Neurochemical anatomy of the zebrafish retina as determined by immunocytochemistry

Abstract

The zebrafish retina is rapidly becoming a major preparation for the study of molecular genetic mechanisms underlying neural development and visual behavior. Studies utilizing retinal mutants would benefit by the availability of a data base on the distribution of neurotransmitter systems in the wild-type fish. To this end, the neurochemical anatomy of the zebrafish retina was surveyed by light microscopic immunocytochemistry. An extensive series of 60 separate antibodies were used to describe the distribution of major transmitter systems and a variety of neuron-associated membrane channels and proteins.These include markers (i.e., antibodies against enzymes, receptors, transporters) for transmitters: GABA, glycine, glutamate, biogenic amines, acetylcholine, cannabinoids and neuropeptides; as well as a sample of voltage-gated channels and synapse associated membrane proteins. Discussion of the comparative localization of these antibodies is restricted to other teleost fishes, particularly goldfish. Overall, there was great similarity in the distribution of the various markers, as might be expected. However, there were some notable differences, including several antibodies that did not label zebrafish at all, even though goldfish retinas that were processed in parallel, labeled beautifully. This survey is extensive, but not exhaustive, and hopefully will serve as a valuable resource for future studies of the zebrafish retina.

Zebrafish: A model system for the study of eye genetics

“…From DiOlistic, fluorescent images and physiological recordings, horizontal cells were classified into three or perhaps four subtypes, H1/H2, H3 and H4 (Song et al, 2007), and whole-cell recording demonstrates many properties similar to other teleost horizontal cells (McMahon, 1994). Amacrine cells were categorized into seven morphological subtypes, and though these morphological classifications are in basic agreement with previous reports using immunolabelling and metabolic signatures, a greater diversity of the amacrine cell subtypes will likely be identified using additional methods (Marc and Cameron, 2001;Yazulla and Studholme, 2001). Bipolar cells have been best characterized in zebrafish using a combination of physiological recordings followed by backfilling with a fluorescent dye or DiOlistic labeling (Connaughton and Nelson, 2000).…”

Electrophysiological evidence of GABAA and GABAC receptors on zebrafish retinal bipolar cells

“…This suggests that glycinergic innervation might be type specific, whereas GABAergic innervation might be universal. The fact that zebrafish bipolar cells receive direct glycinergic inputs agrees with the immunocytochemical localization of glycine-positive processes (Connaughton et al, 1999) and glycine receptor subunits (Yazulla & Studholme, 2001) in both plexiform layers of the zebrafish retina. Glycinergic inputs to bipolar cell dendrites most likely arise from the population of glycine-containing interplexiform cells present in zebrafish (Connaughton et al, 1999;Marc & Cameron, 2001); while presynaptic glycine-containing amacrine cells most likely provide inputs to bipolar terminals.…”

A Role for 1 Tubulin-Expressing Muller Glia in Regeneration of the Injured Zebrafish Retina

“…Immunohistochemistry was performed as described previously (Senut et al, 2004) using the following primary antibodies: rat anti-BrdU (dividing cell marker; 1:250; Harlan Sera-Lab, Loughborough, UK); mouse zpr1 [double-cone photoreceptor marker (Larison and Bremiller, 1990); 1:250; Zebrafish International Resource Center (ZIRC), Eugene, OR]; rabbit anti-GFP (1:1000; Invitrogen, Carlsbad, CA); mouse anti-HuC/D (amacrine cell marker; 1:500; Invitrogen); rabbit anti-protein kinase C (PKC) [ON bipolar cell marker (Yazulla and Studholme, 2001); 1:250; Santa Cruz Biotechnology, Santa Cruz, CA]; mouse anti-tyrosine hydroxylase (TH) (interplexiform cell marker; 1:500; Chemicon, Temecula, CA); mouse anti-zrf1 (glial marker; 1:250; ZIRC); mouse anti-zn5 [retinal ganglion cell (RGC) marker; 1:500; ZIRC]; and mouse anti-glutamine synthetase (GS) (glial marker; 1:500; Chemicon). For BrdU immunostaining, sections were pretreated with 2N HCl for 30 min at 37°C, two 5 min rinses in 0.1 M sodium borate buffer, and three 5 min rinses in PBS, pH 7.4.…”