Calretinin-immunoreactive elements in the retina and optic tectum of the frog, Rana esculenta

Abstract

The frog retina contains numerous 28 kDa calbindin positive elements in its every layer. At the same time, parvalbumin has been observed only in a few elements in the visual system of amphibian species, whilst calretinin immunoreactivity could be detected in salamander retina and the optic tectum of tench. However, the presence and distribution of calretinin have been described to date neither in the retina nor in the other parts of the visual system of anurans. Therefore, the aim of this study is to describe the calretinin immunoreactive elements in the retina and the optic tectum of the frog and to establish whether or not the expression of this calcium-binding protein is transmitter-related and/or cell type specific in these parts of the central nervous system. In the retina, numerous bipolar cells showed calretinin immunoreactivity. The axon terminals of the bipolar cells branched in both the OFF and ON sublayers of the inner plexiform layer. The few labeled amacrine cells were larger than 10 microns in diameter. Over 50% of the cells in the ganglion cell layer contained calretinin. The labeled cells in the ganglion cell layer were of usually 16-22 microns in diameter, although a few smaller cells were also seen. Accordingly, many optic fibers were also labeled. In colocalization experiments, gamma-aminobutyric acid and calretinin were found in partially overlapping amacrine cell populations, cells with the former marker being much more numerous. At the same time, all the gamma-aminobutyric acid positive bipolar cells also contained calretinin. Most of the calretinin positive neurons in the ganglion cell layer however were only single-labeled. Axons of ganglion cells terminated in B, C and F sublayers of layer 9 in the optic tectum. Local tectal neuron populations in layer 4, 6, 8 and 9 were also labeled and a few calretinin positive cells were detected also in layer 2. Approximately 10% of the tectal cells were found to be immunoreactive for calretinin. Layer 4 and 6 cells were mostly large pear-shaped neurons while cells in the 8th layer were small pear-shaped and ganglion cells labeled too. Coexistence of gamma-aminobutyric acid and calretinin was characteristic in cells of the upper tectal layers while they were not detected in neurons of deep layers of the tectum. After monocular enucleation, contralateral to the removed eye, calretinin-immunoreactivity disappeared almost completely from F sublayer and became less pronounced in sublayers B and C after 90 days. Calretinin-immunoreactivity remained mostly unchanged in local tectal cells. The results show that, although its function remains undetermined, calretinin is the major EF-hand calcium-binding protein in the frog retina and optic tectum.