Fig 1.
Sperm motility patterns of bovine spermatozoa cultured in non-capacitation media and capacitation media.
Top left; normal motile sperm, right; sperm exhibiting activated motility, bottom; hyper-activated motility.
Fig 2.
Sperm patterns arising from the chlortetracycline fluorescence assay.
Left; F (uncapacitated)-pattern, central; B (capacitated)-pattern, right; AR (acrosome-reacted)-pattern. Bovine normal motile and activated sperm were incubated in the three types of media and analysed by CTC staining. Bar = 20 μm (B).
Fig 3.
Mitochondrial activity of sperm by JC-1 staining.
Top; Sperm annotated with green staining represent inactive mitochondria with no membrane potential (left: differential interference contrast: DIC, right: JC-1), Bottom; sperm annotated with bright orange staining represent active mitochondria with high membrane potential (left: DIC, right: JC-1). Bar = 20 μm.
Fig 4.
Chromosomal pattern of the bovine embryo at the blastocyst stage.
(A) Typical example in which the number of chromosomes were classified as ‘normal’, 60 (= 2n). (B) Typical example in which the number of chromosomes were classified as ‘Abnormal’, 90 (= 3n). (C) Typical example in which the number of chromosomes were classified as ‘Abnormal’, 30 (= n).
Fig 5.
DNA fragmentation at the pronuclear stage following the injection of activated and hyper-activated sperm.
(A) Positive (below) and negative (above) pattern following TUNEL staining. Differential interference contrast (DIC: left panel), Hoechst staining (central panel), and TUNEL staining (right panel). (B) Rate of DNA fragmentation in the three experimental groups. n = number of embryos. Different letters indicate significant differences as determined by the Chi Square test (P<0.01). Letters indicate significant differences (a-b). This experiment was repeated 3 times. Bar = 20 μm.
Fig 6.
ROS production of sperm (CPS) in non-capacitation media and capacitation media.
Different letters within columns indicate significant differences as determined by one-way ANOVA with Tukey-Kramer analysis (P<0.05). This experiment was repeated 3–4 times. Statistical significance compared to control: a vs b.
Fig 7.
ROS production of one individual motile sperm in non-capacitation media and capacitation media.
Different letters within columns indicate significant differences as determined by one-way ANOVA with Tukey-Kramer analysis (P<0.01). This experiment was repeated 3–4 times. Statistical significance compared to control: a vs b vs c.
Table 1.
The proportion (%) of sperm exhibiting normal, activated and hyper-activated motility in the two media after 0–4 hrs (n = 3–6).
Fig 8.
The proportion of capacitation and acrosome-reacted patterns in bovine sperm.
n = number of sperm. This experiment was repeated 3 times.
Fig 9.
Tyrosine phosphorylation localization in sperm.
Top and bottom images represent sperm cultured in non-capacitation media and capacitation media for 6hrs. Left) differential interference contrast (DIC) and right) immunofluorescence images. This experiment was repeated 3 times. Bar = 100 μm.
Table 2.
The direct effect of motile sperm in denuded oocytes on embryo developments and chromosomal aberration (n = 8).
Table 3.
The effect of type of motility upon embryo developments and chromosomal aberration in bovine embryos after ICSI (n = 8–11).
Fig 10.
The effect of CCCP treatment upon capacitated sperm.
The rate of capacitation and acrosome-reacted patterns of sperm as determined by CTC staining. This experiment was repeated 3 times.
Table 4.
The effect of reduced mitochondria activity upon on embryo developments and chromosomal aberration in bovine embryos after ICSI (n = 11).