Localization of GAD- and GABA-like immunoreactivity in ground squirrel retina: retrograde labeling demonstrates GAD-positive ganglion cells

Abstract

Glutamic acid decarboxylase (GAD)- and γ-aminobutyric acid (GABA)-like immunoreactivity was examined in the retina of the 13-lined ground squirrel ( Spermophilus tridecemlineatus). Labeling was observed in the inner nuclear layer (INL), inner plexiform layer (IPL) and ganglion cell layer (GCL). The immunoreactive cell bodies in the inner third of the INL were 6–13 μm in diameter and, because of their size and location it was considered that these were amacrine cells. Labeling in the IPL was concentrated in 5 bands corresponding to laminae 1a, 1c, 2, 4 and 5. In the GCL a heterogeneous population of neurons exhibited GAD- and GABA-like immunoreactivity. The soma diameters of the GCL cells ranged from 5 to 17 μm. These may represent displaced amcrines and/or ganglion cells. To determine if any of the immunoreactive cells in the GCL were ganglion cells, double labeling experiments were performed using rhodamine latex microspheres (‘beads’) as retrograde neuronal tracers. Rhodamine beads were injected into the superior colliculus, and retinas with retrogradely labeled ganglion cells were subsequently incubated with the anti-GAD antiserum. These experiments revealed a small population of GAD-positive ganglion cells, setting a lower limit for the total number of GABAergic ganglion cells.