Hua Zeng

The genetic effects of the dopamine D1 receptor gene on chicken egg production and broodiness traits.

BackgroundThe elevation of egg production and the inhibition of incubation behavior are the aims of modern poultry production. Prolactin (PRL) gene is confirmed to be critical for the onset and maintenance of these reproductive behaviors in birds. Through PRL, dopamine D1 receptor (DRD1) was also involved in the regulation of chicken reproductive behavior. However, the genetic effects of this gene on chicken egg production and broodiness have not been studied extensively. The objective of this research was to evaluate the genetic effects of the DRD1 gene on chicken egg production and broodiness traits.ResultsIn this study, the chicken DRD1 gene was screened for the polymorphisms by cloning and sequencing and 29 variations were identified in 3,342 bp length of this gene. Seven single nucleotide polymorphism (SNPs) among these variations, including a non-synonymous mutation (A+505G, Ser169Gly), were located in the coding region and were chosen to analyze their association with chicken egg production and broodiness traits in 644 Ningdu Sanhuang individuals. Two SNPs, G+123A and C+1107T, were significantly associated with chicken broody frequency (P < 0.05). Significant association was also found between the G+1065A - C+1107T haplotypes and chicken broody frequency (P < 0.05). In addition, the haplotypes of G+123A and T+198C were significantly associated with weight of first egg (EW) (P = 0.03). On the other hand, the distribution of the DRD1 mRNA was observed and the expression difference was compared between broodiness and non-broodiness chickens. The DRD1 mRNA was predominantly expressed in subcutaneous fat and abdominal fat of non-broodiness chicken, and then in heart, kidney, oviduct, glandular stomach, hypothalamus, and pituitary. In subcutaneous fat and abdominal fat, the level of non-broodiness was 26 to 28 times higher than that of broodiness. In pituitary, it was 5-fold higher. In heart, oviduct, and kidney, a 2-3 times decrease from non-broodiness to broodiness was displayed. In glandular stomach and hypothalamus, the level seen in non-broodiness and broodiness was almost the same.ConclusionThe polymorphisms of the DRD1 gene and their haplotypes were associated with chicken broody frequency and some egg production traits. The mRNA distribution was significant different between broodiness and non-broodiness chickens.

Polymorphisms of Vasoactive Intestinal Peptide Receptor-1 Gene and Their Genetic Effects on Broodiness in Chickens

Broodiness is a polygenic trait controlled by a small number of autosomal genes. Vasoactive intestinal peptide receptor-1 (VIPR-1) gene could be a candidate of chicken broodiness, and its genomic variations and genetic effects on chicken broodiness traits were analyzed in this study. The partial cloning and sequencing of the VIPR-1 gene showed that the average nucleotide diversity was 0.00669 +/- 0.00093 in Red Jungle Fowls (RJF), and 0.00582 +/- 0.00026 in domestic chickens. One hundred twenty-eight variation sites were identified in the 11,136-bp region of the chicken VIPR-1 gene. Twenty variation sites were genotyped using PCR-RFLP or PCR method to analyze average diversity, linkage-disequilibrium pattern, and haplotype structure in RJF, Xinghua chickens, Ningdu Sanhuang chickens, Baier Huang chickens, and Leghorn Layers. The RJF, Xinghua, Ningdu Sanhuang, and Baier Huang exhibited distinct characteristic of decreasing r(2) value over physical distance. Haplotype analyses showed that some variation sites of the 27-kb region from exon 6 to exon 11 could be associated with broodiness. The distribution of genotypic and allelic frequencies, and heterozygosities in the above 5 populations showed that A-284G, A+457G, C+598T, D+19820I, C+37454T, C+42913T, and C+53327T might be associated with broodiness. The 7 sites and the other 4 sites were genotyped in 644 NDH individuals under cage condition and were used for association analyses between each site and chicken broodiness traits. A significant association (P < 0.05) was found between C+598T in intron 2 and broody frequency (%). Another significant association (P < 0.05) was found between C+53327T and duration of broodiness, in which allele C was positive for DB.

Genetic effects of polymorphisms in candidate genes and the QTL region on chicken age at first egg

BackgroundThe age at first egg (AFE), an important indicator for sexual maturation in female chickens, is controlled by polygenes. Based on our knowledge of reproductive physiology, 6 genes including gonadotrophin releasing hormone-I (GnRH-I), neuropeptide Y (NPY), dopamine D2 receptor (DRD2), vasoactive intestinal polypeptide (VIP), VIP receptor-1 (VIPR-1), and prolactin (PRL), were selected as candidates for influencing AFE. Additionally, the region between ADL0201 and MCW0241 of chromosome Z was chosen as the candidate QTL region according to some QTL databases. The objective of the present study was to investigate the effects of mutations in candidate genes and the QTL region on chicken AFE.ResultsMarker-trait association analysis of 8 mutations in those 6 genes in a Chinese native population found a highly significant association (P < 0.01) between G840327C of the GnRH-I gene with AFE, and it remained significant even with Bonferroni correction. Based on the results of the 2-tailed χ2 test, mutations T32742394C, T32742468C, G32742603A, and C33379782T in the candidate QTL region of chromosome Z were selected for marker-trait association analysis. The haplotypes of T32742394C and T32742468C were significantly associated (P < 0.05) with AFE. Bioinformatics analysis indicated that T32742394C and T32742468C were located in the intron region of the SH3-domain GRB2-like 2 (SH3GL2) gene, which appeared to be associated in the endocytosis and development of the oocyte.ConclusionThis study found that G840327C of the GnRH-I gene and the haplotypes of T32742394C-T32742468C of the SH3GL2 gene were associated with the chicken AFE.

Genomic organisation of the chicken ghrelin gene and its single nucleotide polymorphisms detected by denaturing high-performance liquid chromatography.

1. Ghrelin is a novel endogenous ligand for the growth hormone secretagogue receptor (GHS-R) and is expressed primarily in the stomach and hypothalamus with the probable function of stimulating GH secretion and food intake both in mammals and poultry. The complete sequences of ghrelin gene have been reported in humans and mice; however, that of chickens remains unclear. 2. Here, we report the complete sequence of chicken ghrelin gene (submitted to Genbank; accession number AY303688), which consists of 5 exons and 4 introns. As in mice, the first exon of chicken ghrelin gene does not encode any amino acid. 3. Scanning point mutations with denaturing high-performance liquid chromatography (DHPLC) using WAVE® DNA Fragment Analysis Systems and confirmed with direct sequencing for polymerase chain reaction (PCR) products, we analysed the single nucleotide polymorphisms (SNPs) in the entire gene of chicken ghrelin. 4. Results showed that there were 19 SNPs in chicken ghrelin gene, and most of these SNPs were scattered in the 4 introns. In these SNPs, one mutation in exon 5 (A2355G) led to the change of amino acid from glutamine to arginine (Gln 113 Arg): as a result a different ghrelin precursor instead of a mature peptide was produced. In addition, one SNP in 5′UTR (C223G) determined the presence or absence of a potential binding site of transcription factor serum response factor (SRF), which might affect the expression of chicken ghrelin gene. Some of the SNPs detected in the present study could be used in quantitative trait loci (QTL) mapping for growth characters in chickens. 5. Because one SNP is located in a polymorphic site of restriction enzyme PagI of intron 4, it was possible to design a PCR-RFLP procedure and analyse the diversity of 10 chicken populations. Results showed the allelic frequencies of C2100T differ among these breeds, however, no significant difference was observed between imported breeds and Chinese native ones, nor between egg layers and meat type chickens. A chi-square test showed that 4 chicken populations of Beijing Fat, Xinghua, Recessive White and Silky followed the Hardy–Weinberg equilibrium.

The dopamine D2 receptor gene polymorphisms associated with chicken broodiness

Chicken broodiness is a polygenic trait controlled by autosomal genes. Prolactin gene is a candidate of great interest in molecular studies of broodiness. However, another candidate dopamine D2 receptor (DRD2) gene has not been studied extensively. The objective of this study was to analyze the genetic effects of the DRD2 gene on chicken broodiness through linkage disequilibrium analyses, tag SNP selection, genetic diversity observation, 2-tailed test, and association analyses. In this study, we assayed 27 variations of this gene in 456 individuals from 6 chicken populations to observe linkage disequilibrium pattern, the tag SNP, and genetic diversity. Among the 6 populations, Taihe Silkies exhibited no characteristic between the square of the correlation coefficient of gene frequencies (r(2)) and physical distance. The other populations including Red Jungle Fowls, Xinghua chickens, Ningdu Sanhuang chickens (NDH), Baier Huang chickens, and Leghorn layers exhibited conspicuous characteristic of decreasing r(2) value over physical distance. Linkage disequilibrium decayed more rapidly in Red Jungle Fowls, Xinghua, and NDH than in Baier Huang and Leghorn layers. Allelic frequencies and genotype distributions in the 5 populations showed that A-38600G, I-38463D, T-32751C, A-16105G, A-6543G, C-6539T, and A+2794G were possibly associated with broodiness. Besides the above 7 sites, another 2 sites that might be associated with broodiness were screened by 2-tailed test. All 9 sites were used for association analyses with broodiness in 644 NDH chickens. A significant association (P < 0.05) was found between A-16105G and broody frequency (%), and the T+619C in intron 1 was significantly associated with duration of broodiness (P < 0.05). These findings suggested that the DRD2 gene should be included in future genetic studies of chicken broodiness and 2 SNP of A-16105G and T+619C might be markers for breeding against broodiness.

Associations between polymorphisms in the chicken VIP gene, egg production and broody traits

1. In this study, the vasoactive intestinal peptide (VIP) gene was screened for variations by cloning and sequencing to analyse the association with chicken egg production and broodiness traits. 2. Sequencing revealed that 69 polymorphisms were found in a 9305-bp length of the chicken VIP gene, of which 39 were located in introns, 28 in 5′ regulatory region, 1 in exon 6 (synonymous) and 1 in 3′-UTR. The nucleotide diversity corrected for sample size was 1·96 × 10−3. 3. Five polymorphisms, C-3134T, “AGG” indel at −2648 to −2650, C + 338T, G + 780T and A + 4691G, were genotyped in 644 individuals of Ningdu Sanhuang chickens to evaluate their effects on egg production and broodiness traits. Marker-trait association analyses showed that the “AGG” indel was associated with total number of eggs from 90 to 300 d of age and total number of qualified eggs from 90 to 300 d of age. In contrast, C + 338T was associated with duration of broodiness.

Single nucleotide polymorphism (SNP) at the GHR gene and its associations with chicken growth and fat deposition traits.

1. The growth hormone receptor (GHR) plays crucial roles on chicken growth and metabolism. 2. The full cDNA of the chicken GHR gene was scanned for single nucleotide polymorphisms (SNP) by means of denaturing high-performance liquid chromatography (DHPLC). Three SNP, C6540334T, C6542011T and G6631778A, were genotyped in a F2 designed full-sib resource population to analyse their associations with chicken growth and fat deposition traits. 3. Fifty-five SNP and two other variations were identified in the 8908 bp region of the GHR gene. Among the 55 SNP, 10 were located in coding exons (6 resulted in changes of amino acids) and 45 were in non-coding regions (introns, 5′UTR and 3′UTR). The nucleotide diversity (θ), corrected for sample size of chicken GHR gene, is 1·45 × 10−3. Fourteen PCR-RFLP markers were developed in the chicken GHR gene. 4. The G6631778A was associated with body weight at 63 d (BW63), dressed weight (DW) and subcutaneous fat thickness (SFT), BW35 and BW49 (P < 0·01) as well as hatch weight (HW) and BW42 in the male population. However, G6631778A was only associated with BW28 in the female population. G rather than A was dominant for chicken growth and fat deposition. Haplotypes based on the three SNP were associated with BW21, BW70, BW77 and SFT, BW7, BW35, BW42, BW49 and BW56 in males, and associated with BW7 and BW14 in females. For growth in males, the H2 and H6 haplotypes had positive and negative effects, respectively; meanwhile H6 was predominant for fat deposition.

The Effects of Different Sex-Linked Dwarf Variations on Chinese Native Chickens

Abstract Variants in chicken growth hormone receptor (GHR) gene lead to sex-linked dwarf (SLD) chickens, but effects of different variants are distinct. In this study, 11 SLD chicken breeds or strains including 3 Chinese native breeds and 8 breeding strains were studied in order to investigate the effects of different sex-linked dwarf variations on growth performance. The results showed that there were three reasons which could lead to dwarfism in the 11 breeds or strains. Firstly, an about 1.7 kb deletion of growth hormone receptor (GHR) gene leads to dwarfism in Jiangxi dwarf chicken, strains GF24, GF26, N308, N309, and N310. Secondly, a T354C mutation in exon 5 of the GHR gene leads to dwarfism in strains N301 and N305. Thirdly, an unknown variant leads to dwarfism in Guizhou Yellow Dwarf chicken and Yixing Bantam chicken. In addition, all individuals of N303 had the 1.7 kb deletion of the GHR gene, and additionally, some of them also carried the T354C mutation. As far as the performance of individuals were compared among T354C homozygote, deletion homozygote, and heterozygote carrying both T354C and deletion, it was found that the T354Cs impacts on body weight of Chinese chickens were maximum, the body weight of chickens with homozygote T354C was 92.12% of those with heterozygote, and the difference of the body weight between deletion homozygote and heterozygote was not significant. There was no significant difference of shank length among three genotypes.