Regulation of m1 muscarinic receptors and nNOS mRNA levels by autoantibodies from schizophrenic patients.
In this paper we demonstrate that, circulating antibodies from schizophrenic patients interacting with cerebral M1 muscarinic acetylcholine receptors (M1 mAChRs), can act as an inducer of m1 mAChR-mRNA, and neuronal nitric oxide synthase (nNOS) mRNA gene expression of rat frontal cortex. The different signaling pathways involved in the autoantibody's actions, were characterized. As previously reported serum autoantibodies from schizophrenic patients reacted against neural cells surface inhibiting the binding of the specific mAChR radioligand to rat cerebral frontal cortex membrane. Moreover, by ELISA using M1 synthetic peptide (with identical aminoacid sequence to human M1 mAChR) as coating antigen we demonstrated the reactivity against the second extracellular loop of human cerebral M1 mAChR. The corresponding affinity-purified anti M1 peptide IgG (anti M1 peptide IgG) from schizophrenic patients by stimulation of M1 mAChR exerted an increase in m1 mAChR-mRNA and nNOS-mRNA levels, that significantly correlated with the accumulation of phosphoinositides (IPs) and activation of NOS (alpha = 0.05). All these effects were blunted by pirenzepine and mimicked the action of the authentic agonist. Concurrent analysis of the effects of nNOS, phospholipase C (PLC) and calcium/calmodulin (CaM) inhibition on both, m1 mAChR-mRNA and nNOS-mRNA levels, showing that antibody up-regulation mRNA level is under the control of endogenous nitric oxide (NO) signaling system. On the basis of our results, the activation of M1 mAChR by schizophrenic autoantibody appears to induce nNOS-mRNA expression and reciprocally, the activation of NOS up-regulates m1 mAChR gene expression. These results gave support to the participation of an autoimmune process in a particular group of chronic schizophrenic patients.
Ganzinelli
Sabrina
S
Pharmacology Unit, School of Dentistry, University of Buenos Aires and Argentine National Research Council (CONICET), Marcelo T. de Alvear 2142, 4 B, 1122AAH Buenos Aires, Argentina.
Borda
Tania
T
Sterin-Borda
Leonor
L
D000328:Adult; D000704:Analysis of Variance; D000818:Animals; D001323:Autoantibodies / Q000737:chemistry / Q000494:pharmacology*; D015152:Blotting, Northern / Q000379:methods; D002540:Cerebral Cortex / Q000166:cytology / Q000187:drug effects; D002846:Chromatography, Affinity / Q000379:methods; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D007295:Inositol Phosphates / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D018727:Muscarinic Antagonists / Q000493:pharmacokinetics; D019001:Nitric Oxide Synthase / Q000378:metabolism; D052248:Nitric Oxide Synthase Type I / Q000235:genetics / Q000378:metabolism*; D011813:Quinuclidinyl Benzilate / Q000493:pharmacokinetics; D011869:Radioligand Assay / Q000379:methods; D051381:Rats; D017208:Rats, Wistar; D043643:Receptor, Muscarinic M1 / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012559:Schizophrenia / Q000276:immunology*; D014316:Tritium / Q000493:pharmacokinetics
D001323:Autoantibodies; D007295:Inositol Phosphates; D018727:Muscarinic Antagonists; D043643:Receptor, Muscarinic M1; D014316:Tritium; D011813:Quinuclidinyl Benzilate; D019001:Nitric Oxide Synthase; D052248:Nitric Oxide Synthase Type I