Boar spermatozoa encapsulated in barium alginate membranes: a microdensitometric evaluation of some enzymatic activities during storage at 18 degrees C.
The customary dilution of boar semen for subsequent artificial insemination (AI) procedures damages the cell membrane of spermatozoa, resulting in a loss of enzymes and other cytoplasmic contents and acrosomal reactions. We encapsulated non-diluted boar semen in barium alginate membranes to optimize AI procedures and to improve the functional integrity of spermatozoal membranes during storage. The percentage of non-reacted acrosomes (NRA) and measurements of enzyme leakage (cytochrome c oxidase (COX), lactate dehydrogenase (LDH), and glucose-6-phosphate dehydrogenase (G6PDH)) were used as indices of the functional status of diluted, unencapsulated and encapsulated spermatozoa, stored for 72 h at 18 degrees C. Enzymatic activity was assessed in situ by microdensitometry, and non-reacted acrosomes were microscopically determined by staining. The percentage of acrosome integrity and the intracellular enzymatic activities during storage were different for unencapsulated and encapsulated semen. Semen dilution caused a rapid decline in enzymatic activities and concomitant acrosomal reactions. Encapsulated spermatozoa had significantly higher acrosome integrity (77% versus 55%; P < 0.01 after 72 h) and an overall higher in situ enzymatic activity. For cytochrome c oxidase and lactate dehydrogenase the greatest differences between encapsulated and unencapsulated spermatozoa were present after 72 h whereas for glucose-6-phosphate dehydrogenase significant differences were found within 24h of storage. The encapsulation process maintains a better preservation environment for boar spermatozoa and could be a promising, innovative technique to improve storage of these cells.
Faustini
Massimo
M
Department of Veterinary Science and Technology for Food Safety, University of Milan, Milan, Italy
Torre
Maria Luisa
ML
Stacchezzini
Simona
S
Norberti
Roberta
R
Consiglio
Anna Lange
AL
Porcelli
Franca
F
Conte
Ubaldo
U
Munari
Eleonora
E
Russo
Vincenzo
V
Vigo
Daniele
D
D020101:Acrosome Reaction; D000464:Alginates; D000818:Animals; D003720:Densitometry; D003576:Electron Transport Complex IV / Q000032:analysis; D005260:Female; D005954:Glucosephosphate Dehydrogenase / Q000032:analysis; D020723:Glucuronic Acid; D006603:Hexuronic Acids; D007315:Insemination, Artificial / Q000379:methods / Q000662:veterinary; D007770:L-Lactate Dehydrogenase / Q000032:analysis; D008297:Male; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D012996:Solutions; D013048:Specimen Handling / Q000662:veterinary; D013094:Spermatozoa / Q000201:enzymology*; D013696:Temperature; D013997:Time Factors
D000464:Alginates; D006603:Hexuronic Acids; D012996:Solutions; D020723:Glucuronic Acid; D007770:L-Lactate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D003576:Electron Transport Complex IV