Meiotic progression of rat spermatocytes requires mitogen-activated protein kinases of Sertoli cells and close contacts between the germ cells and the Sertoli cells.
Progression of germ cells through meiosis is regulated by phosphorylation events. We previously showed the key role of cyclin dependent kinases in meiotic divisions of rat spermatocytes co-cultured with Sertoli cells (SC). In the present study, we used the same culture system to address the role of mitogen-activated protein kinases (MAPKs) in meiotic progression. Phosphorylated ERK1/2 were detected in vivo and in freshly isolated SC and in pachytene spermatocytes (PS) as early as 3 h after seeding on SC. The yield of the two meiotic divisions and the percentage of highly MPM-2-labeled pachytene and secondary spermatocytes (SII) were decreased in co-cultures treated with U0126, an inhibitor of the ERK-activating kinases, MEK1/2. Pre-incubation of PS with U0126 resulted in a reduced number of in vitro formed round spermatids without modifying the number of SII or the MPM-2 labeling of PS or SII. Conversely, pre-treatment of SC with U0126 led to a decrease in the percentage of highly MPM-2-labeled PS associated with a decreased number of SII and round spermatids. These results show that meiotic progression of spermatocytes is dependent on SC-activated MAPKs. In addition, high MPM-2 labeling was not acquired by PS cultured alone in Sertoli cell conditioned media, indicating a specific need for cell-cell contact between germ cells and SC.
Godet
Murielle
M
Institut de Génomique Fonctionnelle de Lyon, Université de Lyon, Lyon F-69003, France. godet@lyon.inserm.fr
Sabido
Odile
O
Gilleron
Jérôme
J
Durand
Philippe
P
D000375:Aging / Q000502:physiology; D000818:Animals; D018920:Coculture Techniques; D004789:Enzyme Activation; D019084:Fluorescent Antibody Technique, Indirect; D005456:Fluorescent Dyes / Q000378:metabolism; D007150:Immunohistochemistry; D008297:Male; D008540:Meiosis / Q000502:physiology*; D020928:Mitogen-Activated Protein Kinases / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012235:Rhodamines / Q000378:metabolism; D012708:Sertoli Cells / Q000201:enzymology* / Q000502:physiology; D013090:Spermatocytes / Q000166:cytology* / Q000201:enzymology*
D005456:Fluorescent Dyes; D012235:Rhodamines; C009434:tetramethylrhodamine isothiocyanate; D020928:Mitogen-Activated Protein Kinases