Lysophospholipids desestabilize the sperm plasma membrane and promote its fusion with outer acrosomal membranes, by accelerating acrosome reaction (RA). Lysophosphatidylcholine (LC) has been used to induce the RA on capacitated sperms from different mammals. The aim of this work was to evaluate the effect of LC on RA in canine spermatozoa. Different concentrations of LC (0, 100, 200 and 300 μg/mL) were utilized during 15 minutes to induce RA in spermatozoa incubated during 0, 3 and 4 hours in capacitation medium (mCCM). Sperm viability and acrosomal status were determined using double fluorescence Pisum sativum aglutinin with fluorescein isothiocyanate (PSA-FITC) and Hoechst 33258 The analysis of varience (ANOVA) test was utilized for statistical analysis. The sperm viability was significantly reduced with 200 and 300 μg/mL of LC (P<0.05). The live spermatozoa with RA show no statistically significant differences (P>0.05) between groups with 0 and 100 μg/ml of LC incubated to 0 hour (21.0 ± 4.2% vs. 21.0 ± 6.6%), 3 hours (43.8 ± 4.7% vs. 49.1 ± 5.2%) and 4 hours (51.3 ± 14.8% vs. 57.6 ± 9.9%). Nevertheless the percentage of live spermatozoa with RA increased (P<0.05) with incubation at 3 and 4 hours. In conclusion, LC (100 μg/mL) shows no significant effect for inducing the acrosome reaction in canine spermatozoa incubated in mCCM mediun free of glucose.