5’-AMP-Activated Protein Kinase Regulates Goat Sperm Functions via Energy Metabolism In Vitro
| Autor: | Jun Luo, Wenxian Zeng, Zhendong Zhu, Xiaoteng Fan, Yinghua Lv, Wenjing Bai, Rongnan Li, Gongzhen Ma |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Male AMPK Physiology Acrosome reaction AMP-Activated Protein Kinases lcsh:Physiology lcsh:Biochemistry 03 medical and health sciences Capacitation Animals lcsh:QD415-436 Phosphorylation Protein kinase A Inner mitochondrial membrane Sperm motility lcsh:QP1-981 Chemistry Goats Acrosome Reaction Sperm Spermatozoa Cell biology 030104 developmental biology Energy Metabolism Pyruvate kinase |
| Zdroj: | Cellular Physiology and Biochemistry, Vol 47, Iss 6, Pp 2420-2431 (2018) |
| ISSN: | 1421-9778 1015-8987 |
| Popis: | Background/Aims: ATP is essential for mammalian sperm to survive and maintain fertilizing capacity. AMP-activated protein kinase (AMPK) is a sensor of cellular energy status. The aims of the present study were to explore the localization of AMPK in goat sperm and to investigate whether and how AMPK regulates sperm functions in vitro. Methods: Sperm were treated with AMPK modulators (AICAR, metformin and Compound C) during incubation. Sperm motility was assessed with a computer-assisted spermatozoa analysis system (CASA). Membrane integrity, acrosome reaction and mitochondrial membrane potentials were detected by SYBR-14/PI, FITC-PNA and JC-1 staining, respectively. And the lactate content, ATP content, AMPK activity, activity of pyruvate kinase (PK) and lactate dehydrogenase (LDH) were also measured with the commercial assay kits. Immunofluorescence staining was used to analyze the distribution of PK, LDH, AMPK and phospho-Thr172-AMPK in sperm. The role of AMPK was further studied during induction of capacitation and acrosome reaction. Results: We found that AMPKα was localized in the entire acrosomal region, the midpiece and the flagellum, while the phospho-Thr172-AMPK was distributed in the head, the midpiece and flagellum. Activation of AMPK by AICAR and metformin significantly improved sperm motility, membrane integrity and acrosome reaction, largely maintained sperm mitochondrial membrane potentials, lactate content and ATP content, and enhanced the activity of AMPK, PK and LDH, whereas inhibition by Compound C triggered the converse effects. Moreover, PK was localized in the acrosomal area and the midpiece, while LDH was distributed in the tail. Induction of capacitation and acrosome reaction led to AMPK phosphorylation. AMPK phosphorylation regulated the activity of energetic enzymes. Conclusion: This study for the first time provides evidence that AMPK governs goat sperm functions through energy metabolism in vitro. This finding will help to improve assisted reproductive techniques in goats and the other species. |
| Databáze: | OpenAIRE |
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