Journal of Reproduction and Development
Online ISSN : 1348-4400
Print ISSN : 0916-8818
ISSN-L : 0916-8818
Volume 51, Issue 4
August
Displaying 1-15 of 15 articles from this issue
Original Article
  • Jun Babaan FERANIL, Naoki ISOBE, Toshihiko NAKAO
    2005 Volume 51 Issue 4 Pages 419-426
    Published: 2005
    Released on J-STAGE: September 02, 2005
    Advance online publication: April 22, 2005
    JOURNAL FREE ACCESS
    The aim of this study was to investigate the distribution pattern of von Willebrand factor (vWF) and vascular endothelial growth factor (VEGF) in the healthy antral and atretic follicles of Philippine swamp buffaloes (SB) in comparison with Holstein-Friesian cows (HF). Paraffin sections of healthy follicles and atretic follicles at various stages were immunostained with vWF antibody and VEGF antibody. The density of vWF-positive capillary vessels in the theca interna significantly increased as atresia progressed in SB, whereas the density significantly decreased in late atretic follicles compared with advanced ones in HF. On the other hand, the area of vWF-positive capillary vessels in the theca interna significantly increased as atresia progressed in both SB and HF. Immunoreactions of VEGF in the granulosa cells (in all follicle types) were observed in both SB and HF. In the granulosa layer, a reduction in the VEGF immunoreaction was noted as follicles progressed from healthy to advanced atretic follicles in both animals. Granulosa cells (in both SB and HF) showed a higher immunopositive staining than theca cells. In the theca interna, VEGF immunostaining diminished as follicles progressed to the late atretic follicles in both animals. These results indicate that during atresia, changes of vWF expression are the opposite of VEGF expression in SB. Both vWF and VEGF are suggested to be associated with follicular atresia in SB.
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  • Jozsef RÁTKY, Klaus-Peter BRÜSSOW, Istvan EGERSZEGI, Helmu ...
    2005 Volume 51 Issue 4 Pages 427-432
    Published: 2005
    Released on J-STAGE: September 02, 2005
    Advance online publication: June 10, 2005
    JOURNAL FREE ACCESS
    Only a very small amount of physiological data is available about the low fertility (mean litter size is 5.7 ± 0.8) of Hungarian native breed, Mangalica (M), sows. The aim of the present paper is to reveal the differences in preovulatory follicle development and intrafollicular oocyte maturation between M and Landrace (L) gilts, with special reference to the peri- and postovulatory secretion and peripheral concentrations of estradiol-17β (E2), progesterone (P4), and luteinizing hormone (LH). The number of preovulatory follicles was 6.8 ± 1.4 and 19.6 ± 6.6 in M and L gilts, respectively. A lower degree of cumulus expansion and a lower percentage of mature oocytes (TI/M II) was noted in M. Higher LH and E2 peak levels, a longer E2 to LH peak interval, and lower embryo survival was confirmed. Interestingly, despite the lower number of corpora lutea, a higher peripheral blood level of P4 was shown in M than in L gilts. Both diminished follicular development and protracted oocyte maturation may be involved in low fecundity in M, and the present findings may explain these reproductive phenomena.
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  • Enrico VIZZA, Silvia CORRER, Fabrizio BARBERINI, Rosemarie HEYN, Seren ...
    2005 Volume 51 Issue 4 Pages 433-443
    Published: 2005
    Released on J-STAGE: September 02, 2005
    Advance online publication: April 22, 2005
    JOURNAL FREE ACCESS
    In order to understand the 3-D distribution of collagen in relation to vascularization, chorionic villi of human placentae, belonging to normal pregnancies at term, were studied by scanning electron microscopy (SEM) after alkali maceration techniques, and by transmission electron microscopy (TEM). The villous tree appeared made of an uninterrupted structure of collagen fibres. The collagen fibres connected the chorionic villi axis with their basal plates and organised differently according to the various levels of villous branching. The collagen of stem villi showed copious fibres. The external fibres (facing the villous surface) were arranged mainly longitudinally. The central core of the villi (inner fibres) were arranged concentrically around the wall of the fetal vessels. Both external and internal fibres formed stratified lamellae or small parallel bundles. The inner core of stem villi showed small holes housing capillary spaces. Mature intermediate and terminal villi showed a scarce amount of collagen arranged in thin concentric layer within the villous core, surrounding numerous dilated capillary and sinusoid spaces.These observations demonstrated that the extracellular matrix of human chorionic villi is highly compartmentalised and shows a variable structural 3-D distribution depending on the branching level of the villous tree, such a distribution ensures the most favourable microenvironment for feto-maternal exchanges and it is likely able to provide a modulated support to the developing chorionic fetal vessels and trophoblastic layer as well.
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  • Ann-Sofi BERGQVIST, Masaki YOKOO, Renée BÅGE, Eimei SATO, ...
    2005 Volume 51 Issue 4 Pages 445-453
    Published: 2005
    Released on J-STAGE: September 02, 2005
    Advance online publication: April 22, 2005
    JOURNAL FREE ACCESS
    Hyaluronan is involved in fundamental reproductive events such as sperm storage in the female reproductive tract, fertilization, and early embryo development, these functions are presumably mediated by its major cell surface receptor, CD44. The present study was conducted to investigate the presence and localization of CD44 in the bovine oviductal epithelium, using immunohistochemical and Western blot methods on tissue sections and epithelial cell extracts collected from the uterotubal junction (UTJ), isthmus, and ampulla of animals in the oestrus or luteal phase of the oestrous cycle. While positive immunolabelling for CD44 was found on the ad-luminal surface and supra-nuclear region of epithelial cells in all tubal segments investigated, in the UTJ, there were epithelial cells in which the entire cytoplasm positively stained. We found no differences in terms of CD44-positive staining between the different stages of the oestrous cycle. Presence of CD44 was detected by Western blotting in the tubal epithelium as a single band at 200 kDa. Although it appeared in all tubal segments, the expression of CD44 protein was more accentuated in the sperm reservoir (UTJ) than in the other segments. This is the first time CD44 has been detected in the epithelium of the tubal sperm reservoir in cattle, suggesting a pathway for the action of hyaluronan in this segment.
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  • Mohamed S. MEDAN, Gen WATANABE, Kazuaki SASAKI, Nigel P. GROOME, Sayed ...
    2005 Volume 51 Issue 4 Pages 455-463
    Published: 2005
    Released on J-STAGE: September 02, 2005
    JOURNAL FREE ACCESS
    Transrectal ultrasonography of ovaries was performed daily in 6 goats for 3 consecutive estrous cycles. Blood samples collected daily were measured for concentrations of FSH, inhibin A, and estradiol-17β. Follicular and hormonal data were analyzed for associations between the follicular waves and hormonal concentrations. During the interovulatory intervals, follicular growth and regression occurred in a wave like pattern (2-5 waves), and the predominant patterns were three and four follicular waves. In addition, there was no significant difference among the diameters of dominant follicles during the growth phase of the follicular waves. The number of 3 mm follicles peaked on days 0, 7, and 11 in interovulatory intervals that had three follicular waves and on days -1, 5, 11, and 15 in those that had four follicular waves. Plasma concentrations of FSH increased around the day of follicular wave emergence and declined with the growth of follicles. Circulating FSH increased again concomitant with regression of dominant follicles in the anovulatory wave, whereas FSH levels remained low in the ovulatory wave. Inhibin A was negatively correlated with FSH, while it was positively correlated with estradiol-17β, suggesting that inhibin A is a product of healthy growing follicles and that it contributes to the suppression of FSH secretion. In conclusion, the growth of ovarian follicles in goats exhibits a wave-like pattern, and follicular dominance is less apparent in goats. Moreover, inhibin A may be a key hormone for regulation of the follicular wave through suppression of FSH secretion in goats.
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  • Sanjay BHOJWANI, Gabor VAJTA, Henrik CALLESEN, Knut ROSCHLAU, Andreas ...
    2005 Volume 51 Issue 4 Pages 465-475
    Published: 2005
    Released on J-STAGE: September 02, 2005
    JOURNAL FREE ACCESS
    To enable us to handle a large number of oocytes at a given time and to have an increased throughput of cloned embryos, we attempted the Handmade cloning (HMCTM) technique, a zona-free method of bovine somatic cell nuclear transfer. Our objective was to study the developmental competence of the HMCTM derived embryos obtained using different types of somatic cells. A total of 6,874 cumulus-oocyte-complexes were used with either 7th or 11th passage fibroblasts (1st and 2nd groups, respectively), which were prepared from male animals, or granulosa cells (3rd group) as nuclei donors. The average cleavage rate was 65%, accompanied by a blastocyst rate of just 2% for the cleaved products and 5% for the > 8-cell embryos, and there was no significant difference between the three groups. Out of 27 blastocysts recovered, 22 blastocysts were transferred to 22 recipients, resulting in two pregnancies. One pregnancy was lost after the fourth week while the other progressed to full term with the birth of a male calf. This first successful cloning of a male calf with the HMCTM technique in Europe indicates the successful adoption and establishment of this technique in our laboratory, and that this technique can be successful in producing viable embryos.
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  • Kazuyuki KANEKO, Yuumi AKIYA, Haruno SATO, Ayako TANAKA, Hajime AOKI, ...
    2005 Volume 51 Issue 4 Pages 477-482
    Published: 2005
    Released on J-STAGE: September 02, 2005
    Advance online publication: June 10, 2005
    JOURNAL FREE ACCESS
    In order to clarify the breeding capability of male raccoons in Japan, the testes of raccoons, a nuisance animal, collected in Kanagawa Prefecture and Hokkaido were histologically inspected. Furthermore, testosterone concentrations in their blood were measured. The testosterone concentrations increased in winter and the diameter of seminiferous tubules and the spermatogenetic score decreased in summer for the animals captured in Kanagawa Prefecture. For the animals captured in Hokkaido, the diameter of seminiferous tubules did not change and the decrease of the spermatogenetic score in summer was slight. As the above results show, there is a breeding season in male raccoons in Japan, and the reduction of testicular function in summer was greater in animals captured in Kanagawa Prefecture than in animals captured in Hokkaido.
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  • Tomas J. ACOSTA, Masa TETSUKA, Motozumi MATSUI, Takashi SHIMIZU, Bajra ...
    2005 Volume 51 Issue 4 Pages 483-489
    Published: 2005
    Released on J-STAGE: September 02, 2005
    Advance online publication: June 10, 2005
    JOURNAL FREE ACCESS
    The aim of the present in vivo study was to monitor real-time fluctuations of cortisol (Cr) in the wall of preovulatory follicles using a microdialysis system (MDS) implanted in the theca layer as well as changes in ovarian venous plasma (OVP) and jugular venous plasma (JVP). Seven cows were superovulated using FSH and prostaglandin F2α injections. Dialysis capillary membranes were surgically implanted into the theca layer of mature follicles and connected to a microdialysis system. Fractions of the perfusates were collected from Day -1 (Day 0=LH surge) to Day 3. No difference in the concentrations of Cr between JVP and OVP was detected throughout the experiment. Circulating concentrations of Cr ranged from 20 to 35 ng/ml 8 h after surgery in ovulatory and anovulatory cows. In five ovulatory cows, the Cr concentration decreased to basal levels (<10 ng/ml) between 12 and 24 h after surgery, however, two anovulatory cows retained high Cr levels (>10 ng/ml) up to 42 h after surgery. There was a clear increase in the local concentration of Cr from 13.3 ± 2.1 pg/ml at -24 h to 27.5 ± 1.7 pg/ml at 0 h (peak of the LH surge) within the wall of ovulatory follicles. This increase was not detected in anovulatory follicles. This transient increase in Cr occurred only in the follicle wall, but not in the OVP or JVP, indicating that the presence of a local regulatory mechanism for Cr production/conversion in ovulatory follicles, and this mechanism may modulate the inflammatory-like reaction induced by LH surge in the follicle wall. The present results demonstrate that the glucocorticoid environment in the follicular wall adjusts at the local level in bovine ovulatory follicles. This mechanism may protect follicles from the adverse effects of glucocorticoid, and it may prevent excess inflammatory reactions associated with ovulation by temporarily increasing local concentrations of glucocorticoid, thus forming an integral part of the regulatory mechanism in ovarian physiology.
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  • Ki-Doek KIM, Kwang-Sook KI, Hyun-Gu KANG, Ill-Hwa KIM
    2005 Volume 51 Issue 4 Pages 491-498
    Published: 2005
    Released on J-STAGE: September 02, 2005
    Advance online publication: June 10, 2005
    JOURNAL FREE ACCESS
    The objectives of this study were to determine the risk factors for development of postpartum ovarian cysts by evaluating several reproductive factors in individual cows, and to determine the economic impact of ovarian cysts on subsequent reproductive performance in dairy herds in Korea. The data, including cow parity, abnormal puerperium, endometritis, body condition score (BCS), and breeding status were collected from 634 cows in 9 dairy herds. We used logistic regression to evaluate the effects of these factors on ovarian cysts. A stepwise procedure, used to obtain the appropriate model with α=0.05, revealed that cow parity was the most important risk factor for ovarian cyst development within 8 weeks postpartum, while development of endometritis and BCS loss ≥1 from the dry period to 8 weeks postpartum were the most important risk factors for ovarian cyst development beyond 8 weeks postpartum. The occurrence of ovarian cysts beyond 8 weeks postpartum prolonged (P<0.01) the mean intervals from calving to first service (27 days) and conception (77 days), and increased (P<0.05) the culling rate (7.8%), while ovarian cyst development within 8 weeks postpartum did not affect (P>0.05) the mean intervals from calving to first service and conception or the culling rate. The economic loss resulting from the occurrence of ovarian cysts was estimated at approximately 823,996 won ($687) due to effects on the cost of nutrition, average growth of calves, labor and medical costs, and culling. These results suggest that cow parity is correlated with the development of ovarian cysts within 8 weeks postpartum, and endometritis and BCS loss ≥1 from the dry period to 8 weeks postpartum are correlated with the development of ovarian cysts after 8 weeks postpartum, which decreases reproductive performance and results in economic loss in dairy herds in Korea.
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  • Eui-Ju HONG, Kyung-Chul CHOI, Eui-Bae JEUNG
    2005 Volume 51 Issue 4 Pages 499-508
    Published: 2005
    Released on J-STAGE: September 02, 2005
    Advance online publication: June 10, 2005
    JOURNAL FREE ACCESS
    It has been reported that Calbindin-D9k (CaBP-9k) is rapidly and strongly induced by environmental estrogenic compounds, possibly through estrogen receptors (ERα) in the uterus of mammals. CaBP-9k can be evaluated as an early gene marker for assaying estrogenic effects of putative environmental chemicals in the rat uterus. This study was undertaken to investigate CaBP-9k mRNA and protein expression in the postnatal rat uterus following maternal exposure to 17β-estradiol (E2) and bisphenol A (BPA) during the neonatal period. Treatment with a high dose of BPA (600 mg/kg body weight (BW) per day) resulted in a 3-fold increase in CaBP-9k mRNA expression for 3 days, while a single dose of E2 (40 μg/kg BW per day) induced 2-fold increase of this gene in the maternal uterus. In an agreement with maternal CaBP-9k mRNA, postnatal CaBP-9k mRNA in the uterus increased 4-fold when treated with BPA (600 mg/kg BW per day). In addition, treatment with increasing concentrations of BPA resulted in significant increases in CaBP-9k protein in the maternal rat uterus. It is of interest that increasing doses of BPA induced a significant ERα mRNA increase in the postnatal uterus. Furthermore, immunohistochemistry revealed that treatment with BPA induced CaBP-9k protein in the maternal uterus. We demonstrated that maternal exposure to BPA during late pregnancy induced CaBP-9k mRNA and protein in maternal and postnatal rat uteri. These results suggest that rapid absorption and distribution of environmental estrogenic compounds occurs in maternal and neonatal rat uteri and these chemicals can easily pass though the placenta during pregnancy to affect postnatal reproductive functions.
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  • Karin WOLLENHAUPT, Harald WELTER, Klaus-Peter BRÜSSOW, Ralf EINSP ...
    2005 Volume 51 Issue 4 Pages 509-519
    Published: 2005
    Released on J-STAGE: September 02, 2005
    Advance online publication: June 24, 2005
    JOURNAL FREE ACCESS
    The aim of this study was to characterize the effect of ovarian steroids and early gestation on the expression of fibroblast growth factor 7 (FGF-7) and its receptor (FGFR2IIIb) in the porcine endometrium. In Experiment 1, gilts were ovariectomized (OVX) on day 10 of the estrous cycle and treated thereafter with vehicle (VEH), progesterone (P4), estradiol benzoate (EB), or P4 + EB. Days 12 and 20 cyclic gilts (C12 and C20) were used to determine the influence of physiologically low and high plasma estradiol and progesterone concentrations on their expression. In Experiment 2, the expression of FGF-7 and FGFR2IIIB was characterized on days 1 (G 1) and 12 (G 12) of gestation. FGF-7 and FGFR2IIIb mRNA were quantified by quantitative real-time RT-PCR, and localization of FGF-7 protein in steroid-treated and early pregnant gilts was performed by immunohistochemistry. VEH-gilts expressed both FGF-7 and FGFR2IIIB mRNA. We found a significant effect of EB, but no effects of P4 or P4+EB on the mRNA expression of FGF-7. FGFR2IIIb mRNA significantly decreased after the EB and combined P4 + EB treatments, compared to P4 only substituted animals. Day 12 cyclic gilts showed significantly higher FGF-7 and FGFR2IIIb mRNA expression compared with day 20 gilts. Between day 1 and 12 of gestation, FGF-7 mRNA expression differed highly while FGFR2IIIb transcripts only varied significantly. FGF-7 protein was localized in endometrial epithelia, vascular smooth muscle, and the endothelium of different types of blood vessels. Staining was weak in VEH and P4 treated gilts, whereas it was prominent following EB and P4+EB. FGF-7 antibody strongly stained the luminal epithelium on day 12 of gestation. In summary, FGF-7 and FGFR2IIIb mRNA expression is regulated differently by exogenous ovarian steroids, assuming progesterone in connection with a specific amount of 17β-estradiol, whereas the receptor seems to be inhibited by estradiol. Both transcripts coordinately increased during the progesterone dominated phase on day 12 both in cyclic and early pregnant gilts. We conclude that estradiol and progesterone are involved in the regulation of this ligand-receptor system, which might have an important role in preparing endometrial tissue for implantation in gilts.
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Research Note
  • Qiang WENG, Mohamed S. MEDAN, LongQuan REN, Gen WATANABE, Koji Y. ARAI ...
    2005 Volume 51 Issue 4 Pages 521-526
    Published: 2005
    Released on J-STAGE: September 02, 2005
    JOURNAL FREE ACCESS
    The objective of this study was to investigate the cellular immunolocalization of inhibin α and inhibin/activin (βA and βB) subunits in the fetal, neonatal and adult testes of Shiba goats. The testes were obtained from a fetus at 90 days, a neonate at 15 days, and two adult Shiba goats (both of 3 years old). The sections of testes were immunostained by the avidin-biotin-peroxidase complex method (ABC) using polyclonal antisera raised against porcine inhibin α, inhibin/activin βA, and inhibin/activin βB. Inhibin α and inhibin/activin (βA and βB) subunits were expressed in Leydig cells, but not in the Sertoli cells of the fetus with a weak immunostaining. An increase in the number of positive cells and a more intense immunohistochemical signal for inhibin α and inhibin/activin (βA and βB) subunits were observed in the Leydig cells of neonatal testes. Moreover, inhibin α, βA, and βB subunits were expressed in the Sertoli cells and Leydig cells of adult testes, respectively. These results suggest that Shiba goats testes have the ability to synthesize inhibins in the fetus, neonate, and adult, and the cellular localization of inhibin/activin subunits showed age-related changes in fetal, neonatal, and adult testes of Shiba goats.
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  • Johannes KAUFFOLD, Hussein A. H. AMER, Uwe BERGFELD, Frank MÜLLER ...
    2005 Volume 51 Issue 4 Pages 527-532
    Published: 2005
    Released on J-STAGE: September 02, 2005
    Advance online publication: June 24, 2005
    JOURNAL FREE ACCESS
    This study investigated the viability of embryos from non-stimulated 2-3-month-old calves generated in vitro using oocytes from follicles of defined size in terms of their ability to produce full-term pregnancies. Ablation of follicles ≥4 mm was used to induce the emergence of a new follicular wave, and calves (n=3) were laparoscopically punctured three times at 7-day-intervals to recover cumulus-oocyte-complexes (COCs) from follicles >8 (group A) and between 4-8 mm (group B). Calves were aged 49, 56, and 80 days, respectively, at first recovery. Morphologically intact COCs were subjected to in vitro maturation, fertilization, and embryo culture, and compact morulae/blastocysts were transferred on day 7 post-insemination to synchronized virgin heifers. Blood typing was used for maternity analysis. A total of 29 COCs were recovered, 21 cultured, yielding 11 cleaved embryos (52.4%) and 6 compact morulae/blastocysts (28.6%). No differences were observed between groups. Transfer of the 6 embryos to 5 recipients resulted in three pregnancies (one from group A and two from group B). Two normal male offspring (both from group B), with birth weights of 44 and 51 kg, were born, and two donor calves, aged 56 and 59 days, were identified as the dams. In conclusion, the results demonstrate that embryos generated in vitro from oocytes from non-stimulated calves at an age younger than two months are viable in terms of their ability to produce full-term pregnancies, and suggest that the viability of calf embryos is not related to follicle size.
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Technical Note
  • Mikiko TAKA, Hiroshi IWAYAMA, Yutaka FUKUI
    2005 Volume 51 Issue 4 Pages 533-537
    Published: 2005
    Released on J-STAGE: September 02, 2005
    JOURNAL FREE ACCESS
    For developmental competence of porcine embryos in vitro, it is important to improve the culture environment. The present study was performed to evaluate four different culture systems for in vitro matured porcine oocytes following intracytoplasmic sperm injection (ICSI); drop, well and two sizes of the well of the well (WOW) systems (500 and 1,000 μm in diameter). The cleavage rate on Day 2 and the mean cell number in blastocysts on Day 6 were not significantly different among the four treatments. However, the 1,000 μm WOW (24.6%) resulted in a significantly higher (P<0.05) blastocyst rate than those in the other culture systems (12.9, 14.8, and 7.1% for drop, well, and 500 μm WOW, respectively). The present study indicates that the microenvironment created by the 1,000 μm diameter WOW improves blastocyst production of in vitro matured porcine oocytes after ICSI, and that the effectiveness of the WOW system is dependent on the size (diameter) of the WOW.
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  • Katsutoshi NARUSE, Hiroshi ISHIKAWA, Hiro-omi KAWANO, Hideto UEDA, May ...
    2005 Volume 51 Issue 4 Pages 539-546
    Published: 2005
    Released on J-STAGE: September 02, 2005
    Advance online publication: June 10, 2005
    JOURNAL FREE ACCESS
    We introduced a fusion gene of human albumin and enhanced green fluorescent protein (EGFP) into porcine oocytes using the sperm vector method, and produced a piglet that showed clear expression of GFP in the hooves and skin. PCR and Southern blotting analysis of genomic DNA extracted from the piglet's tissues, including the liver, showed that the tissues carried the transgene. RT-PCR analysis demonstrated that both the human albumin and EGFP genes were expressed in the tissues. The fact that human albumin gene was integrated and expressed in the liver of the transgenic pig opened a way for us to achieve our goal, which was the use of transgenic pigs for the bioartificial liver support system.
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