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Cellular Retinol-Binding Protein and Retinoic Acid-Binding Protein in Rat Testes: Effect of Retinol Depletion¹

Departments of Biochemistry and Medicine, Vanderbilt University, Nashville, Tennessee 37232

Testes of rats contain two cellular binding proteins of interest in vitamin A metabolism. One protein binds retinoic acid with high specificity; the other binds retinol with high specificity. When the cellular retinol-binding protein was partially purified from rat testes, it exhibited fluorescence excitation and emission spectra similar to that of all-trans-retinol in hexane. Exposure of this preparation to UV light destroyed this fluorescence but spectra identical to the original were obtained after addition of retinol. Hexane extracts of the binding protein had fluorescence spectra identical to all-trans-retinol, suggesting that this compound is bound to the protein in vivo. Extracts of testes from retinol depleted rats were submitted to gel filtration but failed to show a retinol-like fluorescence at the elution position of retinol binding protein. This fluorescence was observed in the preparations from pair fed control animals. However, after addition of all-trans-retinol to the extracts from the depleted rats, fluorescence at that elution position was observed. This indicates that in testes of retinol depleted rats the cellular retinol binding protein is present but without bound retinol, in contrast to the non-depleted rats where 30–43% of the binding protein had bound retinol. The amounts of cellular retinol binding protein and retinoic acid binding protein in testes, as determined by sucrose gradient centrifugation, were found to be similar for retinol depleted and pair fed control rats.

KEY WORDS: • cellular retinol-binding protein • cellular retinoic acid-binding protein • retinol-depletion

¹ Supported by U.S. Public Health Service Grants No. HD 05384, HD 09195, HL 14 214, HL 15 341.

Manuscript received 1 July 1975.