title: Nicotinic receptor involvement in regulation of functions of mouse neutrophils from inflammatory site.
abstract: Participation of nicotinic acetylcholine receptors (nAChRs) in functioning of polymorphonuclear neutrophils (PMNs) isolated from inflammatory site of mice and expression of different nAChR subunits were studied. Nicotine and acetylcholine (ACh) modified respiratory burst induced by a chemotactic peptide N-formyl-MLF in neutrophils of male (but not female) mice. Antagonists of nAChRs α-cobratoxin (αCTX), α-conotoxins MII and [A10L]PnIA at concentrations of 0.01-5μM, 0.2μM and 1μM, respectively, eliminated nAChR agonist effects. ACh also affected adhesion of PMNs, this effect was also prevented by αCTX (100nM) and MII (1nM). Neutrophils of female mice after chronic nicotine consumption acquired sensitivity to nAChR agonists. Changes of free intracellular Ca(2+) concentration in neutrophils under the action of nAChR ligands were analyzed. In cells with no Ca(2+) oscillations and relatively low resting level of intracellular Ca(2+), nicotine triggered Ca(2+)-spikes, the lag of the response shortened with increasing nicotine concentration. A nicotinic antagonist caramiphen strongly decreased the effect of nicotine. RT-PCR analysis revealed mRNAs of α2, α3, α4, α5, α6, α7, α9, β2, β3, and β4 nAChR subunits. Specific binding of [(125)I]-α-bungarotoxin was demonstrated. Thus in view of the effects and binding characteristics the results obtained suggest a regulatory role of α7, α3β2 or α6* nAChR types in specific functions of PMNs.
authors.affiliation: Institute of Cell Biophysics, Russian Academy of Sciences, Institutskaya str., 3, Pushchino, Moscow region, 142290, Russia. Electronic address: safronova@icb.psn.ru.
authors.forename: Valentina G
authors.initials: VG
authors.lastname: Safronova
authors.affiliation: Institute of Cell Biophysics, Russian Academy of Sciences, Institutskaya str., 3, Pushchino, Moscow region, 142290, Russia.
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Vulfius
authors.affiliation: Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya str., 16/10, Moscow, 117997, Russia.
authors.forename: Irina V
authors.initials: IV
authors.lastname: Shelukhina
authors.affiliation: Institute of Cell Biophysics, Russian Academy of Sciences, Institutskaya str., 3, Pushchino, Moscow region, 142290, Russia.
authors.forename: Valentina N
authors.initials: VN
authors.lastname: Mal'tseva
authors.affiliation: Institute of Cell Biophysics, Russian Academy of Sciences, Institutskaya str., 3, Pushchino, Moscow region, 142290, Russia.
authors.forename: Alexey V
authors.initials: AV
authors.lastname: Berezhnov
authors.affiliation: Institute of Cell Biophysics, Russian Academy of Sciences, Institutskaya str., 3, Pushchino, Moscow region, 142290, Russia.
authors.forename: Eugeniya I
authors.initials: EI
authors.lastname: Fedotova
authors.affiliation: Institute of Cell Biophysics, Russian Academy of Sciences, Institutskaya str., 3, Pushchino, Moscow region, 142290, Russia; Kazan Federal (Volga Region) University, Kremlevskaya str., 18, Kazan, 420008, Russia.
authors.forename: Regina G
authors.initials: RG
authors.lastname: Miftahova
authors.affiliation: Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya str., 16/10, Moscow, 117997, Russia.
authors.forename: Elena V
authors.initials: EV
authors.lastname: Kryukova
authors.affiliation: Institute of Cell Biophysics, Russian Academy of Sciences, Institutskaya str., 3, Pushchino, Moscow region, 142290, Russia.
authors.forename: Andrey A
authors.initials: AA
authors.lastname: Grinevich
authors.affiliation: Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya str., 16/10, Moscow, 117997, Russia.
authors.forename: Victor I
authors.initials: VI
authors.lastname: Tsetlin
mesh_terms: D000109:Acetylcholine / Q000378:metabolism; D000818:Animals; D020013:Calcium Signaling; D002448:Cell Adhesion; D002478:Cells, Cultured; D003039:Cobra Neurotoxin Proteins / Q000494:pharmacology; D005260:Female; D007249:Inflammation / Q000276:immunology*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009240:N-Formylmethionine Leucyl-Phenylalanine; D009504:Neutrophils / Q000276:immunology*; D009538:Nicotine / Q000378:metabolism; D021122:Protein Subunits / Q000235:genetics; D011978:Receptors, Nicotinic / Q000235:genetics / Q000378:metabolism*; D016897:Respiratory Burst
chemical_list: D003039:Cobra Neurotoxin Proteins; D021122:Protein Subunits; D011978:Receptors, Nicotinic; D009240:N-Formylmethionine Leucyl-Phenylalanine; C025154:alpha-cobratoxin; D009538:Nicotine; D000109:Acetylcholine
keywords: Adhesion; Intracellular Ca(2+); Neutrophilic granulocyte; Nicotinic acetylcholine receptors; Nicotinic ligands; Respiratory burst; nAChR subunit mRNAs