RESUMO
We investigated whether cortical glutamatergic and GABAergic release machineries can be differentiated on the basis of the proteins they express, by studying the degree of co-localization of synapsin (SYN) I and II, synaptophysin (SYP) I and II, synaptosomal-associated protein (SNAP)-25 and SNAP-23 in vesicular glutamate transporter (VGLUT) 1-, VGLUT2- and vesicular GABA transporter (VGAT)-positive (+) puncta in the rat cerebral cortex. Co-localization studies showed that SYNI and II were expressed in approximately 90% of VGLUT1+, approximately 30% of VGLUT2+ and 30-50% of VGAT+ puncta; SYPI was expressed in approximately 95% of VGLUT1+, 30% of VGLUT2+, and 45% of VGAT+ puncta; SYPII in approximately 7% of VGLUT1+, 3% of VGLUT2+, and 20% of VGAT+ puncta; SNAP-25 in approximately 94% of VGLUT1+, 5% of VGLUT2+, and 1% of VGAT+ puncta, and SNAP-23 in approximately 3% of VGLUT1+, 86% of VGLUT2+, and 22% of VGAT+ puncta. Since SYPI, which is considered ubiquitous, was expressed in about half of GABAergic axon terminals, we studied its localization electron microscopically and in immunoisolated synaptic vesicles: these studies showed that approximately 30% of axon terminals forming symmetric synapses were SYPI-negative, and that immunoisolated VGAT-positive synaptic vesicles were relatively depleted of SYPI as compared with VGLUT1+ vesicles. Overall, the present investigation shows that in the cerebral cortex of rats distinct presynaptic proteins involved in neurotransmitter release are differentially expressed in GABAergic and in the two major types of glutamatergic axon terminals in the cerebral cortex of rats.
Assuntos
Córtex Cerebral/metabolismo , Ácido Glutâmico/metabolismo , Ácido gama-Aminobutírico/metabolismo , Animais , Córtex Cerebral/ultraestrutura , Espectroscopia de Ressonância de Spin Eletrônica/métodos , Expressão Gênica , Masculino , Terminações Pré-Sinápticas/metabolismo , Terminações Pré-Sinápticas/ultraestrutura , Ratos , Ratos Sprague-Dawley , Sinapsinas/metabolismo , Proteínas Vesiculares de Transporte de Glutamato/metabolismoRESUMO
Using immunocytochemical techniques and confocal microscopy we have studied the localization of the vesicular glutamate transporters (VGLUTs) 1 and 2 in the mammalian cerebral cortex. The cardinal observations gathered to date can be summarized as follows: 1) Many VGLUT1-positive puncta coexpressing synaptophysin-1 outline pyramidal cell somata and proximal dendrites; of these, a sizeable fraction coexpress VGAT, the vesicular transporter for GABA; 2) VGLUT2-positive puncta are also present in layers II-III and some of them coexpress VGLUTI. These findings suggest that in the cerebral cortex of adult rats axon terminals expressing VGLUT1 are heterogeneous.