplified 16S and 18S rRNA genes. Similarity clustering indicated that the intestinal microbiota derived from these four fish species could be divided into four groups based on 16S rRNA gene similarity, whereas the eukaryotic 18S rRNA genes showed no distinct groups. The water sample from the shared environment contained microbiota of an independent group as indicated by both 16S and 18S rRNA genes segments. The bacterial community structures were visualized using rank-abundance plots fitted with linear regression models. Results showed that the intestinal bacterial evenness was significantly different between species (P<0.05) and between species and the water sample (P<0.01). Thirty-five relatively dominant bands in DGGE patterns were sequenced and grouped into five major taxa: Proteobacteria (26), Actinobacteria (5), Bacteroidetes (1), Firmicutes (2), and Cyanobacterial (1). Six eukaryotes were detected by sequencing 18S rRNA genes segments. The present study suggests that the intestines of the four fish larvae, although reared in the same environment, contained distinct bacterial populations, while intestinal eukaryotic microorganisms were almost identical.     

High-fat diet induces aberrant hepatic lipid secretion in blunt snout bream by activating endoplasmic reticulum stress-associated IRE1/XBP1 pathway

This study was conducted to understand the effect of high-fat diet challenge on lipid transport and endoplasmic reticulum stress in blunt snout bream. Ninety fish (average weight: 41.84 ± 0.07 g) were randomly fed a control diet (6% fat) or a high-fat diet (11% fat) for 9 weeks. The growth performance and feed utilization efficiency were evaluated at the end of the trial. The liver samples of both groups were harvested for molecular analysis and histological evaluation. Compared to the Control group, the high-fat diet group showed no effects on either growth performance or energy intake in blunt snout bream. However, high-fat diet resulted in a massive accumulation of lipid and pathological structural alternations, and disrupted expression of lipid transport-related genes and endoplasmic reticulum stress in the liver of the fish. In vitro, after exposure of the isolated primary hepatocytes from blunt snout bream to oleic acid, the cells showed increased intracellular TG accumulation, decreased VLDL secretion, which was attributed to altered expression levels of lipid transport-related genes through the activated IRE1/XBP1 signaling. The oleic acid-induced detrimental effects were alleviated by co-incubating the cells with an IER1 inhibitor, 4μ8c. In conclusion, high-fat diet could lead to aberrant lipid secretion by activating the ER stress-associated IRE1/XBP1 pathway. Inhibiting the activity of IRE1 represents a promising target to rescue the side-effects of high-fat diet on the liver function of blunt snout bream.     

Reproduction mode of an artificial allotetraploid carp (Pisces; Cyprinidae)

Mature eggs of allotetraploid carp were activated by inactive sperm or crossed with normal sperms of common carp (Cyprinus carpio), crucian carp (Carassius auratus), Chinese blunt snout bream (Megalobrama amblycephala), Hemiculter leucisculus and Pseudorasbora parva. Chromosome counts showed that all offspring of these crosses presented a mode number of 200 chromosomes (4n = 200), and their morphological traits are much like maternal. Microsatelite marker and RAPD patterns between allotetraploid maternal and its offspring, reproduced from different paternal species, were identical. Cytological, morphological and molecular evidences suggested that allotetraploid carp female nucleus would not fuse with any male nucleus and its reproduction mode might be gynogenesis and therefore their offspring are retaining their tetraploidy and give origin to clonal individuals.     

氨氮胁迫下不同食性鱼类仔鱼抗氧化和非特异性免疫的差异响应

为探究氨氮胁迫对不同食性鱼类的影响,研究选取不同食性的4种鱼类(鲢Hypophthalmichthys molitrix、草鱼Ctenopharyngodon idella、团头鲂Megalobrama amblycephala和黄颡鱼Pelteobagrus fulvidraco)仔鱼为研究对象,探讨不同浓度氨氮(0、1、2和3 mg/L)短期胁迫(96h)对其生长、抗氧化和非特异性免疫响应的影响及其分子机制。结果显示:(1)氨氮胁迫导致4种仔鱼的体长生长速度呈现剂量依赖性减缓;(2)不同浓度氨氮导致4种仔鱼体内T-AOC、CAT和GPx含量显著降低(P<0.05), 2和3 mg/L氨氮暴露显著降低了草鱼、团头鲂和黄颡鱼仔鱼体内SOD活力(P<0.05),仅检测到黄颡鱼gpx及鲢sod转录水平出现显著性下调(P<0.05);(3)在不同浓度氨氮胁迫下, 4种仔鱼相关免疫基因转录水平均呈现一定的上调,仅鲢il1β转录水平显著下降(P<0.05),相对地草鱼仔鱼LYZ含量显著性下降,黄颡鱼仔鱼LYZ含量在2 mg/L氨氮组显著性上升(P<0.05)。双因素...     

Molecular cloning and function analysis of insulin-like growth factorbinding protein 1a in blunt snout bream(Megalobrama amblycephala)

Insulin-like growth factor-binding protein 1(IGFBP-1), a hypoxia-induced protein, is a member of the IGFBP family that regulates vertebrate growth and development. In this study, full-length IGFBP-1a cDNA was cloned from a hypoxia-sensitive Cyprinidae fish species, the blunt snout bream(Megalobrama amblycephala). IGFBP-1a was expressed in various organs of adult blunt snout bream, including strongly in the liver and weakly in the gonads. Under hypoxia, IGFBP-1a mRNA levels increased sharply in the skin, liver, kidney, spleen, intestine and heart tissues of juvenile blunt snout bream, but recovered to normal levels after 24-hour exposure to normal dissolved oxygen. In blunt snout bream embryos, IGFBP-1a mRNA was expressed at very low levels at both four and eight hours post-fertilization, and strongly at later stages. Embryonic growth and development rates decreased significantly in embryos injected with IGFBP-1a mRNA. The average body length of IGFBP-1a-overexpressed embryos was 82.4% of that of the control group, and somite numbers decreased to 85.2%. These findings suggest that hypoxia-induced IGFBP-1a may inhibit growth in this species under hypoxic conditions.     

Evidence for the Formation of the Male Gynogenetic Fish

The females and unexpected males of gynogenetic red crucian carps (GRCC) with the 1:1 sex ratio were found in the progeny of the distant crossing of red crucian carp (RCC; ♀, 2n?=?100) × blunt snout bream (BSB; ♂, 2n?=?48). The females and males of GRCC were fertile, and they mated each other to generate the red crucian carps (GRCC₁) and another variational gray crucian carps (GGCC). The GRCC and their offspring were proved to be diploids (2n?=?100) with one to three microchromosomes by examining the chromosomal metaphases. The evidences for the male’s genetic effect in GRCC were provided by means of fluorescence in situ hybridization, Sox-HMG DNA markers, and microsatellite DNA markers. The genotypic variances of GRCC resulted in their phenotypic variances which were quite different from their maternal parent. It was concluded that the formation of the male gynogenetic fish in GRCC resulted from the genetic leakage of the paternal fish in the form of the microchromosomes including the paternal male-determining gene. After being activated by the sperm of BSB, which was inactivated and finally degraded but left the microchromosomes, the egg of RCC, in which the 50 chromosomes were spontaneously doubled to 100 chromosomes, developed into the diploid male gynogenetic fish. The formation of the bisexual GRCC and their progeny indicated that the distant hybridization could generate the bisexual diploid gynogenetic fish with genetic variation derived from the paternal fish, which is of great significance in both fish genetic breeding and evolutionary biology.