Disaccharide combinations and the expression of enolase3 and plasma membrane Ca²⁺ATPase isoform in sturgeon sperm cryopreservation

Abstract

Acipenser sinensis and Acipenser dabryanus are critically endangered species, so germplasm conservation via cryopreservation of sperm is necessary. Disaccharides can act as membrane-impermeable cryoprotectants, and enolase3 (ENO3) and plasma membrane Ca ATPase isoform (PMCA2) are proteins associated with sperm quality. We considered seven characteristics of sperm quality in cultured brood stock from A. sinensis and A. dabryanus. We tested use of sucrose or trehalose alone and in combination at different concentrations for cryopreservation of A. dabryanus sperm. A low concentration of sucrose plus trehalose (S T ) was optimal. Mixing of the extender with sucrose, lactose, or trehalose alone or with pairwise mixtures revealed that a mixture of lactose and trehalose (L T ) gave the best results for both A. sinensis and A. dabryanus. Enolase3 and PMCA2 expression levels were measured in cryopreserved A. sinensis sperm via Western blotting. Relative ENO3 and PMCA2 expression levels were examined, and the relationship between disaccharide composition, sperm quality and protein expression was explored in A. sinensis. The results showed that relative ENO3 and PMCA2 expression levels were the highest at L T in cryopreserved A. sinensis sperm. There were significant positive correlations between ENO3 expression and percentage membrane integrity, and between PMCA2 expression and sperm motility parameters (percentage of motile sperm, curvilinear velocity, straight-line velocity and average path velocity; p < .05) in cryopreserved A. sinensis sperm. Our results indicate the optimal disaccharide combination and concentrations for cryopreservation of A. sinensis and A. dabryanus sperm and suggest that ENO3 and PMCA2 expression levels could serve as a valuable indicator of sperm quality in A. sinensis.

A proteomic approach to identifying spermatozoa proteins in Indonesian native Madura bulls

“…This enzyme produces sperm motility energy ( 25 ). Xi et al ( 26 ) found a significant positive correlation between ENO3 and sperm motility parameters in sturgeons. Similarly, He et al ( 25 ) discovered ENO3 in the midpiece and tail of ram sperm, which strongly correlated with sperm motility.…”

Cryopreservation Studies in Aquaculture from Past to Present: Scientific Techniques and Quality Controls for Commercial Applications

“…When the use of sucrose or trehalose alone and in combination with different concentrations was tested for cryopreservation of Dabry's sturgeon (Acipenser dabryanus) sperm, low concentration sucrose plus trehalose (S 15 T 15 ) solution was reported to be the optimal solution. Also, it was reported that mixing of the extender with sucrose, lactose, or trehalose alone or with pairwise mixtures revealed that a mixture of lactose and trehalose (L15T15) gave the best results for both Chinese sturgeon (A. sinensis) and Dabry's sturgeon [47].…”

Role of Mono- and Disaccharide Combination in Cryoprotective Medium for Rooster Semen to Ensure Cryoresistance of Spermatozoa

“…According to the literature, various combinations of non-reducing and reducing saccharides can provide semen with the most effective protection during freezing/thawing [ 24 ]. The effect of a combination of monosaccharides (fructose, galactose, glucose, and xylose) and disaccharides (lactose, trehalose, maltose, and sucrose) on the composition of diluents for semen cryopreservation has been evaluated on various animal species [ 23 , 25 , 26 ].…”