1H NMR metabolic profiling of the cryopreserved spermatozoa of the wild endangered Persian sturgeon (Acipenser persicus) with the use of beta-cyclodextrin as an external cryoprotectant.

Abstract

Persian sturgeon (Acipenser persicus) is an endangered species and genetic resource banking such as gametes and embryo preservation could be one of the most pursued conservation approaches. In this study, deleterious effects of the traditional cryopreservation technique and the effect of different doses of 2-hydroxypropyl-beta-cyclodextrin (HβCD) on thawed spermatozoa quality (motility duration and percentage) of Persian sturgeon (Acipenser persicus) were investigated from metabolic aspects of view. For cryopreserving, semen was diluted with Tris-HCl (100mM) extenders containing 0, 5, 10, and 15mM of HβCD in a ratio of 1:1 (semen/extenders). Semen-extenders were filled into 0.5-mL straws and were frozen with the vapor of liquid nitrogen, and then immersed into liquid nitrogen. Cryopreserved spermatozoa were thawed in water baths in 15s. Two treatments with the highest and the lowest motility percentages (0 and 10mM of HβCD) were chosen to reveal the extremes of the metabolites change range and were objected to 1H NMR spectroscopy. Univariate (ANOVA) and multivariate (PCA) analysis of the obtained metabolic profiles showed significant changes (P < 0.05) in metabolites. The use of 10mM of HβCD was completely successful in the preservation of creatinine, glucose, guanidoacetate, O-phosphocholine, and N, N-dimethylglycine and probably their corresponding biochemical pathways, but it failed to preserve lactate, carnitine, betain, β-alanin, and trimethylamine N-oxide. It was also partially successful in preserving acetate, creatine, creatine phosphate, and glycine, all suggesting how HβCD can be effective as a cryoprotectant.