During transit through the female reproductive tract, sperm encounter metabolites and environmental conditions that modulate various processes leading to fertilization. Intracellular Ca2+ dynamics regulate the acrosome reaction (AR), which involves exocytosis of the acrosomal granule, a prerequisite for successful fertilization. We explored the ability of progesterone, prostanglandin-E1, and GABA to induce Ca2+ mobilization and AR in single human spermatozoa capacitated under external pH (pH(e)) conditions found in different regions of the female reproductive tract (pH(e) 6.5, 7.4 and 8.0). The highest percentage of AR induction, regardless of the inducer, occurred when sperm were capacitated at pH, 7.4. Interestingly, at pH(e) 6.5 a high percentage of cells exhibit Ca2+ oscillations, which prevent AR. These oscillations involve extracellular and intracellular Ca2+ channels. Pharmacological inhibition of Ca2+ oscillations restores the ability of spermatozoa to undergo the AR when exposed to progesterone, even if capacitated at pH(e) 6.5. (C) 2018 Elsevier Inc. All rights reserved.