Quantification and characterization of GABA-ergic amacrine cells in the retina of GAD67-GFP knock-in mice.
OBJECTIVE
Although the presence of gamma-aminobutyrate acid (GABA) in amacrine cells and its co-localization with other neuronal substances is well known, there exists only little information about their quantitative distribution in the mouse eye. The aim of the present study was to characterize GABA-ergic amacrine cells in the retina of the recently introduced glutamate decarboxylase 67-green fluorescent protein (GAD67-GFP) knock-in mouse.


METHODS
Whole mounts of the retina were prepared and the GFP-positive neurons quantified. Immunofluorescence staining was performed with antibodies against GABA, calbindin (CB), calretinin (CR), parvalbumin (PV), choline acetyl transferase (ChAT), tyrosine hydroxylase (TH), vesicular glutamate transporter (VGluT) 1, VGluT2 and VGluT3.


RESULTS
Displaced GABA-ergic amacrine cells in the ganglion cell layer (GCL) showed a density of 1006 +/- 170 cells/mm(2). In the inner nuclear layer (INL), the density of amacrine cells was 8821 +/- 448 cells/mm(2) in the central region and 6825 +/- 408 cells/mm(2) in the peripheral region. GFP-positive amacrine cells co-localized with GABA (99%), CR (INL 18%, GCL 71.3%), CB (INL 6.3%), bNOS (INL 1%, GCL 4%), and ChAT (INL 17%, GCL 92.6%). No co-localization was seen with antibodies against PV, TH, and VGluT 1-3.


CONCLUSIONS
This study presents the first quantitative data concerning the co-localization of GABA-ergic neurons in the mouse retina with various neuronal markers.
May
Christian Albrecht
CA
Department of Anatomy, Carl Gustav Carus Medical Faculty, Technical University Dresden, Dresden, Germany. Albrecht.May@mailbox.tu-dresden.de
Nakamura
Kouichi
K
Fujiyama
Fumino
F
Yanagawa
Yuchio
Y
D025042:Amacrine Cells / Q000166:cytology / Q000201:enzymology*; D026922:Amino Acid Transport Systems, Acidic / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism*; D064032:Calbindin 2; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D005968:Glutamate Decarboxylase / Q000235:genetics* / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D008810:Mice, Inbred C57BL; D008817:Mice, Mutant Strains; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism; D010320:Parvalbumins / Q000378:metabolism; D064030:S100 Calcium Binding Protein G / Q000378:metabolism; D014446:Tyrosine 3-Monooxygenase / Q000378:metabolism; D050598:Vesicular Glutamate Transport Protein 1 / Q000378:metabolism; D050599:Vesicular Glutamate Transport Protein 2 / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000096:biosynthesis*
D026922:Amino Acid Transport Systems, Acidic; D015415:Biomarkers; D064032:Calbindin 2; D002795:Choline O-Acetyltransferase; D005968:Glutamate Decarboxylase; D049452:Green Fluorescent Proteins; D052248:Nitric Oxide Synthase Type I; D010320:Parvalbumins; D064030:S100 Calcium Binding Protein G; D014446:Tyrosine 3-Monooxygenase; D050598:Vesicular Glutamate Transport Protein 1; D050599:Vesicular Glutamate Transport Protein 2; D005680:gamma-Aminobutyric Acid; C576511:Calb2 protein, mouse; C493477:Slc17a6 protein, mouse; C493465:Slc17a7 protein, mouse; C504941:Slc17a8 protein, mouse; C496321:Nos1 protein, mouse; C402565:glutamate decarboxylase 1