Nucleotide polymorphisms, transcriptional analysis, gene expression of the bovine growth hormone secretagogue receptor 1a (GHS-R1a) gene and its genetic association with growth and carcass traits in cattle

Masanori Komatsu, Yuki Fujimori, Tomohito Itoh, Yoichi Sato, Hiroaki Okamura, Motohide Nishio, Osamu Sasaki, Aduli E.O. Malau-Aduli, Hideaki Takahashi, Hisato Takeda, Masahiro Satoh

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Ghrelin - Growth hormone secretagogue receptor 1a (GHS-R1a) is involved in many important functions including growth hormone (GH) secretion and food intake. In this chapter, we explore existing nucleotide polymorphisms, transcriptional analysis, gene expression of the bovine GHS-R1a gene and its genetic association with growth and carcass traits in cattle. Firstly, we evaluated haplotype variety and characterized the microsatellite ((TG)n, 5'-untransrated region (UTR)) and nucleotide polymorphisms of the GHS-R1a gene in cattle. Nucleotide sequencing of this gene (~6 kb) revealed 47 single nucleotide polymorphisms (SNPs), four indels and the microsatellite ((GTTT)n, Intron 1). The 19 haplotypes were constructed from all nucleotide viability patterns and divided into 3 major groups. Four SNPs (L24V, nt456(G>A), D191N and nt667(C>T)) and DelR242 in Exon 1 and a haplotype block of about 2.2 kb (nt667(C>T) ~ nt2884 (A>G)) were identified in Bos taurus breeds. Significant breed differences in allele frequencies of the two microsatellites, nt-7(C>A), L24V, and DelR242 loci were found. A DelR242 was found in the Japanese Shorthorn (frequency: ~ 0.44), Japanese Brown, 5 European cattle breeds, the Philippine native cattle, but none detected in the Japanese Black nor the Mishima Island cattle. Secondly, 5'-rapid amplification of cDNA ends (5'-RACE) and reverse transcription-polymerase chain reaction (RT-PCR) analyses revealed that there were two different kinds of transcripts: spliced, without a microsatellite within 5'-UTR (GHS-R1a); and non-spliced, with the microsatellite (GHS-R1b). Thirdly, we examined age-related changes in the expressions of GHS-R1a and GHS-R1b (the truncated-type receptor) in the arcuate nucleus, pituitary gland and other tissues by real-time RT-PCR in cattle. The GHS-R 1a mRNA expression in the arcuate nucleus of post-weaning calves was more than 10-fold higher than those of pre-weaning calves and cows, and its expression level was the highest in all tissues examined. The GHS-R1a mRNA expression in the pituitary gland of pre-weaning calves was higher than those of post-weaning calves and cows. The GHS-R1b mRNA expression was widespread in all tissues examined and predominantly occurred in the pancreas, pituitary gland, spleen and arcuate nucleus in adult. Fourthly, we carried out a genetic association study between five nucleotide polymorphisms (5'UTR microsatellite ((TG)n), nt-7(C>A), L24V, DelR242 and Intron 1 microsatellite (GTTT)n) of the GHS-R1a gene and growth and carcass traits in 1,285 steers sired by 117 Japanese Black bulls in a progeny testing program. Statistical analysis revealed that the 5'UTR microsatellite locus had a significant additive effect on carcass weight (CW) and average daily gain (ADG). One of the four major microsatellite alleles (19-TG allele) with an allele frequency of 0.145, had a significantly desirable effect on CW and ADG. We proposed a translational hypothesis that the association is due to differences in the secondary structure of GHS-R1b mRNA (the non-spliced type with the 5'UTR microsatellite) among the GH-SR1a gene haplotypes. Finally, we predicted the potential increase in profitability due to increased CW in cow-calf fattening enterprises through planned matings based on DNA testing of the 5'UTR microsatellite. We concluded that the 19-TG allele could potentially be an economically useful nucleotide marker for growth and carcass traits in Japanese Black cattle.

Original languageEnglish
Title of host publicationGhrelin
Subtitle of host publicationProduction, Action Mechanisms and Physiological Effects
PublisherNova Science Publishers, Inc.
Pages1-31
Number of pages31
ISBN (Print)9781619424005
Publication statusPublished - 2012 Feb 1
Externally publishedYes

Keywords

  • Cattle
  • Growth and carcass traits
  • Growth hormone secretagogue receptor (GHS-R)
  • MRNA expression
  • MRNA secondary structure
  • Nucleotide polymorphism
  • Profitability
  • Tissue distribution

ASJC Scopus subject areas

  • Medicine(all)

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    Komatsu, M., Fujimori, Y., Itoh, T., Sato, Y., Okamura, H., Nishio, M., Sasaki, O., Malau-Aduli, A. E. O., Takahashi, H., Takeda, H., & Satoh, M. (2012). Nucleotide polymorphisms, transcriptional analysis, gene expression of the bovine growth hormone secretagogue receptor 1a (GHS-R1a) gene and its genetic association with growth and carcass traits in cattle. In Ghrelin: Production, Action Mechanisms and Physiological Effects (pp. 1-31). Nova Science Publishers, Inc..