动物甜味受体研究进展及猪饲料甜味剂的选择

动物的味觉识别是通过口腔味觉细胞中的味觉受体来启动。味觉受体T1R家族中的T1R2和T1R3以异构体方式发挥甜味识别作用,其中T1R2起主导作用,T1R3对T1R2结合位点构象起调节作用。猪与人在甜味受体基因序列上存在种族差异,导致猪与人在味觉感觉上存在明显不同,对人很甜的一些人工甜味剂(如索马甜、甜蜜素、NHDC、阿斯巴甜等)不能被猪识别,其对猪不能起到甜味作用。     

甜味识别与转导机理

甜味是动物最喜好的基本味感,甜味食物通常能为机体提供所需的碳水化合物。功能学研究表明,甜味是由T1R2和T1R3介导的。它们单独存在时是糖的低亲和力受体;它们以异二聚体存在时,是糖和各种人工甜味剂的高亲和力受体。T1R2和T1R3在甜味感受中发挥着不同的作用。T1R2的N端是某些甜味剂(如阿斯巴甜和莫内林)的重要结合位点,C端跨膜区与G蛋白偶联;T1R3的跨膜区也是某些甜味剂的结合位点,该部位可能在受体的激活过程中发挥着关键作用。甜味受体具有多个结合位点,用以识别结构各异的甜味物质。由于甜味化合物化学构成不同,甜味信息的转导可能存在着不同的路径。     

甜味受体的营养研究进展及其基因表达调控

甜味受体(T1R2/T1R3)属于G蛋白耦联受体(GPCR)C家族的成员,是一种通过非共价键结合且具有7个α螺旋跨膜结构域(TMD)和N末端胞外结构域(NTD)结构的异源二聚体。甜味分子通过与T1R2/T1R3上的关键结合位点相互作用,使T1R2/T1R3由失活状态的收缩构象变为激活状态的展开构象,并经由环腺苷酸(cAMP)途径和三磷酸肌醇/二酯酰甘油(IP3/DAG)途径,最终引起胞内游离钙离子(Ca~(2+))浓度的上升和甜味细胞膜的去极化,产生甜味味觉信号并传导至甜味味觉传入神经丛,最后传导至大脑皮层味觉神经中枢,感知味觉。文章综述了T1R2/T1R3的生物学特征、甜味信号的转导机制、T1R2/T1R3的基因表达调控和反馈调节以及前景展望。     

动物鲜味受体的研究进展及其基因表达调控

动物的鲜味受体包括代谢型谷氨酸受体(m GluR)和味觉受体异源二聚体(T1R1/T1R3),是C型G蛋白偶联受体,N末端捕蝇草模块(VFT)区域可与鲜味配体结合,识别鲜味。本文主要论述了鲜味受体的研究进展、鲜味识别转导机制及鲜味受体基因的表达调控等,以期为相关研究提供参考。     

绵羊味觉受体第一家族基因外显子多态性分析

为了探讨绵羊味觉受体第一家族（taste receptor family 1 member,T1R）基因外显子多态性及其基因型在乌珠穆沁羊和湖羊群体中分布的差异性,试验采用DNA池直接测序及飞行时间质谱（MALDI-TOFMS）法对中国蒙古系两个绵羊品种共172个个体T1Rs基因外显子的遗传变异情况进行分析,利用生物信息学软件预测多态位点对T1Rs基因mRNA二级结构和蛋白质二级结构的影响。结果表明,在两个群体的T1R家族基因中筛查到9个SNPs。独立性卡方检验显示有5个多态位点基因型的分布在两个绵羊群体中存在显著性差异（P<0.05）,分别为TAS1R1基因上的SNP2,TAS1R2基因上的SNP4、SNP7和SNP8,TAS1R3上的SNP10,其中,SNP2、SNP7和SNP10为同义突变;SNP2和SNP10导致相应基因mRNA二级结构和最小自由能的改变,而SNP7仅导致TAS1R2基因最小自由能发生改变;SNP4和SNP8为错义突变,分别导致TAS1R2蛋白质中第379位天冬酰胺变为丝氨酸和第701位苏氨酸变成蛋氨酸,且突变前后受体蛋白的二级结构均发生改变。     

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。     

湖羊消化道各段T1R1、T1R3及PepT1基因表达水平的分析

为分析T1R1(味觉受体1家族Ⅰ型)、T1R3(味觉受体1家族Ⅲ型)及PepT1(Ⅰ型寡肽转运载体)基因对湖羊氨基酸吸收的影响,以6月龄的湖羊为试验材料,采用荧光定量PCR技术对湖羊瘤胃、网胃、瓣胃、皱胃、十二指肠、空肠、回肠、盲肠、结肠和直肠的T1R1、T1R3和PepT1基因的表达量进行相对定量分析。结果表明:T1R1和T1R3基因在检测的各段组织中均有表达,而PepT1基因在结肠中不表达;其中T1R1和T1R3基因在十二指肠中表达量最高,其次是空肠,并且两者差异显著极显著(P0.01);T1R1和T1R3基因在十二指肠(P0.01)和空肠(P0.05)中的表达量与消化道其他部位相比差异极显著或显著;Pep T1基因在空肠中表达量最高,其次是回肠,并且两者差异极显著(P0.01);Pep T1基因在空肠和回肠中的表达量与消化道其他部位相比差异极显著(P0.01)。该结果为进一步研究T1R1、T1R3和Pep T1基因对湖羊氨基酸吸收相关功能影响奠定基础。     

干扰素的新家族--IFN-λs研究现状

干扰素λ(IFN-λ)家族是干扰素的一个新家族,不同于Ⅰ型IFN和Ⅱ型IFN,包括IFN-λ1、IFN-λ2和IFN-λ3(又称IL-29、IL-28A和IL-28B)。IFN-λs的信号转导机制类似于IFN-α,两者都是通过诱导受体异二聚体化,活化Jak-STAT信号通路,进而发挥抗病毒、抗细胞增殖等生物学作用。所不同的是IFN-λs是通过诱导其独特的Ⅱ型细胞因子受体CRF2-12/IFNLR1/IL-28Rα和CRF2-4/IL-10Rβ异二聚体化。IFN是第一个应用于临床的基因工程产品,IFN-α/β在治疗肿瘤和病毒性疾病等方面都取得了显著的疗效。IFN-λs与Ⅰ型IFN有着几乎相同的功能,是否具有独特功能,正在进一步的研究之中。     

昆虫化学感受受体的研究进展

化学感受这一重要的生化过程对于昆虫的生存来说是十分必要的,昆虫的化学感受受体家族由气味(嗅觉)受体和味觉受体组成。嗅觉可以识别挥发性的化学物质,使昆虫发现食物、寻找配偶和逃避敌害;而味觉能识别可溶性刺激物,以引起觅食、交配和产卵等行为。文章主要介绍了昆虫气味受体及味觉受体家族的特性,以及目前对多种昆虫化学感受受体超家族的研究进展。     

肠道蛋白质感应系统与肠道内分泌细胞调控研究进展

肠道上皮细胞的感应系统能够感应肠腔营养物质,影响肠道内分泌细胞(EECs)分泌脑肠肽,从而调节机体生理活动。机体对蛋白胨的感应主要通过小肽转运蛋白(Pep T1)和溶血磷脂酸受体5(LPAR5)发挥作用,对氨基酸的感应主要通过G蛋白偶联受体C家族6组a亚型受体(GPRC6A)、1型味觉受体1和3(T1R1/T1R3)以及雷帕霉素靶蛋白复合体1(m TORC1)信号通路发挥作用。文章对动物肠道蛋白质的感应机制及内分泌调控进行综述,旨在为同行提供参考。     

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).     

苦味受体的生物学特征、信号转导机制及苦味剂和苦味抑制剂对苦味受体的影响

苦味受体(bitter taste receptors,TAS2Rs)是一种G-蛋白偶联受体(GPCR),由30个基因组成的基因家族编码。苦味可使动物远离有毒有害物质,当动物尝到苦味物质时,会刺激舌头的味蕾中味觉受体细胞表达TAS2Rs,进而引发下游一系列信号转导反应,最终通过鼓索神经和舌咽神经将信息整合传到大脑,使动物产生厌恶的感觉,从而选择拒绝摄入这些苦味物质。本文就TAS2Rs的生物学特征、信号转导机制及苦味剂和苦味抑制剂对苦味受体的影响进行简要综述。     

饲粮中添加风味剂对猪采食量的影响及其作用机理

调控猪各个阶段的采食量是动物营养研究的热点问题。风味剂主要有香味剂、甜味剂、鲜味剂,可以改善饲粮的适口性,提高猪对饲粮的喜爱程度,也可以缓解因环境变化、饲料原料改变等应激反应导致的采食量下降。猪的嗅觉系统特别发达,灵敏度很高,味觉也要比人类敏感,因此在饲粮中添加风味剂主要通过刺激猪的嗅觉和味觉来使摄食中枢兴奋,进而促进猪采食。本文综述了饲粮中添加风味剂对猪采食量的影响及其作用机理,为相关试验提供理论依据。     

Lineage‐specific evolution of bitter taste receptor genes in the giant and red pandas implies dietary adaptation

Taste 2 receptors (TAS2R) mediate bitterness perception in mammals, thus are called bitter taste receptors. It is believed that these genes evolved in response to species‐specific diets. The giant panda (Ailuropoda melanoleuca) and red panda (Ailurus fulgens styani) in the order Carnivora are specialized herbivores with an almost exclusive bamboo diet (>90% bamboo). Because bamboo is full of bitter tasting compounds, we hypothesized that adaptive evolution has occurred at TAS2R genes in giant and red pandas throughout the course of their dietary shift. Here, we characterized 195 TAS2R genes in 9 Carnivora species and examined selective pressures on these genes. We found that both pandas harbor more putative functional TAS2R genes than other carnivores, and pseudogenized TAS2R genes in the giant panda are different from the red panda. The purifying selection on TAS2R1, TAS2R9 and TAS2R38 in the giant panda, and TAS2R62 in the red panda, has been strengthened throughout the course of adaptation to bamboo diet, while selective constraint on TAS2R4 and TAS2R38 in the red panda is relaxed. Remarkably, a few positively selected sites on TAS2R42 have been specifically detected in the giant panda. These results suggest an adaptive response in both pandas to a dietary shift from carnivory to herbivory, and TAS2R genes evolved independently in the 2 pandas. Our findings provide new insight into the molecular basis of mammalian sensory evolution and the process of adaptation to new ecological niches.     

迪庆藏猪与野藏杂交猪肌肉全谱游离氨基酸味道强度值比较

本研究旨在探明迪庆藏猪与野猪×迪庆藏猪（野藏杂交猪）肌肉全谱游离氨基酸（FAA）味道强度值（TAV）的差异。选择胎次相同、出生日期相近、体重20 kg左右的迪庆藏猪和野藏杂交猪各12头（公、母各半）,以迪庆藏猪为对照,采用相同饲粮在相同饲养条件下直线育肥,100 kg左右屠宰,采集背最长肌,沸水蒸30 min后,用高效液相色谱-四级杆离子阱串联质谱仪检测其全谱游离氨基酸,计算呈味氨基酸TAV并比较其差异。结果显示,与迪庆藏猪相比,野藏杂交猪肌肉游离氨基酸总鲜味TAV降低16.27%（P<0.05）,总甜味、酸味和苦味TAV分别降低3.50%、11.68%和1.50%,但差异均不显著（P>0.05）;野藏杂交猪肌肉谷氨酰胺（Gln）鲜味TAV、甜味TAV均降低22.89%（P<0.05）,缬氨酸（Val）甜味、苦味TAV均增加36.32%（P<0.05）;天冬氨酸（Asp）、甘氨酸（Gly）、谷氨酸（Glu）与组氨酸（His）等其他游离氨基酸的鲜味、甜味、酸味和苦味TAV与迪庆藏猪间均差异不显著（P>0.05）。以上结果表明,与野猪杂交会降低迪庆藏猪肌肉鲜味TAV,降低Gln的鲜味、甜味TAV,提高Val甜味、苦味TAV,该结果可为迪庆藏猪的开发利用提供依据。     

Correlation between skeletal muscle fiber type and responses of a taste sensing system in various beef samples

The difference of muscle fiber type composition affects several parameters related to meat quality; however, the relationship between muscle fiber types and meat taste is unclear. To elucidate this relationship, we determined the taste of various beef samples using a taste sensor (INSENT SA402B) and analyzed its correlation with different muscle fiber type composition. We used 22 kinds of beef samples and measured nine tastes, including the relative and change of membrane potential caused by adsorption (CPA) values, using six sensors (GL1, CT0, CA0, AAE, C00, and AE1). The taste sensor analysis indicated positive value outputs for the relative C00, AAE, and GL1 values as well as for the CPA value of AAE, which corresponded to bitterness, umami, sweetness, and richness, respectively. We found significant positive correlations of the myosin heavy chain 1 (MyHC1) composition with umami taste, and with richness. This result suggests that high levels of slow MyHC1 can induce strong umami taste and richness in beef. We expect that our results will contribute to the elucidation of the relationship between muscle fiber types and meat palatability.     

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.