Figure 1.
Localization of AVPR2 in epididymal mouse spermatozoa.
Immunohistochemistry staining of mouse caput (A, B, C) and cauda (D, E, F) spermatozoa. B and E are merged images of acrosome (Lectin PNA conjugated Alex 647, red) and nucleus (DAPI, blue). C and F are merged image of AVPR2, acrosome and nucleus. Arrows directed AVPR2 (A, C, D, and F). Images obtained using Nikon TS-1000 microscope with NIS Elements image software (Nikon, Japan). Bar = 10 µm.
Figure 2.
Change of the percentage of motility in various VP treatment concentrations.
Data represent mean ± SEM, n = 6. Values with different superscripts (a,b) were significantly different between control and treatment groups by one-way ANOVA (p<0.05).
Figure 3.
Change of [pH]i and calcium concentration during incubation with dDAVP.
(A) Change of [pH]i in various treatment condition. (B) Change of [Ca2+]i concentration in various treatment condition. Data represent mean ± SEM, n = 4. Values with different superscripts (a,b,c,d) were significantly different between control and treatment groups by one-way ANOVA (p<0.05).
Figure 4.
Effect of dDVAP on sperm capacitation status.
(A) Change of acrosome reacted spermatozoa in various treatment condition. (B) Change of capacitated spermatozoa in various treatment conditions. (C) Change of non-capacitated spermatozoa in various treatment conditions. Data represent mean ± SEM, n = 3. Values with different superscripts (a,b) were significantly different between control and treatment groups by one-way ANOVA (p<0.05).
Figure 5.
Effect of dDAVP on PKA activity and protein tyrosine phosphorylation.
(A) Density of PKA substrates in various treatments (Open Bar: ∼55 kDa, Black Bar: ∼23 kDa, Grey Bar: ∼22 kDa, Stripe Bar: ∼21 kDa, and Dot Bar: ∼18 kDa). Data represent mean ± SEM, n = 3. Values with different superscripts (A,B,a,b,I,II,III) were significantly different between control and treatment groups by One-way ANOVA (p<0.05). (B) Phospho-PKA substrates were probed with anti-phospho-PKA substrates; lane 1: Control, lane 2: 10−11 M dDAVP, lane 3: 10−8 M dDAVP, lane 4: 10−5 M dDAVP. (C) Density of tyrosine phosphorylated protein (∼30 kDa) in various treatments. Data represent mean ± SEM, n = 3. Values with different superscripts (a,b) were significantly different between control and treatment groups by One-way ANOVA (p<0.05). (D) tyrosine phosphorylated proteins were probed with PY 20; lane 1: Control, lane 2: 10−11 M dDAVP, lane 3: 10−8 M dDAVP, lane 4: 10−5 M dDAVP.
Figure 6.
Effect of dDVAP on fertilization and embryo development.
(A) Change of cleavage rate in various treatment conditions. (B) Change of blastocyst rate in various treatment conditions. Data represent mean ± SEM, n = 6. Values with different superscripts (a,b) were significantly different between control and treatment groups by One-way ANOVA (p<0.05).