title: ATP-induced cytoplasmic [Ca2+] increases in isolated cochlear outer hair cells. Involved receptor and channel mechanisms.
abstract: Outer hair cells (OHC) of the mammalian cochlea are thought to preprocess the sound signal by active movements, which can be induced by electrical or chemical stimulation, e.g. depolarization evoked by high [K+] or increased cytoplasmic [Ca2+]. Extracellular ATP has been found to induce cytoplasmic [Ca2+] increases in OHC but involved mechanisms have not been elucidated. Cytoplasmic [Ca2+] was measured in non-enzymatically isolated single OHC using Fura-2 microspectrometry. Results, using ATP/derivatives and other P2-purinergic receptor (P2R) ligands, as well as Ca(2+)-channel blockers and pertussis toxin, revealed several signal transduction pathways that increase cytoplasmic [Ca2+] in OHC: a P2-purinergic receptor (P2R)--G-protein--effector (phospholipase C or an ion channel) system and a voltage-dependent Ca2+ channel. Agonist potency studies denote a pattern analogous to that found in skeletal muscle, i.e. ATP-alpha-S > ATP = 2-methyl-S-ATP >> ADP > alpha,beta-methylene-ATP, but no activation by ADP beta F or UTP, leaving a choice of P2y or P2zR subtypes. The latter possibility gained strength from calculations showing that up to 8% of ATP may have formed the P2zR agonist ATP4- in the experimental medium. Experiments in Ca(2+)-free medium and with pertussis toxin revealed that the main Ca2+ source was intracellular. Pertussis toxin did not affect [Ca2+] increase induced by carbachol. Acetylcholine, administered a few seconds before ATP, did not affect total cytoplasmic [Ca2+] increases. Induced cytoplasmic [Ca2+] increases were high enough (> 500 nM at 50 microM ATP/derivatives) to hyperpolarize the OHC membrane by opening K(+)-channels and decreased little with time.(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Department of Neurochemistry and Neurotoxicology, Stockholm University, Sweden.
authors.forename: R
authors.initials: R
authors.lastname: Nilles
authors.forename: L
authors.initials: L
authors.lastname: Järlebark
authors.forename: H P
authors.initials: HP
authors.lastname: Zenner
authors.forename: E
authors.initials: E
authors.lastname: Heilbronn
mesh_terms: D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D015220:Calcium Channels / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D005260:Female; D016257:Fura-2; D019204:GTP-Binding Proteins / Q000378:metabolism; D006168:Guinea Pigs; D018072:Hair Cells, Auditory, Outer / Q000187:drug effects / Q000378:metabolism*; D066298:In Vitro Techniques; D008024:Ligands; D008297:Male; D037342:Pertussis Toxin; D011983:Receptors, Purinergic / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects; D013050:Spectrometry, Fluorescence; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
chemical_list: D002121:Calcium Channel Blockers; D015220:Calcium Channels; D008024:Ligands; D011983:Receptors, Purinergic; D010566:Virulence Factors, Bordetella; D000255:Adenosine Triphosphate; D037342:Pertussis Toxin; D019204:GTP-Binding Proteins; D002118:Calcium; D016257:Fura-2