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First published online May 24, 2004
Journal of Experimental Biology 207, 2209-2213 (2004)
Published by The Company of Biologists 2004
doi: 10.1242/jeb.01000

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Environmental influence on testicular MAP kinase (ERK1) activity in the frog Rana esculenta

Paolo Chieffi^* and Sergio Minucci

Dipartimento di Medicina Sperimentale, II Università di Napoli, Naples, Italy

View larger version (23K): [in a new window]   Fig. 1. Western blot detection of ERK1 protein in the testicular extracts of R. esculenta at various times, 22°C and 12h:12h photoperiod. (A) Proteins (25 mg/lane) were resolved by SDS-PAGE, transferred to nitrocellulose membranes and then incubated with antibody raised against phospho-ERK1 and ERK1 protein. A specific band was observed of 44 kDa (determined by comparison with comigrating size markers). The blot is representative of three separate assays. (B) The amount of activated ERK1 was quantitated by using ImageQuant 5.2 Program and normalized by total ERK1. The values shown are representive of three separate experiments.

View larger version (14K): [in a new window]   Fig. 2. (A) Mitotic index of primary spermatogonia (I SPG) in the frog R. esculenta testis at various times, 22°C and 12h:12h photoperiod. The I SPG was expressed as the number of metaphases per total primary spermatogonia counted x100 in three randomly chosen sections/animal. Values are means ± S.D. Differences were considered significant at P<0.01. (B) Western blot detection of PCNA protein in the cytosolic testicular extracts of R. esculenta at various times, 22°C and 12h:12h photoperiod. Proteins (40 µg/lane) were resolved by SDS-PAGE, transferred to nitrocellulose membranes and then incubated with antibody raised against PCNA protein; a specific band was observed of 36 kDa (determined by comparison with comigrating size markers). (C) The amount of PCNA was quantitated using ImageQuant 5.2 Program. The values shown are representative of three separate experiments.

View larger version (21K): [in a new window]   Fig. 3. (A) Western blot detection of ERK1 protein in the testicular extracts of R. esculenta at various times, 4°C and 8 h:16 h photoperiod. Proteins (25 mg/lane) were resolved by SDS-PAGE, transferred to nitrocellulose membranes and then incubated with antibody raised against phospho-ERK1 and ERK1 protein. A specific band was observed of 44 kDa (determined by comparison with comigrating size markers). (B) The amount of activated ERK1 was quantitated by using ImageQuant 5.2 Program and normalized by total ERK1. The values shown are representative of three separate experiments.

View larger version (13K): [in a new window]   Fig. 4. (A) Mitotic index of primary spermatogonia (I SPG) in the frog R. esculenta testis at various times, 4°C and 8 h:16 h photoperiod. The I SPG was expressed as the number of metaphases per total primary spermatogonia counted x100 in three randomly chosen sections/animal. Values are expressed as means ± S.D. Differences were considered significant at P<0.01. (B) Western blot detection of PCNA protein in cytosolic testicular extracts of R. esculenta at various times, 4°C and 8 h:16 h photoperiod. Proteins (40 µg/lane) were resolved by SDS-PAGE, transferred to nitrocellulose membrane and then incubated with antibody raised against PCNA protein; a specific band at 36 kDa (by comparison with comigrating size markers). The blot is representative of three separate assays. (C) The amount of PCNA was quantitated by using ImageQuant 5.2 Program. The values shown are representative of three separate experiments.