屡配不孕母牛FOXP3、FSHR、FMR1基因多态性与生殖激素相关性分析

doi:10.11843/j.issn.0366-6964.2024.06.043

Abstract

Abstract:

The polymorphisms of forkhead box protein P3 (FOXP3), follicle stimulating hormone receptor (FSHR) and fragile X mental retardation 1 (FMR1) were studied, and the effects of gene polymorphisms on infertility and hormone levels in cows were discussed. The sequence of the gene mutation site was amplified by PCR, and the PCR results were identified by enzyme digestion and sequencing peak analysis, and the genotypes of the three genes were counted. The intergroup differences in the mutations of the three genes were analyzed. The concentrations of estrogen (E), follicle-stimulating hormone (FSH), prolactin (PRL), testosterone (T) and luteinizing hormone (LH) in the two groups were detected by ELISA. The correlation between gene polymorphism and hormone level was analyzed. Three genotypes of FOXP3 gene were detected: AA, AG and GG; three genotypes of FSHR gene were detected: AA, AC and CC; T base insertion was detected at 302 bp of 5'UTR of FMR1 gene. The results of chi-square adaptability test showed that the 92 377 635 A>G loci of FOXP3 gene and the 31 450 279 A>C loci of FSHR gene were lower than Hardy-Weinberg equilibrium (P < 0.05). Correlation analysis showed that FOXP3 gene G allele and FSHR gene C allele were significantly positively correlated with repeated infertility in cows. There were significant differences in E, FSH, PRL and T hormones between the infertile group and the healthy mother group (P < 0.05). The E hormone concentration of FOXP3 AA and GG genotypes was significantly different from that of AG genotype (P < 0.05). FSHR gene AA, AC genotype E hormone concentration was significantly different from CC genotype (P < 0.05). (concentration) was significantly different from AG genotype (P < 0.05); FSHR gene 31 450 279 A>C locus AA, AC genotype E hormone content (concentration) FOXP3 gene and FSHR gene are polymorphic in Simmental, Brown and Angus populations, which indicates that the gene may be a potential genetic factor affecting the infertility of cows. The G allele of FOXP3 gene was significantly different between the healthy group and the multiple infertility group (P < 0.05), and the C allele of FSHR gene was significantly different between the healthy group and the multiple infertility group (P < 0.05). The G allele of g.92 377 635 A>G locus of FOXP3 gene was significantly positively correlated with multiple infertility in cows (the C allele of PC locus was significantly positively correlated with multiple infertility in cows (P < 0.05, r=0.14).The polymorphism sites of FOXP3 gene and FSHR gene may be used as the marker identification sites of repeated mating infertility in cows, which can be used to screen the potential candidate genetic markers of repeated mating infertility cows, which needs further proof. The G allele of FOXP3 gene was weakly positively correlated with E concentration (P < 0.05, r=0.16), and the T base insertion of FMR1 gene was weakly positively correlated with E concentration (P < 0.05, r=0.187). However, how the variation of FOXP3 gene and FSHR gene regulates the reproductive ability of cows needs further study.

Key words: cattle, infertility cows, gene polymorphism, hormone, relevance analysis

CLC Number:

S857.22

Xuanyi WANG, Yawei SUN, Yuwei LONG, Liying WANG, Yuxin ZHOU, Na LI, Xuelian MA, Hongqiong ZHAO, Gang YAO. Correlation Analysis of FOXP3, FSHR, FMR1 Gene Polymorphisms and Reproductive Hormones in Infertile Cows[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(6): 2727-2740.

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References 41

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基因 Gene	登录号 NCBI ID	引物序列(5′→3′) Primers sequences	退火温度/℃ Annealing temperature	产物长度/bp Product length
FMR1	507482	F: 5′-ACGCGGTTTCACTGTTTACAC-3′	49.8	310
		R: 5′-ACCTTGTAGAAAGCGCCATTG-3′
FOXP3	506053	F: 5′-ATTCTAAGGTTCTGGCATCGT-3′	60.3	618
		R: 5′-GCCAGAGCATATAAGAAACCACT-3′
FSHR	281172	F: 5′-CTGCCTCCCTCAAGGTGCCCCTC-3′	58.0	306
		R: 5′-AGTTCTTGGCTAAATGTCTTAGGGGG

品种 Variety	基因型频率 Gene frequency			基因频率 Genotype frequency		χ²	P值 P value
品种 Variety	AA	AG	GG	A	G	χ²	P值 P value
西门塔尔牛Simmental heifer	0.70	0.27	0.03	0.84	0.16	0.826	0.363
褐牛Brown cattle	0.62	0.30	0.08	0.77	0.23	0.241	0.623
安格斯牛Aberdeen angus	0.55	0.36	0.09	0.73	0.27	5.046	0.025^*

基因Gene	遗传纯和度He	杂合度Ho	有效等位基因数Ne	多态信息含量PIC
FOXP3	0.624	0.376	1.60	0.33

品种 Variety	基因型频率 Gene frequency			基因频率 Genotype frequency		χ²	P值 P value
品种 Variety	AA	AC	CC	A	C	χ²	P值 P value
西门塔尔牛Simmental heifer	0.52	0.35	0.13	0.70	0.30	2.267	0.132
褐牛Brown cattle	0.66	0.30	0.04	0.81	0.19	0.584	0.445
安格斯牛Aberdeen angus	0.75	0.21	0.04	0.86	0.14	7.198	0.007^**

基因Gene	遗传纯和度He	杂合度Ho	有效等位基因数Ne	多态信息含量PIC
FSHR	0.621	0.379	1.61	0.35

Correlation Analysis of FOXP3, FSHR, FMR1 Gene Polymorphisms and Reproductive Hormones in Infertile Cows

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激素 Hormone	FOXP3基因型FOXP3 genotype
激素 Hormone	AA(n=126)	AG(n=61)	GG(n=14)
E	112.84±1.42^a	113.05±1.72^b	107.9±4.05^a
FSH	35.30±0.48	35.46±0.64	31.81±0.82
PRL	235.54±3.24	235.90±4.82	223.26±8.70
T	33.92±0.47	34.83±0.49	32.72±0.95
LH	3371.76±50.46	3451.92±74.14	3190.61±104.33

激素 Hormone	FSHR基因型FSHR genotype
激素 Hormone	AA(n=144)	AC(n=70)	CC(n=18)
E	114.01±1.26^a	112.07±2.04^a	99.65±3.92^b
FSH	35.56±0.47	34.41±0.69	34.51±0.83
PRL	240.01±3.12	228.05±3.87	215.41±18.59
T	34.66±0.37	33.02±0.76	35.04±0.81
LH	3389.64±46.27	3364.35±83.89	3474.08±167.33

激素 Hormone	FMR1基因型FMR1 genotype
激素 Hormone	T碱基插入(n=43)	正常(n=139)
E	114.00±2.71	111.58±1.13
FSH	35.03±0.90	35.22±0.52
PRL	221.89±7.42^a	240.59±3.63^b
T	36.53±1.12	36.00±0.41
LH	3 545.14±147.01	3 438.41.35±58.61

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