Ethane 1,2-dimethane sulphonate is a useful tool for studying cell-to-cell interactions in the testis of the frog, Rana esculenta.
Ethane 1,2-dimethane sulphonate (EDS), a toxin which specifically destroys Leydig cells (LC), has been used to study cellular interactions in the testis of the frog Rana esculenta. Animals received three consecutive EDS injections and were sacrificed on day 4, 8, and 28 from the first injection. No significant morphological differences were observed between present observation and that obtained, in a previous experiment, after four consecutive EDS injections. In fact, on day 4, in the germinal tubules adjacent to apparently normal LC, Sertoli cells surrounding primary spermatogonia (I SPG) show heterochromatic nuclei and loss of cellular adhesion. Interestingly, I SPG surrounded by the heterochromatic Sertoli cells present grossly swollen mitochondria with ballooned cristae. On day 8, sometimes in the interstitium many LC appear strongly damaged and the germinal tubules appear disorganized; the only cell type still distinguishable is the I SPG. On day 28 from the first EDS injection a new population of LC reappear in the interstitium and spermatogenesis normalizes. These data confirm the close relationship between the interstitial and the geminal compartments. Immunocytochemical data obtained using a polyclonal antibody anticonnexin-43 (Cx-43, the most abundant Cx found in mammalian testis) demonstrate the presence of Cx-43 in the frog testis. In particular, Cx-43 is present between LC in the interstitium, between Sertoli and germ cells in the cysts and between Sertoli cells and I SPG. Cx-43 immunopositivity sharply decreases on day 4 from the first EDS injection simultaneously with the loss of cellular adhesion between Sertoli and germ cells. On day 8 and 28 from the first EDS injection Cx-43, immunopositivity is restored and, this data is also supported by Western blot analysis. Our data provide, for the first time, evidence that Cx-43 protein is present in the frog testis and confirm that EDS is a useful tool for studying cellular communication at the paracrine pathway or through direct contact depending on the gap junctional pathway in R. esculenta testis
Palmiero
Carmela
C
Dipartimento di Medicina Sperimentale-Sezione di Fisiologia Umana e Funzioni Biologiche Integrate F Bottazzi, Facoltà di Medicina e Chirurgia, Seconda Università degli Studi di Napoli, via Costantinopoli 16, 80138 Napoli, Italy.
Ferrara
Diana
D
De Rienzo
Gianluca
G
d'Istria
Michela
M
Minucci
Sergio
S
D000818:Animals; D015153:Blotting, Western; D002450:Cell Communication / Q000187:drug effects*; D018031:Connexin 43 / Q000378:metabolism; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D017629:Gap Junctions / Q000187:drug effects / Q000378:metabolism; D007150:Immunohistochemistry; D007985:Leydig Cells / Q000187:drug effects / Q000648:ultrastructure; D008297:Male; D008698:Mesylates; D008854:Microscopy, Electron; D014176:Protein Biosynthesis; D011506:Proteins / Q000032:analysis; D011893:Rana esculenta; D012671:Seminiferous Tubules / Q000166:cytology / Q000187:drug effects / Q000648:ultrastructure; D013737:Testis / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure
D018031:Connexin 43; D008698:Mesylates; D011506:Proteins; C002994:ethylene dimethanesulfonate; D004121:Dimethyl Sulfoxide