Analyses of gene mutations of luteinizing hormone receptor in animals with crptorchidism and development of methods of genetic diagnosis.

隐睾动物黄体生成素受体基因突变分析及基因诊断方法的建立。

• 批准号: 16580265
• 负责人: KAWATE Noritoshi
• 金额: $ 2.37万
• 依托单位: Osaka Prefecture University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16580265/
• 关键词: cryptorchidism; dog; bull; LH receptor; gene polymorphism (SNP); DNA; genetic diagnosis; cDNA; 遺伝子変異

In this research project, we determined cDNA sequences of luteinizing hormone (LH) receptors derived from canine testis. Also, we analyzed the sequences of LH receptors gene in dogs and bulls with cryptorchidism, as compared with normal animals.From total RNA exctraced from normal male dogs, 2123 bp of LH receptor cDNA, except a partial sequence of 5' region, was cloned. In these process, we found a splice variant of the receptor mRNA which shows a deletion of 25 amino acids in the extracellular region. The canine LH receptor cDNA up to the terminator codon shows homologies with porcine, bovine and human sequences of 91.9%, 91.7% and 90.3%, respectively.Total RNA or genomic DNA was extracted from testicular tissues or blood sample of normal (n=46) male and cryptorchid (n=45) dogs, and normal (n=3) and cryptorchid (n=9) bulls. Partial sequences of the LH receptor gene were amplified with RT-PCR or PCR and compared between normal and affected animals.Both of the 1137^<th> and 1194^<th> b … More ases of the canine LH receptor show single nucleotide polymorphisms (SNP) with C, T or C/T for 1137^<th> base and A, G or A/G for 1194^<th> base. There were no different ratios of each SNP in both bases between normal and cryptorchid dogs. However, the ratio of A at the 1194^<th> base in the cryptorchid dogs with testicular retention in the abdoman was significantly lower than that of normal dogs and cryptorchid dogs with retention at the inguinal region. There were 6 haplotypes for 1137^<th> and 1194^<th> bases with T-G, C-A, C-G, T/C-A/G, T/C-G or C-A/G. The C-A type in the cryptorchid dogs with testicular retention in the abdoman was significantly lower than that of normal dogs and cryptorchid dogs with retention at the inguinal region. The 703^<th> base of the bovine LH receptor gene shows SNP (A (Lys), C (Gin) or A/C (Lys/Gin) ). However, there were no marked different ratios of each SNP between normal and cryptorchid dogs.We successfully cloned 2123 bp of cDNA of the canine LH receptor, except partial 5' region from testicular tissues. It is evident, in the present study, that a new type of splicing variant of LH receptor mRNA, which has not been reported in other species, is expressed in canine testis. These studies also suggest that, in crpyorchid dogs with abdominal retention, SNP patterns of the two bases (1137^<th> and 1194^<th>) in the transmembrane region may differ compared with normal and affected animals with inguinal retention. Less

在本研究项目中，我们测定了来自犬睾丸的黄体生成素（LH）受体的cDNA序列。此外，我们还分析了隐睾犬和公牛的LH受体基因序列，并与正常动物进行了比较。从正常公犬的总RNA中克隆出除5′区部分序列外的2123 bp的LH受体cDNA。在这个过程中，我们发现了受体mRNA的剪接变体，在细胞外区域缺失了25个氨基酸。犬LH受体cDNA与猪、牛、人终止密码子的同源性分别为91.9%、91.7%和90.3%。从正常雄性犬（n=46）和隐睾犬（n=45）、正常公牛（n=3）和隐睾犬（n=9）的睾丸组织或血液样本中提取总RNA或基因组DNA。用RT-PCR或PCR扩增LH受体基因的部分序列，比较正常动物和患病动物之间的差异。犬LH受体的1137^< >和1194^< >均存在单核苷酸多态性（SNP）， 1137^< >碱基为C、T或C/T， 1194^< >碱基为A、G或A/G。在正常犬和隐睾犬之间，两个碱基中每个SNP的比例没有差异。然而，睾丸潴留在腹部的隐睾犬，其1194^< >碱基的A比值明显低于正常犬和腹股沟区潴留的隐睾犬。1137^< >和1194^< >碱基有6个单倍型，分别为T-G、C-A、C-G、T/C-A/G、T/C-G和C-A/G。腹部睾丸潴留的隐睾犬的C-A型明显低于正常犬和腹股沟区睾丸潴留的隐睾犬。牛LH受体基因的703^< >碱基显示SNP (A （Lys), C （Gin）或A/C (Lys/Gin)）。然而，每个SNP在正常犬和隐睾犬之间的比例没有显著差异。我们成功地从犬睾丸组织中克隆了2123 bp的LH受体cDNA，但不包括部分5′区。很明显，在本研究中，在犬睾丸中表达了一种新的LH受体mRNA剪接变体，这种变体在其他物种中尚未报道。这些研究还表明，在患有腹股沟潴留的crpyorchid犬中，跨膜区两个碱基（1137^<th>和1194^<th>）的SNP模式可能与正常和患有腹股沟潴留的患病动物不同。少

项目成果

Preventive effects of CIDR-based protocols on premature ovulation before timed-AI in Ovsynch in cycling beef cows

基于 CIDR 的方案对循环肉牛 Ovsynch 中定时 AI 之前过早排卵的预防作用

• 发表时间: 2007
• 期刊: The Veterinary Journal 173
• 作者: Sakase M;Kawate N;Nakagawa C;Fukushima M;Noda M;Takeda K;Ueno S;Inaba T;Kida K;Tamada H;Sawada T
• 通讯作者: Sawada T

Relations between plasma IGF-I concentrations during treatment with CIDR-based or Ovsynch protocol for timed AI and conception in early postpartum Japanese black beef cows.

日本黑肉牛产后早期采用基于 CIDR 或 Ovsynch 方案的定时 AI 治疗期间血浆 IGF-I 浓度与受孕之间的关系。

• 发表时间: 2006
• 期刊: Journal of Reproduction and Devlopment 52・1
• 作者: Kawate N;Sakase M;Seo Y;Fukushima M;Noda M;Takeda K;Ueno S;Inaba T;Kida K;Tamada H;Sawada T
• 通讯作者: Sawada T

Inhibitory effects of CIDR-based ovulation-synchronization protocols on uterine PGF2alpha secretion at the following luteal phase in early postpartum non-cycling beef cows.

基于 CIDR 的排卵同步方案对产后早期非周期肉牛黄体期子宫 PGF2α 分泌的抑制作用。

• 发表时间: 2006
• 期刊: Journal of Reproduction and Devlopment 52-4
• 作者: Sakase M;Kawate N;Nakagawa C;Fukushima M;Noda M;Takeda K;Ueno S;Inaba T;Kida K;Tamada H;Sawada T.
• 通讯作者: Sawada T.

Effect of CIDR-based protocols for timed-AI on the conception rate and ovarian functions of Japanese Black beef cows in the early postpartum period

• DOI: 10.1016/j.theriogenology.2005.02.008
• 发表时间: 2005-09-15
• 期刊: THERIOGENOLOGY
• 影响因子: 2.8
• 作者: Sakase, A;Seo, Y;Kawate, N
• 通讯作者: Kawate, N

獣医繁殖学 第三版 (第7章 発情と排卵の同期化)

兽医生殖，第三版（第七章发情和排卵的同步）

• 发表时间: 2006
• 作者: Tamada H;Tominaga M;Kida K;Kawate N;Inaba T;Matsuyama S;Sawada T;川手憲俊 他 (分担著者)
• 通讯作者: 川手憲俊 他 (分担著者)