Comparative investigation of transporter proteins in normal and cultured rat retina

Comparative investigation of transporter proteins in normal and cultured rat retina

Field: Development and regeneration

Authors: Törngren, Marie
Johansson, Kjell
Bruun, Anitha
Ehinger, Berndt

Address of presenting
author: Dpt of Ophthalmology
Lund University Hospital
S-221 85 Lund, Sweden

E-mail: Marie.Torngren@oft.lu.se

Phone: +46 46 17 27 07

Fax: +46 46 17 27 21

Text of abstract: Purpose. To study the immunohistochemical distribution of various transporter proteins in cultured rat retina and investigate if similar pattern is found in normal retina. We focus on glycine transporter 1 (Glyt-1), GABA transporter 1 (GAT-1) and vesicular acetylcholine transporter (VAChT), which are known to be expressed in neuronal subpopulations that innervate the inner plexiform layer (IPL). Methods. Explants of postnatal day 3 retina were cultured in Dulbeccos Modified Eagles Medium (DMEM) for two weeks in CO₂-incubator at 37°C. The explants were cultured without the retinal pigment epithelium and with the photoreceptor layer facing the membrane of the insert. Tissue from age matched normal rats constituted control retina. The specimens were fixed in 4% paraformaldehyde and processed for immunohistochemistry. Results. Retinas cultured for two weeks showed similar features as normal P14 retinas except that the outer segments of the photoreceptors appeared rudimentary. Both nuclear layers appeared normal in thickness. Due to eunucleation we assume that the ganglion cells degenerate, which may explain the thin ganglion cell layer (GCL). Immunoreactivities to the various transporter proteins investigated here, were present and well developed in cultured and control retinas. Not only subsets of GAT-1 and Glyt-1 immunolabeled amacrine cell somata, but also their processes were distinguished in cultured retina. The entire IPL showed strong GAT-1 and Glyt-1 immunolabeling. Immunoreactivity for VAChT was distributed in two distinct strata in IPL and in some amacrine cell somata. In comparison we found that the immunolabeling pattern displayed by the various antibodies was similar in cultured and control P14 retinas. Conclusion. By means of immunohistochemistry the present investigation confirms that some transporter proteins showed normal distribution and development within the IPL of cultured retinas. We also show that the IPL developed normally despite the absence of retinal pigment epithelium. CR: None, Supported by MRC, EU-BioMed 2.

Keywords: retina, transporter protein, immunohistochemistry, localisation, retina culture

	Comparative investigation of transporter proteins in normal and cultured rat retina
Field:	Development and regeneration
Authors:	Törngren, Marie Johansson, Kjell Bruun, Anitha Ehinger, Berndt
Address of presenting author:	Dpt of Ophthalmology Lund University Hospital S-221 85 Lund, Sweden
E-mail:	Marie.Torngren@oft.lu.se
Phone:	+46 46 17 27 07
Fax:	+46 46 17 27 21
Text of abstract:	Purpose. To study the immunohistochemical distribution of various transporter proteins in cultured rat retina and investigate if similar pattern is found in normal retina. We focus on glycine transporter 1 (Glyt-1), GABA transporter 1 (GAT-1) and vesicular acetylcholine transporter (VAChT), which are known to be expressed in neuronal subpopulations that innervate the inner plexiform layer (IPL). Methods. Explants of postnatal day 3 retina were cultured in Dulbeccos Modified Eagles Medium (DMEM) for two weeks in CO₂-incubator at 37°C. The explants were cultured without the retinal pigment epithelium and with the photoreceptor layer facing the membrane of the insert. Tissue from age matched normal rats constituted control retina. The specimens were fixed in 4% paraformaldehyde and processed for immunohistochemistry. Results. Retinas cultured for two weeks showed similar features as normal P14 retinas except that the outer segments of the photoreceptors appeared rudimentary. Both nuclear layers appeared normal in thickness. Due to eunucleation we assume that the ganglion cells degenerate, which may explain the thin ganglion cell layer (GCL). Immunoreactivities to the various transporter proteins investigated here, were present and well developed in cultured and control retinas. Not only subsets of GAT-1 and Glyt-1 immunolabeled amacrine cell somata, but also their processes were distinguished in cultured retina. The entire IPL showed strong GAT-1 and Glyt-1 immunolabeling. Immunoreactivity for VAChT was distributed in two distinct strata in IPL and in some amacrine cell somata. In comparison we found that the immunolabeling pattern displayed by the various antibodies was similar in cultured and control P14 retinas. Conclusion. By means of immunohistochemistry the present investigation confirms that some transporter proteins showed normal distribution and development within the IPL of cultured retinas. We also show that the IPL developed normally despite the absence of retinal pigment epithelium. CR: None, Supported by MRC, EU-BioMed 2.

Keywords:	retina, transporter protein, immunohistochemistry, localisation, retina culture

Index

Created 2000-03-13

Department of Physiological Sciences, Lund University

Web programming FormOnLine AB