Ultrastructural analysis of the glutamatergic system in the outer plexiform layer of zebrafish retina

Abstract

L-Glutamate, the photoreceptor neurotransmitter, depolarizes horizontal cells and OFF-bipolar cells by ionotropic receptors and hyperpolarizes ON-bipolar cells by metabotropic receptors. Despite extensive light microscopy on the distribution of glutamate receptors in zebrafish retina, there are little ultrastructural data. Given the importance of zebrafish in studies on the genetic manipulation of retinal development and function, precise data on the synaptic neurochemical organization of the zebrafish retina is needed. Immunohistochemical techniques were used to determine the ultrastructural localization of glutamate receptor subunits GluR2, GluR4, NMDA2B (NR2B) and mGluR1alpha in zebrafish outer plexiform layer (OPL). These antibodies were chosen because of an apparent conservation of localization of GluR2, GluR4 and mGluR1alpha in the vertebrate OPL, while there is some support for NMDA receptors in the OPL. GluR2-immunoreactivity (IR) was in all horizontal cell dendrites that invaginated cone pedicles and rod spherules. Three arrangements of dendrites contained GluR-IR in rod spherules: classical-type with GluR2-IR on lateral horizontal cell dendrites, a butterfly-shaped horizontal cell dendrite, and a goblet-shaped dendrite, likely of bipolar cell origin. GluR4-IR was restricted to dendrites of OFF-bipolar cells that innervated rod and cone terminals. NR2B-IR was restricted to a subtype of cone ON-bipolar cell. mGluR1alpha-IR was restricted to ON mixed rod/cone (Mb) bipolar cells whose dendrites innervated rod and cone synaptic terminals. The presence of mGluR1alpha on Mb bipolar cell dendrites is consistent with a role in retrograde endocannabinoid suppression. The subunit composition of glutamate receptors should affect the kinetics and pharmacology of these cells to glutamate receptor activation.