a Molecular Genetics Laboratory, Project Directorate on Cattle, Indian Council of Agricultural Research, Meerut Cantt , Uttar Pradesh , India.
Anim Biotechnol. 2014 Apr 3;25(2):128-38. doi: 10.1080/10495398.2013.836106.
Calcium channel, voltage-dependent, alpha-2/delta subunit 1 (CACNA2D1) gene is considered to be an important noncytokine candidate gene influencing mastitis. Scanty of reports are available until today regarding the role play of CACNA2D1 gene on the susceptibility of bovine mastitis. We interrogated the CACNA2D1 G519663A [A>G] SNP by PCR-RFLP among two hundreds Frieswal (HF X Sahiwal) crossbred cattle of Indian origin. Genotypic frequency of AA (51.5, n=101) was comparatively higher than AG (35, n=70) and GG (14.5, n=29). Association of Somatic cell score (SCS) with genotypes revealed that, GG genotypes showing lesser count (less susceptible to mastitis) compare to AA and AG. Relative expression of CACNA2D1 transcript (in milk samples) was significantly higher among GG than AG and AA. Further we have also isolated blood sample from the all groups and PBMCs were cultured from each blood sample as per the standard protocol. They were treated with Calcium channel blocker and the expression level of the CACNA2D1 gene was evaluated by Real Time PCR. Results show that expression level decline in each genotypic group after treatment and expression level of GG are again significantly higher than AA and AG. Thus, it may be concluded that GG genotypic animals are favorable for selecting disease resistant breeds.
钙通道,电压依赖性,α-2/δ亚基 1(CACNA2D1)基因被认为是影响乳腺炎的重要非细胞因子候选基因。迄今为止,有关 CACNA2D1 基因对奶牛乳腺炎易感性的作用的报道很少。我们通过 PCR-RFLP 在 200 头印度起源的 Frieswal(HF X Sahiwal)杂交牛中检测了 CACNA2D1 G519663A[A>G]SNP。AA(51.5,n=101)基因型的频率明显高于 AG(35,n=70)和 GG(14.5,n=29)。体细胞评分(SCS)与基因型的关联表明,与 AA 和 AG 相比,GG 基因型的计数较少(对乳腺炎的易感性较低)。与 AG 和 AA 相比,GG 基因型的 CACNA2D1 转录本(在牛奶样本中)的相对表达显著更高。此外,我们还从所有组中分离了血液样本,并根据标准方案从每个血液样本中培养 PBMCs。用钙通道阻滞剂处理它们,并通过实时 PCR 评估 CACNA2D1 基因的表达水平。结果表明,在每种基因型组处理后表达水平下降,并且 GG 基因型的表达水平再次明显高于 AA 和 AG。因此,可以得出结论,GG 基因型的动物有利于选择抗疾病品种。