Figure 4.

Abnormal sperm function parameters observed in the tamoxifen-inducible G3PP KO mice. A-L. Various sperm function parameters in icG3PP-KO mice were measured using a computer-assisted sperm analysis (CASA). A. Total motility, B. progressive motility, C. Curvilinear velocity (VCL), D. Average path velocity (VAP), E. Straight-line velocity (VSL), F. Straightness (STR), G. Linearity of forward progression (LIN) and H. Amplitude of lateral head displacement (ALH). I. Total sperm abnormalities, J. Sperm capacitation (%) and K. Spontaneous acrosome reaction (%SAR) were evaluated by determining the percentage of spermatozoa without acrosome by the Giemsa stain as described in Methods. L. Lipid peroxidation product 4-HNE and M. sperm DNA oxidation (8-dOHG) were measured using immunocytochemistry and relative intensity was assessed using ImageJ software. Data shown in panels A-M represent mean ± SEM and were analyzed by one-way ANOVA with Tukey's test. ∗∗p < 0.01 and ∗∗∗p < 0.001. “&” means lower compared to all other genotypes, whereas “#” means higher compared to all other genotypes. N. Membrane potential measured in sperm using JC-1 dye and flow cytometry. O. Reactive Oxygen Species (ROS) in the sperm were measured with DCFDA dye and flow cytometry. Data shown in panels N and O represent mean ± SEM and were analyzed by unpaired two-tailed t-test (∗p < 0.05 and ∗∗∗p < 0.001).