Scientia Agricultura Sinica ›› 2022, Vol. 55 ›› Issue (22): 4539-4549.doi: 10.3864/j.issn.0578-1752.2022.22.017

• ANIMAL SCIENCE·VETERINARY SCIENCE • Previous Articles     Next Articles

Association of Levels of Cholecystokinin A Receptor Expression and Sequence Variants with Feed Conversion Efficiency of Lueyang Black-Boned Chicken

WANG ZhePeng(),ZHOU WenXin,HE JunXi,HU QiaoYan,ZHAO JiaYue   

  1. College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi
  • Received:2021-09-03 Accepted:2021-11-24 Online:2022-11-16 Published:2022-12-14

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Abstract:

【Background】 Increased appetite and feed conversion efficiency forms a physiological basis underlying high growth of modern commercial broiler chickens. Cholecystokinin A receptor (CCKAR) is a key receptor for mediating CCK signals and involving in regulation of appetite and digestion. In view of the important role of CCK signals in the control of energy balance, the levels of CCKAR expression are closely related to the growth of chickens. However, the mechanism by which the abundance of CCKAR affects the growth of chickens remains unclear. 【Objective】 The aim of this study was to elucidate the relationship of levels of CCKAR expression and sequence variants with feed conversion efficiency, which would promote the understanding of genetic basis of the growth traits and breeding for improvement of feed efficiency in the chicken. 【Method】The feed to meat conversion ratio of Lueyang black-boned chickens (rooster n=62, pullet n=67) from 12 to 16 weeks of age and the feed to egg conversion ratio from 26 to 30 weeks of age (n=150) were individually measured. Two synonymous mutations (g.73206714T>C and g.73209189C>T) in the CCKAR gene were genotyped by using PCR-RFLP. Association of two SNP with the feed to meat conversion ratio was tested by using two-way analysis of variance with sex treated as a fixed effect. Association of two SNP with the feed to egg conversion ratio was tested by using one-way analysis of variance. Disrupting effect of g.73206714T>C on transcription factor binding sites was predicted in silico by using JASPAR database. Ten birds that were heterozygotes (CT) at the g.73206714T>C locus were selected to analyze imbalance of CCKAR expression between T and C alleles. The difference of allele-specific expression was detected by using PCR-RFLP. The Levels of CCKAR expression were detected by qPCR in the hypothalamus, pituitary, pancreas, gallbladder and duodenum of Lueyang black-boned chickens (n=30). The correlation of levels of CCKAR expression with food intake and feed to meat conversion ratio was analyzed. 【Result】 Sex had a significant (F=29.44，P<0.0001) effect on the feed to meat conversion ratio, and the feed efficiency (5.41±0.76) of roosters was significantly superior to pullets (6.62± 1.00). Of two SNP, only g.73206714T>C was significantly associated with the feed to meat conversion ratio. The feed to meat conversion ratio (5.18±0.63) of the TT genotype was significantly lower than ones of CC (6.37±1.1) and CT (6.17±0.95) genotypes. The feed to meat conversion ratio had no significant difference between CC and CT. There was no significant interaction effect between genotypes of g.73206714T>C and sex. G.73206714T>C potentially disrupted binding sites of 28 transcription factors. Allele-specific expression assay showed that the abundance of transcripts with the T allele was higher than one of transcripts with the C allele, supporting that differential expression of CCKAR gene between alleles might likely contribute to genetic association of g.73206714 with the feed to meat conversion ratio. Average feed to egg conversion ratio of Lueyang black-boned chickens was 2.85±0.41 at the 26-30 weeks of age. Both g.73206714T>C and g.73209189C>T had no significant effect on the feed to egg conversion ratio. The levels of CCKAR expression in the pancreas were the highest among five tissues, which showed a negative correlation (r=-0.45, P=0.017) with feed intake. But there was no significant correlation between levels of CCKAR expression and feed intake in other tissues. In the pancreas (r=-0.41, P=0.03) and pituitary (r=-0.57, P=0.0018), the higher levels of CCKAR expression was, the lower the feed to meat conversion ratio was. But in other tissues, there was no the negative correlation.【Conclusion】 This study found that increased levels of CCKAR was associated with high feed efficiency. The T allele of g.73296714T>C was associated with high expression activity of CCKAR gene, and with high feed efficiency.

Key words: chicken, growth, CCKAR, feed conversion efficiency, appetite

Fig. 1

Distribution of feed to meat conversion ratio of Lueyang black-bone chicken from 12 to 16 weeks of age"

Fig. 2

Association of CCKAR sequence variants with feed conversion ratio of Lueyang black-boned chicken Low cases on the boxplots indicate the results of multiple comparison. The same letters or the boxplots without letters represent there is no significant difference among genotypes. Different letters represent significant (P<0.05) difference between two genotypes"

Fig. 3

Allele-specific expression of CCKAR gene detected by PCR-RFLP G.73206714T>C was used as a marker to analyze allele-specific expression difference of CCKAR gene. RT-PCR products from hypothalamus, pituitary, pancreas, gallbladder and duodenum of 10 heterozygous (CT) birds were digested using MnlI restriction enzyme. Digested products were separated using 3% agarose gel electrophoresis. The T allele will produce a 138 bp band, and the C allele will produce a 109 bp and a 29 bp bands. The 29 bp band is invisible on the agarose gel. PCR products that were amplified using genomic DNA were used as the control to validate results of allele-specific expression detected by PCR-RFLP"

Fig. 4

Comparison of levels of CCKAR expression in five tissues Low cases on the boxplot indicate results of multiple comparison of CCKAR expression levels in five tissues. The same letters represent that levels of CCKAR expression have no significant difference between two tissues. Difference letters mean that there is a significant difference between two tissues"

Fig. 5

Correlation of levels of CCKAR expression with food intake and feed to meat conversion ratio of Lueyang black-boned chicken ADFI_16wk represents average daily food intake at the 16 weeks of age. FCR_16wk is the feed to meat conversion ratio at the 16 weeks of age. Numbers on the each picture are the coefficients of correlation and P values of significant test"

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