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Localization of proline-like immunoreactivity in young rat retinal neurons
S Nishikawa1, S Ishiguro, M Tamai
1Department of Ophthalmology, Tohoku University School of Medicine, Sendai, Japan.
Purpose:
To localize proline in the rat retina.
Method:
We prepared antibodies raised against proline coupled to bovine serum albumin (BSA) with glutaraldehyde. To confirm the specificity of the antiserum, crossreactivity with other amino acids was determined using an immunodot procedure. Rat eyes were enucleated on postnatal day 21. Immunohistochemistry was performed on semi-thin sections using gold-labelled secondary antibodies and a silver-enhancement technique.
Results:
Immunodots revealed that the antiserum was specific for proline. Amino acids from retinal extracts were separated on gel, transferred to BSA-glutaraldehyde treated filters, and stained with antibodies to determine if our antibody reacted only with proline. Staining was most intense on postnatal day 21 and disappeared in the adult retina. In addition to Muller cells, horizontal cells, amacrine cells, and some cells in the ganglion cell layer were labelled.
Conclusions:
Antibodies raised against proline revealed a transient immunoreactivity in young rat neurons. Our findings strongly indicate that the retina may express a high level of proline in the developmental stages. This result may provide a clue for clarifying the role of proline in the retina.