Metabolic changes in droplet vitrified semen of wild endangered Persian sturgeon Acipenser persicus (Borodin, 1997).
Comparative quantitative metabolite profiling can be used for better understanding of cell functions and dysfunctions in particular circumstances such as sperm banking which is an important approach for cryopreservation of endangered species. Cryopreservation techniques have some deleterious effects on spermatozoa which put the obtained results in controversy. Therefore, in the present study, quantitative1H NMR (Nuclear Magnetic Resonance) based metabolite profiling was conducted to evaluate metabolite changes related to energetics and some other detected metabolites in vitrified semen of critically endangered wild Acipenser persicus. The semen was diluted with extenders containing 0, 5, 10, and 15 μM of fish antifreeze protein (AFP) type III as a cryoprotectant. Semen-extenders were vitrified and stored for two days. Based on post-thaw motility duration and motility percentage assessments, two treatments with 10 μM and 0 μM of AFP had the highest and the lowest motility percentages respectively and they were objected to1H NMR spectroscopy investigations in order to reveal the extremes of the metabolites dynamic range. Univariate (ANOVA) and multivariate (PCA) analysis of the resulting metabolic profiles indicated significant changes (P > 0.05) in metabolites. The level of some metabolites including acetate, adenine, creatine, creatine phosphate, lactate, betaine, sarcosine, β-alanine and trimethylamine N-oxide significantly decreased in vitrified semen while some others such as creatinine, guanidinoacetate, N, N-dimethylglycine, and glycine significantly increased. There were also significant differences between vitrified treatments in levels of creatine, creatine phosphate, creatinine, glucose, guanidinoacetate, lactate, N, N-dimethylglycine, and glycine, suggesting how fish AFP type III can be effective as a cryoprotectant.
Department of Fisheries Sciences, Faculty of Natural Resources, University of Tehran, PO Box 31585-4314, Karaj, Iran.
Amirreza
A
Abed-Elmdoust
Department of Fisheries Sciences, Faculty of Natural Resources, University of Tehran, PO Box 31585-4314, Karaj, Iran. Electronic address: hfarahmand@ut.ac.ir.
Hamid
H
Farahmand
Department of Fisheries Sciences, Faculty of Natural Resources, University of Tehran, PO Box 31585-4314, Karaj, Iran.
Bagher
B
Mojazi-Amiri
Department of Fisheries Sciences, Faculty of Natural Resources, University of Tehran, PO Box 31585-4314, Karaj, Iran.
Gholamreza
G
Rafiee
Department of Fisheries Sciences, Faculty of Natural Resources, University of Tehran, PO Box 31585-4314, Karaj, Iran.
Ruhollah
R
Rahimi
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D021343:Antifreeze Proteins, Type III / Q000494:pharmacology; D003404:Creatinine / Q000378:metabolism; D015925:Cryopreservation; D003451:Cryoprotective Agents / Q000494:pharmacology; D005399:Fishes / Q000378:metabolism*; D005947:Glucose / Q000378:metabolism; D019344:Lactic Acid / Q000378:metabolism; D008297:Male; D008744:Methylamines / Q000378:metabolism; D012661:Semen; D012662:Semen Preservation; D013081:Sperm Motility / Q000502:physiology; D058989:Vitrification
D000596:Amino Acids; D021343:Antifreeze Proteins, Type III; D003451:Cryoprotective Agents; D008744:Methylamines; D019344:Lactic Acid; D003404:Creatinine; C005855:trimethyloxamine; D005947:Glucose
(1)H NMR spectroscopy; Fish antifreeze protein; Persian sturgeon semen; Quantitative metabolite profiling; Vitrification