in blocking solution containing 1 bovine serum albumin bsa 0 4 triton x 100 and 1 5 normal donkey serum nds tissue sections were incubated in primary antibody see below overnight on the following day tissue sections were washed in kpbs containing 0 2 triton x 100 and incubated for two hours in the appropriate fluorophore conjugated secondary antibody sections were washed again incubated in 4 u2032 6 diamidine 2 phenylindole dapi molecular probes carlsbad calif for 5 minutes and washed a final time in kpbs prior to being cover slipped with vectashield fluorescence mounting medium vector burlingame calif labeling patterns were imaged with a fluorescence capable microscope nikon eclipse e800 equipped with a digital camera and prepared using adobe photoshop cs version 8 0 and macromedia freehand mx version 11 0 for the macintosh primary antibodies used in this study included the following rabbit anti protein kinase c alpha isoform pkc sigma st louis mo guinea pig anti vesicular 11 glutamate transporter 1 vglut1 chemicon international inc temecula calif rabbit anti microtubule associated protein 2 map2 chemicon international inc temecula calif rabbit anti glutamine synthetase gs sigma st louis mo and rabbit anti glial fibrillary acidic protein gfap dakocytomation carpinteria calif secondary antibodies included fluorescein isothiocyanate fitc conjugated donkey anti guinea pig igg jackson immunoresearch west grove pa or fitc conjugated donkey anti rabbit igg.