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Figure 3


Fig. 3. Odorant receptor stimulation results in phosphorylation and nuclear translocation of MAPK in human spermatozoa. (A) p42/44 and p38 MAPK are phosphorylated in response to bourgeonal stimulation of human sperm. Western blot analysis of total cell lysate of untreated human sperm cells (0) and cells that were treated with bourgeonal for 5 and 30 minutes. Lysates were probed for the presence of phosphorylated p44/42 MAPK and phosphorylated p38 MAPK using antibodies that specifically recognize the phosphorylated form of the proteins. Detection of the total amounts of p42/44 and p38 are shown as control. (B,C) p42/44 MAPK and p38 MAPK signals and signal from the unphosphorylated forms were quantified by densitometry. The amount of each phosphorylated MAPK was normalized to the total MAPK level in three independent experiments. Error bars represent s.e.m. (D) Phosphorylated p42/44 MAPK is present in the nuclei of bourgeonal-treated human sperm; detection of the total amount of p42/44 in the nuclear fraction is shown as a control. (E) The signal from phosphorylated p42/44 MAPK in the nuclear fraction was quantified by densitometry and normalized to a total amount p42/44 MAPK levels in two independent experiments. Error bars represent s.e.m.





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