Determination and cellular localization of adenylyl cyclase isozymes expressed in embryonic chick heart.
Determination and cellular localization of adenylyl cyclase isozymes expressed in embryonic chick heart.
复制标题
鸡胚心脏中表达的腺苷酸环化酶同工酶的测定和细胞定位。
DOI：
10.1016/0014-5793(95)01084-r
复制
发表时间：
1995
期刊：
影响因子：
3.5
通讯作者：
Green,RD
中科院分区：
文献类型：
--
作者：
Yu,HJ;Unnerstall,JR;Green,RD
已有报道哺乳动物心脏表达AC同工酶(V和VI型)，并被~lt；μM[Ca~(2+)]抑制；禽类心脏表达腺酰环化酶活性，被~lt；μM[Ca~(2+)]抑制。我们使用逆转录聚合酶链式反应(RT-PCR)来确定在新鲜分离的心室肌细胞中存在V型和VI型AC mRNAs。随后的核糖核酸酶保护分析表明，V型信号是VI型同工酶的4-5倍。采用高比活性cRNA探针原位杂交和鸡抗肌球蛋白抗体免疫细胞化学相结合的方法，探讨V型和VI型AC信号的细胞来源。这些研究表明，心肌细胞同时含有V型和VI型同工酶的信息，但AC V是主要的同工酶。有趣的是，虽然V型ACmRNA似乎主要定位于心肌细胞，但在非心肌细胞中ACVI型mRNA的信号强于心肌细胞。
Mammalian heart has been reported to express AC isozymes (types V and VI) that are inhibited by < μM [Ca2+]; avian heart has been reported to express adenylyl cyclase activity that is inhibited by < μM [Ca2+]. We have used reverse transcription polymerase chain reaction (RT-PCR) to determine that type V and VI AC mRNAs are present in freshly isolated ventricular myocytes. Subsequent RNase protection assays revealed that that the type V signal is 4–5 times that for the type VI isozyme. In situ hybridization with high specific activity cRNA probes combined with immunocytochemistry with a chick anti-myosin antibody was used to probe the cellular origins of type V and type VI AC signals. These studies show that myocytes contain messages for both the type V and VI isozymes but that AC V is the major isoform. Interestingly, while the type V AC mRNA appears to be localized primarily, if not exclusively, in myocytes, the signal for type AC VI mRNA in non-myocytes is stronger than in myocytes.