title: Pre-synaptic nicotinic receptors evoke endogenous glutamate and aspartate release from hippocampal synaptosomes by way of distinct coupling mechanisms.
abstract: OBJECTIVE
  The present work aimed to investigate whether and through which mechanisms selective α7 and α4β2 nicotinic receptor (nAChR) agonists stimulate endogenous glutamate (GLU) and aspartate (ASP) release in rat hippocampus.
  
  
  METHODS
  Rat hippocampal synaptosomes were purified on Percoll gradients and superfused in vitro to study endogenous GLU and ASP release. The synaptosomes were superfused with selective α7 and α4β2 nAChR agonists and antagonists. The excitatory amino acid (EAA) content of the samples of superfusate was determined by HPLC after pre-column derivatization and separation on a chromatographic column coupled with fluorimetric detection.
  
  
  RESULTS
  Choline (Ch), a selective α7 receptor agonist, elicited a significant release of both GLU and ASP which was blocked by the α7 receptor antagonist methyllycaconitine (MLA), but was unaltered by the α4β2 receptor antagonist dihydro-β-erythroidine (DHβE). The stimulant effect of Ch was strongly reduced in a Ca(2+) -free medium, was not inhibited by Cd(2+) and tetrodotoxin (TTX), but was antagonized by dantrolene, xestospongin C and thapsigargin. 5-Iodo-A-85380 dihydrochloride (5IA85380), a selective α4β2 receptor agonist, elicited EAA release in a DHβE-sensitive, MLA-insensitive fashion. The 5IA85380-evoked release was dependent on extracellular Ca(2+) , blocked by Cd(2+) and TTX, but unaffected by dantrolene.
  
  
  CONCLUSIONS
  Our study shows for the first time that rat hippocampal synaptosomes possess α7 and α4β2 nAChR subtypes, which can enhance the release of endogenous GLU and ASP via two distinct mechanisms of action. These results extend our knowledge of the nicotinic modulation of excitatory synaptic transmission in the hippocampus.
authors.affiliation: Department of Experimental Medicine, Section of Pharmacology and Toxicology Center of Excellence for Biomedical Research National Institute of Neuroscience, University of Genoa, Viale Cembrano 4, Genoa, Italy.
authors.forename: S
authors.initials: S
authors.lastname: Zappettini
authors.forename: M
authors.initials: M
authors.lastname: Grilli
authors.forename: A
authors.initials: A
authors.lastname: Salamone
authors.forename: E
authors.initials: E
authors.lastname: Fedele
authors.forename: M
authors.initials: M
authors.lastname: Marchi
mesh_terms: D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D018698:Glutamic Acid / Q000378:metabolism*; D006624:Hippocampus / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D018722:Nicotinic Agonists / Q000494:pharmacology; D018733:Nicotinic Antagonists / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011978:Receptors, Nicotinic / Q000187:drug effects / Q000378:metabolism*; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism; D064569:alpha7 Nicotinic Acetylcholine Receptor
chemical_list: C579165:Chrna7 protein, rat; D018722:Nicotinic Agonists; D018733:Nicotinic Antagonists; D011978:Receptors, Nicotinic; D064569:alpha7 Nicotinic Acetylcholine Receptor; C403531:nicotinic receptor alpha4beta2; D001224:Aspartic Acid; D018698:Glutamic Acid; D002118:Calcium