α-黑素细胞刺激素及黑皮质素受体-4在能量平衡和繁殖中的功能研究

阿片黑素促皮质激素原(POMC)是黑皮质激素的前体物质,在机体中广泛分布。近年来,POMC及其衍生物黑素细胞刺激素(MSH)和黑皮质素受体-4(MC4R)在能量稳态和繁殖活动中的作用备受关注。研究发现,POMC神经元及其加工后的产物对摄食和体重的调节具有重要作用。此外,POMC神经元在下丘脑中与GnRH神经元紧密接触,由其产生的神经递质和神经肽可能对GnRH神经元具有重要的输入作用,从而直接或间接地调控下丘脑–垂体–性腺轴(HPG)中繁殖相关激素的合成并影响生殖活动。本文综述了黑皮质素系统对能量稳态和繁殖活动的调控作用以及目前MC4R在硬骨鱼类中的药理学研究,以期其能够在动物的生长发育和繁殖中具有应用前景。     

大口黑鲈生长激素促分泌素cDNA结构和早期发育阶段表达谱分析

研究采用RT-PCR、RACE和PCR技术从大口黑鲈胃组织中克隆得到ghrelin基因的cDNA。该基因全长434bp,其中开放阅读框(open reading frame,ORF)321bp,编码107个氨基酸的前肽,20个氨基酸的成熟肽位于第27位到第46位氨基酸处。前肽和成熟肽的氨基酸与已报导的舌齿鲈进行同源性分析,相似性分别为85%和90%。为进一步了解ghrelin基因在鱼体早期发育阶段的表达,本实验采用实时定量PCR(real-time PCR)方法检测了ghrelin基因在大口黑鲈胚胎和仔鱼发育过程的表达谱。结果显示,ghrelin基因在受精卵时期就有少量表达,但直到体节出现之前表达量均较低。出膜第4天ghrelin基因出现大量表达,第12天ghrelin基因表达量更显著增加,出膜第4天和第12天的表达量分别是受精卵时期表达量的206.77倍和531.20倍。出膜第4天正是大口黑鲈仔鱼开口觅食期,仔鱼消化系统初步发育成型,由完全利用卵黄囊营养转为从外界觅食阶段,到出膜第12天,仔鱼消化系统已发育完善,完全依靠外界营养提供能量,由ghrelin基因的表达量变化可推测其可能参与了鱼类早期发育阶段的摄...     

Leptin in fish: possible role in sexual maturation in male Atlantic salmon

Leptin plays an important role in energy homeostasis and reproductive function in mammals, while its function in fish is still poorly understood, especially its role in reproduction. In the present study, leptin gene expression and circulating leptin plasma levels were measured during sexual maturation in male Atlantic salmon parr. In maturing male salmon, higher hepatic leptin (lepa1) gene expression levels were observed during mid-spermatogenesis compared to immature fish. An upregulation of leptin during sexual maturation has also been observed in a few other teleost species. The physiological significance of elevated leptin levels during the maturational process in teleosts remains to be explored.     

The influence of dietary cation-anion differences on acid-base balance,food intake,growth and nutrient utilisation of juvenile African catfish Clarias gariepinus (Burchell)

A study was conducted to examine the effect of dietary cation-anion difference (CAD, Na⁺+K⁺-Cl^–, mEq kg ^–1) on acid-base balance, food consumption and growth of juvenile African catfish, Clarias gariepinus. Four dietary CAD levels (-100, 100, 500, 700 mEq kg^–1) were established by altering levels of NaHCO₃, CaCl₂ and NH₄Cl in the diets. The group fed the diet of excess anions (CAD: -100 mEq kg^–1) had low food consumption and growth rate. Food consumption and growth increased linearly with an increase in dietary CAD within the range -100 to 700 meq. kg^–1. Blood pH was not significantly influenced by dietary CAD. A quadratic relationship between Na⁺, Cl^– and CAD level in plasma and dietary CAD was found. Body composition of fish and nitrogen/energy utilisation efficiency differed between dietary CAD treatments. The results suggest that dietary CAD influence acid-base regulation and thus the energy costs for maintaining homeostasis. The increased food intake and growth performance of C. gariepinus at higher CAD levels may be related to a reduced maintenance costs for homeostasis.     

Oxidative effect of l -carnitine on energy metabolism in diploid and triploid rainbow trout ( Oncorhynchus mykiss ): impact on metabolites

l-Carnitine plays a key role in the regulation of energy metabolism and growth in fish. The aim of this study was to determine the effects of dietary l-carnitine levels on carnitine homeostasis and energy metabolism in diploid and triploid trout (Oncorhynchus mykiss). Diploid and triploid trout (29.5 ± 0.6 and 31.8 ± 0.5 g, respectively) were fed with three diets supplemented with 15, 200 or 530 mg l-carnitine/kg, respectively, for 56 days. Compared to 15 mg l-carnitine, total carnitine content increased in the liver by 182% in diploid and by 154% in triploid trout fed 530 mg l-carnitine. In muscle, total carnitine content increased by 60 and 73% in diploid and by 34 and 75% in triploid with the increase in dietary l-carnitine levels. l-Carnitine caused no significant change in the plasma concentrations of metabolites like proteins (ammonia, urea and protein), lipid (triglycerides), carbohydrate (glucose, lactate) and enzyme activities (lipase, lactate, alanine transaminase, lactic acid dehydrogenase). Triploid trout had significantly lower plasma ammonia (P = 0.003), lipase (P = 0.005) and triiodothyronine (T3; P = 0.003) levels than diploid trout. In conclusion, ploidy significantly affected the energy metabolism in rainbow trout, dietary l-carnitine levels altered the l-carnitine homeostasis, but not influence nutritional metabolism.     

温度和盐度对中华圆田螺和铜锈环棱螺标准代谢的影响

利用室内模拟实验研究了环境因子(温度和盐度)对中华圆田螺(Cipangopaludina cathayensis)和铜锈环棱螺(Bellamya aeruginosa)标准代谢的影响。结果表明,在10~35℃范围内,两种螺的耗氧率、CO2排放率和排氨率均随温度的升高而升高,至30℃达到最大值。从Q10值来看,铜锈环棱螺的代谢水平对水温的响应较中华圆田螺更为敏感。呼吸熵和氧氮比结果表明,25℃时两种螺体内的蛋白质消耗最少,由此初步判断二者的最适温度在25℃左右。盐度升高至1.8时两种螺的耗氧率等代谢指标显著提高,同时二者对温度变化的敏感性降低。但从动物呼吸熵和氧氮比的测定结果来看,该盐度下机体仍维持在相对正常的代谢水平。温度和盐度对中华圆田螺代谢活动的影响有显著的交互作用,但对铜锈环棱螺的影响不显著。此外,两种螺的代谢率都存在明显的昼夜变化规律,夜晚的代谢率要显著高于白天。     

黄条鰤 npy基因克隆及其对饥饿再投喂的应答特性

为探究神经肽Y (neuropeptide Y, NPY)在黄条鰤 (Seriola aureovittata)摄食调控中的作用及机制,本研究采用同源克隆的方法获得了黄条鰤 npy基因的开放阅读框(ORF)序列,并利用实时荧光定量PCR技术分析了npy基因的组织分布以及其对饥饿再投喂的应答特性。黄条鰤 npy基因ORF序列长度为300 bp,编码99个氨基酸的前体蛋白,其中包括28个氨基酸的信号肽、36个氨基酸的成熟肽。氨基酸序列同源性比对发现,黄条鰤 npy编码的氨基酸序列与斑马鱼(Danio rerio)等其他硬骨鱼高度保守(>90%);系统进化树分析表明,黄条鰤 npy与高体鰤 (Seriola dumerili)的关系最近。npy mRNA在所检测的12种组织中均有表达,其中,在脑组织表达量最高,在垂体和胃中表达量次之。在饥饿再投喂实验中,饥饿刺激了npy mRNA的表达,特别是饥饿21 d时,实验组垂体npy mRNA表达量显著高于对照组,再投喂7 d后恢复到对照组水平。上述结果表明,npy可能参与了黄条鰤的摄食调控,在饥饿代谢补偿机制中发挥了重要作用。     

Health of farmed fish: its relation to fish welfare and its utility as welfare indicator

This brief review focuses on health and biological function as cornerstones of fish welfare. From the function-based point of view, good welfare is reflected in the ability of the animal to cope with infectious and non-infectious stressors, thereby maintaining homeostasis and good health, whereas stressful husbandry conditions and protracted suffering will lead to the loss of the coping ability and, thus, to impaired health. In the first part of the review, the physiological processes through which stressful husbandry conditions modulate health of farmed fish are examined. If fish are subjected to unfavourable husbandry conditions, the resulting disruption of internal homeostasis necessitates energy-demanding physiological adjustments (allostasis/acclimation). The ensuing energy drain leads to trade-offs with other energy-demanding processes such as the functioning of the primary epithelial barriers (gut, skin, gills) and the immune system. Understanding of the relation between husbandry conditions, allostatic responses and fish health provides the basis for the second theme developed in this review, the potential use of biological function and health parameters as operational welfare indicators (OWIs). Advantages of function- and health-related parameters are that they are relatively straightforward to recognize and to measure and are routinely monitored in most aquaculture units, thereby providing feasible tools to assess fish welfare under practical farming conditions. As the efforts to improve fish welfare and environmental sustainability lead to increasingly diverse solutions, in particular integrated production, it is imperative that we have objective OWIs to compare with other production forms, such as high-density aquaculture. However, to receive the necessary acceptance for legislation, more robust scientific backing of the health- and function-related OWIs is urgently needed.     

Changes in the cerebral phosphotransfer network impair energetic homeostasis in an aflatoxin B<Subscript>1</Subscript>-contaminated diet

The phosphotransfer network system, through the enzymes creatine kinase (CK), adenylate kinase (AK), and pyruvate kinase (PK), contributes to efficient intracellular energetic communication between cellular adenosine triphosphate (ATP) consumption and production in tissues with high energetic demand, such as cerebral tissue. Thus, the aim of this study was to evaluate whether aflatoxin B₁ (AFB₁) intoxication in diet negatively affects the cerebral phosphotransfer network related to impairment of cerebral ATP levels in silver catfish (Rhamdia quelen). Brain cytosolic CK activity decreased in animals fed with a diet contaminated with AFB₁ on days 14 and 21 post-feeding, while mitochondrial CK activity increased, when compared to the control group (basal diet). Also, cerebral AK and PK activity decreased in animals fed with a diet contaminated with AFB₁ on days 14 and 21 post-feeding, similarly to the results observed for cerebral ATP levels. Based on this evidence, inhibition of cerebral cytosolic CK activity is compensated by stimulation of mitochondrial CK activity in an attempt to prevent impairment of communication between sites of ATP generation and ATP utilization. The inhibition of cerebral AK and PK activity leads to impairment of cerebral energy homeostasis, decreasing the brain’s ATP availability. Moreover, the absence of a reciprocal compensatory mechanism between these enzymes contributes to cerebral energetic imbalance, which may contribute to disease pathophysiology.     

Mitochondrial changes in fish cells in vitro in response to serum deprivation

Fish Physiology and Biochemistry - Mitochondria are critical to cellular activity that implicated in expansive networks to maintain organismal homeostasis under external stimuli of nutrient...     

The feedback regulation of carbohydrates intake on food intake and appetite in grass carp (Ctenopharyngodon idella)

Fish Physiology and Biochemistry - Improving carbohydrate utilization can contribute to sustainability of aquaculture. In order to explore the feedback mechanism of glucose homeostasis in fish, one...     

Overview of Fish Growth Hormone Family. New Insights in Genomic Organization and Heterogeneity of Growth Hormone Receptors

Growth hormone (GH), prolactin (PRL) and somatolactin (SL) are single chain proteins structurally and functionally related. Fish PRL and GH receptors (PRLR, GHR) have been characterized in several fish species. There is limited evidence of fish PRLR isoforms, but emerging data support the existence of different GHR variants. In gilthead sea bream, black sea bream, turbot and fugu, but not in zebrafish, GHR has retained an exclusive fish intron (10/10A). In gilthead sea bream and turbot, this intron is not alternatively spliced, but the black sea bream intron is either removed or retained during mRNA processing, resulting in a long GHR isoform with a 31 amino acid insertion that does not alter the open reading frame. This or any other GHR variant are not found in gilthead sea bream, but a truncated anchored form has been reported in turbot. The latter GHR isoform comprises extracellular and trans-membrane domains, the first 28 amino acids of the intracellular domain and 21 divergent amino acids before a stop codon. This GHR variant is the result of alternative splicing, being the 3′ UTR and the divergent sequence identical to the sequence of the 5′ end of the 9/10 intron. The physiological significance of different fish GHR isoforms remains unclear, but emerging data provide suitable evidence for season and nutrition related changes in the somatototropic axis activity. The up-regulation of circulating GH together with the decrease of plasma titres of insulin-like growth factor-I (IGF-I), an altered pattern of serum IGF binding proteins and a reduced expression of hepatic IGF-I and GHRs represent a mechanism conserved through vertebrate evolution. It secures the preferential utilization of mobilized substrates to maintain energy homeostasis rather than tissue growth. Somatolactin also changes as a function of season, ration size, dietary amino acid profile and dietary protein source creating opposite plasma GH and SL profiles. There is now direct evidence for a lipolytic effect of fish SL, acting at the same time as an inhibitory factor of voluntary food intake. Indeed, long-term feeding restriction results in the enlargement of the summer GH peak, whereas the SL rise coincident with shortened day length is delayed in juvenile fish until late autumn. These findings agree with the idea that SL may act as a marker of energy surplus, priming some particular process such as puberty onset. However, it remains unclear whether SL works through specific receptors and/or dimers or heterodimers of GH and PRL receptors.     

cDNA cloning and expression analysis of glutaredoxin 3 in black tiger shrimp Penaeus monodon

Aquaculture International - Glutathione (Grx) is a glutathione-dependent oxidoreductase belonging to the thioredoxin (Trx) superfamily. It maintains redox homeostasis in organisms and plays a...     

In silico and gene expression analysis of the acute inflammatory response of gilthead seabream (Sparus aurata) after subcutaneous administration of carrageenin

Fish Physiology and Biochemistry - Inflammation is one of the main causes of loss of homeostasis at both the systemic and molecular levels. The aim of this study was to investigate in silico the...     

大口黑鲈leptin A基因的组织表达及其对急性低氧胁迫的响应

瘦素(leptin)是一种重要的激素,参与调节动物的摄食、繁殖和能量消耗,其可以通过抑制食欲和促进能量代谢的方式维持机体能量稳态。大多数鱼类具有双重瘦素基因。在本研究中,利用PCR技术克隆了大口黑鲈leptin A基因的编码区,其开放阅读框(ORF)为486 bp,编码161个氨基酸蛋白。通过与其他物种进行同源性比对,发现鲈形目的 leptin基因的保守性较高,一致性高达91.46%,在大口黑鲈leptin A中几乎不存在基因特异性突变位点,而且大口黑鲈leptin A氨基酸序列与鳜同源性最高(92.59%),与花鲈(89.51%)、布氏鲳鲹(87.04%)、斜带石斑鱼(83.87%)的同源性次之。组织分布结果显示,leptin A基因在肝脏中的表达量最高,在心脏、头肾、脑、中肾、肠、脾脏、鳃、肌肉和性腺中的表达量均较低。此外,对大口黑鲈进行不同程度急性低氧胁迫[重度低氧(1.2±0.2)mg/L和中度低氧(3.5±0.2)mg/L],发现低氧胁迫初期时,严重低氧组和中度低氧组的leptin A的表达量都升高,显著高于对照组Leptin A的表达。这表明急性低氧能够诱导大口黑鲈leptin A在肝脏中的表达。综上所述,大口黑鲈leptin A基因与鳜leptin A基因的同源性最高,leptin A主要在大口黑鲈肝脏中显著表达,急性低氧胁迫能显著诱导leptin A的表达。     

急性胁迫对团头鲂铁稳态及其相关基因表达的影响

为研究急性应激反应对鱼体铁含量及铁稳态相关基因的影响,以团头鲂为研究对象,通过腹腔注射皮质醇来模拟急性应激,在注射后不同时间点对团头鲂的血液、肠和肝脏进行取样,采用分光光度法测定血清和肝脏中的铁含量,采用实时荧光定量PCR对铁稳态相关基因的表达量进行检测。结果显示,皮质醇注射后血浆皮质醇显著升高,血浆中铁含量在4、8、10和12 h显著高于对照组,而肝脏中的铁含量在4、8、10和12 h时显著低于对照组。团头鲂肝脏中铁调素(hep)基因表达量显著增加,并分别在8和10 h达最大值,随后有所下降。肠道和肝脏中转铁蛋白基因(Tf)的表达量均呈先升高后下降的趋势。研究表明,急性应激下机体细胞内储存铁大量释放到体液中,造成病原感染和体内增殖风险增加;hep及Tf上调表达降低细胞内铁释放和促进体液铁向胞内蓄积,在鱼体内铁稳态的调控和固有免疫反应中发挥着重要的作用。     

建鲤生长激素促泌素受体1基因的特性及其与增重相关SNP位点的筛选

GHS-R基因在哺乳类为候选的肥胖和生长数量性状位点。本实验使用RT-PCR及PCR的方法,从建鲤(Cyprinus carpio var.jian)分离到2个jlGHS-R1s,称为jlGHS-R1a和jlGHS-R1b(分别简称1a和1b)。jlGHS-R1s的阅读框编码360个氨基酸,两者间相似性高达96%;另外还存在2个jlGHS-R1s的转录变体(分别简称1a’和1b’),选择性切割了翻译起始碱基后第490-680 nt的191 bp片段,该片段两端碱基分别为gt-ag,这种选择性切割导致翻译提前终止,仅翻译184个氨基酸,包括前面3个半跨膜区,这与已报道的罗非鱼等的保留内含子的转录变体不同。jlGHS-R1s的阅读框被1个内含子分隔,该内含子位于第260个氨基酸密码子的第1和第2个碱基之间,1a和1b的内含子长度分别为676 bp和885 bp。使用分段PCR在建鲤群体的1a和1b基因上共找到32个SNP位点,构建PCR-RFLP法对其中9个SNPs进行基因型检测,结果发现各种基因型在322尾建鲤中均有出现,但存在着明显的偏分布。与增重的相关分析表明,位点1a-I_C386T和1b-E1_G159T与雌雄鱼鱼种和成鱼阶段增重分别呈极显著(P<0.01)和显著(P<0.05)相关,CC型和GG型个体增重最快,另外有5个位点则与其中的某阶段或者某性别的增重相关,为了确定上述所得标记的适用性,选取其中4个标记在另外7个家系共610尾鱼中检测,结果 C386T和G159T还是与增重呈极显著(P<0.01)和显著(P<0.05)相关,可用于建鲤分子育种。     

Energy balance of juvenile <Emphasis Type="Italic">Cyprinus carpio</Emphasis> after a short-term exposure to sublethal water-borne cadmium

Stress in fish can be assessed by means of a bioenergetic approach, based on the evaluation of changes in their physiological parameters. The objective of this study was to determine the impact of sublethal water-borne cadmium (Cd) on the energetic balance of juvenile Cyprinus carpio under laboratory conditions after a short-term exposure. Fish were exposed to a concentration of Cd (0.15 mg Cd l⁻¹) for 2 weeks. This concentration is environmentally realistic since it is usually found, even at higher values, in heavily polluted periurban water bodies of Argentina. No mortality was recorded among the animals used in the experiments. Food intake, food assimilation and assimilation efficiency, fecal production, liver glycogen content, oxygen consumption, oxygen extraction efficiency, specific metabolic rate, ammonia excretion and ammonia quotient (AQ), condition factor, and liver somatic index were determined. The overall balance was expressed as the scope for growth (SFG). The morphological indices and the liver glycogen content of Cd-exposed fish showed no significant differences when compared to those of controls. There was a significant decrease in the food intake, fecal production, and food assimilation rates as well as in AQ; the SFG exhibited a highly significant decrease. The remaining parameters (assimilation efficiency, oxygen consumption, oxygen extraction efficiency, specific metabolic rate, and ammonia excretion) increased after the exposure to Cd. We concluded that the sub-chronic exposure of Cyprinus carpio to a sublethal concentration of Cd causes important alterations in the energy-related homeostasis of fish. Most of the responses are indicative of physiological adaptations to compensate an increased energy requirement due to the impairments caused by the metal.     

Molecular characterization and expression analysis of two Na+/H+ antiporter genes in the marine red alga Porphyra yezoensis

Na⁺/H⁺ antiporters are known to play a crucial role in pH and Na⁺ homeostasis. In the present study, we characterized the molecular structures and expression patterns of two Na⁺/H⁺ antiporters from the marine red alga Porphyra yezoensis (designated PySOS1 and PyNhaD). The full-length cDNAs of PySOS1 and PyNhaD were 5122 and 1804?bp, and contained open reading frames (ORFs) of 4773 and 1275?bp, respectively. The deduced amino acid sequences showed high similarity to SOS1 and NhaD from the higher plant Arabidopsis thaliana. PySOS1 and PyNhaD contained conserved sequences found in the cation?Cproton antiporter. Quantitative real-time PCR analysis revealed that both antiporter genes were expressed in both the gametophyte and sporophyte of P. yezoensis. In addition, mRNA expression of PySOS1 and PyNhaD was simultaneously upregulated by light irradiation, suggesting that coordinated activity between the two is important in pH and Na⁺ homeostasis under light conditions. Moreover, the expression levels of both genes were partially reduced by the photosynthetic inhibitors DCMU and DBMIB, suggesting that upregulation is linked to photosynthesis-related metabolism. These findings provide an initial step towards understanding Na⁺/H⁺ antiporters in marine red algae.     

Energy structure and efficiency of a typical Chinese integrated fish farm

Integrated fish culture is an ancient practice in China and South East Asia but the structure, function and management of such systems have been little understood scientifically. This paper describes the energy input-output structure and its efficiency in marketable fish ponds at Nanhui Fish Farm in the Changjiang Delta of the People's Republic of China. In 1983–1984, the photosynthetic solar energy of the grow-out ponds was 1.15% of the total solar radiation. The net conversion efficiencies of biological energy, biological-industrial energy and total energy were 8.74, 8.43 and 3.40% respectively. The energy consumptions per tonne of fish were 46.68, 48.35 and 119.87 GJ for biological, biological-industrial and total energy respectively. About 60, 39 and 1% of the energy consumed came from solar radiation, biological energy and industrial energy respectively. Compared with 1982, the conversion efficiency in 1984 increased by 23.3%, which resulted from the modification to an aquaculture, animal husbandry and agriculture complex ecosystem, and from the improvement of the aquaculture system by increasing the proportion of grazing fish and decreasing the proportion of filtering fish as well as by increasing the input of compounded feeds. One of the remaining problems is the ecological energy trap of the soil at the bottom of the ponds.