期刊
JOURNAL OF MOLECULAR AND CELLULAR CARDIOLOGY
卷 119, 期 -, 页码 64-74出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.yjmcc.2018.04.010
关键词
-
资金
- AHA [13PRE16500029, 15PRE25860042]
- [NHLBIR01HL124091]
- [NIMHR01MH065531]
- [NHLBI5R37HL076795]
Dysregulation of L-type Ca2+ channels (LTCCs) underlies numerous cardiac pathologies. Understanding their modulation with high fidelity relies on investigating LTCCs in their native environment with intact interacting proteins. Such studies benefit from genetic manipulation of endogenous channels in cardiomyocytes, which often proves cumbersome in mammalian models. Drosophila melanogaster, however, offers a potentially efficient alternative as it possesses a relatively simple heart, is genetically pliable, and expresses well-conserved genes. Fluorescence m situ hybridization confirmed an abundance of Ca-alD and Ca-alT mRNA in fly myocardium, which encode subunits that specify hetero-ohgomenc channels homologous to mammalian LTCCs and T-type Ca2+ channels, respectively. Cardiac-specific knockdown of Ca-alD via interfering RNA abolished cardiac contraction, suggesting Ca-alD (i.e. AID) represents the primary functioning Ca2+ channel in Drosophila hearts. Moreover, we successfully isolated viable single cardiomyocytes and recorded Ca2+ currents via patch clamping, a feat never before accomplished with the fly model. The profile of Ca2+ currents recorded in individual cells when Ca2+ channels were hypomorphic, absent, or under selective LTCC blockage by nifedipine, additionally confirmed the predominance of AID current across all activation voltages. T-type current, activated at more negative voltages, was also detected. Lastly, AID channels displayed Ca2 +-dependent inactivation, a critical negative feedback mechanism of LTCCs, and the current through them was augmented by forskolin, an activator of the protein kinase A pathway. In sum, the Drosophila heart possesses a conserved compendium of Ca2+ channels, suggesting that the fly may serve as a robust and effective platform for studying cardiac channelopathies.
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