In vivo light-induced and basal phospholipase C activity in Drosophila photoreceptors measured with genetically targeted phosphatidylinositol 4,5-bisphosphate-sensitive ion channels (Kir2.1).

The Journal of Biological Chemistry
R C HardiePadinjat Raghu

Abstract

The phosphatidylinositol 4,5-bisphosphate (PIP(2))-sensitive inward rectifier channel Kir2.1 was expressed in Drosophila photoreceptors and used to monitor in vivo PIP(2) levels. Since the wild-type (WT) Kir2.1 channel appeared to be saturated by the prevailing PIP(2) concentration, we made a single amino acid substitution (R228Q), which reduced the effective affinity for PIP(2) and yielded channels generating currents proportional to the PIP(2) levels relevant for phototransduction. To isolate Kir2.1 currents, recordings were made from mutants lacking both classes of light-sensitive transient receptor potential channels (TRP and TRPL). Light resulted in the effective depletion of PIP(2) by phospholipase C (PLC) in approximately three or four microvilli per absorbed photon at rates exceeding approximately 150% of total microvillar phosphoinositides per second. PIP(2) was resynthesized with a half-time of approximately 50 s. When PIP(2) resynthesis was prevented by depriving the cell of ATP, the Kir current spontaneously decayed at maximal rates representing a loss of approximately 40% loss of total PIP(2) per minute. This loss was attributed primarily to basal PLC activity, because it was greatly decreased in norpA mutants lack...Continue Reading

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Citations

Feb 3, 2011·Pflügers Archiv : European journal of physiology·Roger C Hardie
Apr 16, 2008·The Journal of General Physiology·Stephen J Tucker, Thomas Baukrowitz
Oct 16, 2012·Science·Roger C Hardie, Kristian Franze
Feb 5, 2013·Journal of Cell Science·Sukanya SenguptaRoger C Hardie
Sep 23, 2006·The Journal of Physiology·Roger C Hardie
Apr 11, 2012·Biochimica Et Biophysica Acta·Padinjat RaghuNaresh Babu Naidu Mallampati
Nov 5, 2014·Journal of Pharmacological Sciences·Masayuki X Mori, Ryuji Inoue
Mar 4, 2015·Biochimica Et Biophysica Acta·Gerald R V Hammond, Tamas Balla
Jul 12, 2005·Current Biology : CB·Yuchun GuRoger C Hardie
Nov 28, 2012·Journal of Physiology, Paris·Anne-Kathrin WarzechaJan Grewe
Mar 24, 2018·Journal of Cell Science·Che-Hsiung LiuRoger C Hardie
Jun 25, 2019·Frontiers in Cell and Developmental Biology·Rajan ThakurPadinjat Raghu
Nov 29, 2017·Journal of Cell Science·Shweta YadavPadinjat Raghu
Oct 21, 2015·Journal of Cell Science·Roger C HardieSukanya Sengupta
May 11, 2021·Frontiers in Pharmacology·Martin Kruse, Rayne J Whitten
Jan 11, 2007·Seminars in Cell & Developmental Biology·Padinjat Raghu
Apr 14, 2009·Cell Calcium·Padinjat Raghu, Roger C Hardie

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