爱拔益加肉仔鸡肠道PepT1、B0 AT和EAAT3 mRNA的表达差异与发育规律

本试验旨在研究爱拔益加(AA)肉仔鸡肠道小肽转运载体1(PepT1)、B0系统中性氨基酸转运载体(B0AT)、兴奋性氨基酸转运载体3(EAA T3)mRNA的表达差异与发育规律.试验选取1日龄雄性AA肉仔鸡120只,随机分为8个组,于1、7、14、21、28、35、42日龄,每个组选取1只肉仔鸡,采用实时荧光定量PCR技术检测不同肠段Pep T1、B0AT、EAAT3 mRNA的相对表达量.结果显示:1)肉仔鸡不同肠段PepT1、B0AT、EAAT3 mRNA的相对表达量差异显著(P＜0.05).其中,PepT1 mRNA的相对表达量在十二指肠最高,空肠次之,回肠最低,B0AT、EAA T3mRNA的相对表达量在回肠最高,空肠次之,十二指肠最低.2)肉仔鸡不同日龄Pep T1、B0AT、EAAT3 mRNA的相对表达量存在极差异显著(P＜0.01);在不同肠段上,PepT1、B0AT、EAA T3mRNA的相对表达量随日龄的变化均表现为前期升高,后期平稳的趋势.由此可见,肉仔鸡肠道参与小肽及氨基酸吸收的转运载体PepT1、B0AT、EAAT3 mRNA表达具有肠段及日龄的差异性.     

T1R1和T1R3在从江香猪附睾发育中的表达模式

为研究味觉受体第一家族亚型1（T1R1）和3（T1R3）在从江香猪附睾发育过程中的表达模式,探讨味觉受体在哺乳动物雄性生殖机能中可能发挥的作用及潜在医学价值,本试验以从江香猪附睾组织为研究对象,分析附睾发育4个关键时期：初情前（15 d）、初情时（30 d）、初情后（60 d）和性成熟期（180 d）T1R1与T1R3的差异表达。采用实时荧光定量PCR、免疫组织化学（IHC）和Western blot检测两个味觉受体在不同日龄从江香猪附睾组织中转录、翻译水平的变化及其分布情况。RT-qPCR结果表明：TAS1R1与TAS1R3 mRNA在从江香猪附睾初情前（15 d）至性成熟期（180 d）表达量逐渐增加,且任意两个时期间差异极显著（P<0.01）。Western blot结果显示,T1R1/T1R3蛋白在180 d表达量最高,在15 d表达量最低,两者之间差异显著（P<0.05）,平均表达丰度依次为180 d > 30 d > 60 d > 15 d。IHC结果显示,T1R1和T1R3蛋白在各日龄组从江香猪附睾组织均有分布,其中T1R1蛋白主要在上皮细胞膜上,尤其是基细胞和窄细胞;而T1R3蛋白主要在微绒毛、环状空泡和精子呈强阳性表达。综上,本研究发现不同日龄从江香猪附睾的T1R1/T1R3表达从15 d逐渐增加,至性成熟达到峰值,这一表达变化与附睾上皮基细胞和窄细胞及微绒毛的T1R1/T1R3的差异表达有关,这些特殊的表达模式与附睾生理功能存在时间关联,故推测T1R1和T1R3参与附睾内精子成熟和储存的调节过程。     

蛋鸡小肠肽转运载体PepT1表达差异性的评定

本试验旨在研究蛋鸡十二指肠、空肠、回肠小肽转运载体mRNA表达的差异性.选用相同背景的健康成年蛋鸡10只,从十二指肠、空肠、回肠组织提取RNA样品,采用相对定量RT-PCR,对蛋鸡小肠各段肽转运载体mRNA表达的差异性进行评定.结果表明,蛋鸡空肠PepT1 mRNA的表达水平显著高于十二指肠(P＜0.05),与回肠比差异不显著(P＞0.05),十二指肠PepT1 mRNA的表达水平与回肠之间无显著差异(P＞0.05).     

湖羊和徐淮山羊CaSR基因组织表达水平的初步分析

为分析钙敏感受体(CaSR)基因对湖羊和徐淮山羊消化吸收的作用,以6个月龄的湖羊和徐淮山羊为试验材料,利用荧光定量PCR SYBR GreenⅡ荧光染料法,对湖羊和徐淮山羊瘤胃、网胃、瓣胃、皱胃、十二指肠、空肠、回肠、盲肠、结肠和直肠10个组织中CaSR基因的表达水平进行相对定量分析。结果表明:在湖羊的样品组织中,CaSR基因在十二指肠中表达量最高,其次是回肠,两者的表达量差异达到显著水平(P0.05);CaSR基因在瘤胃、网胃、瓣胃、皱胃、空肠、盲肠、结肠、直肠中的表达量显著低于在十二指肠及回肠中的表达量(P0.05)。在徐淮山羊的组织样中,CaSR基因在回肠中的表达量最高,其次是十二指肠,且二者差异显著(P0.05);另外,该基因在回肠和十二指肠的表达量显著高于在瘤胃、网胃、瓣胃、皱胃、空肠、直肠中的表达量(P0.05),盲肠和结肠中该基因的表达量最低。通过试验说明CaSR在不同组织里的表达具有特异性,在不同的物种间主要表达组织也不同,表明湖羊和徐淮山羊具有不同的消化吸收方式。     

香猪睾丸间质细胞TAS1R3基因干扰后对自噬相关因子的影响

旨在探究TAS1R3基因通过mTOR对从江香猪自噬相关基因表达的影响。本试验选择30日龄健康雄性从江香猪3头，采集睾丸组织并培养出睾丸间质细胞。将构建成功的干扰载体和空载体转染至细胞中，利用qRT-PCR检测其对TAS1R3基因表达影响，运用qRT-PCR和Western blot检测TAS1R3基因干扰后对味觉受体第一家族一号受体(TAS1R1)、细胞外信号调节激酶1(ERK1)、细胞外信号调节激酶2(ERK2)、雷帕霉素靶蛋白(mTOR)、BECN1和微管相关蛋白1轻链3β(MAP1LC3B)表达的影响。结果表明，当TAS1R3基因被干扰后，相较于shRNA-NC对照组，TAS1R1、ERK1、ERK2、mTOR基因mRNA表达量均极显著降低(P<0.01),自噬因子BECN1和MAP1LC3B表达量极显著升高(P<0.01)。Western blot结果表明当TAS1R3基因被干扰后，与shRNA-NC对照组相比，蛋白T1R3、T1R1、p-mTOR、p-S6K1、p-ERK12均极显著降低(P<0.01),而自噬蛋白Beclin 1表达量极显著升高(P<...     

日粮赖氨酸对肉鸡生长发育和肠道蛋白质转运载体基因表达的影响

试验旨在评估日粮赖氨酸缺乏或过量对肉鸡生长发育和肠道蛋白质转运载体基因表达的影响。采用单因素试验设计,选取360只1日龄AA肉鸡雏随机分成3个处理,每个处理6个重复,每个重复20只鸡,分别饲喂赖氨酸水平为0.60%(赖氨酸缺乏组,LL组)、1.00%(赖氨酸适量组,ML组)、1.40%(赖氨酸过量组,HL组)的日粮,以ML组为对照组,LL组和HL组为试验组。试验期为3周。结果表明:(1)与ML组相比,LL组肉鸡胸肌重、腿肌重极显著降低,料重比极显著升高(P0.01)。(2)与ML组相比,LL组极显著上调肉鸡十二指肠Pep T1、B0AT、EAAT3基因表达,极显著下调CAT1基因表达(P0.01);HL组极显著下调Pep T1、B0AT、EAAT3基因表达(P0.01),显著下调CAT1基因表达(P0.05)。(3)与ML组相比,LL组极显著下调肉鸡空肠Pep T1、B0AT、EAAT3、CAT1基因表达(P0.01);HL组极显著上调Pep T1、B0AT基因表达,极显著下调EAAT3、CAT1基因表达(P0.01)。(4)与ML组相比,LL组极显著上调肉鸡回肠Pep T1、B0AT基因表达,极显著下调EAAT3基因表达(P0.01),显著下调CAT1基因表达(P0.05);HL组极显著上调EAAT3基因表达(P0.01),显著上调CAT1基因表达,显著下调B0AT基因表达(P0.05)。试验揭示赖氨酸缺乏主要通过下调肉鸡空肠蛋白质转运载体基因表达量,而过量主要通过下调肉鸡十二指肠蛋白质转运载体基因表达量,从而抑制肉鸡肠道蛋白质转运吸收,降低蛋白质沉积效率,进而可能严重阻碍肉鸡生长发育。     

肉鸡肠道PepT1 mRNA表达的肠段差异性与发育性变化

选用遗传背景相同的1日龄父母代雄性Arbor Acre(AA)鸡和父母代雄性岭南黄肉雏鸡各120羽,采用相对定量RT-PCR方法,以30 d时岭南黄鸡肠道样品为模板,研究肉鸡肠道寡肽转运载体1(Peptide transporter 1,PepT1)mRNA表达的肠段差异性;以AA肉鸡和岭南黄鸡十二指肠和空肠样品为模板,研究不同品种肉鸡肠道PepT1 mRNA表达的发育性变化。结果显示:(1)岭南黄肉鸡肠道PepT1 mRNA的表达丰度从十二指肠、空肠、回肠到结直肠依次降低,其中十二指肠显著高于结直肠(P<0.05);(2)AA鸡和岭南黄肉鸡PepT1 mRNA在十二指肠及空肠中的表达具有相同的发育模式,16 d表达丰度最高,16~44 d逐渐下降,58 d略微回升;不同基因型之间PepT1 mRNA丰度比较,AA鸡和岭南黄肉鸡两品种间PepT1 mRNA的表达没有显著差异(P>0.05)。以上结果说明:(1)随着肠道空间位置的后移,岭南黄肉鸡肠道PepT1 mRNA的表达丰度逐渐降低,其中十二指肠的表达丰度显著高于结直肠(P<0.05);(2)不同品种肉鸡十二指肠及空肠PepT1 mRNA的表达具有相同的发育模式,且同日龄两品种间的表达丰度未见明显差异,表明PepT1 mRNA表达受到发育阶段的调控,但品种间具有稳定性。     

不同精粗比对犊牛组织中PepT1和PHT1及PHT2表达的影响

研究不同精粗比全价颗粒饲料对犊牛不同组织中PepT1、PHT2和PHT1 mRNA表达的影响。试验选取日龄相近、体重相近的中国荷斯坦断奶公犊牛12头,分为4组,每组3头,各试验组精粗比分别为75:25(Ⅰ)、70:30(Ⅱ)、65:35(Ⅲ)和60:40(Ⅳ),预试期14 d,正试期56 d。结果表明:脾脏中PHT1mRNA表达量最高,肺中的PHT2mRNA表迭量最高,瘤胃中的PepT1mRNA表达量最高,而它们在心脏中表达量均最低;试验Ⅰ和Ⅱ组肝脏、皱胃、瓣胃和瘤胃中的PHT1 mRNA表达量极显著高于试验Ⅳ组(P0.01),而试验Ⅲ组网胃的PHT1 mRNA表达量极显著高于其他各组(P0.01);试验Ⅲ组犊牛肝脏和网胃以及试验Ⅱ组皱胃中PHT2 mRNA表达量极显著高于其他各组(P0.01),试验Ⅰ和Ⅳ组瘤胃和瓣胃中PHT2 mRNA表达量极显著高于其他2组(P0.01);试验Ⅳ组犊牛肝脏中Pep T1 mRNA表达量极显著高于其他各组(P0.01),试验Ⅰ组瘤胃、网胃和瓣胃中PepT1 mRNA表达量极显著高于试验Ⅲ和Ⅳ组(P0.01)。结果表明,不同精粗比全价颗粒饲料对不同组织的PepT1、PHT1和PHT2表达具有一定的影响作用,其中高精粗料促进其表达。     

高、低淀粉饲粮对山羊小肠甜味受体信号通路基因表达的影响

本试验旨在探讨高、低淀粉饲粮对山羊小肠黏膜结构、肠道激素浓度、甜味受体及其通路元件基因表达的影响。试验选择体况相似的雄性湘东成年黑山羊20只,随机分为2组,每组10只,分别饲喂高淀粉(50.4%,HS组)和低淀粉饲粮(13.1%,LS组)。试验期38 d。结果表明:1)与LS组相比,HS组空肠和回肠的隐窝深度增加(P0.05),十二指肠和空肠的绒毛高度降低(P0.05);回肠黏膜中胆囊收缩素(CCK)(P0.05)和胰高血糖素样肽-2(GLP-2)(P0.05)及回肠内容物中胰高血糖素样肽-1(GLP-1)(P0.05)和酪酪肽(Pyy)(P0.05)的浓度均增加,但回肠黏膜和回肠内容物中胃泌素抑制肽(GIP)的浓度降低(P0.05)。2)2组间Ⅰ型味觉受体3(T1R3)、α-味移导素(α-gustducin)、瞬时受体电位离子通道蛋白M型5(TRPM5)、葡萄糖转运蛋白2(GLUT2)和钠/葡萄糖共转运载体1(SGLT1)在十二指肠、空肠和回肠中的基因表达量无显著差异(P 0.05),但HS组空肠α-gustducin基因表达量显著低于LS组(P 0.05)。3)相关性分析显示,回肠内容物葡萄糖浓度与回肠α-gustducin、TRPM5和SGLT1的基因表达量之间呈显著正相关(0.05 r0.30,P 0.05),而与空肠甜味受体关键元件T1R3与SGLT1的基因表达量之间的相关系数为0.93(P0.01),与空肠TRPM5基因表达量之间的相关系数为0.66(P0.01),与空肠GLUT2基因表达量之间的相关系数为0.48(P0.01)。综上所述,相比粗蛋白质和粗纤维,淀粉是甜味信号通路元件以及葡萄糖转运载体的主要影响因素,但高淀粉饲粮对发育成熟的山羊肠道黏膜结构及功能影响较小。     

仔猪断奶后1周内血清生化指标、十二指肠形态结构及空肠中钠/葡萄糖共转运载体1与二肽转运载体1基因表达量的变化

本试验旨在探讨仔猪在断奶前及断奶后1周内血清生化指标、十二指肠形态结构及空肠中钠/葡萄糖共转运载体1(SGLT1)和二肽转运载体1(PepT1)基因表达量的变化规律。试验选取20日龄仔猪55头,断奶前1天屠宰5头,余下50头仔猪在21日龄断奶,并分为5组(圈),每组10头仔猪,分别在断奶后第2、4和8天从每组各挑选1头仔猪屠宰。结果表明:1)与断奶前相比,血清碱性磷酸酶活性在断奶后第2天显著降低(P0.05),在断奶后第8天极显著降低(P0.01),血清钾离子含量在断奶后第4和8天显著增加(P0.05),血清尿素氮含量在断奶后第2天极显著增加(P0.01),在断奶后第8天又显著降低(P0.05),血清免疫球蛋白G含量显著降低(P0.05)。2)与断奶前相比,十二指肠绒毛高度在断奶后第2天极显著降低(P0.01),断奶后第4天显著降低(P0.05),隐窝深度在断奶后第2和4天均显著增加(P0.05),两者比值在断奶后第2和4天均显著降低(P0.05)。3)与断奶前相比,空肠中SGLT1基因表达量在断奶后均显著降低(P0.05),空肠中PepT1基因表达量在断奶后第4和8天均显著降低(P0.05)。由此表明,断奶1周内对仔猪生理机能造成了一定的应激;断奶降低了仔猪血清免疫球蛋白G含量,改变了仔猪十二指肠形态结构,降低了空肠中SGLT1和PepT1基因表达量,以断奶后第2和4天应激最强。     

Expression Patterns of T1R1 and T1R3 in the Congjiang Xiang Pig during Epididymal Development

The aim of this experiment was to study the expression pattern of taste receptor family 1 subtypes 1 (T1R1) and 3 (T1R3) during epididymal development of Congjiang Xiang pig, and to explore the possible role of these taste receptors in mammalian male reproductive function and its potential medical value. In this study, the differential expressions of T1R1 and T1R3 in epididymis at 4 key developmental periods (neonatal (15 d), peri-puberty (30 d), puberty (60 d) and sexual maturity (180 d)) of Congjiang Xiang pigs were analyzed. RT-qPCR, immunohistochemistry (IHC) and Western blot were used to detect the changes and distribution of the two taste receptors in epididymis of Congjiang Xiang pigs at different ages. The results of RT-qPCR showed that the expression of TAS1R1 and TAS1R3 mRNA increased gradually from neonatal (15 d) to sexual maturity (180 d), and there was a significant difference between each period (P<0.01). The results of Western blot showed that the expression of T1R1/T1R3 protein was the highest on the 180 d and the lowest on the 15 d. The average protein abundance of T1R1/T1R3 was as follows: 180 d > 30 d > 60 d > 15 d. The results of IHC showed that T1R1 and T1R3 proteins were distributed in the epididymis of Congjiang Xiang pigs at 4 periods, in which T1R1 protein was mainly concentrated in epithelial cell membrane, especially in basal and narrow cells, while T1R3 protein was strongly positive in stereocilia, annular vacuoles and spermatozoa. In summary, the expression of T1R1/T1R3 in the epididymis of Congjiang Xiang pigs increased gradually from 15 d to the peak of sexual maturation, which was related to the differential expression of T1R1/T1R3 in epithelial basal cells, narrow cells and stereocilia of epididymis. These special expression patterns were time related to the physiological function of epididymis, so it is speculated that T1R1/T1R3 are involved in the regulation of sperm maturation and storage in epididymis.     

Influences of non‐nutritive sweeteners on ovarian and uterine expression of T1R2 and T1R3 in peripubertal female guinea pigs

The underlying mechanism of taste receptor type 1 subunit 2 (T1R2) and taste receptor type 1 subunit 3 (T1R3) in the hormonal and reproductive system is still elusive. A low or a high dose of sweetness equivalent to that sodium saccharin (SS, 1.5 or 7.5 mM) and rebaudioside A (RA, 0.5 or 2.5 mM) was administered to young female guinea pigs for 28 consecutive days from the age of 28 days. Our results indicated that the sweet taste receptor subunit T1R2 was markedly expressed in the ovary and uterus of guinea pigs, whereas the T1R3 protein was expressed at a lower level. We elucidated that low‐dose (1.5 mM) SS increased body and ovary weight associated with elevated ovarian expression of T1R2 in guinea pigs, unlike the high‐dose (7.5 mM) SS, which suppressed the ovarian expression of T1R2 and resulted in certain adverse effects on ovarian and uterine morphology. Furthermore, high‐dose (2.5 mM) RA increased the number of corpus luteum and elevated uterine expression of T1R2, whereas low‐dose (0.5 mM) RA induced increased secretion of serum progesterone. Therefore, our findings suggest that we should pay more attention to the potential adverse effects, including increases in ovary weight, morphology changes, and increased progesterone that result from the dose‐dependent regulation of T1R2 by non‐nutritive sweeteners (NNS) in the ovaries and uteri of peripubertal females.     

哺乳动物味觉感受器T1Rs受体家族及信号转导机制的研究进展

味觉受体第一家族（T1Rs）是一类能感知甜味和鲜味的受体家族,它包括T1R1,T1R2,T1R3三个成员。T1R2＋T1R3以异二聚体形式共表达参与甜味识别,而T1R1＋T1R3也以异二聚体形式共表达参与鲜味识别。良好的味觉能促进牛的食欲,促进生长发育,但目前关于牛的T1Rs受体家族的研究并不多见,有必要对这个受体家族做进一步探索。本文从甜味和鲜味两个方面对这一受体家族的分子结构及信号转导机制的最新研究进展做简要综述。     

The association of bovine T1R family of receptors polymorphisms with cattle growth traits

The three members of the T1R class of taste-specific G protein-coupled receptors have been proven to function in combination with heterodimeric sweet and umami taste receptors in many mammals that affect food intake. This may in turn affect growth traits of livestock. We performed a comprehensive evaluation of single-nucleotide polymorphisms (SNPs) in the bovine TAS1R gene family, which encodes receptors for umami and sweet tastes. Complete DNA sequences of TAS1R1-, TAS1R2-, and TAS1R3-coding regions, obtained from 436 unrelated female cattle, representing three breeds (Qinchuan, Jiaxian Red, Luxi), revealed substantial coding and noncoding diversity. A total of nine SNPs in the TAS1R1 gene were identified, among which seven SNPs were in the coding region, and two SNPs were in the introns. All five SNPs in the TAS1R2 gene and all three SNPs in the TAS1R3 gene were identified in the coding region. Four SNPs (TAS1R1 g.5081C>T, TAS1R1 g.5110C>A, TAS1R2 g.288A>G, TAS1R2 g.2552T>C) were significantly associated with body height of Qinchuan cattle (P<0.05). The heterozygous genotypes of the four SNPs showed a molecular heterosis on cattle heights at hip cross and sacra. The individuals with different genotypic combinations of the four SNPs had significant association with heights at hip cross and sacra (P<0.05).     

Expression of the sweet receptor protein, T1R3, in the human liver and pancreas

The expression of T1R3, a taste receptor essential for the perception of sweetness and umami-taste, was examined by immunohistochemistry to determine whether and where it may be localized in the liver and pancreas. In the liver, both immunopositive and immunonegative reactions were detected; bile ducts and intercalated portions of the bile ductules were immunopositive to T1R3, while arterioles and venules were immunonegative in interlobular connective tissue. In the hepatic lobule, all other cells including liver cells (hepatocytes) and bile capillaries were immunonegative. In the pancreas, all endocrine portions of the pancreas were immunonegative to T1R3. Within the exocrine portions, immunopositive reactions were detected in excretory duct cells, intercalated cells, and centroacinar cells. In contrast, acinar cells were immunonegative, as were vessels, lymph capillaries, nerve fibers, and connective tissue cells in the exocrine portions. The restricted localization of T1R3 in the duct cells of the liver and pancreas in the present study may indicate that T1R3 is involved in monitoring changes in the makeup of bile and pancreatic juices in the hepatic and pancreatic duct systems.     

MC1R基因的研究进展

MC1R基因作为控制毛色的重要候选基因之一,不仅在黑色素合成过程中起着重要的调控作用,参与毛色的形成,还对人畜疾病、生长性状、行为有着不同程度的影响,因而引起相关学者的广泛关注。为加深对MC1R基因的认识和了解,作者对MC1R基因结构、功能、作用机理、基因的定位、多态性检测及其在人畜方面的研究进展进行综述,并提出新的设想。     

Research Advances on MC1R Gene

MC1R gene, as one of the important candidate genes for controlling coat color, not only plays an important role in the process of melanin synthesis, but also participates in the formation of coat color and also has different effects on human and animal diseases, growth traits and behaviors. The study of MC1R gene has attracted the attention of scholars.In order to deepen the understanding of MC1R gene, the structure, function, mechanism, gene locus, polymorphism analysis and research progress of human and animal of MC1R gene were reviewed and new ideas were put forward in this review.