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. 2017 Dec 21;64(2):117–127. doi: 10.1262/jrd.2017-148

Fig. 4.

Fig. 4.

Plasma membrane integrity, acrosomal status, and capacitation status of frozen-thawed spermatozoa from the parental strains. (A) Spermatozoa stained with PI and PNA conjugated with Alexa Fluor 488. The PI(+) pattern indicates spermatozoa with damaged plasma membranes. The PI(–)PNA(+) pattern indicates live spermatozoa ready to undergo an acrosome reaction (bar = 10 μm). (B) The rates of the PI(+) sperm in the parental strains (See PI(–) in Table 1). (C) The rates of the PI(–)PNA(+) sperm staining pattern in the parental strains (* P < 0.05; ** P < 0.01 by Tukey-Kramer test). See Table 1 for the result of two-way ANOVA. (D) Spermatozoa stained with CTC. The F pattern has uniform fluorescence over the sperm head (uncapacitated). The B pattern shows fluorescence on the anterior portion of the head and a dark band over the postacrosomal region (arrow) (capacitated but not acrosome-reacted). The AR pattern shows no or little fluorescence over the whole surface of the head (capacitated and acrosome-reacted). Spermatozoa with the B and AR patterns are considered to be capacitated. (E) The rates of capacitated spermatozoa in the parental strains (** P < 0.01 by Tukey-Kramer test). See Table 1 for the result of two-way ANOVA.