Growth Hormone Gene Polymorphism Research Articles

Short Communication: Identification of Growth Hormone gene polymorphism for beef cattle in Pesisir Selatan District, West Sumatra, Indonesia

Putra DE, Sumadi, Kanazawa T, Hartatik T. 2016. Short Communication: Identification of Growth Hormone genepolymorphism for beef cattle in Pesisir Selatan District, West Sumatra, Indonesia. Biodiversitas 17: 711-715. This study aimed to determine gene polymorphism of growth hormone of domestic cattle in Pesisir Selatan District of West Sumatra. The research was conducted at Laboratory of Animal Genetics and Breeding Faculty of Animal Science, UGM from August 2013 to January 2014. Blood samples were collected from 66 individuals consist of 15 Pesisir cattle, 15 SimPes cattle, 15 SimPO cattle, 15 Bali cattle and 6 PO cattle. DNA was extracted from each blood samples after SDS-proteinase K digestion, and used for PCR-amplification for a region of growth hormone gene (211 bp), and then the PCR products were analyzed for restriction fragment length polymorphism (RFLP) using AluI restriction enzyme. The results showed that GH gene of Pesisir, PO and Bali cattle are monomorphic, which frequencies of L allele was1.00 and V allele was 0.000 while these LL genotype was 1.000. Frequency of L and V alleles in SimPO and SimPes cattle were 0.634, 0.366 and 0.700, 0.300, respectively. SimPO and SimPes cattle were polymorphic, LL and LV of SimPO cattle was 0.733 and 0.267 as well as SimPes cattle which LL and LV was 0.600 and 0.400, respectively. The correlation between genotype and the performance (body weight and body size) was not significant. The present study indicates that polymorphism of growth hormone gene in AluI site could not yet be used as a molecular marker for body weight and body size of beef cattle.

English

Growth hormone (GH) gene has been described as a candidate gene for marker-assisted selection in different farm animals. The present study was designed to identify the polymorphism in GH gene and its association with variation of wool traits in Egyptian sheep breeds. Wool and blood samples were collected from 42 animals including two breeds (Barki and Rahmani) and one crossbred (Rahmani x Awase). Measurements of wool traits were analyzed and involved staple strength (Str), staple length (STL), fiber diameter (FD) and clean fleece yield (CFW). DNA was extracted from blood samples and a 365-bp fragment from exon V was amplified by polymerase chain reaction (PCR). Single strand conformation polymorphisim (SSCP) analysis showed two conformational patterns. The pattern I was recorded to be more frequent (83.3, 92.86 and 90%) than pattern II (16.7, 7.14 and 10%) in Barki, Rahmani and crossbred, respectively. The sequence analysis showed one single nucleotide polymorphism (C/T). The pattern I (allele T) has been found to affect CFW and FD than pattern II (allele C). Whereas, C allele was more pronounced for Str and STL. These traits are the most important parameters determining commercial values of wool that are preferred for clothing or carpets industry. The nucleotide sequences of C and T alleles were submitted to GenBank and have the accession numbers: KT250511 and KT250512, respectively. In conclusion, the present results provide evidence that there is a single nucleotide polymorphism within GH gene in Egyptian sheep breeds. This mutation was found to have some effects on wool traits. Therefore our data show interesting prospects in future selection programs for improving wool industry. Key words: Sheep, wool, growth hormone (GH) gene, polymorphism, single strand conformation polymorphism (SSCP).

Polymorphism in Growth Hormone Gene and its Association with Growth Traits in Siniperca chuatsi

Growth hormone (GH) is a candidate gene for growth traits in fish. In this study, we assessed associations between single nucleotide polymorphisms (SNPs) in GH gene with growth traits in 357 Siniperca chuatsi individuals using high-resolution melting. Two SNPs were identified in GH gene, with one mutation in exon 5 (g.5045T>C), and one mutation in intron 5 (g.5234T>G). The corrections analysis of SNPs with the four growth traits was carried out using General Linear Model (GLM) estimation. Results showed that both of them were significantly associated with growth performance in S. chuatsi. For g.5234T>G, it was significantly associated with body weight (P＜0.01), body length (P＜0.05), body depth (P＜0.01), and body width (P＜0.01), and the individuals of genotype GG grew faster than those of genotypes TT and TG (P＜0.05). A further diplotype-trait association analysis confirmed that in fish with H3H2 (TC-GG) diplotype body weight, body length, and body width was greater than in those with other diplotypes (P＜0.05). These results demonstrated GH gene SNPs could be used as potential genetic markers in future marker assisted selection of S. chuatsi.

Parental contribution and growth hormone gene polymorphism associated with growth phenotypes of red sea bream Pagrus major in mass production: A case study

Red sea bream is one of the most important aquaculture fish species in Japan. To improve the productivity of this fish during seed production, improved growth traits and reduced size variation are needed. In this study, we assessed parental contribution of fast- and slow-growing individuals observed in two different rearing phases in a mass production lot: (1) 50 dph reared in a tank and (2) 200 dph reared in a net cage. We also assessed GH gene (pmaGH) polymorphisms based on a previously developed minisatellite DNA marker. Specific broodstock individuals were significantly associated with fast- or slow-growing individuals at 50 dph and 200 dph. Significant differences in pmaGH minisatellite allele frequencies were observed between fast- and slow-growing groups at 50 dph in the frequency of two alleles (pmaGH-740 and pmaGH-900, respectively). Combining the results of DNA parentage analysis and pmaGH minisatellite allele analysis, one dam and two sires, possessing pmaGH-740, were significantly associated with the slow-growing groups. These results suggest that the minisatellite marker of pmaGH could be a useful tool for growth selection of this fish species.

ASSOCIATIONS BETWEEN POLYMORPHISM OF GROWTH HORMONE GENE WITH MILK PRODUCTION, FAT AND PROTEIN CONTENT IN FRIESIAN HOLSTEIN CATTLE

The aim of the research was to determine the associations between polymorphism of the bovine growth hormone (GH) gene (Leu/Val) and milk production of Friesian Holstein Cattle. A total of 62 cows which consist of two Friesian Holstein cattle groups (from New Zealand=19 heads and Australia=43 heads) were used for the research. We performed the Polymerase Chain Reaction and followed by Restriction Fragment Length Polymorphism using AluI restriction enzyme. The genotype was observed base on the target gene GH 211 bp in the part of intron 4 and exon 5 of growth hormone gene. The frequencies of genotypes LL were found higher than genotype LV in both groups. Friesian Holstein cattle from New Zealand showed the genotype LL and LV as 84% and 16%, respectively. Friesian Holstein cattle from Australia show the genotype LL and LV as 79% and 21%, respectively. The association between Leu/Val polymorphism on milk production, fat and protein content in both groups did not show the significant effect. Base on two groups of the origin of cattle, the result showed the significant different on fat and protein content of milk. Fat and protein contents of milk were higher in breed of FH imported from Australia compared to those in breed of FH imported from New Zailand.

English

Growth hormone (GH) is an anabolic hormone synthesized and secreted by the somatotroph cells of the anterior lobe of the pituitary gland in a circadian and pulsatile manner, the pattern of which plays an important role in postnatal longitudinal growth and development, tissue growth, lactation, reproduction as well as protein, lipid and carbohydrate metabolism. The aim of this study was to detect the genetic polymorphism of GH gene in five camel breeds reared in Egypt which are Sudany, Somali, Mowaled, Maghrabi and Falahy, using polymerase chain reaction- restriction fragment length polymorphism (PCR-RFLP) technique. Also, this work aimed to identify the single nucleotide polymorphism (SNP) between different genotypes detected in these camel breeds. The amplified fragment of camel GH at 613-bp was digested with the restriction enzyme Msp I and the result reveals the presence of three different genotypes; CC, CT and TT in tested breeds. Significant differences were recorded in the genotype frequencies between these camel breeds. The result shows that the Maghrabi breed that is classified as a dual purpose camels had higher frequency for allele C (0.75) than those in the other tested four breeds. The sequence analysis declared the presence of a SNP (C264T) in the amplified fragment which is responsible for the elimination of the restriction site C^CGG and consequently the appearance of two different alleles C and T. The nucleotide sequences of camel GH alleles T and C were submitted to nucleotide sequences database NCBI/Bankit/GenBank and have accession numbers: KP143517 and KP143518, respectively. It is concluded that only one SNP C→T was detected in GH gene among the five tested camel breeds reared in Egypt and this nucleotide substitution can be used as a marker for the genetic biodiversity between these camel breeds. Also, due to the possible association between allele C and higher growth rate, we can used it in marker assisted selection (MAS) for camels in breeding program as a way for enhancement of growth trait in camel breeds reared in Egypt. Key words: Camel breeds in Egypt, GH , PCR, RFLP, SNPs.

Polymorphism and association of growth hormone gene with growth traits in Sirohi and Barbari breeds of goat.

Aim:The aim was to study the polymorphism of exon 2 and exon 3 of growth hormone (GH) gene, to test the polymorphic variants for Hardy–Weinberg equilibrium and to investigate association of these polymorphisms with chest girth and paunch girth in Sirohi and Barbari breeds of goat.Materials and Methods:A total of 80 kids involving forty each of Sirohi and Barbari breeds of goat were included in the study. A good quality genomic DNA isolated from the whole blood using standard protocol were used for polymerase chain reaction (PCR) amplification and products obtained on restriction digestion of amplicon with enzyme HaeIII were separated on 2% agarose gel, and documented in a gel doc system. The chest girth and paunch girth of kids at birth and weekly intervals up to 4 weeks of age and subsequently at 2 months, 3 months and 6 months of age were recorded. Allele frequency and genotype distribution of polymorphism were tested for Hardy–Weinberg equilibrium by program me Genepop package. Association between different genetic variants on chest girth and paunch girth were analyzed by least squares analysis employing suitable statistical model.Results:The PCR product of genomic DNA isolated from kids of Sirohi and Barbari breeds of goat on digestion with the restriction enzyme HaeIII revealed two genotypic variants viz., AB and BB. None of the two breeds was in Hardy–Weinberg equilibrium for these variants. The least squares analysis of variance revealed non-significant effect of GH genotype and breed × genotype interaction on chest girth and paunch girth from birth to 180 days of age. The effect of breed was highly significant (p<0.01) at all ages.Conclusion:The present study showed that both the breeds were polymorphic at the exon 2 and exon 3 loci of GH gene under study with respect to HaeIII restriction endonuclease. None of the breeds was in Hardy–Weinberg equilibrium for this region of GH gene. In the present study, no significant association between GH genotype and chest girth and paunch girth could be established but comparatively higher chest girth and paunch girth were observed for AB genotype across the breeds.

Examination of Growth Hormone (GH) Gene Polymorphism and its Association with Body Weight and Selected Body Dimensions in Ducks.

The main objective of the study was to assess the polymorphism in intron 2 of the GH gene and its association with some morphological traits (body weight--BW, length of trunk with neck--LTN, length of trunk--LT, chest girth--CG, length of breast bone--LBB, length of shank--LS). Polymorphism in intron 2 of the GH gene was evaluated for four duck populations (Pekin ducks AF51, Muscovy ducks from a CK and CRAMMLCFF mother and Mulard ducks). Genetic polymorphism was determined with the PCR-RFLP method using the BsmFI restriction enzyme. In the studied duck sample two alleles (GH(C) and GH(T)) and three genotypes (GH/TT, GH/CT, GH/CC) were found at locus GH/BsmFI. In both groups of Muscovies and in Mulards the dominant allele was GH(T). On the contrary in Pekin ducks AF51, the frequency of both alleles was found to be similar. The most frequent genotype in the examined ducks was GH/TT. In Pekin ducks AF51 three genotypes were observed, while in Mulard ducks and in male Muscovy ducks from a mother marked as CK, two genotypes (GH/TT and GH/CT) were identified. Muscovy duck females from a CK mother and all males and females of Muscovy duck from a CRAMMLCFF mother were monomorphic with only the GH/TTgenotype detected. The results showed that males of Pekin duck AF51 with the GH/TT genotype were characterized by higher (P < 0.01) BW value than those with the GH/CC and GH/CTgenotype. In females of Pekin ducks AF51, this same trend was observed; individuals with GH/TT genotype were superior (P < 0.05 and P < 0.01) to birds with two other detected genotypes in respect to BW, CG, LBB and LS. In the case of Mulards, ducks with the GH/TT genotype were distinguished by higher values of all evaluated traits compared to ducks with GH/CT and GH/CC genotypes, however most of the recorded differences were not significant. The only trait markedly impacted (P < 0.05) by the polymorphism of the GH gene intron 2 was the LS value in males.

The sheep growth hormone gene polymorphism and its effects on milk traits

Growth hormone (GH) is encoded by the GH gene, which may be single copy or duplicate in sheep. The two copies of the sheep GH gene (GH1/GH2-N and GH2-Z) were entirely sequenced in one 106 ewes of Sarda breed, in order to highlight sequence polymorphisms and investigate possible association between genetic variants and milk traits. Milk traits included milk yield, fat, protein, casein and lactose percentage. We evidenced 75 nucleotide changes. Transcription factor binding site prediction revealed two sequences potentially recognised by the pituitary-specific transcription factor POU1FI at the GH1/GH2-N gene, which were lost at the promoter of GH2-Z, which might explain the different tissues of expression of GH1/GH2-N (pituitary) and GH2-Z (placenta). Significant differences in milk traits were observed among genotypes at polymorphic loci only for the GH2-Z gene. Sheep with homozygote genotype ss748770547 CC had higher fat percentage (P < 0.01) than TT. SNP ss748770547 was part of a potential transcription factor binding site for C/EBP alpha (CCAAT/Enhancer Binding Protein), which is involved in the regulation of adipogenesis and adipoblast differentiation. SNP ss748770547, located in the GH2-Z gene 5' flanking region, may be a causal mutation affecting milk fat content. These findings might contribute to the knowledge of the sheep GH locus and might be useful in selection processes in sheep.

Growth hormone gene polymorphism and its association with body length and withers height in sirohi and barbari goats

An investigation was undertaken to study growth hormone (GH) gene polymorphism and its association with body length and withers height at different ages in kids of Sirohi and Barbari goats. Exon-2 and exon-3 regions of GH gene were amplified in 40 kids from the breeds. The RFLP pattern for HaeIII showed the presence of two genotypic variants (AB and BB) with their respective frequencies of 0.82 and 0.18 in Sirohi and 0.90 and 0.10 in Barbari goats. The frequencies of A and B alleles were respectively, 0.41 and 0.59 in Sirohi and 0.45 and 0.55 in Barbari breed. None of the two breeds was in Hardy-Weinberg equilibrium for these variants. The least squares analysis of variance revealed non-significant effect of GH genotype and breed x genotype interactions on body length and withers height from birth to 180 days.

Polymorphism of growth hormone (GH) Gene in Vembur and Kilakarsal Breeds of Sheep

Part of exon 2, intron 3 and part of exon 3 (422 bp), exon 4 (214 bp) and part of intron 4, exon 5 and part of 3'UTR (365 bp) of growth hormone (GH) gene in Vembur and Kilakarsal sheep were screened by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) to analyse the genetic variants. MspI-RFLP of GH gene (422, 214 and 365 bp fragments) revealed absence of polymorphism in all the samples of both the breeds studied indicating no variation exists in the studied regions of GH gene.

Growth hormone (GH), prolactin (PRL), and diacylglycerol acyltransferase (DGAT1) gene polymorphisms in Turkish native cattle breeds

Abstract: The aim of this study was to determine the genetic diversity of 4 native Turkish cattle breeds, based on the growth hormone (GH), prolactin (PRL), and diacylglycerol acyltransferase (DGAT1) genes. In order to study the polymorphisms in these genes, the polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method was performed. A 329-bp fragment and a 223-bp fragment of GH, a 156-bp fragment of PRL, and a 411-bp fragment of DGAT1, thus 4 loci of 3 genes, were amplified via PCR. These fragments were then restricted with the enzymes MspI, Alu1, RsaI, and CfrI, respectively. In this study, 2 types of alleles, ( + ) and ( - ) for the GH-MspI, L and V for the GH-Alu1, A and B for the PRL, and K and A for the DGAT1 loci, were observed. The results of the present study will contribute to the polymorphism data on the world's cattle breeds. Furthermore, the above-mentioned allele frequencies of Turkish native breeds are evaluated in relation to their genetic relatedness, and to infer their milk production properties on the basis of the available literature. In turn, these results can be utilized for future breeding programs of dairy cattle in Turkey.

Determination of the AluI polymorphism effect of bovine growth hormone gene on carcass traits in Zavot cattle with analysis of covariance

The aim of this study was to determine the genotypes of bovine growth hormone and to investigate their association with daily weight gain in the Zavot cattle breed. The bovine growth hormone (GH) genotypes of 45 male Zavot cattle were identified by PCR-RFLP. The effect of the bovine growth hormone gene on daily weight gain was estimated from a dataset obtained from cattle up to the age of 9 months. The mean differences of carcass weight or carcass yield among the genotype groups were assessed by analysis of covariance (ANCOVA). Two alleles for bovine growth hormone locus, L and V, were observed in Zavot cattle. The frequency of the L and V alleles was found to be 0.767 and 0.233, respectively. The genotype effect on live weight was not found to be significant (P > 0.05). The difference between the means of genotype groups was not found to be significant for carcass weight and carcass yield (P > 0.05). Further research on the association between bovine growth hormone gene polymorphism and yield traits in different cattle breeds should be conducted.

Association of the bovine growth hormone gene with Holstein cattle reproductive parameters

ABSTRACTObjective. To determine the association of intron 3 polymorphism of the bovine growth hormone (BGH) gene, with age at first service, first birth, first postpartum service and second birth in a population of Holstein cows in the state of Antioquia. Materials and methods. The study was conducted on 408 Holstein cows in 8 herds. Genotyping was performed using PCR-RFLP technique with DNA extracted from peripheral blood by the salting out technique. The Phenotypic information used was collected for 4 years, from a dairy production control program. To determine the association between characteristics and gene polymorphism, parametric statistical analyzes were performed as generalized linear models and linear regression analysis. Results. Allele frequency (positive) was 0.91 and was 0.09 for (negative) allele. Genotypic frequencies were 0.77, 0.2 and 0.03 for (positive/positive), (positive/negative) and (negative/negative) respectively. There were significant differences between the mean of age at first service, age at first birth, age at first postpartum service and age at second birth. For all of the (negative/negative) genotype characteristics there were greater ages for each event. Conclusions. These results suggest that intron-3 polymorphism of the BGH gene is associated with reproductive traits, facilitating the selection of individuals with favorable genotypes for use in breeding programs.

Genetic polymorphisms in growth hormone gene in Patanwadi, Marwari and Dumba breeds of sheep

Genetic polymorphisms in growth hormone gene in Patanwadi, Marwari and Dumba breeds of sheep

Bovine growth hormone gene polymorphism affects stress response in Japanese Black cattle.

We investigate the associations between growth hormone (GH) gene polymorphism and behavioral and physiological responses to stressors and learning ability in Japanese Black cattle. Flight distance test was conducted in the first experiment. Steers with haplotype C of GH gene polymorphism avoided human approaches at a significantly greater distance than ones without haplotype C (C: 1.9 ± 0.9, non-C: 1.0 ± 0.2 m, P < 0.05). An open-field test was conducted in the second experiment. Behavioral responses did not differ significantly between steers with and without haplotype C. Increases of heart rates to dropping of iron pipes was significantly higher in steers with haplotype C (C:161.7 ± 21.8, non-C:130.7 ± 31.3%, P < 0.05). Despite basal serum concentrations not being different between steers with and without haplotype C, serum cortisol in blood sampling immediately after severe confinement in a race tended to be higher in steers with haplotype C (P = 0.1). The maze test was conducted as the third experiment. There was no difference in performance in the maze test between steers with and without haplotype C. It is concluded that genetic polymorphism of GH may affect stress responses through GH concentration in steers.

Single nucleotide polymorphisms in growth hormone gene and their association with growth traits in Siniperca chuatsi (Basilewsky).

Growth hormone (GH) has been considered as a candidate gene for growth traits in fish. In this study, polymorphisms of the GH gene were evaluated for associations with growth traits in 282 Siniperca chuatsi individuals. Using directly sequencing, four single nucleotide polymorphisms (SNPs) were identified in GH gene, with two mutations in intron 4 (g.4940A>C, g.4948A>T), one mutation in exon 5 (g.5045T>C) and one in intron 5 (g.5234T>G). Notably, three of them were significantly associated with growth performance, particularly for g.4940A>C which was highly correlated with all the four growth traits. In conclusion, our results demonstrated that these SNPs in GH gene could influence growth performance of S.chuatsi and could be used for marker-assisted selection (MAS) in this species.

GHgene polymorphisms and expression associated with egg laying in muscovy ducks (Cairina moschata)

Accumulated evidence suggests that the growth hormone (GH) gene plays a physiological role in the control of reproductive function. Here, we examined the correlation between egg-laying traits and GH gene polymorphisms and expression patterns in the muscovy duck (Cairina moschata). PCR single-strand conformation polymorphism was used to identify polymorphisms in intron 3 of GH. One single nucleotide polymorphism (g.3270 A > G) was detected by sequencing, and the frequencies of the A and G alleles in the population were 0.65 and 0.35, respectively. A comparison test showed that the AA genotype group had more consecutive laying days and more eggs at 300 days than the GG genotype group (P < 0.05); however, there was no significant difference for the age at first laying (P > 0.05). Such a significant correlation between GH polymorphisms and egg-laying performance suggested that GH could be a candidate locus affecting the laying trait in muscovy duck. Furthermore, real-time fluorescent quantitative PCR demonstrated that GH is expressed in all selected tissues, but is highly expressed in the hypothalamic-pituitary-gonadal axis and heart. This unique expression pattern suggested that GH may exert its local physiological function through the autocrine or paracrine pathway during gonad development and growth in the muscovy duck. The data presented in this paper revealed GH polymorphisms and expression patterns in the muscovy duck and indicated a potential regulatory effect of GH on reproduction.

Association of growth hormone gene polymorphism with growth and fatness traits in Arian broilers

The aim of the study was to investigate the associations of HG gene polymorphism on chicken growth and body composition traits in Arian broilers. Blood samples randomly collected before slaughter from four different commercial Arian broiler lines (A, B, C and D; 400 samples) at 6 weeks of age. Genomic DNA was extracted and a fragment of 776 bp in size amplified using PCR-RFLP method. To determine the restriction site in intron 1 of GH gene of the animals, the MspI endonuclease and the resultant digested products were run on 2% agarose gel studied. Birds were slaughtered at 6 weeks of age and body composition was determined. The treatment of the fragment at GH loci with MspI restriction enzyme was revealed A1, A2 and A3 alleles. According to the results, recorded allele was: A1 allele (414, 217, 125 bp); A2 allele (125, 147, 237, 267 bp) and for allele A3 (237, 539 bp), respectively. The comparison of least square means of different genotypes indicated that polymorphism in the GH gene was significantly associated with body weight, tsmurd kci weight and percentage of drumstick weight at 6 weeks of age (P<0.05). According to the results, HG genes could be candidate gene that can affect some body composition traits in Arian broiler chickens.

Polymorphism of growth hormone gene and its association with seminal and sexual behavioral traits in crossbred cattle

The decline in the male reproductive ability in terms of sexual behavior and seminal traits might lead to nonavailability of required number of bulls in a progeny testing program. The present study was conducted in 493 crossbred cattle (Bos taurus × Bos indicus) bulls to study polymorphisms of growth hormone (GH) gene and its association with seminal and sexual behavioral characteristics. A 428-base pair fragment of GH gene spanning over the fourth exon, fourth intron, and fifth exon was amplified and digested with AluI restriction enzyme. Bulls were found to be polymorphic, with two variants, LL and LV, and higher genotypic frequency for LL being 0.88. Twelve nucleotide changes and a single nonsynonymous substitution of Leucine by Valine were observed from GH1 (L) to GH2 (V). Statistical analysis revealed that the genotype of the GH gene had a significant effect on libido score, reaction time, Flehmen response, requirement of mounting stimulus, sperm mass activity, number of semen doses per collection, individual fresh sperm motility, postthaw sperm motility, acrosome integrity, hypo-osmotic swelling test, live and dead count, total morphological abnormality, and head abnormality of sperm in crossbred bulls. Growth hormone gene might be considered a candidate gene for seminal and sexual behavioral traits in crossbred cattle.