Abstract
ObjectiveThis study investigated the effects of 5-aminolevulinic acid (5-ALA) on the motility parameters, mitochondrial membrane depolarization, and ATP levels in chicken sperm.MethodsThe pooled semen from Barred Plymouth Rock males was used. In the first experiment, the semen was diluted 4-times with phosphate-buffered saline (PBS (−)) containing various concentrations (0, 0.01, 0.05, and 0.1 mM) of 5-ALA, and then the sperm motility parameters after incubation were evaluated by computer-assisted sperm analysis (CASA). In the second experiment, the semen was diluted 4-times with PBS (−) containing 0.05 mM 5-ALA, and then sperm mitochondrial membrane depolarization and ATP levels after 1.5 h of incubation were analyzed with the MitoPT® JC-1 Assay and ATP Assay kits, respectively. In the third experiment, the semen was removed from the seminal plasma and resuspended with the mediums of PBS (−), PBS (−) supplemented with CaCl2 and MgCl2 (PBS (+)) + 5-ALA, PBS (+) + caffeine, and PBS (+) + caffeine + 5-ALA. Then, the sperm motility parameters after incubation were evaluated by CASA. In the last experiment, the semen was treated with the mediums of PBS (−), PBS (−) + 5-ALA, 5.7% glucose, 5.7% glucose + 5-ALA after removing the seminal plasma, and then the sperm motility parameters were evaluated by CASA.ResultsThe addition of 0.05 mM 5-ALA significantly increased the chicken sperm motility, progressive motility, linearity, average path velocity, curvilinear velocity, straight-line velocity, and the wobble. The sperm mitochondrial membrane depolarization was also increased by the 5-ALA treatment. The 5-ALA treatment decreased the sperm ATP levels. Both the caffeine treatment and glucose treatment decreased the sperm motility during incubation period.Conclusion5-ALA might increase sperm mitochondrial membrane depolarization to utilize the ATP for enhancing sperm movement.
Highlights
In the present study, we aimed to examine the effects of 5-aminolevulinic acid (5-ALA) on the sperm motility parameters, mitochondrial membrane depolarization, and ATP levels in the chicken
To investigate whether 5-ALA improve chicken sperm motility by enhancing ATP production via glycolysis pathway or not, we compared the roles of 5-ALA in phosphate-buffered saline (PBS) (-) medium and the 5.7% glucose medium
Sperm motility, progressive motility, LIN, VAP, VCL, VSL, and WOB were significantly increased by the 0.05 mM 5-ALA treatment after 1.5 h of incubation (Experiment 1); we investigated whether the chicken sperm mitochondrial membrane depolarization was altered by 5-ALA
Summary
This study investigated the effects of 5-aminolevulinic acid (5-ALA) on the motility parameters, mitochondrial membrane depolarization, and ATP levels in chicken sperm. The semen was diluted 4-times with phosphate-buffered saline (PBS (-)) con taining various concentrations (0, 0.01, 0.05, and 0.1 mM) of 5-ALA, and the sperm motility parameters after incubation were evaluated by computer-assisted sperm analysis (CASA). The semen was diluted 4-times with PBS (-) containing 0.05 mM 5-ALA, and sperm mitochondrial membrane depolarization and ATP levels after 1.5 h of incubation were analyzed with the MitoPT® JC-1 Assay and ATP Assay kits, respectively. The semen was treated with the mediums of PBS (-), PBS (-) + 5-ALA, 5.7% glucose, 5.7% glucose + 5-ALA after removing the seminal plasma, and the sperm motility parameters were evaluated by CASA. Conclusion: 5-ALA might increase sperm mitochondrial membrane depolarization to utilize the ATP for enhancing sperm movement
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