Molecular Detection Of Programmed Cell Death (Apoptosis In Fresh And Cryopreserved Buffalo Sper Matozoa)
By: Daulat Raheem Khan
| Prof. Dr. Nasim Ahmad.
Contributor(s): Dr. Irshad | Dr. Muhammad Amir Saeed | Faculty of Veterinary Sciences.
Material type: 
BookPublisher: 2006Subject(s): Department of TheriogenologyDDC classification: 0933,T Dissertation note: The objectives of present study were a) to validate annexin V/Pl assay, for buffalo sperm, using graded doses of camptothecin through fluorescent microscopy (Exp. 1) and b) to determine the effect of stages of cryopreservation on apoptosis (PS externalization, using annexin V/PT assay), motility and plasma membrane integrity in buffalo sperm (Exp.2). In the first experiment graded doses of camptothecin were used (n=2) in different aliquots of sperm suspension (1x106/mL) for induction of apoptosis. Higher dose levels of camptothecin (10.0 iM and 20 jiM) resulted in inducing apoptosis (P<O.05) compared to the lower (5tM) dose or control. In the second experiment semen samples (n=9, from three bulls) were cryopreserved using vapor freezing. Annexin V/PI assay for apoptosis, visual assessment for percentage motility and hypo-osmotic swelling test for plasma membrane integrity were employed at various stages of cryopreservation (i.e. fresh, after equilibration and post thaw). The mean percentage of apoptotic, necrotic and viable sperm did not differ between fresh and after equilibration stages. However, freezing and thawing increased (P<0.05) the percentage of apoptotic sperm (25.4±0.6 vs. 36.5±1 .9) while decreased (P<0.05) the necrotic (29.7±0.7 vs. 35.1±1.2) and viable sperm (37.2±1.3 vs. 32.8±1.9, (P<0.07). Similarly, freezing and thawing decreased (P<0.05) the mean percentage motility and plasma membrane integrity, compared to other stages. Based upon the difference in initial and post thaw values of two variables (percent motility and percent apoptosis) it is concluded that apoptosis contributes more than 50% sperm motility loss during the process of freezing and thawing in buffalo.
| Item type | Current location | Collection | Call number | Status | Date due | Barcode | Item holds |
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Thesis
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UVAS Library Thesis Section | Veterinary Science | 0933,T (Browse shelf) | Available | 0933,T |
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The objectives of present study were a) to validate annexin V/Pl assay, for buffalo sperm, using graded doses of camptothecin through fluorescent microscopy (Exp. 1) and b) to determine the effect of stages of cryopreservation on apoptosis (PS externalization, using annexin V/PT assay), motility and plasma membrane integrity in buffalo sperm (Exp.2). In the first experiment graded doses of camptothecin were used (n=2) in different aliquots of sperm suspension (1x106/mL) for induction of apoptosis. Higher dose levels of camptothecin (10.0 iM and 20 jiM) resulted in inducing apoptosis (P
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