Biomedical subjects

Mariko Shirota

Publications and source records attributed to Mariko Shirota.

11 recordsLinked to original sources

PubMed2006

Internal dose-effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in gonadotropin-primed weanling rat model.

Single sc injection of 5 IU equine chorionic gonadotropin (eCG) induces ovulation in weanling female rats 3 days later. It has been shown that treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) 24 h before eCG injection reduces eCG-stimulated ovarian hypertrophy and inhibits ovulation. The present study intended to compare internal dose-effects of TCDD between these endpoints and representative endpoints for TCDD toxicity, such as weights of the liver and thymus, in weanling female rats given orally 0, 1, 4 or 16 microg/kg TCDD 24 h before eCG injection on postnatal day 25. Measurement of plasma TCDD concentrations by ELISA at 6, 72 and 96 h after TCDD revealed that significant levels of TCDD were maintained in systemic circulation until 96 h (on the day of induced ovulation) with the highest level at 6 h after TCDD treatment. Ovarian TCDD concentrations varied similarly and tended to be higher than those in the thymus at all time points, whereas hepatic concentrations of TCDD were the highest among the tissues. Although > or = 4 microg/kg TCDD affected the weights of the thymus and liver, no differences were observed in ovarian weights at any time point or in ovulation between corn oil-treated and TCDD-treated groups. Furthermore, ovarian levels of representative mRNAs in follicles were not affected by TCDD treatment. Since TCDD increased the amount of cytochrome P450 1A1 mRNA in the ovary, the administered TCDD stimulated the aryl hydrocarbon receptor-signaling pathway. From these results, we concluded that thymus weights of weanling female rats responded to TCDD at a lower internal dose as compared with that ovarian hypertrophy and follicular growth from early antral stage to ovulation would respond to.

Animals↗

PubMed2006

Effects of vertically transferred 3,3',4,4',5-pentachlorobiphenyl (PCB-126) on the reproductive development of female rats.

In order to study the effects of vertically transferred coplanar polychlorinated biphenyls on female reproductive development, female rat offspring from dams of Sprague-Dawley strain, which received daily oral administration of vehicle (corn oil) or 1 or 3 microg/kg of 3,3',4,4',5-pentachlorobiphenyl (PCB-126) from 2 weeks prior to mating with intact males until 20 days after delivery were examined from birth until puberty. Hepatic expression of the aryl hydrocarbon receptor (AhR)-inducible enzyme cytochrome P450 1A1 (CYP1A1) was detected in all offspring from PCB-126-exposed dams, indicating vertical transfer of PCB-126. Furthermore, quantification of ovarian mRNAs encoding CYP1A1, AhR and ARNT demonstrated that the ovary equipped the AhR-signaling system through which transcription of the CYP1A1 gene was enhanced in a dose-dependent manner. Exposure to PCB-126 retarded the growth of offspring in both exposed groups, while the viability of the neonates of the exposed groups was comparable to that of the oil-exposed controls. The exposure to 3 mug/kg/day reduced the ovarian weight on postnatal day (PND) 24, with atresia of most of the antral follicles and delayed vaginal opening. Exposure to 1 microg/kg/day did not produce such effects; however, both doses of PCB-126 induced external urogenital anomalies, such as vaginal thread and hypospadias, in all of the PCB-126-exposed female offspring. These results indicate that vertically transferred PCB-126 is potent enough to exert a direct effect on the ovary and adversely affect female puberty by altering the morphological and functional development of the female reproductive system.

Animals↗

PubMed2006

New approach to in situ quantification of ovarian gene expression in rat using a laser microdissection technique: relationship between follicle types and regulation of inhibin-alpha and cytochrome P450aromatase genes in the rat ovary.

The recently developed laser microdissection (LMD) technique makes it possible to quantify local gene expression in the target cells of various tissues. Using the LMD technique, this study aimed at comparing the amounts of mRNAs encoding the inhibin-alpha subunit and cytochrome P450 aromatase (P450(arom)) in granulosa cells between preantral and antral follicles in immature rat ovaries. Serial frozen sections of the ovaries from 24-day-old female Wistar rats were made and 30 healthy preantral (100-200 microm maximum diameter) and ten healthy antral ( > 300 microm maximum diameter) follicles were selected in each ovary based on morphological examinations, including immunohistochemistry for inhibin-alpha, in sections adjacent to those used for LMD. The amounts of mRNAs encoding inhibin-alpha subunit and P450(arom) were quantified by real-time polymerase chain reaction (PCR). While the amount of P450(arom) mRNA in the granulosa cell layers from the antral follicles was about 12-times higher than that in the preantral follicles, no difference in the amount of inhibin-alpha mRNA was found between these two types of follicles. Thus, the LMD technique allowed the in situ quantification of gene expression in the ovary and revealed that each granulosa cell expresses a stable amount of inhibin-alpha subunit mRNA independently of antral formation in immature rat ovaries.

Animals↗

PubMed2006

Immunological characteristics of Hatano high-and low-avoidance rats.

Hatano high- and low-avoidance (HAA and LAA) rats have been genetically selected on the basis of their two-way active avoidance behavior, and have been shown to differ in other behavioral and hormonal parameters. Since close interconnections among the nervous, endocrine and immune systems have been well documented, these two strains might possess differences in aspects of immunological action. In Experiment 1, plasma levels of IgG, IgM, complement 3 (C3), classical pathway hemolytic complement (CH50) and beta(2)-microglobulin were compared between males of the two strains at 5 and 24 weeks of age. Plasma levels of IgG and CH50 were lower in LAA than HAA rats at 5 weeks of age, whereas those differences disappeared at 24 weeks of age. There were no differences between the two strains in plasma levels of IgM, C3 and beta(2)-microglobulin. In Experiment 2, antibody production to sheep red blood cells (SRBC) and mitogen-induced lymphocyte proliferation were compared between 12-week-old males of the two strains. Antibody responses in the PFC assay, plasma anti-SRBC-IgM levels and spleen weights were higher in LAA than HAA rats. LPS-induced lymphocyte proliferation was greater in LAA than HAA rats. It was concluded that HAA rats show earlier development of immunological development, but that antibody production and mitotic response of B lymphocytes may be more pronounced in adult LAA than HAA rats. The strain differences observed in the immunological response may indicate the usefulness of using Hatano rats in studies of behavioral-immunological relationships.

Animals↗

PubMed2005

Reduction of primordial follicles caused by maternal treatment with busulfan promotes endometrial adenocarcinoma development in donryu rats.

Ovarian dysfunction leading to hormonal imbalance plays a crucial role in uterine carcinogenesis in rats as well as women. However, the effects of a reduction in primordial follicles at birth on uterine adenocarcinoma development have hitherto not been determined. The present study was therefore conducted using female Donryu rats, a high incidence rat strain of uterine adenocarcinoma. The animals were maternally exposed to 2.5 or 5.0 mg/kg of busulfan on gestation day 14 to reduce primordial follicles, and were then initiated by intrauterine treatment with N-ethyl-N'-nitro-N-nitrosoguanidine at 11 weeks of age. Both busulfan treatment doses caused earlier occurrence of persistent estrus, with dose-dependence as compared to controls. At 15 months of age, the rats were euthanized. The incidence of uterine adenocarcinomas and multiplicity of uterine neoplastic lesions were significantly increased by the 5.0 mg/kg, but not the 2.5 mg/kg busulfan treatment. Morphologically, the ovaries exposed to busulfan treatment exhibited severe atrophy, with few or no follicles and corpus lutea. Serum 17beta-estradiol (E2), progesterone, and inhibin levels were significantly decreased in the busulfan treatment groups, with a clear dose-relation. Interestingly, only the 5.0 mg/kg busulfan treatment elevated the E2/progesterone ratio. These results provide evidence that the reduction of primordial follicles promotes uterine adenocarcinoma development in rats in association with an earlier occurrence of the persistent estrus status.

Adenocarcinoma↗

PubMed2005

Influence of di-(2-ethylhexyl)phthalate on fetal testicular development by oral administration to pregnant rats.

Influence of di-(2-ethylhexyl)phthalate (DEHP) on testicular development was studied by oral administration of DEHP at doses of 500 and 1000 mg/kg/day to pregnant rats on gestational days (G) 7 to 18. Ethinyl estradiol (EE) at dose levels of 0.25 and 0.5 mg/kg/day was used as a reference substance. Each 5-6 pregnant rats were sacrificed and their fetuses were examined on G12, 14, 16, 18 and 20. Fetal deaths averaging 20-36% were observed at every examination in the group receiving 1000 mg/kg of DEHP. Increases of fetal deaths over 50% were also observed in the reference group that received 0.5 mg/kg of EE. Microscopic examination of the fetal testis in groups treated with DEHP revealed degeneration of germ cells in G16 fetuses and localized proliferation or hyperplasia of interstitial cells in G18 and 20 fetuses. Germ cells having more than two nuclei were observed in a few cases including the control testes of G14 fetuses. These multinucleated cells were observed frequently in G20 fetuses treated with DEHP. Examination of testes of naturally delivered offspring of dams treated with 1000 mg/kg of DEHP at 7 weeks of age revealed scattered atrophy or dilatation of seminiferous tubules. Another experiment was carried out to confirm the dose of DEHP affecting testicular development and spermatogenesis. DEHP was given to pregnant rats at doses of 125, 250 and 500 mg/kg/day during G7-18. Similar histopathological changes were observed in fetal testes of the group exposed to 500 and 250 mg/kg of DEHP, but not in those exposed to 125 mg/kg. In postnatal examinations, however, no abnormality was found in the testes at 5 and 10 weeks after birth in any of the treated groups. Furthermore, no abnormal findings were observed in the function of sperm, sperm counts and sperm morphology in the offspring of the group treated with DEHP during the fetal period at 10 weeks of age. Thus, 125 mg/kg/day is considered the no-observed-effect-level of DEHP on testicular development of rats by exposure in utero during the period of organogenesis.

Administration, Oral↗

PubMed2002

Detection of gene expression of vascular endothelial growth factor and flk-1 in the renal glomeruli of the normal rat kidney using the laser microdissection system.

The recent development of the laser microdissection (LMD) technique enables one to target particular tissues or cells for gene or protein analyses. The purpose of this study was to detect local mRNA expression of vascular endothelial growth factor (VEGF) and its receptor, flk-1, in the glomeruli of normal rat kidneys using the LMD system. Frozen sections of the kidney of 8-week-old male Wistar rats were made. The glomeruli were dissected from the frozen sections with the LMD system, and total RNA was extracted from 200 glomeruli in each kidney. Reverse-transcription polymerase chain reaction (RT-PCR) revealed the local mRNA expression of three isoforms of VEGF, flk-1 and GAPDH in the glomeruli. Moreover, the real-time PCR was performed to evaluate the experimental condition for quantification of VEGF and flk-1 mRNA expression using this system, and the results showed that at least 10 glomeruli might be needed for quantifying local VEGF mRNA expression. However, cDNA from 200 glomeruli was not enough for quantitative evaluation of flk-1 mRNA with this system. These results demonstrate the reproducibility of the analysis of mRNA expression in the renal glomeruli using the LMD system and also suggest that the application of the LMD technique will provide information to further our understanding of the mechanisms involved in kidney diseases.

Angiogenesis Inducing Agents↗

PubMed2002

Maternal behavior, milk ejection, and plasma hormones in Hatano high- and low-avoidance rats.

Hatano high- and low-avoidance (in a two-way active avoidance task) animals (HAA and LAA, respectively) were successfully selected from a Sprague-Dawley rat population. Pup growth of both strains was dependent on the maternal strain in a cross-fostering study. To determine whether there are strain-specific differences between HAA and LAA in maternal care, both strains of dams were subjected to a test battery as measured by nest building, home cage behavior, pup retrieval, and milk ejection tests. In addition, changes in plasma concentrations of lactotropic and corticotropic hormones such as prolactin, ACTH, and corticosterone were examined during lactation. The test battery indicated that the dams of both strains built good nests and spent an identical amount of time with their offspring. However, LAA dams showed a prolonged latency time for pup retrieval and often left pups outside the nest until the end of the test period. LAA dams also showed a decreased amount of milk ejection, whereas no strain differences were observed in milk ejection after oxytocin treatment. During lactation, a lesser increase in plasma concentrations of prolactin and a greater increase in ACTH were found in LAA dams. There were no differences between the two strains in plasma concentrations of corticosterone. These results clearly demonstrated decreases in maternal behavior and milk ejection in LAA as compared to HAA dams. The present results also suggest that maternal motivation and mechanisms responsible for maternal hormones related to suckling are involved in the degree of pup growth.

Animals↗

PubMed2002

Strain differences in the spontaneous incidence of sperm morphological abnormalities in Hatano rats.

Two inbred strains of Hatano rats, which had been bred selectively on the basis for shuttlebox avoidance responses (high- and low-avoidance animals; HAA and LAA), were different in sperm motility. We have found that there are strain differences in the spontaneous incidence of sperm morphological abnormalities. The percentages of abnormalities were extraordinarily higher in the LAA rats, whose sperm motility was lesser. Since the high sperm abnormality was an exceptional case in rats, these rats may be useful models for sperm quality.

Animals↗

PubMed2002

Reproductive endocrinology in Hatano high- and low-avoidance rats during the estrous cycle.

The high- and low-avoidance animals (HAA and LAA rats) were originally selected from Sprague-Dawley rats for their shuttle-box task. Reproductive endocrinology during the estrous cycle was compared between HAA and LAA rats. All HAA rats showed a regular 4-d estrous cycle, whereas most LAA rats (70.8%) showed a regular 5-d estrous cycle. The peak level of preovulatory luteinizing hormone (LH) surge level was significantly lower in LAA rats than in HAA rats on the day of proestrus. In contrast, the peak level of prolactin surge on the day of proestrus was significantly higher in LAA rats than in HAA rats. Plasma concentrations of follicle-stimulating hormone (FSH) and estradiol-17beta were significantly lower in LAA rats as compared with HAA rats at 12 h on the day of estrus and from 24 h on the day of diestrus to 18 h on the day of proestrus. On the other hand, plasma concentrations of progesterone were significantly higher in LAA rats compared with HAA rats on the day of diestrus. The number of antral follicles (300-600 microm in diameter) at 12 h on the day of proestrus was significantly fewer in LAA rats than in HAA rats. The size and number of corpus luteum at 12 h on the day of estrus were significantly greater in LAA rats than in HAA rats. These results clearly demonstrated that apparent differences are observed in reproductive endocrinology between two Hatano strains. These strain differences probably originated from neural regulation of pituitary hormones.

Animals↗

PubMedDate not supplied

Differences in sperm motion between high- and low-shuttlebox avoidance rats (Hatano strains).

Sperm from the caudal epididymis of 2 inbred strains of Sprague-Dawley (SD) rats, selected on the basis of their high- or low-shuttlebox avoidance responses, were analyzed for motion characteristics by a computer-assisted sperm motion analysis (CASA) system. Sperm motion in high-avoidance animals (HAA) was characterized by high velocities, high amplitude of lateral head displacement (ALH), and low beat cross frequency (BCF). Conversely, sperm from low-avoidance animals (LAA) displayed low velocities, low ALH, and high BCF. These characteristics in sperm motion were not changed by washing. Furthermore, after treatment with alpha-chlorohydrin (aCH) as a male antifertility agent affecting rat epididymal sperm motion, sperm velocities in HAA rats were significantly reduced to levels similar to those in untreated LAA rats. However, ALH and BCF in HAA rats treated with aCH were different from those in untreated LAA rats. Sperm adenosine triphosphate (ATP) content was higher in HAA than in LAA rats, correlating with values of their sperm velocities. These data suggest there are apparent strain differences in sperm motion between HAA and LAA rats and that these differences are dependent on factors, including sperm energy production.

Adenosine Triphosphate↗