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Phosphatidylinositol 4,5‐bisphosphate (PIP2) modulates afterhyperpolarizations in oxytocin neurons of the supraoptic nucleus

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The Journal of Physiology

Published online on

Abstract

Key points Afterhyperpolarizations (AHPs) generated by repetitive action potentials in supraoptic magnocellular neurons regulate repetitive firing and spike frequency adaptation but relatively little is known about PIP2’s control of these AHPs. We examined how changes in PIP2 levels affected AHPs, somatic [Ca2+]i, and whole cell Ca2+ currents. Manipulations of PIP2 levels affected both medium and slow AHP currents in oxytocin (OT) neurons of the supraoptic nucleus. Manipulations of PIP2 levels did not modulate AHPs by influencing Ca2+ release from IP3‐triggered Ca2+ stores, suggesting more direct modulation of channels by PIP2. PIP2 depletion reduced spike‐evoked Ca2+ entry and voltage‐gated Ca2+ currents. PIP2 appears to influence AHPs in OT neurons by reducing Ca2+ influx during spiking. Abstract Oxytocin (OT)‐ and vasopressin (VP)‐secreting magnocellular neurons of the supraoptic nucleus (SON) display calcium‐dependent afterhyperpolarizations (AHPs) following a train of action potentials that are critical to shaping the firing patterns of these cells. Previous work demonstrated that the lipid phosphatidylinositol 4,5‐bisphosphate (PIP2) enabled the slow AHP component (sAHP) in cortical pyramidal neurons. We investigated whether this phenomenon occurred in OT and VP neurons of the SON. Using whole cell recordings in coronal hypothalamic slices from adult female rats, we demonstrated that inhibition of PIP2 synthesis with wortmannin robustly blocked both the medium and slow AHP currents (ImAHP and IsAHP) of OT, but not VP neurons with high affinity. We further tested this by introducing a water‐soluble PIP2 analogue (diC8‐PIP2) into neurons, which in OT neurons not only prevented wortmannin's inhibitory effect, but slowed rundown of the ImAHP and IsAHP. Inhibition of phospholipase C (PLC) with U73122 did not inhibit either ImAHP or IsAHP in OT neurons, consistent with wortmannin's effects not being due to reducing diacylglycerol (DAG) or IP3 availability, i.e. PIP2 modulation of AHPs is not likely to involve downstream Ca2+ release from inositol 1,4,5‐trisphosphate (IP3)‐triggered Ca2+‐store release, or channel modulation via DAG and protein kinase C (PKC). We found that wortmannin reduced [Ca2+]i increase induced by spike trains in OT neurons, but had no effect on AHPs evoked by uncaging intracellular Ca2+. Finally, wortmannin selectively reduced whole cell Ca2+ currents in OT neurons while leaving VP neurons unaffected. The results indicate that PIP2 modulates both the ImAHP and IsAHP in OT neurons, most likely by controlling Ca2+ entry through voltage‐gated Ca2+ channels opened during spike trains.