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. 2012 Jan 15;125(2):473–485. doi: 10.1242/jcs.093344

Fig. 4.

Fig. 4.

Decrease in [Na+]i is regulated by a cAMP-PKA-dependent pathway. Sperm from the cauda epididymus were placed in medium without HCO−3, which does not support capacitation, loaded for 30 minutes with CoroNa Red and washed as described. Aliquots of sperm suspension were incubated in the media described below for an additional hour. (A–C) Two-dimensional dot plots of PI versus CoroNa Red fluorescence of sperm incubated in non-capacitating medium (A, NON), in medium that supported capacitation (B, CAP) and, in order to directly stimulate PKA, sperm were also incubated in medium that did not support capacitation, i.e. with cAMP agonists (1 mM db-cAMP in combination with 100 μM IBMX; C, cAMP). (D) Quantification of CoroNa Red fluorescence of cells in A–C. Values are means ± s.e.m., n=4 (***P≤0.001). (E) Western blots, using anti-PKAS-P (left panel) and anti-Y-P (right panel) antibodies, of sperm extracts incubated as in A–C.