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Flow Cytometric Determination of the Proportions of X- and Y-Chromosome-Bearing Sperm in Samples of Purportedly Separated Bull Sperm^1,2,

Lawrence Livermore National Laboratory, University of California, Livermore 94550³; Oklahoma State University, Stillwater 74078^,3 and US Department of Agriculture, Beltsville, MD 207055

Abstract

A rapid assay for determining the proportions of X- and Y-chromosome-bearing sperm in semen samples would benefit research aimed at sex ratio control through sperm separation. It also would be of value for quality control should a separation technique be developed. Flow cytometric methods capable of measuring sperm DNA content precisely enough to resolve and quantify the X and Y populations in many mammalian species have been developed. They are effective for fresh and cryopreserved sperm of most domestic animals. Results are reported of flow cytometric analyses of bull sperm samples from seven commercial and academic sources after processing with procedures purported to separate the X and Y populations. In no case was enrichment of either sperm population observed. Breeding trials carried out by the sources of two of the sets of samples showed these procedures were ineffective in altering the sex ratio.

¹ Work performed under the auspices of the U.S. Department of Energy by the Lawrence Livermore National Laboratory under contract number W-7405-ENG-48 and Oklahoma State Univ. (Cooperative Agreement 12-14-7001-114 with USDA, Beltsville, MD).

² We thank the various sources that supplied semen samples for testing. We appreciate the cooperation of Drs. J. J. Sullivan and M. M. Pace of American Breeders Service, DeForest, WI, who supplied cryopreserved bull semen for use in development and testing of our techniques and for measurement standards.

³ Biomedical Sci. Div., P.O. Box 5507, L-452.

⁴ Dept. of Physiol. Sci., College of Vet. Med.

⁵ Reprod. Lab., Anim. Sci. Institute, Agr. Res. Center, ARS.

⁶ This document was prepared as an account of work sponsored by an agency of the U.S. Government. Neither the U.S. Government nor the Univ. of California nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product, or process disclosed, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial products, process, or service by trade name, trademark, manufacturer, or otherwise, does not necessarily constitute or imply its endorsement, recommendation, or favoring by the U.S. Government or the Univ. of California. The views and opinions of authors expressed herein do not necessarily state or reflect those of the U.S. Government thereof, and shall not be used for advertising or product endorsement purposes.