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SEPARATION OF BOVINE X- FROM Y-CHROMOSOME BEARING SPERMATOZOA USING MONOCLONAL H-Y ANTIBODIES
Abstract
Sixty-three samples of semen were collected from 11 bulls using an electroejaculator. The samples were washed and the sperm concentration was determined with a hemacytometer. The spermatozoa were treated with monoclonal H-Y antibody, fluorescein conjugated goat-anti mouse F(ab)2 fragments, and the percentage of spermatozoa labeled (fluorescing) was determined by analyzing 300 spermatozoa/sample under a fluorescent microscope. The percentages of spermatozoa labeled ranged from 40-65 for the treated samples and from 1-15 for the corresponding controls. Labeling was confined to the post-acrosomal region except when the acrosome was disrupted then, the acrosomal area was also labeled. A fluorescence activated cell sorter (FACS) was used to confirm the percentage of labeled spermatozoa. The FACS was also used to separate labeled from non-labeled spermatozoa. Control, labeled and nonlabeled samples from 3 bulls were prepared using a Feulgen staining process and the DNA content of each cell was analyzed using a scanning microdensitometer. Results obtained from analysis of control samples showed two distinct peaks. The peak with the smaller amount of DNA was assumed to represent Y-bearing spermatozoa and the other X-bearing spermatozoa. The mean difference between the 2 peaks was 3.95% of the 3 bulls. Results obtained from analysis of labeled samples showed 24:76, 12:88, and 23:77 while results obtained from analysis of non-labeled samples showed 74:26, 65:35, and 77:23 ratios of X- to Y-bearing spermatozoa for bulls 9, 10, and 11, respectively. The proportion of X- to Y-bearing spermatozoa in the control samples was not different (P > .5) among bulls while they were different (P < .001) between the treated and the corresponding control samples. These results were interpreted to indicate that monoclonal H-Y antibody (MoAb2) could be used to increase the proportion of X-bearing spermatozoa in a bovine semen sample since the H-Y antigen is apparently expressed on the surface membrane of the haploid Y-bearing spermatozoa.
Subject Area
Livestock
Recommended Citation
ALI, JAWAD ISMAIL, "SEPARATION OF BOVINE X- FROM Y-CHROMOSOME BEARING SPERMATOZOA USING MONOCLONAL H-Y ANTIBODIES" (1986). ETD collection for University of Nebraska-Lincoln. AAI8624577.
https://digitalcommons.unl.edu/dissertations/AAI8624577