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Biology of Reproduction, Vol 15, 79-83, Copyright © 1976 by Society for the Study of Reproduction

Comparison of Several Extraction Procedures for Boar Spermatozoal Acrosin

L. A. JOHNSON 1, V. G. PURSEL 1, N. CHANEY 1, , and D. L. GARNER 1

1 Reproduction Laboratory, Animal Physiology and Genetics Institute, U.S.D.A., Agricultural Research Center, Beltsville, Maryland 20705


Six procedures to extract acrosin from boar spermatozoa were compared. A procedure in which a combination of Hyamine 2389, glycerol, and freeze-thawing (HGF) was used, yielded an extract with significantly higher specific enzyme activity (Units/mg protein) than the other five procedures. The HGF procedure and two procedures in which Orvus ES Paste (OEP) and acetic acid were used as extractants, yielded extracts with the highest total enzyme activity. The amount of protein extracted by the procedures in which OEP and acetic acid were used was significantly greater than the amount of protein extracted by the HGF procedure. A procedure involving a simple acetic acid extraction was least effective in extracting acrosin from boar sperm. Storage of HGF acrosomal extract for 14 days at -196°C resulted in a higher enzyme activity than storage for 14 days at 5°C (P<0.05). In extracts from OEP-acetic acid procedures, enzyme activity was greater after storage for 14 days at 5°C than at -196°C. The correlation between enzyme activity (mU/106 sperm) and specific enzyme activity (Units/mg protein) within procedures was positive (P<0.001), when values were combined across length and temperature of storage.

Note:
ACKNOWLEDGMENTS The skillful technical assistance of Ms. Gwen Sowa and Mr. Kenneth Bender is gratefully acknowledged; thanks are due to Dr. Bernard Weinland for aid in the statistical analyses. This work was supported in part by a grant, No. HD07481, from the National Institutes of Health to Duane L. Garner. Mention of a trade name, proprietary product, or specific equipment does not constitute a guarantee or warranty by the USDA and does not imply its approval to the exclusion of other products that may be suitable.

Submitted on December 9, 1975
Accepted on March 15, 1976






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Copyright © 1976 by the Society for the Study of Reproduction.