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| Objective 1: To study the inheritance of semen quality, freezability and fertility of crossbred bulls based on available information |
| Approach to research: |
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Routine semen analysis and biochemical (HOS, MBRT) parameters will be evaluated for different crossbred bulls (Frieswal, Karan Fries and Vrindavani) |
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Seasonal variation in semen quality (biochemical parameters) will be investigated |
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Pedigree information of semen quality, and freezability fertility of crossbred bulls and bull wise rejection rate will be collected |
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Familial relationship on basic information of inheritance of semen quality, freezability and fertility of crossbred bulls as well as bull-wise rejection rate will be calculated using appropriate statistical procedure |
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| Activities under the objective: |
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Collection of pedigree information on bull wise rejection rate (Frieswal, Karan Fries and Vrindavani) (to be done by PDC, NDRI & IVRI respectively) |
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Collection of semen (at 30±3 months) from bulls and evaluation of semen for routine (volume, concentration, initial progressive motility, post thaw motility, livability) and biochemical (HOS, MBRT) parameters in two different seasons (to be done by PDC, NDRI & IVRI) |
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| Objective 2: To study the global gene expression of sperm of crossbred bulls for identifying the upregulated and downregulated genes |
| Approach to research: |
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Micro array analysis to study the global gene expression of crossbred bulls |
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Validation of micro array results by Real Time PCR |
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| Activities under the objective: |
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Categorization of bulls based on semen parameters into two groups (good and poor) ) ( to be done by PDC, NDRI & IVRI) |
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Collection of testicular biopsy material after completion of their target semen doses in two different seasons ) ( to be done by PDC, NDRI & IVRI) |
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Micro array experiments for global expression profiling of testicular genes ) ( to be done by PDC & IVRI) |
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Validation of micro array results by Real Time PCR to study some prominent upregulated and downregulated genes (as revealed by Microarray analysis) involved in important pathways (motility & fertilizing ability, energy metabolism, membrane integrity, functional ion channels etc.) ( to be done by PDC & IVRI) |
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| Objective 3: To study the effect of crossbred semen on embryonic development with special reference to Y chromosome microdeletion and epigenetic modification) ( to be done by NDRI) |
| Approach to research: |
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Standard IVF protocol will be used to generate embryos using semen samples from bulls (good and poor in terms of produced semen quality) under test |
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Y-chromosomal micro-deletions will be performed by normal / multiplex (at least duplex) PCR amplification of Y chromosomal DNA |
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DNA will be isolated from sperm and testicular samples as above and from different embryonic stages to study imprinting status of H19 and SNRPN genes by using bisulfite-sequencing approach |
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| Activities under the objective: |
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Detection of Y chromosome microdeletions in selected crossbred bulls |
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IVF of cattle oocytes using sperm samples from bulls (good and poor motility status) under test |
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Imprinting status of two important genes (H-19 and SNRPN) in the testicular tissues and ejaculated spermatozoa of bulls varying in initial sperm motility |
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Establishing the fertilizing ability of spermatozoa (following IVF) from bulls under study and its relationship with observed Y chromosome microdeletion and methylation status of genes under study |
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