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. 2022 May 18;154(6):e202113074. doi: 10.1085/jgp.202113074

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© 2022 Jensen et al.

This article is available under a Creative Commons License (Attribution 4.0 International, as described at https://creativecommons.org/licenses/by/4.0/).

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Figure 8. — Recovery of PtdIns(4,5)P₂ after depletion. KCNQ2/3 current suppression and recovery are recorded to reflect depletion and recovery of PtdIns(4,5)P₂. Recovery half times are indicated. (A) Diagram of PtdIns(4,5)P₂ resynthesis showing also phosphatases and the voltage-sensitive VSP enzyme. (B) Depletion and recovery after a short application (25 s) of Oxo-M agonist (10 μM). The experiment is similar to that in Fig. 3 except that rather than using a neuron with endogenous receptors and channels, this one uses tsA201 cells transfected with M₁ receptors and KCNQ2 and KCNQ3 channel subunits. PIP₂, PtdIns(4,5)P₂. From Horowitz et al. (2005). (C) A tsA201 cell transfected with channels and VSP (no receptors) is depolarized in a three-step protocol starting at 0 s. The first 1-s step to −20 mV activates KCNQ2/3 current, the next 1-s step to 100 mV activates VSP briefly, and the third step to −20 mV monitors current recovery during the resynthesis of PtdIns(4,5)P₂. The olive trace shows a cell that had also been transfected with PIP 5-kinase. From Falkenburger et al. (2010a).