Sperm Motility: Conclusion 2

Interestingly, we also observed a more marked aquaporin 1a and aquaglyceroporin immunostaining at the plasma membrane in activated spermatozoa compared with nonactivated spermatozoa, which may suggest a possible recruitment of aquaporins into the plasma membrane from intracellular vesicles during the hyperosmotic shock. This mechanism might be similar to that reported for AQP1 in rat peritoneal…

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Sperm Motility: Conclusion

Water and solute transport through most AQPs is blocked by mercurial compounds, and in some cases, this inhibition can be reversed by the reducing agent ME. In mammalian AQP1, the Cys189 (close to the second NPA motif) is the Hg2+-sensitive site, although in other AQPs, as in rat and killifish (Fundulus heteroclitus) Aqp3 and and…

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Sperm Motility: DISCUSSION

In the hindgut, however, two additional reactive bands of approximately 27 and 25 kDa were detected, which probably correspond to degradation products. Protein extracts from water-injected oocytes did not show any reactive band, indicating the specificity of the aquaglyceroporin antiserum (Fig. 3A, lane 1). Western blot analysis, carried out using an anti-aquaporin 1a antiserum, identified…

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Sperm Motility: Aquaglyceroporin and Aquaporin

Aquaporin mRNA Expression in Gilthead Sea Bream Spermatozoa In the gilthead sea bream, the cDNAs encoding two Aqp1-like aquaporins (S. aurata aquaporin 1a and S. aurata aquaporin 1b) and one aquaglyceroporin have been reported. In the testis, aquaporin 1a but not aquaporin1b is expressed, and therefore the expression of aquaporin1a as well as of aquaglyceroporin…

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Sperm Motility: Statistical Analysis

The horseradish peroxidase-conjugated secondary antibody, diluted 1:100 in PBS and 0.1% Tween-20, was applied to the slides and incubated for 60 min at room temperature. The reaction was made visible by using a mixture of 0.06% 3,30-diaminobenzidine (DAB) in 0.05 M Tris-HCl buffer, pH 7.6, and 0.03% hydrogen peroxide for 20 min in the dark,…

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Sperm Motility: Western Blot Analysis

To extract proteins, the pellet was resuspended in lysis buffer (50 mM Tris-HCl, pH 7.4; 1% Nonidet P-40; 0.25% sodium deoxycholate; 150 mM NaCl; 1 mM ethylenediaminetetraacetic acid [EDTA]; 1 mM PMSF; 1 ig/il leupeptin; 1 mM Na3VO4; and 1 mM NaF). The suspension was spun at 18 000 X g for 15 min at…

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Sperm Motility: Computer-Assisted Sperm Analysis

Sperm movement was videotaped using a Nikon Alphaphot 2 microscope with a 20X negative phase objective and a Sony CCD black and white video camera (SSC-M188CE). Gilthead sea bream milt was diluted 1:100 in nonactivating medium (NAM; in mg/ml: NaCl 3.5, KCl 0.11, MgC^ 1.23, CaCl2 0.39, NaHCO3 1.68, glucose 0.08, and bovine serum albumin…

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