Characterization and Gene Expression Profiling of Epididymal Sperm Collected from Dead Mithun (Bos Frontalis) Bulls and its Preservation
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Title |
Characterization and Gene Expression Profiling of Epididymal Sperm Collected from Dead Mithun (Bos Frontalis) Bulls and its Preservation
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Creator |
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M. Mondal KK. Baruah A. Chatterjee MK. Ghosh |
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Subject |
Epididymal sperm; Mithun (Bos frontalis); Cryopreservation; Conservation; Fertility; Gene expression.
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Description |
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Conserving the mithun (Bos frontalis), a rare bovine of south-east Asia, could benefit from effective ex situ genetic management and breeding programmes including the use of assisted reproduction. Longterm storage of epididymal sperm collected from dead mithun bulls by cryopreservation, with high survival rate, is essential for the establishment of genetic resource banks of this rare species. A study was therefore conducted to (i) characterize the epididymal sperm collected from dead mithun bulls, (ii) investigate the effectiveness of epididymal sperm for cryopreservation and (iii) to study the expression pattern of genes related to motility (TSSK6 and ADAM5P ) and fertility (PRM1, PRM2, PRM3, Tnp1 and Tnp2) in mithun epididymal sperm. For the purpose, sperm collected from caudal epididymis of eight dead mithun bulls were evaluated for concentration, progressive motility, morphological abnormalities, live sperm counts, acrosome integrity, membrane stability (hypo-osmotic swelling test; HOST) and DNA integrity. Epididymal sperm were cryopreserved using tris-egg yolk-glycerol diluent in liquid nitrogen. Post-thaw quality of the cryopreserved sperm in terms of progressive motility, morphological abnormalities, live cell counts, acrosome integrity, membrane stability and DNA integrity were assessed. The RNA extracted from fresh and post-thawed cryopreserved epididymal sperm was reverse transcribed to cDNA and expressions of the genes related to motility and fertility were determined by RT-PCR. The progressive motility, live cell counts, morphological abnormalities and acrosome integrity (normal) of fresh sperm were 89.9±2.7, 88.7±6.7, 7.8±1.07 and 95.3±7.8%, respectively. Fresh sperm that responded to HOST were 88.3±7.2%, and 89.5±6.4% fresh sperm had intact DNA. Following cryopreservation, the percentage of progressive motility (fresh vs frozen-thawed), live cell counts, morphological abnormalities, acrosome integrity, membrane stability and DNA integrity were found to decrease significantly (P Not Available |
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Date |
2019-03-20T09:31:26Z
2019-03-20T09:31:26Z 2013 |
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Type |
Research Paper
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Identifier |
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2231-1238 http://krishi.icar.gov.in/jspui/handle/123456789/17552 |
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Language |
English
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Relation |
Not Available;
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Publisher |
Research India Publications
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