Characterization of cocaine
binding sites in the rat testes
by
Li H, George VK, Crossland WJ,
Anderson GF, Dhabuwala CB
Department of Urology,
School of Medicine,
Wayne State University,
Detroit, Michigan, USA.
J Urol 1997 Sep; 158(3 Pt 1):962-5

ABSTRACT

PURPOSE: An estimated 29 million individuals use cocaine in the United States. Studies have shown a high affinity for dose dependent binding of cocaine in the testes. Recent work done in our laboratory has shown that chronic administration of cocaine to male rats has an adverse effect on fertility and spermatogenesis by producing extensive morphological changes in the testes, leading to reduction in sperm production. As a first step toward understanding this process, we characterized and identified the pharmacological properties of [3H]cocaine binding sites in the testes. MATERIALS AND METHODS: Crude membranes from the testes were prepared from 35 days old male Sprague-Dawley rats. [3H]cocaine binding was measured by using the method of Madras et al. (1989) with modifications. The data from saturation binding assays were analyzed by Inplot (GraphPad Software, San Diego, CA) to determine the Kd and Bmax. RESULTS: Specific binding of [3H]cocaine was linearly dependent on membrane protein concentrations ranging from 0.2 to 8 mg./ml. The pooled data from three independent experiments revealed a mean affinity of 36 +/- 2.0 nM and Bmax of 1.84 +/- 0.13 pmol/mg. The present study demonstrates that testicular tissue has receptor protein that binds [3H]cocaine saturably and specifically. Competition displacement experiments revealed a shallow displacement curve for (-)cocaine and Win 35,428 with r2 = 0.96, indicative of multiple binding components. Computer analysis confirmed that a two component binding model was preferred statistically over a one component model in all three experiments (p < 0.001). CONCLUSION: The results from these studies suggest that the testicular tissue contains a protein that binds [3H]cocaine in a saturable and specific manner. It has a different sensitivity from the [3H]cocaine binding protein in the brain and placenta. Further clarification of the relationship between cocaine and its recognition site is necessary to understand the mechanism of testicular damage after cocaine exposure.

Oocytes
Spermatogenesis
Dopaminergic flies?
Dopaminergic agents
Cocaine-bound sperm
The coke-craving brain
Cocaine and depression
Cocaine and the lonely rat
Prenatal cocaine exposure
Cocaine and human spermatozoa

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