鳜慢肌胚后发育与生长特征

鱼类快肌和慢肌分别占据骨骼肌的不同位置,表现不同的生长发育特征。为了解鳜(Sinipercachuatsi)慢肌纤维的胚后发育特征,本研究通过制作孵化后1～33日龄鳜个体的石蜡切片,采用慢肌特异抗体的免疫组织化学染色,观察了背鳍起点处躯干横切面慢肌的发育变化特征,并利用图像分析软件统计慢肌纤维的数目和面积。结果表明,孵化后鳜仔鱼慢肌位于水平肌隔附近,呈楔形,向背、腹两侧生长。孵化后1～9日龄为单层肌纤维,11日龄发育为多层肌纤维,19日龄覆盖侧线附近,33日龄延伸至背侧第2背肌节、腹侧腹部肌肉2/3处,并在水平肌隔和侧线处分别形成两个肌群。位于骨骼肌最外层的扁平状表层细胞,可能为慢肌增生生长的主要来源。躯干单侧慢肌肌纤维数目由孵化后6个增加至315个,总面积从13.18μm2增加到7 839.58μm2,孵化后13日龄的增生生长占优势,其他发育阶段,肥大生长一直占主导优势。     

翘嘴鳜miR-222的表达特征

为了解翘嘴鳜miR-222的时空表达规律, 研究利用实时荧光定量PCR的方法检测miR-222在翘嘴鳜不同组织、胚胎发育及胚后发育中的相对表达丰度。研究结果显示, miR-222在肌肉相关的组织中表达较高, 特别是在成年翘嘴鳜的白肌中表达最高; 胚胎发育阶段结果显示, miR-222在胚胎发育的2细胞期就有表达, 而表达量在心动期达到最高。不同组织及不同发育阶段的差异性表达结果表明, miR-222很可能参与调控鳜鱼肌肉的生长发育。为研究合成代谢过程中miR-222在肌肉生长调控中的表达规律, 通过对翘嘴鳜幼鱼在饥饿一周后饱食一餐的实验处理下, 利用实时荧光定量的方法测定miR-222在骨骼肌中的相对表达变化。结果显示, miR-222的表达量在恢复喂食后的1h显著上升(P0.05), 表明miR-222很可能是调节鱼类骨骼肌生长过程中, 参与快速应答信号系统的一类miRNA。研究为miR-222在鱼类发育中的调控作用提供一些理论依据。       

翘嘴鳜miR-222的表达特征（英文）

为了解翘嘴鳜mi R-222的时空表达规律,研究利用实时荧光定量PCR的方法检测mi R-222在翘嘴鳜不同组织、胚胎发育及胚后发育中的相对表达丰度。研究结果显示,mi R-222在肌肉相关的组织中表达较高,特别是在成年翘嘴鳜的白肌中表达最高;胚胎发育阶段结果显示,mi R-222在胚胎发育的2细胞期就有表达,而表达量在心动期达到最高。不同组织及不同发育阶段的差异性表达结果表明,mi R-222很可能参与调控鳜鱼肌肉的生长发育。为研究合成代谢过程中mi R-222在肌肉生长调控中的表达规律,通过对翘嘴鳜幼鱼在饥饿一周后饱食一餐的实验处理下,利用实时荧光定量的方法测定mi R-222在骨骼肌中的相对表达变化。结果显示,mi R-222的表达量在恢复喂食后的1h显著上升(P0.05),表明mi R-222很可能是调节鱼类骨骼肌生长过程中,参与快速应答信号系统的一类mi RNA。研究为mi R-222在鱼类发育中的调控作用提供一些理论依据。     

三种鳜鱼骨骼肌生长及相关调控基因表达比较

通过制作骨骼肌石蜡切片研究了鳜、斑鳜及斑鳜(♀)×鳜(♂)杂交一代不同发育时期(孵化后10D、20D、30D、60D、90D)骨骼肌肌纤维的生长特征,同时,利用q PCR技术分析了肌肉中myostatin、Myo D、myogenin三个基因m RNA的表达变化。结果表明,3种鳜鱼全长均随日龄呈增大趋势,种间生长差异显著,鳜生长最快,斑鳜最慢,杂交一代介于斑鳜和鳜之间。不同发育时期,3种鳜鱼骨骼肌肌纤维数目较为接近,增生生长种间无明显差异,而肌纤维平均直径大小与其生长快慢间呈正相关,表明不同鳜鱼种间生长差异主要由肌肉肥大生长引起的。鳜、斑鳜和杂交一代myostatin m RNA在D20出现表达高峰,之后表达量呈下降趋势。Myo D在D10、D20表达水平较高,后期表达水平下降;Myogenin在D20时表达量最低,之后,鳜表达量升高最为明显,杂交一代次之,斑鳜保持平稳。不同种类鳜鱼肌肉生长差异与3种基因表达差异间存在一定相关性。     

铜鱼和圆口铜鱼的早期发育

本文比较系统地观察了长江和汉江铜鱼和圆口铜鱼鱼卵、鱼苗的胚胎发育,记述了两种铜鱼胚胎发育的各个阶段的形态特征,比较两种铜鱼早期发育的相似性状和主要差别。这些差别主要是：雏形胚体出现的迟早、眼基和肌节出现的先后、眼睛大小、胸鳍长短以及颌须出现的迟早和长短等,依据这些差异,可以把两种铜鱼在早期发育阶段准确地区分开来。本文还对长江兴修水利枢纽与两种铜鱼资源的关系进行了调查和分析。     

铜鱼和圆口铜鱼的早期发育

本文比较系统地观察了长江和汉江铜鱼和圆口铜鱼鱼卵、鱼苗的胚胎发育,记述了两种铜鱼胚胎发育的各个阶段的形态特征,比较两种铜鱼早期发育的相似性状和主要差别。这些差别主要是:雏形胚体出现的迟早、眼基和肌节出现的先后、眼睛大小、胸鳍长短以及颌须出现的迟早和长短等,依据这些差异,可以把两种铜鱼在早期发育阶段准确地区分开来。本文还对长江兴修水利枢纽与两种铜鱼资源的关系进行了调查和分析。       

猪的骨骼肌生长发育研究进展

瘦肉率对生猪产业来说是一个极其重要的经济指标,而这一指标完全取决于骨骼肌的生长发育。因此,猪骨骼肌生长发育机理的研究是十分必要的。然而,在早期由于各种因素的限制,猪骨骼肌单个基因的研究一直进展缓慢;相反,以小鼠为模型,其骨骼肌单基因的功能研究却取得了较大进展。在这一时期,影响肌决定和肌分化的基因,如MRFs家族和MEF2家族相继被发现,这些基因在猪的肌肉发育中也发挥着同样的作用。然而,这些结果并不能很好地揭示骨骼肌发育过程中复杂的基因间互作关系。随着近年来芯片和测序技术的不断发展,更多人试图从整个转录谱的水平来阐述猪肌肉发育的分子机理,并且也取得了较大的进展。为了对猪骨骼肌生长发育有一个更为清晰的认识,该文将以目前猪骨骼肌生长发育研究结果为基础,同时结合模式动物小鼠骨骼肌单基因的研究成果,对猪的骨骼肌生长发育分子调控机理进行详细的阐述。     

皖西白鹅胚胎中、后期肌组织发育变化的观察

分别于17、23、29胚龄各取6枚胚胎发育正常的种蛋,分离鹅胚的心脏、肌胃及大腿骨骼肌,制作石蜡切片,HE染色,显微观察、摄影.观察皖西白鹅胚胎中、后期肌肉组织发育的变化.结果表明,17d时的骨骼肌、平滑肌、心肌的发育程度均较低,没有形成正常的肌纤维形态,之后呈现随龄性增长.骨骼肌和平滑肌在17 d～23 d期间是以肌纤维数量的增加为主,即成肌细胞的分裂增殖,23 d～29 d 期间成肌细胞分化融合形成肌纤维;心脏成纤维细胞增殖速度较快,17 d心脏肌纤维已经排列紧密基本吻合成网状;17 d后主要是心肌纤维的伸长和成熟化的过程.说明胚胎期心肌组织的生长发育要早于骨骼肌与平滑肌.     

鳜肌肉生长抑制素Myostatin cDNA克隆与组织表达分析

利用RT-PCR和cDNA末端快速扩增法(RACE)克隆了鳜(Siniperca chuatsi)肌肉生长抑制素(myostatin,MSTN)cDNA序列,并分析了该基因的结构特征和亲缘关系。鳜MSTN cDNA序列全长2627bp,包括5′端非翻译区117bp、3′端非翻译区1376bp和开放阅读框(ORF)1134bp,共编码377个氨基酸,含22个氨基酸的信号肽。鳜MSTN具有脊椎动物MSTN的共同序列特征,含有1个蛋白酶水解位点RARR和9个保守的半胱氨酸残基。脊椎动物MSTN氨基酸序列的亲缘关系分析表明,鳜与其他鱼类聚为一支。RT-PCR分析表明,鳜MSTN在成体不同组织中的表达情况不同,其中,卵巢、肾、眼、肌肉、心、脑、皮肤和胃中有表达,肝胰脏未见表达。     

浅水湖泊人工繁育鳜放养规格对其成活、生长和产量的影响

文章采用标志放流回捕方法评估了人工繁育鳜放流规格对其成活、生长和产量的影响。2012年5—6月选取3种不同大小规格的人工繁育鳜鱼苗各800尾, 中等规格鳜苗种和大规格鳜苗种分别采用剪尾鳍和编码金属标标记后放入试验湖泊小赛湖, 2012年12月试验湖泊逐渐降低水位, 回捕收集放流鳜并鉴定身份。结果显示3种规格放流鳜的总成活率为40.5%, 其中大规格放流鳜的成活率为63%, 明显高于中规格放流鳜(39.1%)和小规格放流鳜(19.5%)。放流鳜的终末体长和体重均随放流规格的增加而增加, 且3种规格放流鳜的终末体长和体重均呈现显著差异。放流鳜的产量和净收益在大规格处理组最大, 在小规格处理组最小。结果表明鳜放流规格对鳜放流成功与否有重要影响, 建议长江中游湖泊鳜的最佳放流规格应不小于50 mm。     

Myostatin-deficient medaka exhibit a double-muscling phenotype with hyperplasia and hypertrophy, which occur sequentially during post-hatch development

Myostatin (MSTN) functions as a negative regulator of skeletal muscle mass. In mammals, MSTN-deficient animals result in an increase of skeletal muscle mass with both hyperplasia and hypertrophy. A MSTN gene is highly conserved within the fish species, allowing speculation that MSTN-deficient fish could exhibit a double-muscled phenotype. Some strategies for blocking or knocking down MSTN in adult fish have been already performed; however, these fish show either only hyperplastic or hypertrophic growth in muscle fiber. Therefore, the role of MSTN in fish myogenesis during post-hatch growth remains unclear. To address this question, we have made MSTN-deficient medaka (mstnC315Y) by using the targeting induced local lesions in a genome method. mstnC315Y can reproduce and have the same survival period as WT medaka. Growth rates of WT and mstnC315Y were measured at juvenile (1–2 wk post-hatching), post-juvenile (3–7 wk post-hatching) and adult (8–16 wk post-hatching) stages. In addition, effects of MSTN on skeletal muscle differentiation were investigated at histological and molecular levels at each developmental stage. As a result, mstnC315Y show a significant increase in body weight from the post-juvenile to adult stage. Hyper-morphogenesis of skeletal muscle in mstnC315Y was accomplished due to hyperplastic growth from post-juvenile to early adult stage, followed by hypertrophic growth in the adult stage. Myf-5 and MyoD were up-regulated in mstnC315Y at the hyperplastic growth phase, while myogenin was highly expressed in mstnC315Y at the hypertrophic growth phase. These indicated that MSTN in medaka plays a dual role for muscle fiber development. In conclusion, MSTN in medaka regulates the number and size of muscle fiber in a temporally-controlled manner during posthatch growth.     

Hyperplasia and hypertrophy in the growth of skeletal muscle in juvenile Atlantic salmon, Salmo salar L.

Both red and white muscle fibre numbers in juvenile Atlantic salmon increased gradually with fish length throughout the freshwater growth period. Mean fibre area increased as fish grew to 6.5 cm f.l. , but thereafter was unrelated to fish length. Hyperplasia was most obvious when fish were growing fastest, and was the dominant growth process in fish over 6.5 cm f.l. Hypertrophy was most important when growth was slow, as in autumn and winter.
Mean white fibre area was significantly smaller in deep muscle than at medial and superficial sites. Total cross-sectional area of red, white and total trunk muscle increased with fish length. The ratio of red: white cross-sectional area increased with fish length to a plateau at about 10% after 6.5 cm f.l.     

Muscle growth in yolk-sac larvae of the Atlantic halibut as influenced by temperature in the egg and yolk-sac stage

Atlantic halibut eggs and yolk-sac larvae were incubated at 1, 5 and 8° C. Eggs incubated at 8° C gave slightly shorter larvae at hatching with a significantly smaller total cross-sectional area of white muscle fibres than eggs incubated at 5° C. Transport of eggs 2 days prior to hatching gave significantly longer larvae at hatching with a significantly larger red fibre cross-sectional area than when eggs were transported shortly after the blastopore closure. A higher survival until 230 degree days after hatching was also observed in the former group. All eggs incubated at 1° C died before hatching and all larvae incubated at 1° C died before 45 degree days after hatching. From hatching until 230 degree days the total white cross-sectional area increased threefold in all temperature groups. The increase in white cross-sectional area was entirely due to hypertrophy between hatching and 150 degree days (10 mm L _S). Recruitment of new white fibres increased in germinal zones at the dorsal, ventral and lateral borders of the myotome from 150 degree days onwards, but at 230 degree days (12–13 mm L _S) the recruitment fibre zone constituted <10% of the total white cross-sectional area. Larval incubation at 8° C gave slightly longer larvae with a significantly larger cross-sectional area of recruitment fibres at 230 degree days than incubation at 5° C. The larval group incubated at 8° C also had a significantly lower survival until 230 degree days than did the 5° C group. Incubation temperature regimes did not affect the volume density of myofibrils in the axial muscle fibres at 230 degree days. Thus hypertrophy is the predominant mechanism of axial white muscle growth in Atlantic halibut yolk-sac larvae and an increased rearing temperature during the yolk-sac stage increases white muscle fibre hyperplasia.     

Growth and multiplication of white skeletal muscle fibres in carp larvae in relation to somatic growth rate

A morphometric analysis of white axial muscle of common carp Cyprinus carpio was undertaken in order to quantify increase in fibre size, fibre nuclei and fibre number in relation to somatic growth rate during early life. In fast-growing carp larvae fed zooplankton, length and height of fibres from the central part of dorsolateral muscle increased at the same rate (0.75) relative to the total length of the larvae during the first 2 weeks of feeding. During this period, the number of nuclei per fibre increased threefold while the number of nuclei per unit fibre surface remained constant. In fast-growing larvae fed a formulated diet, the total cross-sectional area of one epaxial quadrant of white muscle and the total area of white fibres increased at almost the same rate (3.15; 3.23) relative to larval total length during the first 28 days of exogenous feeding. The total number of white fibres increased faster (2.07) relative to the total length of larvae than the mean area of white fibres (1.16). Hyperplasia accounted for 64% of muscle growth in these larvae. The proportion of fibres with a width < 10 μm decreased from 72% at first feeding to 14% 28 days later, while the proportion of fibres with a width >20 μm which was 0% at first feeding increased up to 34% in the same time. The recruitment of new white fibres seemed to be almost the same in the whole muscle quadrant at first feeding and 18 or 28 days later but, 8 days after first feeding, a transient significant recruitment of new fibres was shown at the apex of the myotome. Comparisons between fast- and slow-growing groups of larvae showed that for a given larval total length: (1) the mean width of central white fibres was higher and the proportion of central fibres with a width <10 μm was lower in slow-growing larvae than in fast-growing ones; (2) the total number of white fibres was lower for a higher total cross-sectional area of white muscle in slow-growing larvae than in fast-growing ones. These results suggest that, in Cyprinus carpio larvae, slow-growing conditions are related to a decreased contribution of hyperplasia to muscle growth.     

Hyperplastic and hypertrophic growth of lateral muscle in blackspot seabream Pagellus bogaraveo from hatching to juvenile

To understand better the growth mechanisms in the economically important fish Pagellus bogaraveo, in terms of muscle fibre hyperplasia v. hypertrophy, the lateral muscle of this fish was studied morphometrically from hatching to juvenile comparing rostral and caudal locations. Fish were sampled at 0, 5, 23, 40, 70, 100, 140 and 180 days. Fibre types were first identified by succinate dehydrogenase (SDH) and immunostaining with a polyclonal antibody against fish slow myosin (4–96). Morphometric variables were then measured in transverse body sections, at both post‐opercular and post‐anal locations, to estimate the following variables: total muscle area [A (muscle)], total fibre number [N (fibres)], fibre number per unit area of muscle [N_A(fibres, muscle)] and cross‐sectional fibre area [ (fibres)] of the two main muscle fibre types (white and red). Overall, growth throughout the various stages resulted from increases both in the number and in the size of muscle fibres, paralleled by an expansion of the [A (muscle)]. Nonetheless, that increase was not significant between 0–5 days on one hand and 100–140 days, on the other hand. On the contrary, the [N_A(fibres, muscle)] declined as the body length increased. Analysis of the muscle growth kinetics suggested that, within the important time frame studied, hyperplasia gave the main relative contribution to the increase of white muscle [A (white muscle)], whereas red muscle [A (red muscle)] mainly grew by hypertrophy, with both phenomena occurring at a faster pace posteriorly in the body. Finally, when comparing rostral and caudal locations, a greater [N (fibres)] and [A (muscle)] of the posterior white and red fibres were the consistent features. It was also observed that the proportion of the cross‐sectional area of the myotomal muscle comprised of white muscle was greater in the anterior part of the fish.     

Regionalized adaptations and muscle fiber proliferation in stretch-induced enlargement

The relative contribution of increases in fiber area to stretch-induced muscle enlargement was evaluated in the slow tonic fibers of the anterior latissimus dorsi of adult Japanese quails. A weight corresponding to 10% of the bird's body mass was attached to one wing. Thirty days of stretch in 34 birds averaged 171.8 +/- 13.5% increase in muscle mass and 23.5 +/- 0.8% increase in muscle fiber length. The volume density of noncontractile tissue increased in middle and distal regions of stretch-enlarged muscles. Mean fiber cross-sectional area increased 56.7 +/- 12.3% in the midregion of stretched muscles. Further analysis indicated slow beta-fiber hypertrophy occurred in proximal, middle, and distal regions; however, fast alpha-type fiber hypertrophy was limited to middle regions of stretched muscles. Stretched muscles had a significant increase in the frequency of slow beta-fibers that were less than 500 microns 2 in all regions and fast alpha-type fibers in middle and distal regions. Total fiber number was determined after nitric acid digestion of connective tissue in 10 birds. Fiber number increased 51.8 +/- 19.4% in stretched muscle. These results are the first to clearly show that muscle fiber proliferation contributes substantially to adult skeletal muscle stretch-induced enlargement, although we do not know whether the responses of the slow tonic anterior latissimus dorsi might be similar or different from mammalian twitch muscle.     

鳜碱性肌球蛋白轻链基因cDNA的克隆及其发育表达分析

肌球蛋白轻链是构成鱼类肌纤维主要组成部分,在鱼类肌肉生长和收缩过程中具有重要作用。鳜鱼具有生长快、肉质细嫩、味道鲜美、营养成分高等优良的性状。研究通过构建鳜肌肉组织cDNA文库分离到两个碱性肌球蛋白轻链基因,即MLC1和MLC3基因。序列分析显示MLC1和MLC3基因cDNA序列全长分别为1237bp和1070bp,分别编码192和150个氨基酸,除去MLC1N端多出的42个氨基酸残基,MLC1与MLC3氨基酸序列同源性为80.3%。通过PROSITEtools软件预测显示两种轻链都具有两个保守的EF-手相结构,其中第二个EF-hand结构除前三个氨基酸外同源性达100%。鱼类MLC3轻链N端没有高等脊椎动物MLC3特有标志序列。采用实时荧光定量PCR方法对鳜鱼MLC1和MLC3发育性表达分析表明,在原肠期开始有低量表达,与原肠期、尾芽期和肌肉效应期相比,心搏期和仔鱼期MLC1和MLC3表达量显著升高。研究结果首次提供了鳜肌肉组织肌球蛋白主要结构基因的分子生物学信息以及它们在鳜肌肉组织发生和功能的相关性。       

Influence of feed ration size on somatic and muscle growth in landlocked dwarf and farmed Atlantic salmon Salmo salar

We examined the possible adaptation of the dwarf Bleke population of Atlantic salmon Salmo salar from Lake Byglandsfjord in southern Norway to limited food resources. The growth performance and muscle development in juvenile Bleke and farmed S. salar under satiated or restricted (50%) feeding were examined for 10 months, starting 3 weeks after first-feeding stage. Four-thousand fish were divided into four replicated groups and random samples of 16–40 fish per group were measured six times during the experiment. The two strains showed no significant difference in mean body mass when fed restricted ration, but the individual variation was considerably higher in the farmed fish. Both Bleke and farmed S. salar grew significantly faster when fed to satiation, but the farmed S. salar showed much higher gain in mass and were three times heavier (201.5 g vs 66.7 g) and possessed twice as many fast muscle fibres (179,682 vs 84,779) compared with landlocked S. salar after 10 months. Farmed fish fed full ration displayed both hypertrophic and hyperplasic muscle growth, while the increased growth in Bleke S. salar was entirely associated with a larger fibre diameter. The landlocked Bleke strain has apparently adapted to low food availability by minimising the metabolic costs of maintenance and growth through reduced dominance hierarchies and by an increase in average muscle fibre diameter relative to the ancestral condition.