Light-mediated activation reveals a key role for protein kinase A and sarcoma protein kinase in the development of sperm hyper-activated motility
| Autor: | Debby Ickowicz, Haim Breitbart, Adrian Shulman, Rachel Lubart, S. Shahar, Amir Wiser |
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| Rok vydání: | 2011 |
| Předmět: |
Male
Light Motility DNA Fragmentation Biology Capacitation Cyclic AMP Humans Calcium Signaling education Protein kinase A Sperm motility education.field_of_study Hyperactivation Rehabilitation Obstetrics and Gynecology Soluble adenylyl cyclase Cyclic AMP-Dependent Protein Kinases Spermatozoa Sperm Cell biology src-Family Kinases Reproductive Medicine Sperm Motility DNA fragmentation Reactive Oxygen Species Sperm Capacitation |
| Zdroj: | Human Reproduction. 26:2274-2282 |
| ISSN: | 1460-2350 0268-1161 |
| Popis: | background: Hyper-activated motility (HAM) is part of the sperm capacitation process, which is necessary for fertilization. In this study, we investigated the effect of visible light on sperm motility and hyperactivation and evaluated pathways mediating these effects. methods: Human sperm (1 × 10 7 cells/ml) in capacitation media were irradiated for 3 min with 40 mW/cm 2 visible light (400–800 nm with maximum energy at 600 nm). Sperm motility was assessed and analyzed by computer-assisted sperm analysis. The involvement of sperm capacitation factors was investigated as follows. The generation of reactive oxygen species (ROS) was measured using 20,70dichlorofluorescein diacetate. Protein kinase A (PKA) and sarcoma protein kinase (Src) activity were measured using western blot analysis and inhibited using 50 mM H89 and 10 mM PP2, respectively. Soluble adenlyl cyclase was inhibited using 20 mM 2-OH-Estradiol. The intracellular concentration of free Ca 2+ was assessed using the fluorescent calcium indicator, Fluo-4/AM. Sperm DNA fragmentation was determined using the sperm chromatin dispersion test. results: Light irradiation of human sperm caused a significant increase in hyper-HAM but not total motility. The production of ROS and activation of soluble adenylyl cyclase and PKA mediated the effect of light on HAM. Light irradiation also activated Src, and inhibition of Src significantly reduced the effect of light on HAM. Light irradiation caused a rapid increase in intracellular Ca 2+ concentration and the increase in HAM was significantly reduced when voltage-dependent-Ca 2+ -channel activity was blocked or when Ca 2+ -deficient medium was used. conclusions: Light irradiation of human sperm for a short time causes a significant increase in HAM in a mechanism mediated by ROS production, activation of PKA, Src and Ca 2+ influx. |
| Databáze: | OpenAIRE |
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