Phosphatidylinositol 4,5-bisphosphate regulates inspiratory burst activity in the neonatal mouse preBötzinger complex.
Neurons of the preBötzinger complex (preBötC) form local excitatory networks and synchronously discharge bursts of action potentials during the inspiratory phase of respiratory network activity. Synaptic input periodically evokes a Ca(2+)-activated non-specific cation current (I(CAN)) postsynaptically to generate 10-30 mV transient depolarizations, dubbed inspiratory drive potentials, which underlie inspiratory bursts. The molecular identity of I(CAN) and its regulation by intracellular signalling mechanisms during inspiratory drive potential generation remains unknown. Here we show that mRNAs coding for two members of the transient receptor potential (TRP) family of ion channels, namely TRPM4 and TRPM5, are expressed within the preBötC region of neonatal mice. Hypothesizing that the phosphoinositides maintaining TRPM4 and TRPM5 channel sensitivity to Ca(2+) may similarly influence I(CAN) and thus regulate inspiratory drive potentials, we manipulated intracellular phosphatidylinositol 4,5-bisphosphate (PIP(2)) and measured its effect on preBötC neurons in the context of ongoing respiratory-related rhythms in slice preparations. Consistent with the involvement of TRPM4 and TRPM5, excess PIP(2) augmented the inspiratory drive potential and diminution of PIP(2) reduced it; sensitivity to flufenamic acid (FFA) suggested that these effects of PIP(2) were I(CAN) mediated. Inositol 1,4,5-trisphosphate (IP(3)), the product of PIP(2) hydrolysis, ordinarily causes IP(3) receptor-mediated I(CAN) activation. Simultaneously increasing PIP(2) while blocking IP(3) receptors intracellularly counteracted the reduction in the inspiratory drive potential that normally resulted from IP(3) receptor blockade. We propose that PIP(2) protects I(CAN) from rundown by interacting directly with underlying ion channels and preventing desensitization, which may enhance the robustness of respiratory rhythm.
Crowder
Erin A
EA
Department of Applied Science, McGlothlin-Street Hall, The College of William and Mary, Williamsburg, VA 23187-8795, USA.
Saha
Margaret S
MS
Pace
Ryland W
RW
Zhang
Honglu
H
Prestwich
Glenn D
GD
Del Negro
Christopher A
CA
D000200:Action Potentials / Q000187:drug effects / Q000502:physiology; D000818:Animals; D000831:Animals, Newborn; D004594:Electrophysiology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics; D007002:Hypoglossal Nerve / Q000502:physiology; D001239:Inhalation / Q000502:physiology; D007668:Kidney; D051379:Mice; D008810:Mice, Inbred C57BL; D009046:Motor Neurons / Q000187:drug effects / Q000502:physiology; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000494:pharmacology*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction
D019269:Phosphatidylinositol 4,5-Diphosphate; D012313:RNA; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases