Faculty Profiles - FUNAHASHI Hiroaki

MISC - FUNAHASHI Hiroaki

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HISTONE-H1 KINASE (H1K) ACTIVITY AND GLUTATHIONE (GSH) CONTENT OF PORCINE OOCYTES DURING PRONUCLEAR FORMATION AFTER IN-VITRO FERTILIZATION

H FUNAHASHI, TT STUMPF, TC CANTLEY, NH KIM, BN DAY

BIOLOGY OF REPRODUCTION 50 ( Supplement 1 ) 128 - 128 1994

Effects of sodium chloride concentration in maturation media on histone H1 kinase(H1K)activity and cytoplasmic maturation of porcine oocytes.(共著)

Journal of Reproduction and Fertility 72 ( Abstract Series 13 ) 38 1994

Histone H1 kinase activity after electrical activation of in vitro matured porcine oocytes.(共著)

Journal of Animal Science 72 ( Supplement 1 ) 74 1994

EFFECTS OF ELECTRICAL-STIMULATION BEFORE OR AFTER IN-VITRO FERTILIZATION ON SPERM PENETRATION AND PRONUCLEAR FORMATION OF PIG OOCYTES

H FUNAHASHI, TT STUMPF, SL TERLOUW, BN DAY

MOLECULAR REPRODUCTION AND DEVELOPMENT 36 ( 3 ) 361 - 367 1993.11

Language：English Publisher：WILEY-LISS

The effects of exposure of pig oocytes to an electrical pulse on sperm penetration and pronuclear formation were determined before or after in vitro fertilization (IVF). After in vitro maturation (IVM) or after collection from oviducts of unmated gilts, pig oocytes either were not exposed or were exposed to an electrical pulse (a 10 sec pulse at 4.0 V mm-1 AC followed by a 30 musec pulse at 120 V mm-1 DC), followed 30 min later by IVF. The incidence of male pronuclear formation of both IVM and in vivo-matured oocytes at 12 hr after insemination was decreased from 59% and 100%, respectively, to 2% and 36%, respectively, by the electrical pulse, but the penetration rates (88-100%) and polyspermic rates (79-100%) were not affected by exposure to an electrical pulse. Similarly, when pig IVM oocytes were exposed to an electrical pulse at 6 hr after insemination, electrical activation did not decrease penetration rates (93% vs. 90%), polyspermic rates (83% vs. 91%), or number of spermatozoa in penetrated oocytes (4.0 +/- 0.5 vs. 4.6 +/- 0.5) but did decrease the rate of male pronuclear formation from 58% to 18%. When oocytes were examined at 6 hr after insemination, 75% of them had been penetrated and resumed meiotic progression, but all sperm heads in penetrated oocytes were fully condensed or only partially decondensed. The percentage of penetrated eggs with multiple female pronuclei was increased when oocytes were exposed to an electrical pulse in all experimental series. In summary, electrical activation of pig oocytes before or just after IVF does not prevent sperm penetration but does inhibit male pronuclear formation and increases the formation of multiple female pronuclei. (C) 1993 Wiley-Liss, Inc.

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EFFECTS OF ELECTRICAL-STIMULATION BEFORE OR AFTER IN-VITRO FERTILIZATION ON SPERM PENETRATION AND PRONUCLEAR FORMATION OF PIG OOCYTES International journal

H FUNAHASHI, TT STUMPF, SL TERLOUW, BN DAY

MOLECULAR REPRODUCTION AND DEVELOPMENT 36 ( 3 ) 361 - 367 1993.11

Language：English Publisher：WILEY-LISS

DOI： 10.1002/mrd.1080360312

EFFECTS OF FOLLICULAR-FLUID AT FERTILIZATION IN-VITRO ON SPERM PENETRATION IN PIG OOCYTES International journal

H FUNAHASHI, BN DAY

JOURNAL OF REPRODUCTION AND FERTILITY 99 ( 1 ) 97 - 103 1993.9

Language：English Publisher：J REPROD FERTIL INC

The effects of porcine follicular fluid (PFF) on sperm penetration of pig oocytes and on prevention of polyspermy were examined and characteristics of spermatozoa exposed to PFF were determined. The addition of PFF at the level of 1 and 10% to the prefertilization and fertilization media decreased penetration rates and the mean number of spermatozoa in penetrated eggs regardless of the origin of PFF. In the presence of BSA, supplementation of 0.1% PFF to prefertilization and fertilization media and 1% PFF to prefertilization media did not decrease the penetration rates but did increase monospermic penetration to 54 and 68%, respectively. When PFF was added to prefertilization media, the number of spermatozoa binding to the zona and the percentage of acrosome-intact spermatozoa decreased with increased PFF concentration (from 43.1 +/- 2.8 and 73.1 +/- 4.9% to 7.2 +/- 1.3 and 15.7 +/- 15.4%, respectively). At the end of prefertilization incubation, sperm agglutination was observed and the degree depended on PFF concentration. Supplementation of fetal calf serum to prefertilization and fertilization media blocked the effects of PFF on sperm penetration and binding of spermatozoa to the zona. These results indicate that the prefertilization incubation of porcine spermatozoa in suitable concentrations of porcine follicular fluid will effectively reduce both the number of spermatozoa that attach to the surface of pig eggs and the incidence of polyspermy.

DOI： 10.1530/jrf.0.0990097

EFFECTS OF FOLLICULAR-FLUID AT FERTILIZATION IN-VITRO ON SPERM PENETRATION IN PIG OOCYTES

H FUNAHASHI, BN DAY

JOURNAL OF REPRODUCTION AND FERTILITY 99 ( 1 ) 97 - 103 1993.9

Language：English Publisher：J REPROD FERTIL INC

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EFFECTS OF THE DURATION OF EXPOSURE TO HORMONE SUPPLEMENTS ON CYTOPLASMIC MATURATION OF PIG OOCYTES INVITRO International journal

H FUNAHASHI, BN DAY

JOURNAL OF REPRODUCTION AND FERTILITY 98 ( 1 ) 179 - 185 1993.5

Language：English Publisher：J REPROD FERTIL INC

The effect of hormone supplements on cytoplasmic maturation in vitro was examined by incubating oocyte-cumulus complexes in a medium with PMSG (10 iu ml-1), hCG (10 iu ml-1) and oestradiol (1 mug ml-1) for various periods and then transferring them to medium without added hormones for the remainder of the maturation period. Exposure of oocyte-cumulus complexes to hormone supplements for 2 h improved only germinal vesicle breakdown and maturation rates compared with complexes not exposed to added hormones. The removal of hormone supplements at 20 h after the start of culture enhanced the ability of oocytes to form male pronuclei 10 to 12 h after insemination. Further, the effects of transfer of intact, and oocytectomized oocyte-cumulus complexes to hormone-free medium at 20 h on cumulus expansion were examined. The diameter and morphology of the intact oocyte-cumulus complexes were improved after the removal of oocyte-cumulus cell complexes from hormonal exposure. The responses of oocytectomized oocyte-cumulus complexes to hormone were similar to those of intact oocyte-cumulus complexes with the exception of corona radiata expansion. The results suggest that the removal of hormone supplements from maturation media at 20 h after culture enhanced cytoplasmic maturation and cumulus expansion. Further, cumulus expansion does not appear to depend on intercellular communication between cumulus cells and oocytes. Oocytectomy did influence expansion of the corona radiata during culture.

DOI： 10.1530/jrf.0.0980179

EFFECTS OF THE DURATION OF EXPOSURE TO HORMONE SUPPLEMENTS ON CYTOPLASMIC MATURATION OF PIG OOCYTES INVITRO

H FUNAHASHI, BN DAY

JOURNAL OF REPRODUCTION AND FERTILITY 98 ( 1 ) 179 - 185 1993.5

Language：English Publisher：J REPROD FERTIL INC

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EFFECTS OF DIFFERENT SERUM SUPPLEMENTS IN MATURATION MEDIUM ON MEIOTIC CYTOPLASMIC MATURATION OF PIG OOCYTES International journal

H FUNAHASHI, BN DAY

THERIOGENOLOGY 39 ( 4 ) 965 - 973 1993.4

Language：English Publisher：BUTTERWORTH-HEINEMANN

The temporal progression of meiotic and cytoplasmic maturation of pig oocytes cultured in a medium supplemented with 0.4% polyvinylalcohol (PVA), 10% fetal calf serum (FCS), 10% newborn piglet serum (NPS), 10% porcine follicular fluid (PFF) or 10% porcine seminal fluid (PSF) was examined after 20, 30, 40 and 50 hours of culture. There were no differences in germinal vesicle breakdown (GVBD) among FCS and NPS supplements. After 20 hours of culture, the frequency for GVBD was higher (P < 0.05) in FCS and NPS (54% and 52%, respectively) than in PVA (32%) and PFF (33%) culture media but were not different at 40 and 50 hours of culture. Supplementation with PSF resulted in a rapid chromosome condensation of pig oocytes after 20 hours culture, but all GVBD oocytes stopped developing at the condensed germinal vesicle stage. Oocytes were not penetrated by spermatozoa when inseminated following 20 hours of culture, while high penetration (87 to 100%) and polyspermy rates (86 to 100%) were consistently obtained in all the supplement groups when inseminated after 30, 40 or 50 hours of culture. Male pronuclear formation rates at 10 to 12 hours after insemination, following a 50-hour culture period in FCS and NPS, were 28 and 28%, respectively, in comparison with 54% in PVA and 59% in PFF. The results indicate that supplementing maturation media with serum such as FCS and NPS reduced the ability of pig oocytes to form a male pronucleus, and further suggest that the detrimental effects may be due to accelerated progression of maturation events.

DOI： 10.1016/0093-691X(93)90433-6

GLUCOSE REQUIREMENT AT DIFFERENT DEVELOPMENTAL STAGES OF INVITRO FERTILIZED BOVINE EMBRYOS CULTURED IN SEMI-DEFINED MEDIUM International journal

JH KIM, H FUNAHASHI, K NIWA, K OKUDA

THERIOGENOLOGY 39 ( 4 ) 875 - 886 1993.4

Language：English Publisher：BUTTERWORTH-HEINEMANN

Three experiments were conducted in which 2-cell bovine embryos were prepared from oocytes, obtained from abattoir ovaries, by in-vitro maturation for 22 to 24 hours, followed by exposure to spermatozoa for 8 hours and culture for 40 hours within the cumulus. The cumulus cells were then removed, and the cleaved embryos were cultured for a further 120 hours or longer, in the presence or absence of glucose, pyruvate and lactate. Very few embryos developed in the complete absence of energy substrates. Lactate and pyruvate, alone or combined, supported development to the 8-cell stage, but pyruvate was required to support development to the morula stage (Experiment 1). When present throughout culture or when added at 48 or 96 hours postinsemination, 5.56 mM glucose was detrimental to development (Experiments 1 and 2). However, when added at 120 hours postinsemination, 5.56 mM glucose improved development to the blastocyst and expanded blastocyst stages, compared with no glucose or 11.12 mM glucose (Experiment 3).

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EFFECTS OF DIFFERENT SERUM SUPPLEMENTS IN MATURATION MEDIUM ON MEIOTIC CYTOPLASMIC MATURATION OF PIG OOCYTES

H FUNAHASHI, BN DAY

THERIOGENOLOGY 39 ( 4 ) 965 - 973 1993.4

Language：English Publisher：BUTTERWORTH-HEINEMANN

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GLUCOSE REQUIREMENT AT DIFFERENT DEVELOPMENTAL STAGES OF INVITRO FERTILIZED BOVINE EMBRYOS CULTURED IN SEMI-DEFINED MEDIUM

JH KIM, H FUNAHASHI, K NIWA, K OKUDA

THERIOGENOLOGY 39 ( 4 ) 875 - 886 1993.4

Language：English Publisher：BUTTERWORTH-HEINEMANN

DOI： 10.1016/0093-691X(93)90425-5

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Assessment of porcine oocytes using brilliant cresyl blue.(共著)

Theriogenology 39 ( 1 ) 214 1993

Premature chromosomal condensation of nuclei transplanted into in vitro matured porcine oocytes.(共著)

Biology of Reproduction 48 ( Supplement 1 ) 175 1993

Pronuclear formation and in vitro development folowing artificial activation of pig oocytes matured different culture media.(共著)

Biology of Reproduction 48 ( Supplement 1 ) 171 1993

Effects of hormonal conditions during last 20 h of in vitro maturation on cytoplasmic maturation and vitro fertilization of pig oocytes.(共著)

Journal of Animal Science 71 ( Supplement 1 ) 70 1993

Pig oocyte activation and processing of transplanted nuclei.(共著)

39 ( 1 ) 329 1993

Processing of nuclei transplanted into in vitro matured porcine oocytes.(共著)

Theriogenology 39 ( 1 ) 322 1993

Effects of artificial activation before in vitro fertilization on sperm penetration and pronuclear formation in pig oocytes.(共著)

Theriogenology 39 ( 1 ) 222 1993

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