Chromosomal analysis of bovine spermatozoa by in vitro fertilization and its application for separation of X- and Y-bearing spermatozoa.
| Project/Area Number |
63560298
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Applied veterinary science
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| Research Institution | Hokkaido University |
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Principal Investigator |
HISHINUMA Mitsugu (1989) Faculty of Veterinary Medicine, Hokkaido University, Instructor, 獣医学部, 助手 (30183578)
高橋 芳幸 (1988) 北海道大学, 獣医学部, 助教授 (70167485)
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| Co-Investigator(Kenkyū-buntansha) |
TAKAHASHI Yoshiyuki Faculty of Veterinary Medicine, Hokkaido University, Assistant Professor, 獣医学部, 助教授 (70167485)
菱沼 貢 北海道大学, 獣医学部, 助手 (30183578)
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| Project Period (FY) |
1988 – 1989
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| Project Status |
Completed (Fiscal Year 1989)
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1989: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1988: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Keywords | In vitro fertilization / Cow / Spermatozoa / Chromosome / Hamster |
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| Research Abstract |
1. Fertilization rate of hamster oocytes inseminated with Ca-Ionophore A23187 (IA) treated spermatozoa
Zona-free hamster oocytes were inseminated with bovine spermatozoa preincubated with 0.1 muM IA for 30-60 sec. When the oocytes were inseminated with spermatozoa treated in a medium with caffeine, the fertilization rates were different depending on the sire. However, when the spermatozoa treated in a medium with 15 mM theophyllin were used, constantly high fertilization rate (more than 80%) was obtained. A sperm concentration of 2-5x10^7/ml gave high fertilization rate.
2. Detection of metaphase plate of bovine spermatozoa
When 0.5 muM IA treated spermatozoa were incubated with hamster oocytes for 0.5-1.0 hour, fertilization and sex determination rates were 80-90% alid 25-35%, respectively. When a number of penetrated sperm increased, the development rate of sperm to metaphase plate decreased. A higher number of metaphase plate was obtained when two spermatozoa penetrated as compared to
… More
monospermy (70.6% VS 57.9%). This suggests that it is possible to improve the detection rate by incubating the oocyte and spermatozoa under the condition in which 1-3 spermatozoa penetrate.
3. Detection of Y-bearing spermatozoa and the separation of X- and Y-bearing spermatozoa by Parcoll density gradient centrifugation
F-body was not detected in bovine sperm and lymphocyte by the quinacrine mustard fluorescence technique. Interspecific in vitro fertilization has been carried out using the spermatozoa obtained from each fraction of various Parcoll density gradients. on account of variation in activity and concentration of sperm, stable fertilization rate has not been obtained yet. This study is still being carried out.
4. Chroinosomal examination of bovine spermatozoa by interspecific in vitro fertilization
Spermatozoa collected from one sire showed hyperploidy (31,Y) in a metaphase plate. This suggests that this technique can be used for the study on chromosome aberration of bovine spermatozoa. Appearance rate of chromosome aberration using spermatozoa from various sires is being researched. Less
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Report
(3 results)
Research Products
(11 results)
