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BOR - Papers in Press, published online ahead of print October 30, 2002.
Biol Reprod 2002, 10.1095/biolreprod.102.007831
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Submitted May 28, 2002
Returned for revision June 14, 2002
Accepted October 16, 2002

Testis


Ontogenesis of Leptin Receptor in Rat Leydig Cells

Massimiliano Caprio 1, Elisa Fabbrini 1, Giulia Ricci 2, Sabrina Basciani 1, Lucio Gnessi 1, Mario Arizzi 1, Anna R. Carta 3, Massimo U. De Martino 1, Andrea M. Isidori 1, Giovanni V. Frajese 1, Andrea Fabbri 1*
1 Universita di Roma
2 Seconda Universita di Napoli
3 Universita di Cagliari

* To whom correspondence should be addressed. E-mail: a_fabbri{at}hotmail.com.

Abstract

There are still many controversies about the role of leptin in reproductive function and sexual development. We recently demonstrated that leptin receptors are expressed in rodent Leydig cells and that leptin has inhibitory effects on hCG-stimulated testosterone production by adult rat Leydig cells in culture. In this study we evaluated the expression of leptin receptor (Ob-R) in rat testes from gestational to adult age, in comparison with the pattern of expression of relaxin-like factor (RLF), a specific marker of Leydig cells differentiation status. Immunohistochemical analysis showed that in prenatal life Ob-R immunoreactivity was absent at early embryonic age (E14.5) and appeared at a late embryonic age (E19.5); in post-natal life, immunoreactivity was evident only after sexual maturation (35, 60, and 90 days old), whereas it was absent in testes from sexually immature rats (7, 14, and 21 days old). Immunoreaction was always confined to Leydig cells and no signal of Ob-R was detected within the tubules. The pattern of expression of Ob-R during testicular development was similar with that of RLF immunoreactivity, which was present in mature fetal as well as adult-type Leydig cells. In contrast with the findings in the testis, in the hypothalamus the immunohistochemical pattern of Ob-R was very similar between pre- and postpubertal life. RT-PCR studies showed that Ob-R expression was present in embryonic, prepubertal and adult rat testes; semiquantitative analysis showed that mRNA levels were much higher in late vs early embryonic testes, as well as in mature adults vs sexually immature testes, with a gradual increase from younger to older ages. Functional studies showed that while leptin (150 ng/ml) significantly inhibited hCG-stimulated testosterone production in adult rat Leydig cells (46% reduction; p>0.01), it did not modify prepubertal rat Leydig cells steroidogenic function in vitro. In conclusion we showed that in rat testis Ob-R expression is characteristic of mature Leydig cells (fetal and adult-type) and it is functional in adult but not prepubertal life.



Key words: Testis • Leptin • Leptin receptor • Leydig cells • Testosterone


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