C型利钠肽及其受体在水牛卵泡中的表达分析

为了探明C型利钠肽（C-type natriuretic peptide,CNP）及其受体（natriuretic peptide receptor,NPR）在水牛卵泡中的表达模式,本研究首先采用实时荧光定量PCR技术检测水牛卵巢中利钠肽家族主要成员A型、B型、C型利钠肽（ANP、BNP、CNP）及其Ⅰ型、Ⅱ型受体（NPR1、NPR2）的mRNA表达情况,然后利用免疫组化技术对水牛卵泡中CNP及NPR2蛋白进行定位,最后利用实时荧光定量PCR技术检测颗粒细胞和卵丘细胞中CNP及NPR2的mRNA表达规律。结果显示,水牛卵巢主要表达CNP及NPR2,且在各级卵泡中均有表达,其中,CNP主要在壁层颗粒细胞中表达,NPR2主要在卵丘细胞中表达;颗粒细胞上CNP mRNA表达水平显著高于卵母细胞周围的卵丘细胞（P<0.05）,而卵丘细胞上NPR2 mRNA表达水平显著高于颗粒细胞（P<0.05）。综上所述,在利钠肽主要家族成员和受体中,CNP和NPR2在水牛卵巢中呈现强表达,CNP主要在壁层颗粒细胞中表达,而NPR2主要在卵母细胞周围的卵丘细胞中表达。     

口蹄疫病毒整联蛋白受体的研究进展

从组织分布、各毒株利用率、配体结合区介导感染的能力及β亚基胞质区作用等方面论述了4种整联蛋白αvβ1、αvβ3、αvβ6和αvβ8的研究进展，旨在阐明各整联蛋白决定口蹄疫病毒宿主组织嗜性的作用，并为口蹄疫病毒致病机制的研究提供理论依据。     

Trade-off expression of melatonin receptor subtypes (Mel1a and Mel1b) and androgen receptor in lung of a tropical bird,Perdicula asiatica

The role of circulatory steroid hormone along with melatonin in lung of any seasonally breeding bird has never been explored so far. This could be interesting because steroid hormones are immunosuppressive while melatonin is immunostimulatory in nature. In our present study, we report the effect of exogenous melatonin and testosterone on expression of melatonin receptor subtypes (Mel_1a and Mel_1b) and androgen receptor in lung of a tropical bird Perdicula asiatica. Birds were collected from vicinity of Varanasi and acclimatized in laboratory with sufficient food and water. The birds were treated with melatonin and testosterone at dose of 25 µg/100 g B.wt./day and 1 mg/100 g B.wt./day, respectively, for 28 days. At the end of the experiment, the birds were sacrificed and lung tissue and blood sample were collected for immunohistochemistry, Western blot analysis and hormonal assay. Testosterone treatment increased circulatory testosterone and upregulated expression of androgen receptors whereas downregulated expression of melatonin receptor subtypes Mel_1a and Mel_1b. Melatonin administration increased peripheral melatonin and upregulated expression of melatonin receptor subtypes Mel_1a and Mel_1b while downregulated androgen receptor. Thus, our results suggest that a trade-off relationship between melatonin and testosterone exists in regulation of their receptors in lung of Perdicula asiatica.     

草原红牛不同组织雄激素受体基因表达量检测及比较研究

采用实时荧光定量RT-PCR技术检测雄激素受体基因(Androgen receptor gene,AR)在草原红牛公牛和母牛多种组织中的表达情况,探讨该基因在草原红牛公牛和母牛不同组织中的表达差异.结果表明,AR基因在所检测的公牛和母牛的心、肝、脾、肺、肾、肠、胃及睾丸(卵巢)等8种组织中均有表达,且肝脏中的表达量高于其他组织;草原红牛公牛和母牛相同组织闻比较结果显示:AR基因在草原红牛公牛肝脏中表达量与母牛相比较差异不显著,其他不同组织间表达量均有显著差异(P<0.05),且公牛各组织中的表达量均高于母牛.本试验为深入研究AR基因的生物学功能及了解该基因对不同性别个体肉质性状的形成的作用机制奠定了基础.     

孕酮处理对蛋鸡蛋壳钙化过程中CaBP-d28k基因表达的影响

实验旨在研究孕酮对蛋鸡CaBP-d28k基因表达的影响。对产蛋高峰期蛋鸡进行孕酮和孕酮受体拮抗剂(RU-486)处理,利用实时荧光定量PCR技术(QRT-PCR),测定在蛋壳形成过程中输卵管子宫部钙结合蛋白(CaBP-d28k)、孕酮受体(PGR)的表达量;同时对血液中钙离子和孕酮浓度变化进行测定。结果表明:孕酮处理后,血液中钙离子浓度显著降低(P<0.05),输卵管子宫部CaBP-d28k表达量也显著降低(P<0.05);孕酮受体拮抗剂处理后,血液中钙离子浓度显著升高(P<0.05),输卵管子宫部CaBP-d28k表达量极显著升高(P<0.01)。结果提示,在蛋壳钙化过程中,孕酮对CaBP-d28k表达和钙离子转运起负调控作用。     

Polymorphism in promoter region of growth hormone receptor is associated with potential production capacity of insulin‐like growth factor‐1 in pre‐pubertal Holstein heifers

Insulin‐like growth factor‐1 (IGF‐1) is one of the important factors for growth, milk production and reproductive functions and mainly released from the liver in response to growth hormone (GH) via GH receptor (GHR) in cattle. Recently, some single nucleotide polymorphisms (SNPs) were identified in the bovine GHR gene. Some GHR‐SNPs were shown to be related to plasma IGF‐1 concentration in cattle. Hence, the capacity to IGF‐1 production in the liver might be affected by GHR‐SNP and associated with performance in the future. This study examined whether GHR‐SNP is associated with IGF‐1 production in the liver of pre‐pubertal heifers. In 71 Holstein calves, blood samples for genomic DNA extraction were obtained immediately after birth. To genotype the GHR‐SNPs in the promoter region, polymerase chain reaction (PCR) products were digested with restriction enzyme NsiI (cutting sites: AA, AG and GG). All heifers at 4 months of age were intramuscularly injected with 0.4 mg oestradiol benzoate. Blood samples were obtained from the jugular vein just before (0 h) and 24 h after injection. The number of AA, AG and GG at the NsiI site was 0, 17 and 54 respectively. In AG and GG, plasma GH concentrations were higher pre‐injection than 24 h post‐injection (p < 0.01). Moreover, plasma GH concentrations in AG post‐injection were higher than in GG (p < 0.05). In contrast, the GG genotype exhibited higher plasma IGF‐1 concentrations in pre‐injection than post‐injection (p < 0.01), although oestradiol did not change IGF‐1 concentration in the AG genotype. We conclude that the GG polymorphism in the promoter region of GHR is associated with a higher potential capacity of IGF‐1 production in the liver of cattle.     

不同处理方法对受体牛同期发情效果的比较研究

[目的]为了进一步促进人工授精技术广泛应用以及提高胚胎移植效率,为酒泉本地胚胎移植选择高效的同期发情方法。[方法]本试验分别采用PRID＋PGF2α法和PGF2α＋PGF2α法对胚胎移植受体牛做同期友情处理。[结果]表明：两种方法处理受体牛,其同期发情率和移植妊娠率之间差异无统计意义（P〉0．05）,但是用PRID＋PGF2α法处理受体牛同期发情率和移植妊娠率均高于PGF2α＋PGF2α法。[结论]在奶牛养殖及生产上,在考虑移植成本和生产效益的情况下,在农区大面积对牛只进行同期发情技术处理,应优选PGF2α＋PGF2α法处理受体牛。     

Toll-like receptor signaling is changed in ovine lymph node during early pregnancy

Toll-like receptors (TLRs) participate in regulation of adaptive immune responses, and lymph nodes play key roles in the initiation of immune responses. There is a tolerance to the allogenic fetus during pregnancy, but it is unclear that expression of TLR signaling is in ovine lymph node during early pregnancy. In this study, lymph nodes were sampled from day 16 of nonpregnant ewes and days 13, 16, and 25 of pregnant ewes, and the expressions of TLR family (TLR2, TLR3, TLR4, TLR5 and TLR9), adaptor proteins, including myeloid differentiation primary-response protein 88 (MyD88), tumor necrosis factor receptor associated factor 6 (TRAF6), and interleukin-1-receptor-associated kinase 1 (IRAK1), were analyzed through real-time quantitative polymerase chain reaction, Western blot, and immunohistochemistry analysis. The results showed that mRNA and protein levels of TLR2, TLR3, TLR4, TRAF6, and MyD88 were upregulated in the maternal lymph node, but TLR5, TLR9, and IRAK1 were downregulated during early pregnancy. In addition, MyD88 protein was located in the subcapsular sinus and lymph sinuses. Therefore, it is suggested that early pregnancy induces changes in TLR signaling in maternal lymph node, which may be involved in regulation of maternal immune responses in sheep.     

绵羊瘦素受体基因部分序列测定及其变异位点分析

试验旨在检测绵羊瘦素受体(leptin receptor,LEPR)基因序列突变位点,为进一步分析LEPR基因多态性与绵羊生长性状的关系奠定基础。选用美利奴羊与阿华西羊杂交群体作为试验材料,利用RT-PCR、测序等方法测定了4只绵羊的LEPR基因cDNA部分序列及内含子7序列,同时利用PCR-RFLP分析两个位点在群体(229只)中的多态性。测定出LEPR基因cDNA序列长2608 bp(包含外显子2-16完整序列及外显子1和17的部分序列)和LEPR第7内含子序列全长160 bp,共发现5个核苷酸变异位点,第2外显子中2个(T240C和A279G),第10、14外显子中各1个(A1683G,T2373C),内含子7中1个(1285+A73G),外显子中的4个变异位点均未引起编码氨基酸的改变。在研究群体中,A279G与A1683G中A的频率分别为0.415、0.467,后者处于非平衡状态,GG和AA是主要的单倍型。不同物种间序列一致性分值均比较高,但LEPR mRNA区序列一致性比内含子高,基于LEPR mRNA序列构建的系统发育树更符合实际情况,且可靠性值更高。结果表明,绵羊LEPR基因的保守性较强,突变形式主要是转换,A279G和A1683G可能是突变热点。