P.A. and A.A. contributed equally to this work.
Regulation of glutamate release by heteromeric nicotinic receptors in layer V of the secondary motor region (Fr2) in the dorsomedial shoulder of prefrontal cortex in mouse
Article first published online: 19 MAR 2013
Copyright © 2013 Wiley Periodicals, Inc.
Volume 67, Issue 6, pages 338–357, June 2013
How to Cite
Aracri, P., Amadeo, A., Pasini, M. E., Fascio, U. and Becchetti, A. (2013), Regulation of glutamate release by heteromeric nicotinic receptors in layer V of the secondary motor region (Fr2) in the dorsomedial shoulder of prefrontal cortex in mouse. Synapse, 67: 338–357. doi: 10.1002/syn.21655
- Issue published online: 17 APR 2013
- Article first published online: 19 MAR 2013
- Accepted manuscript online: 20 FEB 2013 03:20AM EST
- Manuscript Accepted: 15 FEB 2013
- Manuscript Received: 13 OCT 2012
- Italian Ministry for University and Scientific Research (PRIN 2005)
- University of Milano-Bicocca (FAR)
- Telethon, Italy . Grant Number: GGP12147
- Fondazione Banca del Monte di Lombardia
- nicotinic acetylcholine receptors;
We studied how nicotinic acetylcholine receptors (nAChRs) regulate glutamate release in the secondary motor area (Fr2) of the dorsomedial murine prefrontal cortex, in the presence of steady agonist levels. Fr2 mediates response to behavioral situations that require immediate attention and is a candidate for generating seizures in the frontal epilepsies caused by mutant nAChRs. Morphological analysis showed a peculiar chemoarchitecture and laminar distribution of pyramidal cells and interneurons. Tonic application of 5 µM nicotine on Layer V pyramidal neurons strongly increased the frequency of spontaneous glutamatergic excitatory postsynaptic currents. The effect was inhibited by 1 µM dihydro-β-erythroidine (which blocks α4-containing nAChRs) but not by 10 nM methyllicaconitine (which blocks α7-containing receptors). Excitatory postsynaptic currents s were also stimulated by 5-iodo-3-[2(S)-azetidinylmethoxy]pyridine, selective for β2-containing receptors, in a dihydro-β-erythroidine -sensitive way. We next studied the association of α4 with different populations of glutamatergic terminals, by using as markers the vesicular glutamate transporter type (VGLUT) 1 for corticocortical synapses and VGLUT2 for thalamocortical projecting fibers. Immunoblots showed higher expression of α4 in Fr2, as compared with the somatosensory cortex. Immunofluorescence showed intense VGLUT1 staining throughout the cortical layers, whereas VGLUT2 immunoreactivity displayed a more distinct laminar distribution. In Layer V, colocalization of α4 nAChR subunit with both VGLUT1 and VGLUT2 was considerably stronger in Fr2 than in somatosensory cortex. Thus, in Fr2, α4β2 nAChRs are expressed in both intrinsic and extrinsic glutamatergic terminals and give a major contribution to control glutamate release in Layer V, in the presence of tonic agonist levels. Synapse 00:000–000, 2013. © 2013 Wiley Periodicals, Inc.