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Title: | Characterization and Gene Expression Profiling of Epididymal Sperm Collected from Dead Mithun (Bos Frontalis) Bulls and its Preservation |
Other Titles: | Not Available |
Authors: | Not Available M. Mondal KK. Baruah A. Chatterjee MK. Ghosh |
ICAR Data Use Licennce: | http://krishi.icar.gov.in/PDF/ICAR_Data_Use_Licence.pdf |
Author's Affiliated institute: | ICAR::National Dairy Research Institute ICAR::National Research Centre on Mithun |
Published/ Complete Date: | 2013 |
Project Code: | Not Available |
Keywords: | Epididymal sperm; Mithun (Bos frontalis); Cryopreservation; Conservation; Fertility; Gene expression. |
Publisher: | Research India Publications |
Citation: | Not Available |
Series/Report no.: | Not Available; |
Abstract/Description: | 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<0.01) with a motility recovery rate of 75±8.5%. Transcripts encoding a serine/threonine testis-specific protein kinase (TSSK6) and a metalloproteinase non coding RNA (ADAM5P ) were found to be associated with high-motility status (P<0.01) and their relative expressions were higher in fresh than frozen-thawed sperm. There were no differences in relative expressions for the transcripts related to fertility (PRM1, PRM2, PRM3, Tnp1 and Tnp2) between fresh and frozen-thawed sperm indicating no change of fertility of mithun epididymal sperm after cryopreservation. Taken together, results of our study provide a foundation of collection and cryopreservation of epididymal sperm from dead mithun bulls. Furthermore, the post-thaw quality in terms of percentage of progressive motility, live cell counts, morphological abnormalities, acrosome integrity, membrane stability and DNA integrity were sufficient for genetic improvement of this rare species through AI and conservation of the valuable germplasm for future. |
Description: | Not Available |
ISSN: | 2231-1238 |
Type(s) of content: | Research Paper |
Sponsors: | Not Available |
Language: | English |
Name of Journal: | International Journal of Biotechnology and Bioengineering Research. |
Volume No.: | Volume 4, Number 6 (2013) |
Page Number: | 535-542 |
Name of the Division/Regional Station: | Not Available |
Source, DOI or any other URL: | http://www.nrcmithun.res.in/GlimpseOfRP/9.pdf |
URI: | http://krishi.icar.gov.in/jspui/handle/123456789/17552 |
Appears in Collections: | AS-NRCM-Publication |
Files in This Item:
File | Description | Size | Format | |
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Characterization and Gene Expression Profiling of Epididymal Sperm Collected from Dead Mithun (Bos Frontalis) Bulls and its Preservation.pdf | 455.02 kB | Adobe PDF | View/Open |
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