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The ability of freeze-dried bull spermatozoa to induce calcium oscillations and resumption of meiosis
Abdalla, H
Hirabayashi, A
Hochi, S
Bovine spermatozoa
Calcium oscillations
Freeze-drying
Homogenous ICSI
Interspecies ICSI
The objective was to investigate the ability of freeze-dried (FD) bull spermatozoa to induce calcium oscillations in mouse oocytes and meiosis resumption in in vitro-matured bovine oocytes after intracytoplasmic sperm injection (ICSI). Bull spermatozoa were freeze-dried and stored for 1 y at +25, +4, or -196 degrees C. In the first experiment, rehydrated sperm heads were microinseminated into hybrid mouse oocytes loaded with fluo-3/AM, and the kinetics of intracellular calcium concentration was monitored for I h. Repetitive increases of intracellular calcium concentration were recorded in the majority of injected oocytes, with exception of a few oocytes injected with FD sperm heads stored at +4 degrees C (11%) and +25 degrees C (8%) that exhibited a single increase or no response (non-oscillated). The proportion of oocytes that oscillated with high frequency (>= 10 spikes/h) was higher in the non-dried control group (79%; P < 0.05) than in the FD groups (58, 55, and 54% for storage at -196, +4, and +25 degrees C, respectively). In the second experiment, control and FD spermatozoa were microinseminated into in vitro-matured, denuded bovine oocytes. The oocytes were fixed and stained 12 h after ICSI. A higher proportion of bovine oocytes injected with control spermatozoa (70%; P < 0.05) resumed meiosis than those injected with +25, +4 and -196 degrees C stored FD spermatozoa (53, 48, and 57%, respectively). The proportion of ICSI oocytes that developed to the pronuclear stage (complete activation) was higher in the control group (64%; P < 0.05) than those in all the FD groups (34, 27, and 28% for storage at -196, +4, and +25 degrees C, respectively). Thus, the ability of bull spermatozoa to induce frequent intracellular calcium spikes in mouse oocytes was impaired by the process of freeze-drying, without differences among storage at +25, +4 or -196 degrees C, probably resulting in a lower proportion of bovine oocytes that resumed meiosis and/or developed to the pronuclear stage.
Article
THERIOGENOLOGY. 71(3):543-552 (2009)
journal article
ELSEVIER SCIENCE INC
2009-02
application/pdf
THERIOGENOLOGY
3
71
543
552
0093-691X
AA00862972
https://soar-ir.repo.nii.ac.jp/record/13411/files/THE71_3_543.pdf
eng
18845331
http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?CMD=search&DB=pubmed&term=18845331
10.1016/j.theriogenology.2008.08.021
https://doi.org/10.1016/j.theriogenology.2008.08.021