• Journal of Veterinary Clinics
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• v.19 no.3
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• pp.277-282
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• 2002

This study was designed to evaluate the effects of anesthetics on waveform of SEPs and to authorize possible anesthetic protocol for measurement of the somatosensory evoked potentials (SEPs). Thirteen anesthetic methods were used. The SEPs were recorded on two channels (between the 5th and 6th lumbar vertebra as the channel 1 and between the 11th and 12th thoracic vertebra as the channel 2) following stimulation of posterior tibial nerve. ID analyze SEPs wave, latency and conduction velocity were measured. Among thirteen anesthetic methods, standard SEPs waveforms were observed in dogs anesthetized with following six methods: Acepromazine + Thiepfntal Na + Isoflurane, Acepronazine + Propofol + Isoflurane, Diazepam + Xylazine, Xylazine + Ketamine, Acepromazine + Propofol infusion and Propofol infusion. Above six methods could be used with sufficient anesthetic depth. The differences of latency and conduction velocity among six groups were minimal compared to general waveform of SEPs. These results indicate that the six anesthetic methods can be used for recording SEPs in the dog. In particular, Diazepam + Xylazine and XylaBine + Ketamine as injectable anesthesia are considered more convenient than other four methods in veterinary medicine.

• Development and Reproduction
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• v.4 no.2
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• pp.161-173
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• 2000

The aim of this study was to assess the effect of the frequency of the L$N_2$ infusion on the ultrastructure, metabolism and development of the embryo after freezing and thawing by computerized cell freezer. Two-cell embryos of ICR mouse were randomly allocated into fresh (control), high-frequency freezing (group 1) and low-frequency freezing (group 2). For fresh and frozen-thawed intact 2-cell embryos, total ceil number in the blastocyst was counted by fluorescent microscope after Hoechst 33258 staining. Relative amount of $H_2O$$_2$ was measured by DCHFDA. Intracellular location and membrane potential of the mitochondria were evaluated by staining with rhodamine 123 and JC-1. The structure of actin filament was also evaluated by confocal microscope. DNA fragmentation was assessed by TUNEL method after development into blastocyst. The survival rate of intact embryo was higher in group 1 than group 2 (50.7％ vs. 34.6％ respectively, p<0.05). The blastocyst developmental rate was significantly low in group 2 (86.7％, 76.7％ vs. 44.0％ for control, group 1 and group 2 respectively, p<0.05). Total cell number in the blastocyst was also significantly lower in group 2 than control (79.5$\pm$12.9, 71.6$\pm$8.0, and 62.5$\pm$4.7 for control, group 1 and group 2 respectively, p<0.05). The relative amount of $H_2O$$_2$ was higher in group 2 than other groups (15.3$\pm$3.0, 16.6$\pm$1.6 vs. 23.4$\pm$1.8, p<0.05). After JC-1 staining, relative intensity of mitochondria with high membrane potential was significantly lower in group 2 than control and group 1 (17.2$\pm$3.8, 17.4$\pm$1.3 vs. 13.2$\pm$2.0, p<0.05). In group 2, partial deletion and aggregation of the actin filament was found. DNA fragmentation rate was also hieher for group 2 versus other groups (30.8％, 36.0％ vs. 65.6％, p<0.05). The frequency of the L$N_2$ infusion is an important factor for the development of frozen-thawed mouse embryo. High-frequency infusion may prevent damages of cytoskeleton and mitochondria in the embryo probably by preventing the temperature fluctuation during dehydration phase. We speculate that the application of high-frequency infusion method in human embryo may be promising.