Flow cytometric evaluation of spermatozoan nuclear chromatin structure and viability Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/1544bs84v

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  • I. Frozen bovine semen samples were measured by flow cytometry (FCM) to determine if FCM analyses could be useful for sperm quality evaluation and prediction of bull fertility. Fluorescent dyes utilized were acridine orange, to measure sperm nuclear chromatin heterogeneity by quantifying susceptibility of the chromatin to denaturation, and rhodamine 123, to measure proportion of viable sperm by quantifying mitochondrial membrane potentials. Increasing chromatin heterogeneity was negatively correlated with fertility rating (FR) of the bulls. Proportion of live sperm was positively correlated with FR and negatively correlated with heterogeneity of the chromatin. Multiple regression equations for prediction of bull fertility were developed using using FCM measurements and light microscopic evaluation of sperm quality. Results indicate FCM analyses of sperm chromatin structure and viability are useful for detection of low fertility sires and prediction of young sires' future reproductive performance. II. Genetic factors affecting epididymal sperm head morphology, sperm nuclear chromatin structure and proportions of testicular cell types were estimated using a full diallel cross of four inbred lines of mice. Lines were chosen to provide a wide range in levels of morphologically abnormal sperm cells (ABN) scored by light microscopy. Sperm and testicular cells were measured by flow cytometry (FCM) after staining with acridine orange. Measurements collected by FCM included percentage testicular tetraploid, diploid and haploid cells, and heterogeneity of sperm chromatin structure, evaluated by susceptibility of the DNA to denaturation. Statistical analyses provided estimates of line, heterosis, maternal and reciprocal effects and general and specific combining abilities on body and testis weights, ABN and FCM-derived measurements. Heterosis and line effects were significant for testis weight, sperm chromatin heterogeneity and ABN. A positive correlation was obtained between ABN and heterogeneity of sperm chromatin. Flow cytometric staining patterns of sperm provide information on the alteration of chromatin structure associated with increased proportions of abnormal sperm.
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