College of Animal Science, Henan Agricultural University, Hengzhou 450002, China.
College of Animal Science, Henan Agricultural University, Hengzhou 450002, China.
Poult Sci. 2022 Jun;101(6):101893. doi: 10.1016/j.psj.2022.101893. Epub 2022 Apr 6.
The cytidine diphosphate diacylglycerol synthases (CDSs) gene encodes the cytidine diphosphate-diacylglycerol (CDP-DAG) synthase enzyme that catalyzes the formation of CDP-diacylglycerol from phosphatidic acid. At present, there are no reports of CDS2 in birds. Here, we identified chicken CDS2 transcripts by combining conventional RT-PCR amplification, 5' rapid amplification of cDNA ends (RACE), and 3' RACE, explored the spatio-temporal expression profiles of total CDS2 and the longest transcript variant CDS2-4, and investigated the effect of exogenous insulin on the mRNA level of total CDS2 via quantitative RT-PCR. Four transcripts of chicken CDS2 (CDS2-1, -2, -3, and -4) were identified, which were alternatively spliced at the 3'-untranslated region (UTR). Both total CDS2 and CDS2-4 were prominently expressed in adipose tissue, and exhibited low expression in liver and pectoralis of 49-day-old chickens. Regarding the spatio-temporal expression patterns of CDS2 in chicken, total CDS2 exhibited a similar temporal expression tendency with a high level in the later period of incubation (embryonic day 19 [E19] or 1-day-old) in the brain, liver, and pectoralis. While CDS2-4 presented a distinct temporal expression pattern in these tissues, CDS2-4 levels peaked at 21 d in the brain and pectoralis, while liver CDS2-4 mRNA levels were highest at the early stage of hatching (E10). Total CDS2 (P < 0.001) and CDS2-4 (P = 0.0090) mRNA levels in the liver were differentially regulated throughout the development of the chicken. Total CDS2 levels in the liver of Silky chickens were higher than that of the broiler in the basal state and after insulin stimulation. Exogenous insulin significantly down-regulated the level of total CDS2 at 240 min in the pectoralis of Silky chickens (P < 0.01). In conclusion, chicken CDS2 isoforms with variation at the 3'-UTR were identified, which was prominently expressed in adipose tissue. Total CDS2 and CDS2-4 presented distinct spatio-temporal expression patterns, that is they were differentially regulated with age in brain, liver, and pectoralis. Insulin could regulate chicken CDS2 levels in a breed- and tissue-specific manner.
磷酸胞苷二酰甘油合成酶(CDSs)基因编码磷酸胞苷二酰甘油(CDP-DAG)合酶,该酶催化从磷酸脂酸形成 CDP-二酰基甘油。目前,在鸟类中尚无 CDS2 的报道。在这里,我们通过结合常规 RT-PCR 扩增、5'快速扩增 cDNA 末端(RACE)和 3'RACE,鉴定了鸡 CDS2 转录本,探讨了总 CDS2 和最长转录变体 CDS2-4 的时空表达谱,并通过定量 RT-PCR 研究了外源性胰岛素对总 CDS2mRNA 水平的影响。鉴定了鸡 CDS2 的 4 个转录本(CDS2-1、-2、-3 和-4),它们在 3'非翻译区(UTR)处发生可变剪接。总 CDS2 和 CDS2-4 在脂肪组织中表达明显,而在 49 日龄鸡的肝脏和胸肌中表达水平较低。关于鸡 CDS2 的时空表达模式,总 CDS2 在脑、肝和胸肌中的表达在后期孵化(胚胎第 19 天[E19]或 1 天龄)具有相似的时间表达趋势,具有高水平。而 CDS2-4 在这些组织中呈现出明显不同的时间表达模式,CDS2-4 水平在脑和胸肌中在 21 天达到峰值,而肝脏 CDS2-4mRNA 水平在孵化早期(E10)最高。在鸡的发育过程中,总 CDS2(P<0.001)和 CDS2-4(P=0.0090)mRNA 水平在肝脏中均受到差异调控。基础状态和胰岛素刺激后,丝光鸡肝脏中的总 CDS2 水平均高于肉鸡。外源性胰岛素显著下调丝光鸡胸肌中总 CDS2 的水平(P<0.01)在 240 分钟时。总之,鉴定了在 3'UTR 处具有变异的鸡 CDS2 同工型,其在脂肪组织中表达明显。总 CDS2 和 CDS2-4 呈现出明显不同的时空表达模式,即它们在脑、肝和胸肌中随年龄而差异调控。胰岛素可以以品种和组织特异性的方式调节鸡 CDS2 水平。