梅山与长大母猪下丘脑-垂体-卵巢轴Kiss1和GPR54基因的表达差异

研究早熟与晚熟品种母猪下丘脑-垂体-卵巢轴Kiss1和GPR54基因的表达差异。选用8头梅山母猪与12头长大(LY)母猪为研究对象,梅山母猪于70 d和100 d屠宰,LY母猪于70、100 d和199 d屠宰,收集血清及下丘脑、垂体、卵巢组织样品。ELISA检测血清瘦素(Leptin)和雌二醇(E2)水平,PCR克隆梅山与LY母猪Kiss1基因编码序列,实时荧光定量PCR检测母猪下丘脑、垂体、卵巢组织Kiss1和GPR54基因表达水平。结果表明:梅山与LY母猪Kiss1基因编码序列相似性为100%;梅山与LY母猪初情期下丘脑Kiss1基因表达量极显著高于垂体与卵巢(P<0.01),卵巢GPR54基因表达水平显著高于下丘脑与垂体(P<0.05)。梅山母猪下丘脑-垂体-卵巢轴Kiss1基因表达水平都显著高于相同日龄和初情期LY母猪(P<0.05),梅山母猪血清Leptin水平极显著高于相同日龄LY母猪(P<0.01)。而E2水平显著高于100 d与初情期LY母猪(P<0.01)。Leptin与下丘脑Kiss1和GPR54基因表达呈显著正相关(P<0.05),而E2仅与下丘脑Kiss1基因表达有极显著正相关关系(P<0.01);梅山与LY母猪Kiss1基因编码区序列相似性为100%,梅山母猪下丘脑-垂体-卵巢轴上kiss1基因表达量较LY母猪高,主要原因可能是初情日龄早,下丘脑Kiss1基因表达水平的高低与血清Leptin和E2浓度密切相关。     

Effect of supplements during the cold season on the reproductive system in prepubertal Tibetan sheep ewes

We examined the development of the reproductive system in prepubertal Tibetan sheep ewes when fed only oat hay (CON) or supplemented with either lick blocks (BS) or concentrate feed (CS) during the cold season. The average daily gain of the CS ewes was greater than that of the BS ewes (P  < 0.05), which was greater than that of the CON ewes. The same pattern was observed in the number of ovarian follicles (P  < 0.001), that is, CS > BS > CON. Serum concentrations of gonadotropin‐releasing hormone, follicle‐stimulating hormone, luteotrophic hormone, estradiol and progesterone in the CS and BS groups were higher than in the CON group (P  < 0.05). The messenger RNA (mRNA) expression of KiSS‐1, GPR54 (G protein‐coupled receptor 54), ERα (estradiol receptor α) in the hypothalamic anteroventral periventricular area of the CS group were higher than in both the BS and CON groups (P  < 0.05), while the BS group was higher than in the CON group (P  < 0.05). Similar differences among groups were observed for gonadotropin‐releasing hormone receptor mRNA expression in the pituitary, follicle‐stimulating hormone receptor and luteinizing hormone receptor mRNA expression in the ovary. These results indicated that the KiSS1/GPR54 system was more active with nutrition or trace mineral supplementation during the cold season. The system stimulated the hypothalamic–pituitary?gonadal axis and enhanced folliclar development in prepubertal Tibetan sheep ewes. We concluded that energy, protein and trace minerals supplements could improve the reproductive performance of Tibetan sheep on the Qinghai‐Tibetan plateau.     

金黄地鼠Kcnq1基因的分子克隆与鉴定以及在多种器官组织中的表达差异分析

对金黄地鼠Kcnq1基因进行分子克隆与鉴定以及在多种器官组织的表达差异进行分析,旨在研究Kcnq1基因在地鼠各组织器官中的功能。提取金黄地鼠心脏组织总RNA,根据大鼠Kcnq1的保守序列区域设计引物TK1,用RT-PCR的方法从金黄地鼠心脏组织中扩增出Kcnq1cDNA片段,并且以心脏cDNA片段为模板,将纯化后的cDNA在T4连接酶的作用下与pMD18-T载体特异性连接,转化感受态大肠杆菌DH5α中,筛选重组子并酶切鉴定,将鉴定后的重组子进行DNA测序。提取心、肝、脾、肺、肾各组织总RNA,并反转录,将各组织cDNA做荧光定量检测,检测各组织表达量的差异。结果显示克隆出金黄地鼠Kcnq1基因477bp的部分片段长度,推测出编码的159个氨基酸。与大鼠等物种Kcnq1基因比对,核苷酸和氨基酸序列均具有较高的同源性。荧光定量结果显示,Kcnq1在金黄地鼠心脏中表达量最高,在肺脏和肾脏中均有较高表达,在脾脏中低度表达,在肾脏中基本不表达。该研究结果为深入研究金黄地鼠Kcnq1基因功能奠定了基础。     

金黄地鼠Kcnq1基因的分子克隆与鉴定以及在多种器官组织中的表达差异分析

本研究对金黄地鼠Kcnq1基因进行分子克隆与鉴定以及在多种器官组织的表达差异进行分析,旨在研究Kcnq1基因在地鼠各组织器官中的功能.提取金黄地鼠心脏组织总RNA,根据大鼠Kcnq1的保守序列区域设计引物TK1,用RT-PCR化后的cDNA在T4连接酶的作用下与pMD18-T载体特异性连接,转化感受态大肠杆菌DH5a中,筛选重组子并酶切鉴定,将鉴定后的重组子进行DNA测序.提取心、肝、脾、肺、肾各组织总RNA,并反转,将各组织cDNA做荧光定量检测,检测各组织表达量的差异.结果,克隆出金黄地鼠Kcnq1基因477 bp的部分片段长度,推测出编码的159个氨基酸.与大鼠等物种Kcnq1基因比对,核苷酸和氨基酸序列均具有较高的同源性.荧光定量结果显示,Kcnq1在金黄地鼠心脏中表达量最高,在肺脏和肾脏中均有较高表达,在脾脏中低度表达,在肾脏中基本不表达.该研究结果为深入研究金黄地鼠KCNQ1基因功能奠定了基础.     

Neurokinin‐1 receptor expression and antagonism by the NK‐1R antagonist maropitant in canine melanoma cell lines and primary tumour tissues

We interrogated the neurokinin‐1 receptor (NK‐1R)/substance P (SP) pathway in canine melanoma tumour tissues and cell lines. NK‐1R messenger RNA (mRNA) and protein expression were observed in the majority of tumour tissues. Immunohistochemical assessment of archived tissue sections revealed NK‐1R immunoreactivity in 11 of 15 tumours, which may have diagnostic, prognostic and therapeutic utility. However, we were unable to identify a preclinical in vitro cell line or in vivo xenograft model that recapitulates NK‐1R mRNA and protein expression documented in primary tumours. While maropitant inhibited proliferation and enhanced apoptosis in cell lines, in the absence of documented NK‐1R expression, this may represent off‐target effects. Furthermore, maropitant failed to suppress tumour growth in a canine mouse xenograft model derived from a cell line expressing mRNA but not protein. While NK‐1R represents a novel target, in the absence of preclinical models, in‐species clinical trials will be necessary to investigate the therapeutic potential for antagonists such as maropitant.     

Oxyntomodulin reduces expression of glucagon‐like peptide 1 receptor in the brainstem of chickens

Oxyntomodulin (OXM) is a peptide released from the gut and attenuates food intake by acting on hypothalamus. However, its role at the molecular level is not well studied. In the first section of this study, we analysed the effect of OXM on food intake behaviour after injecting into the lateral ventricle of chickens. The outcome showed that food intake decreased significantly after administering 4 nmol of OXM. In the second part, the expression of glucagon‐like peptide 1 receptor (GLP‐1R) in the brainstem was analysed by real‐time RT‐PCR. The results showed that expression of GLP‐1R was reduced to 27% and 16% at 30 and 90 mins after injection of OXM respectively. In saline‐injected chickens, no reduction in GLP‐1R was seen. It can be concluded that OXM has a down regulatory effect on the responding receptor, GLP‐1R and OXM in chicks has the same reductive effect on food intake as in the mammals.     

Differential gene expression of Eph‐ephrin A1 and LEPR‐LEP with high or low number of embryos in pigs during implantation

The objective of this study was to ascertain whether mRNA and protein expressions of implantation‐related genes (erythropoietin‐producing hepatocellular receptor–ligand A1, Eph‐ephrin A1 and leptin receptor–leptin, LEPR‐LEP) differed between pigs with high and low number of embryos, and whether these differences in gene expression might affect embryo implantation. Experimental pig groups (n = 24) for high and low number of embryos were prepared by altering the number of eggs ovulated in pre‐pubertal gilts treated with 1.5 × (High) or 1.0 × (Low) PG600 ([400 IU PMSG + 200 IU hCG]/dose, AKZO‐NOBEL). Gilts expressing oestrus were artificially inseminated twice and maintained in breeding and gestation until the reproductive tract was collected on day 22 of pregnancy. At slaughter, the reproductive tracts from each pregnant gilt from each treatment were immediately processed to collect samples for RNA and protein analysis. Within each gilt, three conceptus points were sampled, one from each horn and then a random conceptus within the tract. At each conceptus point, endometrial attachment site, chorion–allantois and embryo were collected and immediately frozen in liquid nitrogen. Number of corpus luteum (CL) (35.4 vs. 12.6) and total embryo number (18.8 vs. 10.2) were greater in the high‐embryo compared to the low‐embryo group, respectively (p < .05). Real‐time qPCR results showed that Eph‐ephrin A1 mRNA expression was less in the high‐embryo (p < .05) compared to the low‐embryo group. In addition, Western blotting analysis indicated that Eph‐ephrin A1 and LEP protein expression at endometrial attachment site in high‐embryo was less (p < .05) compared to low‐embryo group. It was also noted that mRNA expression of Eph‐ephrin A1 and LEPR‐LEP was greater in pregnant than non‐pregnant gilts (p < .05). Moreover, mRNA expression of Eph‐ephrin A1 (p < .05) and LEPR‐LEP was greatest at endometrial attachment site among all three tissues. There was a positive correlation between expressions of Eph‐ephrin A1, LEPR‐LEP and embryo length with the correlation coefficient 0.31–0.59. For Eph‐ephrin A1, the highest correlation coefficient appeared between Eph A1 expression and normal embryo number, between ephrin A1 expression and embryo length. For LEPR‐LEP, the highest correlation coefficient appeared between LEPR‐LEP expression and ovary weight (0.79 for both, p < .05), followed by embryo length and weight. The results of this study suggest that low expression of Eph‐ephrin A1 and LEPR‐LEP is somehow related to increased embryo number during implantation and that endometrial attachment site might be the main target tissue of these gene products. Yet, the increased expression of Eph‐ephrin A1 and LEPR‐LEP appeared associated with increased embryo growth (length and weight) and ovary weight, Eph‐ephrin A1 and LEPR‐LEP might play roles in the regulation of embryo implantation in pigs.     

The effect of M‐phase stage‐dependent kinase inhibitors on inositol 1,4,5‐trisphosphate receptor 1 (IP3R1) expression and localization in pig oocytes

At fertilization, inositol 1,4,5‐trisphosphate receptor type 1 (IP₃R1) has a crucial role in Ca²⁺ release in mammals. Expression levels, localization and phosphorylation of IP₃R1 are important for its function, but it still remains unclear which molecule(s) regulates IP₃R1 behavior in pig oocytes. We examined whether there was a difference in localization of IP₃R1 after in vitro or in vivo maturation of pig oocytes. In mouse oocytes, large clusters of IP₃R1 were formed in the cortex of the oocyte except in a ring‐shaped band of cortex adjacent to the spindle. However, no such clusters of IP₃R1 were observed in pig oocytes and there was no difference in its localization between in vitro and in vivo matured oocytes. We next tried to clarify which factor(s) regulates IP₃R1 localization, phosphorylation and expression using M‐phase stage‐dependent kinase inhibitors. Our results show that treatments with roscovitine (p34^cdc2 kinase inhibitor) or U0126 (mitogen‐activated protein kinase inhibitor) did not affect IP₃R1 expression or localization in pig oocytes, although the latter strongly inhibited phosphorylation. However, treatment with BI‐2536, an inhibitor of polo‐like kinase 1 (Plk1), dramatically decreased the expression level of IP₃R1 in pig oocytes in a dose‐dependent manner. From these results, it is suggested that Plk1 is involved in the regulation of IP₃R1 expression in pig oocytes.     

Seasonal and age effects on energy requirements in domestic short‐hair cats (Felis catus) in a temperate environment

There is little information known about the energy requirements of cats in temperature climates. Energy requirement of domestic short‐haired cats was determined using three groups of mixed gender – old kept outside (approximately 9.9 years of age; 4.8 kg; n = 9), young kept outside (approximately 3.1 years of age; 3.9 kg; n = 8) or young kept inside (approximately 3.1 years of age; 3.9 kg; n = 8). Cats were housed individually for 5 weeks during summer (18.5 ± 0.5 °C) and winter (8.5 ± 0.4 °C) and were fed a commercially available maintenance diet ad libitum. In both periods, energy expenditure was determined from the rates of ²H and ¹⁸O elimination for blood H₂O over a 12 day period, from a doubly labelled water bolus ²H₂O (0.7 g/kg BW) and H₂¹⁸O (0.13 g/kg BW) administered intravenously. During the summer period, macronutrient digestibility was determined. Older cats had a reduction (p < 0.05) in apparent digestibility of dry matter (approximately 9%), energy (approximately 8%) and protein (6%). There was a significant effect of age and season on energy intake and energy expenditure. While lean mass was affected by age and season, there was no effect of age or season on energy expenditure when expressed as a proportion of lean mass. Possible seasonal differences in nutrient digestibility may explain these results.     

Effects of short photoperiod on energy intake,thermogenesis, and reproduction in desert hamsters (Phodopus roborovskii)

Desert hamsters (Phodopus roborovskii) are the least known species in the genus Phodopus with respect to ecology and physiology, and deserve scientific attention, particularly because of their small body size. Here, the responses of energy metabolism and reproductive function to short photoperiods in desert hamsters were investigated. Male and female desert hamsters were acclimated to either long day (LD) (L:D 16:8 h) or short day (SD) photoperiods (L:D 8:16 h) for three months, and then the females were transferred back to an LD photoperiod for a further five months, while at the end of the SD acclimation the males were killed and measurements were taken for serum leptin as well as molecular markers for thermogenesis. We found that like the other two species from the genus Phodopus, the desert hamsters under SD decreased body mass, increased adaptive thermogenesis as indicated by elevated mitochondrial protein content and uncoupling protein‐1 content in brown adipose tissue, and suppressed reproduction compared to those under LD. However, different from the other two species, desert hamsters did not show any differences in energy intake or serum leptin concentration between LD and SD. These data suggest that different species from the same genus respond in different ways to the environmental signals, and the desert adapted species are not as sensitive to change in photoperiod as the other two species.     

Effect of dexamethasone on the expression of atrogin‐1/MAFbx in chick skeletal muscle

Expression of atrogin‐1/MAFbx, a muscle‐specific E3 ubiquitin ligase, is high under catabolic conditions, that result in muscle atrophy. Messenger RNA (mRNA) expression of atrogin‐1/MAFbx is increased by the glucocorticoid dexamethasone in mammalian skeletal muscle. This study investigated the effects of dexamethasone on expression of atrogin‐1/MAFbx in skeletal muscle of neonatal chicks and in chick myotubes. Chicks were given a single intraperitoneal injection of dexamethasone at a concentration of 10 mg/kg body weight. Twenty‐four hours after dexamethasone administration, the Pectoralis muscle weight of chicks was decreased. mRNA expression of atrogin‐1/MAFbx in skeletal muscle of chicks was significantly increased by dexamethasone administration. Expression of other proteolytic‐related genes (20S proteasome C2 subunit, m‐calpain large subunit, and cathepsin B) in skeletal muscle of chicks was not increased by dexamethasone administration. Chick myotubes were incubated with dexamethasone (1, 10 or 100 µmol/L) for 6 h. Expression of atrogin‐1/MAFbx mRNA in chick myotubes was increased in the presence of all concentrations of dexamethasone. However, expression of other proteolytic‐related genes (20S proteasome C2 subunit, m‐calpain large subunit and cathepsin B) in chick myotubes was not affected by dexamethasone treatment. These results indicate that dexamethasone enhances atrogin‐1/MAFbx expression in chick skeletal muscle, resulting in increased muscle atrophy.     

肉鸡胫骨软骨发育不良早期钙黏蛋白1(CDH1)差异表达研究

为研究福美双诱发肉鸡胫骨软骨发育不良(Tibial Dyschondroplasia,TD)早期钙黏蛋白1(CDH1)的差异表达,基础日粮中添加福美双,在试验第1、2、6天,对试验鸡进行剖杀,迅速采取胫骨生长板故入4％多聚甲醛溶液于4℃固定,做CDH1免疫组化分析;提取对照组和饲喂福美双组的生长板总RNA,采用Real-time PCR对CDH1 基因进行差异表达验证.结果钙黏蛋白1 (CDH1/E-cadherin)基因在TD生长板表达上调,在对照和TD软骨生长板,其蛋白合成主要在前肥大、肥大区软骨细胞质,增殖区软骨细胞无表达,钙化区软骨细胞表达少,在饲喂福美双第1、2、6天其表达增加与定量PCR变化结果相一致,且在饲喂福美双第2、6天呈明显高表达.结果表明CDH1 参与细胞黏附、血管入侵及调节Wnt/β-cat的信号传递,和其它分子共同调节软骨内骨化.     

Association of A‐FABP gene polymorphism and mRNA expression with intramuscular fat content (IMF) in Baicheng‐You chicken

This study aims to assess the association of polymorphisms and mRNA expression of adipocyte‐type fatty acid‐binding protein (A‐FABP) with intramuscular fat (IMF) in the breast muscle (BM) and leg muscle (LM) of Baicheng‐You chickens (BYCs). A total of 180 chickens, including sixty black Baicheng‐You chickens (BBYCs), sixty silky Baicheng‐You chickens (SBYCs) and sixty white Baicheng‐You chickens (WBYCs), were reared from 1 to 120 day. A polymerase chain reaction–single‐strand conformation polymorphism strategy (PCR‐SSCP) was used to detect the polymorphism of the A‐FABP gene in the first exon, and the C51T silent mutational site was found. The IMF content with the AA genotype was significantly higher than that with the AG genotype (p = 0.0473) in the LM of WBYC. Thus, this site could be taken as a molecular marker in selecting a higher IMF content of LM in WBYC. A‐FABP gene mRNA expression in the BM and LM of BYCs was detected, and a significant positive correlation was observed in the LM of WBYC. These findings provide fundamental data that might be useful in further study of the role of the A‐FABP gene in IMF content and fatty metabolism in chickens.     

Genetic variation at the alpha‐1‐fucosyltransferase (FUT1) gene in Asian wild boar and Chinese and Western commercial pig breeds

Escherichia coli F18 bacteria producing enterotoxins and/or shigatoxin (ETEC/STEC) are main pathogens that cause oedema disease and postweaning diarrhoea in piglets, and alpha‐1‐fucosyltransferase (FUT1) gene has been identified as a candidate gene for controlling the expression of ETEC F18 receptor. The genetic variations at nucleotide position 307 in open reading frame of FUT1 gene in one wild boar breed and 20 western commercial and Chinese native pig breeds were investigated by polymerase chain reaction–restriction fragment length polymorphism. The results showed that the genetic polymorphisms of the FUT1 locus were only detected in western pig breeds and the Chinese Taihu (including Meishan pig, Fengjing pig and Erhualian pig), Huai and Lingao pig breeds; only Duroc and Pietrain possessed the resistant AA genotype, while the wild boar and other Chinese pig breeds only presented the susceptible genotype GG. The results indicated that Chinese native pig breeds lack genetic factors providing resistance to ETEC F18 bacteria. The resistant allele to ETEC F18 might originate from European wild boar. It was inferred that oedema and postweaning diarrhoea caused by ETEC F18 have close relationship with the growth rate, which can explain why on the contrary Chinese native pig breeds have stronger resistance to oedema and postweaning diarrhoea in piglets compared with western pig breeds.     

On the winter enhancement of adaptive humoral immunity: hypothesis testing in desert hamsters (Phodopus roborovskii: Cricetidae,Rodentia) kept under long‐day and short‐day photoperiod

We tested the winter immunity enhancement hypothesis (WIEH) on male desert hamsters (Phodopus roborovskii) kept under long‐day (LD) and short‐day (SD) photoperiods. We assumed that under SD in a laboratory, the adaptive humoral immune responsiveness to the antigenic challenge would be enhanced due to the lack of winter physical stressors and food shortages and/or because of the action of an endogenous winter bolstering mechanism, while under LD the immune responsiveness would be suppressed by the activity of the reproductive system. The results support the WIEH in part. We did not find a difference in antibody production in response to sheep erythrocytes between SD and LD hamsters, but SD males had the lower number of granulocytes and the higher number of lymphocytes in white blood cell counts. Reproductive activity was lower in SD males. These males demonstrated an increase in their mass‐specific resting metabolic rate, their mass‐specific maximal metabolic rate and their level of cortisol. The result of a generalized linear model analysis indicates the negative effect on secondary immunoresponsiveness to sheep erythrocytes of mid‐ventral gland size, the organ characterizing individual reproductive quality, and designates a tradeoff between antibody production and reproductive effort. The mass‐independent maximal metabolic rate also negatively affected antibody production, indicating a tradeoff between maximal aerobic performance and the adaptive immune function. The higher stress in SD males seems to be the most likely reason for the lack of the effect of daylight duration on antibody production.     

Influences of protein ingestion on glucagon‐like peptide (GLP)‐1‐immunoreactive endocrine cells in the chicken ileum

Influences of a specific dietary nutrient on glucagon‐like peptide (GLP)‐1‐containing cells in the chicken intestine are not yet clear. Significance of dietary protein level on GLP‐1‐containing cells in the chicken ileum was investigated. Chickens fed control or experimental diets of varying protein levels were examined using immunohistochemical and morphometrical techniques. We show that the protein ingestion had an impact on the activities of GLP‐1‐immunoreactive cells in the chicken ileum. Weight gains declined with decreasing dietary crude protein (CP) levels, but no significant differences were detected in the daily feed intake and villous height. GLP‐1‐immunoreactive cells with a round or oval shape were frequently observed in the lower CP level groups (4.5% and 0%). Frequencies of occurrence of GLP‐1‐immunoreactive cells were 41.1 ± 4.1, 38.5 ± 4, 34.8 ± 3.1 and 34.3 ± 3.7 (cells/mm², mean ± SD) for dietary CP level of 18%, 9%, 4.5% and 0% groups, respectively and significant differences were recognized between the control and lower CP level groups (P < 0.05). Multiple regression analysis indicated a significant correlation between the daily protein intake and frequencies of occurrence of GLP‐1‐immunoreactive cells. The protein ingestion is one of the signals that influence GLP‐1‐containing cells in the chicken small intestine.     

Effects of short-term infusion with propionate on the mRNA expression of a putative G-protein coupled receptor 41 (GPR41) in adipose tissue of goats

Short chain fatty acids (SCFA) represent the main source for energy supply in ruminants. Propionate up-regulates leptin synthesis through the G protein-coupled receptor 41 (GPR41) in mice but the importance of the GPR41 in ruminants is not yet clarified. Here we characterise the short-term effects of intravenously infused propionate on a putative GPR41 mRNA in goat adipose tissue. Castrated male goats (Capra hircus) received propionate infusion or NaCl solution with equivalent sodium content for 260 min. A putative GPR41 mRNA was quantified in subcutaneous and perirenal adipose tissue by real-time RT-PCR. The mRNA concentration of the putative GPR41 mRNA increased (p = 0.029) in subcutaneous but not in perirenal adipose tissue (p = 0.756) of propionate-infused animals versus the NaCl group. We hypothesise that the differential response of the putative GPR41 mRNA in subcutaneous versus perirenal adipose tissue towards short-term propionate infusion could be involved in a differential nutrient sensing of SCFA in the two adipose depots of goats.     

Functional expression of the free fatty acids receptor-1 and -4 (FFA1/GPR40 and FFA4/GPR120) in bovine endometrial cells

Veterinary Research Communications - Endometrial epithelial cells play a key defensive role as part of the innate immune response of cow uterus. An association between risk of acquiring infectious...     

Leucine regulates slow‐twitch muscle fibers expression and mitochondrial function by Sirt1/AMPK signaling in porcine skeletal muscle satellite cells

A previous study demonstrated that leucine upregulates the slow myosin heavy chain mRNA expression in C2C12 cells. However, the role of leucine in slow‐twitch muscle fibers expression and mitochondrial function of porcine skeletal muscle satellite cells as well as its mechanism remain unclear. In this study, porcine skeletal muscle satellite cells cultured in differentiation medium were treated with 2 mM leucine for 3 days. Sirt1 inhibitor EX527, AMPK inhibitor compound C, and AMPKα1 siRNA were used to examine its underlying mechanism. Here we showed that leucine increased slow‐twitch muscle fibers and mitochondrial function‐related gene expression, as well as increased succinic dehydrogenase (SDH) and malate dehydrogenase (MDH) activities. Moreover, leucine increased the protein levels of Sirt1 and phospho‐AMPK. We also found that AMPKα1 siRNA, AMPK inhibitor compound C, or Sirt1 inhibitor EX527 attenuated the positive effect of leucine on slow‐twitch muscle fibers and mitochondrial function‐related gene expression. Finally, we showed that Sirt1 was required for leucine‐induced AMPK activation. Our results provide, for the first time, evidence that leucine induces slow‐twitch muscle fibers expression and improves mitochondrial function through Sirt1/AMPK signaling pathway in porcine skeletal muscle satellite cells.