Journal of Reproduction and Development
Online ISSN : 1348-4400
Print ISSN : 0916-8818
ISSN-L : 0916-8818
Volume 41, Issue 1
February
Displaying 1-16 of 16 articles from this issue
Original Articles
  • Kaoru TAKAGI, Kiyoshi TSUCHIYA, Shigehisa TSUMAGARI, Yuzou KITAZIMA, M ...
    1995 Volume 41 Issue 1 Pages 1-5
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    Application of DNA fingerprinting in evaluating parent/offspring relationship (pedigree) in Thoroughbred horses was attempted with Myo probe. From blood samples of 22 Thoroughbred horses (4 sires, 9 dams and 9 offsprings), DNA was isolated prior to digestion with the restriction endonuclease, Hinf I. A total of 7-16 (11.5 ± 2.1) bands of the DNA fragments were detected with the use of 32P-labeled Myo probe. Of these, 2 bands of all DNA samples coincided at 2.0 and around 3.8 kb. Among 9 parent/offspring pairs, bands from the offsprings well coincided with those of either the sire or dam, advocating that the application of Myo probe for verifying the parent/offspring relationship by the DNA fingerprinting method was possible. Furthermore, to enhance the reliability of parent/offspring verification in Thoroughbred horses by the DNA fingerprinting method, use of numerous probes and restriction endonuclease is indispensable.
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  • Hiroshi IMAHIE, Eimei SATO, Yutaka TOYODA
    1995 Volume 41 Issue 1 Pages 7-14
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    Progesterone induced in vitro parthenogenetic activation of ovulated mouse oocytes. Progesterone (100 μM, 31.4 μg/ml) induced a high rate (90%) of activation and progression of the second meiotic division from metaphase to anaphase/telophase and pronuclear formation. Oocytes degenerated at progesterone concentrations of 150 μM and above, whereas 50 μM concentrations or less induced quite little or no activation. Aging of the oocytes prior to treatment and a longer treatment period had significant effects on activation. Oocytes treated with 100 μM progesterone at 16, 18, 20, or 22 h after hCG injection for 6 h showed 50, 85, 89 and 98% activation, respectively, while the effect on activation was less significant when these oocytes were treated for 2 or 4 h. Few oocytes (0-4%) were activated in controls that were incubated in the presence of solvent (0.5% DMSO) only. The presence of cumulus cells did not exert significant effects on the incidence of activation by progesterone. Other steroids tested were cholesteryl sodium sulfate, pregnenolone, 17α-hydroxy-20β-dihydroprogesterone, 17α-hydroxyprogesterone, 20α-hydroxy-progesterone, 20β-hydroxyprogesterone, androstenedione, testosterone, and estradiol-17β. Both 20α-hydroxyprogesterone and 20β-hydroxyprogesterone induced activation, but the remainder induced low or no activation. When culture was continued after progesterone exposure, 60% of oocytes developed to the 2-cell stage, and a small proportion (3%) developed to the blastocyst stage. These results show that when progesterone and its metabolites are present at high concentrations (100 μM) for a long period (6 h), they have the unique property of inducing in vitro parthenogenetic activation in mouse oocytes.
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  • Shinichiro KOYAMA, Tsutomu HASHIZUME, Shinichi OHASHI, Shigeto KANEMAT ...
    1995 Volume 41 Issue 1 Pages 15-20
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    The effect of pituitary adenylate cyclase-activating polypeptide (PACAP) on the release of luteinizing hormone (LH) from cultured bovine anterior pituitary cells was determined and compared with that of luteinizing hormone-releasing hormone (LHRH). PACAP 27 significantly increased the release of bovine LH at a dose as low as 10-14 M. PACAP 38 also significantly increased the release of LH and the minimal effective dose was 10-12 M. Maximal LH release induced by PACAP 27 (10-7 M) and PACAP 38 (10-7 M) were 40.5% and 71.6%, respectively of control levels. LHRH significantly increased the release of LH at a dose as low as 10-14 M. The increase in LH reached a maximum level which was 3-fold above the basal level at the highest concentration of LHRH tested (10-7 M, P<0.001). LH release induced by PACAP 27, PACAP 38, and LHRH was in a dose-dependent manner. The results suggest that PACAP 27, PACAP 38, and LHRH stimulate the release of LH in the cattle by acting directly at anterior pituitary cells.
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  • Shiro KURUSU, Yumi HIRANO, Saori KAWAGISHI, Tsuzumi KITABATAKE, Akinor ...
    1995 Volume 41 Issue 1 Pages 21-28
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    To investigate the role of ovarian endogenous eicosanoids in regulation of luteal function, we examined effects of local and chronic administration of eicosanoid synthesis inhibitors on plasma progesterone levels in immature pseudopregnant rats. Injections of drugs in 20 μl oil were performed into the ovarian bursal cavity and repeated three times every 36 h. As compared with the control (oil-injected) group, the phospholipase A2 inhibitor dexamethasone (DEX) at the dose of 62.5 μg/ovary delayed functional luteolysis for 3 days by treatments from day 6 of pseudopregnancy (D6). Indomethacin (IND) also extended luteal activity for 5 days after treatment from D6, while nordihydroguaiaretic acid (NDGA) extended it for only 1 day. The extension of pseudopregnancy by each drug was not so evident with treatments from D2 or D4 as compared to that from D6. From D6, imidazole also delayed luteolysis, while a doubled dose of NDGA did not. Systemic administration of DEX or IND increased plasma progesterone levels more rapidly and significantly than local treatments but were less effective in delaying luteolysis. These results suggest that ovarian endogenous eicosanoids, mainly prostanoids, in the late luteal phase are involved in functional luteolysis in the present model.
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  • Takahito HARA, Satoshi TANAKA, Hiroshi SATO, Motoharu SEIKI, Hideki KO ...
    1995 Volume 41 Issue 1 Pages 29-39
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    The matrix metalloproteinase-11 (MMP-11) is a member of the family of matrix metalloproteinases, and recently discovered by Bassett et al. (1990) in placentae and invasive mammary tumors of humans. Although propositions have been made suggesting that MMP-11 might play a crucial role in the metastatic invasion of cancer cells into the surrounding tissue, neither the enzymatic characteristics nor precise biological functions of MMP-11 have been clearly elucidated. Since trophoblast cells of mammalian embryos exhibit different degree of invasiveness during placentation in different groups of animals, it will be of interest to examine whether MMP-11 is present in animals which develop placentae of non-invasive type. We demonstrated that MMP-11 genes are present in the genome of the Shiba goat which develops placentae of non-invasive type (epitheliochorial or syndesmochorial), and determined a partial sequence of MMP-11 cDNA of the Shiba goat. MMP-11 genes are strongly expressed in the uterus of immature Shiba goats under the influence of ovarian estrogens (following PMSG and hCG treatments). In contrast to the human placenta where MMP-11 genes are abundantly expressed, the caprine placenta (Day 72 and Day 112 of gestation) contains MMP-11 mRNA, if at all, only at extremely low levels. The reason underlying the difference in the patterns of expression of MMP-11 genes in human and caprine placentae remains yet to be clarified. The zoo blot analysis of genomic DNA of different species of animals demonstrated that MMP-11 is distributed probably ubiquitously in mammals, although no solid biological functions have so far been assigned to the enzyme.
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  • Tsutomu HASHIZUME, Shigeto KANEMATSU
    1995 Volume 41 Issue 1 Pages 41-46
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    The objectives of research were to characterize the effects of prostaglandins (PG) on the release of growth hormone (GH), prolactin (PRL), and luteinizing hormone (LH) in cultured bovine anterior pituitary cells. PGE2 significantly increased the media concentrations of GH by 141% (P<0.01) and 27% (P<0.01) at the concentrations of 10-7 and 10-8 M, respectively, compared with controls. The PGE2 significantly increased media concentrations of PRL compared with controls at the concentrations of 10-7 and 10-8 M by 120% (P<0.01) and 22% (P<0.05), respectively. PGE2 also significantly increased LH release at the concentration of 10-7 M by 22% (P<0.05), but not at a concentration of 10-8 M. PGF significantly increased the release of LH compared with controls at the concentrations of 10-8 and 10-9 M by 15% (P<0.05) and 19% (P<0.05), respectively, while PGF failed to stimulate the release of GH and PRL. In contrast to PGE2 and PGF, PGD2 had no effect on the release of GH, PRL, and LH. These results suggest that PGE2 is the most potent prostaglandin among the PGs for the release of GH, PRL, and LH by acting directly on bovine pituitary cells.
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  • Sang-Yong KIM, Yasunari KAWASHIMA, Jutaro TAKAHASHI, Yasuhisa YASUDA
    1995 Volume 41 Issue 1 Pages 47-55
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    The possible application of a computer-based color image processor system (SPICCA-2, Nippon Avionics & Co., Ltd., Tokyo, Japan) for the quantitative description of coat-color patterns in artificially produced aggregation chimeras of mice was investigated. The system is capable of extracting information essential to the quantitative description of coat-color patterns from sampled images of the pelts. In the present series of experiments, coat-color patterns of 6 BALB/c ⇔ C57BL/6 chimeras were quantitatively analyzed by means of the SPICCA-2 system. In those chimeras, the proportion of two allogeneic components, BALB/c and C57BL/6, was relatively well balanced between the left and the right halves of the pelt. However, some chimeras tended to have an unbalanced pigment distribution between the anterior and posterior regions. The results are in essential agreement with the findings reported earlier by Tachi et al. (1991) on C3H/HeN ⇔ BALB/cA chimeras.
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  • Yukio KANAI, Junko YAMASAKI, Yukari TAKEUCHI, Nobuyo KOIKE, Mayumi FUJ ...
    1995 Volume 41 Issue 1 Pages 57-62
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    Electrophysiological recording of the hypothalamic GnRH pulse generator activity was attempted in 3 long-term castrated male goats in which electrodes had been implanted in the medial basal hypothalamus (MBH). While multiple unit activity (MUA) signals from the electrode were continuously recorded, each animal received subcutaneous implants containing testosterone for 7 days (Experiment 1). They were similarly treated with estradiol alone, estradiol plus testosterone, and testosterone alone for 7 days each consecutively (Experiment 2). These steroidal treatments enabled to maintain the plasma levels of testosterone and estradiol within physiological ranges at 2-4 ng/ml and 3-5 pg/ml, respectively. Blood samples for the determination of pulsatile LH secretion were collected at 6-min intervals for 4 h on the last day of each treatment period. The short-lived sudden increase in MUA spikes (MUA volley) was detected being always preceding the occurrence of the LH pulse in the peripheral blood, and this synchrony of MUA volley with the LH pulse was constant in all the animals regardless of steroidal treatments. Testosterone treatment resulted in the increase of the MUA volley intervals from 27 min to 43 min (Experiment 1). Treatment with estradiol in Experiment 2 also resulted in prolonged MUA volley intervals similar to those of testosterone treatment and there was no additive or synergistic effect between these two steroids. The present study demonstrated that the electrophysiological manifestation of the hypothalamic GnRH pulse generator could be successfully monitored in male goats as reported in females, and it is suggested that estradiol may exert a negative feedback effect on the GnRH pulse generator activity in a similar manner to that of testosterone but at much lower doses in male goats.
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  • Osman Valli PATEL, Toru TAKAHASHI, Makoto HIRAKO, Tsuneo TOMIZUKA, Tos ...
    1995 Volume 41 Issue 1 Pages 63-70
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    Peripheral plasma progesterone concentrations were monitored throughout gestation in Holstein cows after non-surgical transfer of in-vitro fertilized embryos. Cows (n=12) were divided into 2 groups; Group 1=single embryo, Group 2=twin embryos. To induce multiple corpora lutea prior to the embryo transfer, 3 cows in each group (n=6) were randomly treated (treated cows) with either porcine follicle stimulating hormone (FSH) or pregnant mare serum gonadotropin (PMSG) followed by prostaglandin F (PGF), commencing in the mid-luteal phase of the cycle preceding pregnancy. In the remaining cows (n=6) estrus was synchronized by using PGF (untreated cows). Treated cows had a significantly higher plasma progesterone concentration throughout gestation than untreated cows (P<0.001). Treated singleton cows had higher plasma progesterone concentration in the first trimester than in the later trimesters (P>0.05). Treated twin-bearing cows had a significantly higher plasma progesterone concentration in the first and the third trimester compared to the second trimester (P<0.001). There was no significant (P>0.05) difference in plasma progesterone concentration throughout gestation between untreated singleton and twin-bearing cows. A distinct undulation in the plasma progesterone profile was seen in all the groups between days 20 to 25. To conclude, the plasma progesterone secretory profile was indistinguishable between untreated singleton and twin-bearing cows, and multiple corpora lutea induced by gonadotropin administration significantly boosted the progesterone pool throughout gestation.
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  • Yukio TSUNODA, Yoko KATO
    1995 Volume 41 Issue 1 Pages 71-75
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    To improve the developmental ability of enucleated mouse oocytes that received male fetal germ cells at 15.5 days post coitum (G0 stage), the effects of parthenogenetic activation with electrical stimulation and pretreatment of germ cells with PDGF or FGF, competent factors for quiescent cells, were examined. Preliminary evaluations revealed that the best conditions for electrical activation of unfertilized eggs were 100 μsec direct current pulses at 150 V/mm followed by 2 pulses of 50 V/mm at 20 min intervals (100% for activation and 94% for development to blastocysts). Fusion rates were the same for PDGF, FGF and control groups (35 to 46%). The activation rate in the PDGF group was significantly higher (90%) than that of controls (78%). The proportion of activated eggs that developed to morulae and blastocysts was high in the PDGF group but was not significantly different from that in controls (82 vs 68%). One live fetus with 32 somites was obtained on day 10.5 of pregnancy after the transfer of eggs reconstituted with PDGF treated germ cells. Repeated electrical parthenogenetic stimulation and pretreatment of fetal germ cells with PDGF appeared to enhance the development of reconstituted eggs.
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  • Ken NAKADA, Jinji MIZUNO, Koichi SHIRAISHI, Kenji ENDO, Shunichi MIYAZ ...
    1995 Volume 41 Issue 1 Pages 77-84
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    A long-lasting series of transient rises in intracellular Ca2+ concentration ([Ca2+]i) were recorded with Ca2+ imaging during fertilization of bovine eggs matured in vivo or in vitro. The first Ca2+ response could be recorded in zona pellucida-free eggs within 1 min after sperm-egg contact, when a preceding slow [Ca2+]i rise reached 150-170 nM. The response had the peak of 565 ±32 nM (n=15; maximum 780 nM) and the duration of 4-6 min, being significantly larger and longer than any succeeding response (359 ± 12 nM, n=68; 2 min). The series of Ca2+ transients occurred at constant intervals of 19-20 min without attenuation in the magnitude for more than 9 h. Ca2+ transients became smaller progressively during pronuclear migration and nuclear envelope breakdown, and no response occurred in 2-cell embryo. In unfertilized eggs, injection of inositol 1, 4, 5-trisphophate (InsP3) induced a Ca2+ transient and the sulfhydryl reagent thimerosal caused repetitive Ca2+ transients. These responses were markedly inhibited or blocked by preinjection of a monoclonal antibody to the mouse InsP3 receptor. However, sperm-induced responses were not blocked in 2/3 of eggs examined. Another mechanism may be involved in addition to InsP3-induced Ca2+ release from intracelluar stores.
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  • Kazuei ITO, Yoshimi IKEMIZU, Jutaro TAKAHASHI, Yasuhisa YASUDA, Kyoko ...
    1995 Volume 41 Issue 1 Pages 85-92
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    The objective of the present investigation was to purify early pregnancy factor (EPF)-like substance(s) from a pregnant bovine ovarian extract and in vitro fertilized ovum culture medium (IVF-OCM). Both the ovarian extract and IVF-OCM had EPF-like activity. These EPF-like substance(s) bound with CM-Sepharose and CNBr-activated Sepharose coupled with anti-EPF IgG but did not bind with a HPLC-DEAE column. The EPF-like substance(s) purified from the ovary and IVF-OCM had EPF-like activity (RIT≥5) at M.W. 17-38 KD and M.W. 21-30 KD, respectively. SDS-PAGE analysis revealed that the ovarian extract and IVF-OCM positive fractions had a band of 21.5 KD similar to that of EPF purified from pregnant bovine serum. Antiserum to HPLC fraction III (IVF-OCM origin, RIT≥5) decreased the RIT value of pregnant bovine serum from 6 to 3 and reacted with a band of 21.5 KD protein purified from bovine pregnant serum. Therefore, these two EPF-like substance(s) and EPF from pregnant serum may be similar substances.
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Research Notes
  • Masao YAMAMOTO, Masato OOE, Mamoru KAWAGUCHI, Tatsuyuki SUZUKI
    1995 Volume 41 Issue 1 Pages 93-96
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    The objective of this investigation was to determine the appropriate dose of refined follicle stimulating hormone (FSH-R) dissolved in polyvinylpyrrolidone (PVP) and administered as a single intramuscular (i.m.) injection for superovulation in cows. Seventy Holstein parous cows were alotted to 4 treatment groups and were given 20 mg (A), 30 mg (B), 40 mg (C) or 50 mg (D) FSH-R in 10 ml of 30% PVP. Standing estrus was observed in groups A (1/1), B (9/20), C (6/8) and D (4/5) at 40-60 h after the first PGF treatment. However, 2/8 in group C and 1/5 in group D showed standing estrus before 40 h, and 11/20 in group B showed estrus at 60-72 h after PGF treatment. There were no significant differences in the mean number of recovered ova/embryos among groups B, C and D. However, the mean number of transferable embryos in groups B (6.2 ± 3.8) and C (7.1 ± 5.9) was significantly higher (P<0.05) than in group D (1.0 ± 0). The rate of unfertilized eggs was significantly higher (P<0.01) in group D (82.6%) than in groups B (18.7%) and C (38.5%). A single injection of 40 mg FSH-R in PVP was effective for superovulation in Holstein cows.
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  • Kaoru TAKAGI, Kiyoshi TSUCHIYA, Shigehisa TSUMAGARI, Masatoshi TAKEISH ...
    1995 Volume 41 Issue 1 Pages 97-101
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    DNA fingerprints were prepared from DNA extracted from 32 Beagles, and 6-13 (9.3 ± 1.9) clear bands were detected in all samples. Based on these bands, the frequency of appearance of a band at the same position by chance in any 2 unrelated dogs was calculated as 0.49 ± 0.13. Although the allelic frequency “q” of 0.29 showed a higher value than that of 0.15 in humans, this figure was similar to that 0.27 reported by Jeffreys (1985) in dogs. The number of bands with chance agreement in dogs without blood relation were conformed to a binomial distribution with an appearance frequency of 0.49. According to these findings, the probability of the occurrence by chance of agreement in all bands in unrelated dogs was estimated as 0.499.3=1.3 × 10-3.
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  • Sueo NIIMURA, Misa HOSOE
    1995 Volume 41 Issue 1 Pages 103-108
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    The distributional changes of cortical granules (CGs) in bovine oocytes during maturation and fertilization in vitro were histochemically examined with lectin. Lens culinaris agglutinin (LCA)-reactive CGs were densely distributed as irregular aggregates of particles throughout the peripheral cytoplasm of every oocyte immediately after collection, while a few were individually dispersed (type I). As the stages of oocyte maturation proceeded, the aggregated CGs collapsed (type II), and the CGs became distributed individually in the cortical cytoplasm (type III). When matured oocytes were inseminated, the CGs completely disappeared in 40, 48 and 69% of oocytes at 3, 6 and 12 h after insemination (type V), respectively. However, type IV oocytes with very few CGs in the inner cytoplasm were always present in about 10% at any time after insemination.
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Technical Note
  • Takashige OTOI, Ken YAMAMOTO, Nobuyuki KOYAMA, Shoichi HAYASHI, Tatsuy ...
    1995 Volume 41 Issue 1 Pages 109-111
    Published: 1995
    Released on J-STAGE: November 30, 2001
    JOURNAL FREE ACCESS
    Bovine follicular oocytes from ovaries of pubertal heifers or slaughtered adult cows were matured, fertilized with frozen-thawed spermatozoa and cultured with cumulus cells. Although cleavage and development rates to the blastocyst stage of oocytes from pubertal heifers were significantly lower than those from slaughtered adult cows, the number of blastocysts per ovary was similar in the two groups. These findings show that oocytes from pubertal heifers are inferior in quality than those from slarughtered adult cows. To confirm the full developmental capacity of oocytes from pubertal heifers, 6 blastocysts were transferred to 3 recipient cows, and one cow became pregnant.
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