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BOR - Papers in Press, published online ahead of print April 12, 2006.
Biol Reprod 2006, 10.1095/biolreprod.106.051730
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Submitted February 13, 2006
Returned for revision March 14, 2006
Accepted March 24, 2006

Mechanisms of Hormone Action


Endocannabinoid System in Frog and Rodent Testis: Type-1 Cannabinoid Receptor and Fatty Acid Amide Hydrolase Activity in Male Germ Cells

Gilda Cobellis , Giovanna Cacciola , Donatella Scarpa , Rosaria Meccariello , Rosanna Chianese , Maria Fosca Franzoni , Ken Mackie , Riccardo Pierantoni *, and Silvia Fasano

* To whom correspondence should be addressed. E-mail: riccardo.pierantoni{at}unina2.it.

Abstract
N-arachidonoylethanolamine (anandamide, AEA) is the main endocannabinoid described to date in the testis. It exerts its effects through the activation of G-protein coupled cannabinoid receptors (CNR). However, the activity of AEA in controlling male reproduction is still poorly known. Here we provide direct evidence on the presence of the "endocannabinoid system", constituted by type-1 cannabinoid receptor (CNR1) and fatty acid amide hydrolase (FAAH), in the frog, Rana esculenta, testis demonstrating its expression in tubular compartment. In fact, during the annual reproductive cycle, both proteins increase in September when the appearance of spermatids (SPT) occurs. Immunocytochemistry confirms their localization in germ cells and , in particular, in elongated SPT. Signals are still present in spermatozoa (SPZ), as demonstrated by Western blot analysis. Furthermore, the activation of CNR1 reduces sperm motility. Comparative research, carried out using mouse and rat SPZ, definitely indicates that the endocannabinoid system operates in SPZ of phylogenetically distant species. A conserved physiological role of endocannabinoid system in controlling the inhibition of sperm motility is suggested.

Key words: Male Reproductive Tract • Testis • Sperm • Sperm motility and transport • Spermatogenesis


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