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Biomedical subjects

A Miyake

Publications and source records attributed to A Miyake.

At least 127 records · Page 7Linked to original sources

Involvement of epidermal growth factor in inducing adiposity of age female mice.

Aged mice exhibit an increase in their body weight (BW), which is associated with fat deposit increase. Epidermal growth factor (EGF) concentration in the submandibular gland also increases with aging. We examined the effects of elevated EGF on the adiposity of aged female mice. Studies were started in two groups of animals consisting of sham-operated (n = 10) and sialoadenectomized (n = 10, Sx; surgical removal of the submandibular glands) mice at 8 weeks of age. Body weight gain and food intake were measured throughout 78 weeks of age in these two groups. Body weight was significantly less in the Sx group throughout 78 weeks, while food intake was not changed by Sx after 12 weeks of age. To examine further if EGF plays a role in the induction of adiposity in aged female mice, sham-operated animals were given 100 microliters anti-EGF rabbit antiserum (anti-EGF group, n = 5) or normal rabbit serum (control group, n = 5) every 3 days, and Sx animals were given 5 micrograms/day EGF (Sx+EGF group, n = 5) or saline (Sx group, n = 5) from 78 weeks of age for 3 weeks. At 81 weeks of age, all animals of these four groups were killed, and carcass fat deposition and fat cell sizes were measured. Although the relative weights (weight ratio to BW) of the liver and kidney were not changed by Sx and anti-EGF treatment, the relative weights of mesenteric and subcutaneous fat tissues and adipocyte weights were significantly decreased in Sx and anti-EGF groups compared with the control group. Moreover, both acyl-CoA synthetase (ACS) and lipoprotein lipase (LPL) mRNA levels were significantly decreased by Sx or anti-EGF administration in mesenteric and subcutaneous fat tissues. On the other hand, EGF administration to Sx animals had no effect on BW, fat tissues and adipocyte weights, and ACS and LPL mRNA levels. The results, however, were consistent with the fact that adipose tissue EGF receptors were down regulated in Sx mice. These findings suggest that EGF may play a role in the induction of adiposity in aged female mice.

Adipose Tissue↗

Activin inhibits but inhibin activates mouse placental lactogen-II secretion.

The regulation of mouse placental lactogen (mPL)-I and mPL-II secretion by activin and inhibin and the expression of activin and inhibin subunit mRNAs in the mouse decidua were examined. Activin-A at a concentration of 10 nM/l significantly inhibited mPL-II secretion by placental cells from days 9 and 12 of pregnancy. However, activin-A did not affect mPL-I secretion by cells from days 7 and 9 of pregnancy nor mPL-II secretion by cells from day 7 of pregnancy. By contrast, 10 nM/l inhibin activated mPL-II secretion by cells from day 12 of pregnancy. These effects of activin and inhibin on mPL-II secretion were dose-dependent. Follistatin, which binds to activin and blocks its bioactivity, completely eliminated the inhibitory effect of activin on mPL-II secretion. Incubation of placental cells from day 12 of pregnancy with activin-A resulted in a significant reduction of the mPL-II mRNA level assessed by Northern blot analysis. Northern blot analysis using poly(A)+RNA extracted from the decidua indicated that mouse decidua, as well as the placenta, express all activin and inhibin subunits and that their gene expressions increased during gestation. The expression of these mRNAs in the decidua was much higher than those in the placenta. These findings suggest that activin and inhibin regulate mPL-II secretion and suggest the presence of an autocrine or paracrine regulation of mPL-II secretion in mouse placenta by activin and inhibin after mid-pregnancy in vivo.

Activins↗

Inhibition of mouse placental lactogen-II release from placental cells by interleukin-1 after mid-pregnancy.

The effects of interleukin (IL)-1 and granulocyte-macrophage colony stimulating factor (GM-CSF), which are present in the mouse placenta, on the secretion of mouse placental lactogen (mPL)-1 and mPL-II by placental cells were tested in vitro. IL-1 alpha and IL-1 beta, 2.5 nmol/l each, significantly inhibited mPL-II secretion by cells from days 9 and 12 of pregnancy, but did not affect mPL-II secretion by cells from day 7 of pregnancy or mPL-I secretion by cells from days 7, 9 or 12 of pregnancy. GM-CSF had no effect on mPL-I and mPL-II secretion by cells from days 7, 9 or 12 of pregnancy. The inhibitory effects of IL-1 alpha and IL-1 beta on mPL-II secretion were completely eliminated by the addition of antibodies to IL-1 alpha and IL-1 beta respectively. Western blot analysis for mPL-II indicated that IL-1 alpha significantly reduced the intensity of the mPL-II band. Steady-state levels of mPL-II mRNA, assessed by Northern blot analysis, were reduced by incubation of placental cells from day 12 of pregnancy with 2.5 nmol/l IL-1 alpha for 5 days. Co-incubation of 0.25 pmol/l IL-1 alpha, 25 pmol/l IL-6, and 25 pmol/l tumor necrosis factor-alpha, each of which did not significantly inhibit mPL-II secretion by itself, together inhibited mPL-II secretion. These results suggest that IL-1, but not GM-CSF, is a potent inhibitor of mPL-II secretion after mid-pregnancy, and that the combined action of cytokines can inhibit mPL-II secretion.

Animals↗

Menstrual cycle specific expression of epidermal growth factor receptors in human fallopian tube epithelium.

We studied the expression of epidermal growth factor (EGF) receptor protein and messenger RNA (mRNA) in human fallopian tubes at three stages of the menstrual cycle: early follicular (n = 3), late follicular (n = 3) and luteal (n = 3). Immunohistochemical studies in the ampullary portion of the tubes showed that specific staining was localized to the epithelium and the vascular endothelium. Staining of the epithelium was intense at the late follicular and luteal stages, while it was weak at the early follicular stage. 125I-EGF binding study in the tubal plasma membranes revealed a class of high-affinity EGF receptors. Although dissociation constants were similar between the stages, numbers of binding sites at the late follicular and luteal stages were significantly (P < 0.01) greater than those at the early follicular stage. Western blotting showed that tubal plasma membranes contain M(r) 170,000 EGF receptor protein. The amounts were significantly (P < 0.01, n = 3) greater at the late follicular and luteal stages than those at the early follicular stage. Reverse transcription and polymerase chain reaction (RT-PCR) revealed that EGF receptor mRNA was expressed in all the 9 RNA samples (n = 3 for each stage) from the tubal ampullary portion. The amounts were significantly (P < 0.01, n = 3) greater at the late follicular and luteal stages than those at the early follicular stage (by a competitive PCR). Increase in the amounts of EGF receptor protein and mRNA occurred in association with an increase in serum oestradiol but not progesterone levels. Next we examined whether EGF receptor and its ligands (EGF and transforming growth factor alpha) are directly induced by oestrogen. We found that specific staining for EGF receptor and its ligands in the tubal epithelium was detected (by immunohistochemistry) in postmenopausal women with oestrogen replacement (n = 3), but not in subjects without oestrogen replacement (n = 3). These results suggested that EGF receptors in the human tubal epithelium are expressed in relation to specific stages of the menstrual cycle and that the expression may be induced by oestrogen.

Base Sequence↗

A case of nonmetastatic trophoblastic disease followed by magnetic resonance imaging.

The consecutive findings of the magnetic resonance (MR) imaging in a patient with nonmetastatic trophoblastic disease are reported. On MR image, there are noted two kinds of MR findings which suggest the existence of trophoblastic disease; one was a typical hypervascular mass of heterogeneous signal intensity within myometrium, and the other was the increase in myometrial and parametrial flow void. The former finding appeared only for a short period while the disease was highly active. The latter finding well paralleled the serum human chorionic gonadotropin (hCG) level, and the remarkable flow void, indicating dilatated blood vessels, disappeared with the complete remission of the disease. These findings suggest that MR imaging may be useful for diagnosing and following gestational trophoblastic disease.

Adult↗

Molecular cloning of human 5-hydroxytryptamine3 receptor: heterogeneity in distribution and function among species.

The 5-hydroxytryptamine3 receptor 5-HT3R has been implicated in gut and cardiac motility and in behavioral disorders. Characteristics of 5-HT3Rs appear to be heterogeneous among species, but human 5-HT3R cDNA has not been identified. We isolated a cDNA encoding 5-HT3R from human hippocampus. The mouse 5-HT3R gene has been reported to generate two alternative splicing isoforms that differ by six amino acids. All of our isolated human clones corresponded to the shorter isoform. Amino acid identities with mouse neuroblastoma N1E-115 and rat brain 5-HT3Rs were 84% for each. Southern blot analysis of human genomic DNA suggested that our cloned transcript encoded a human counterpart for the rodent 5-HT3Rs. This gene was assigned to chromosome 11 using polymerase chain reaction analysis of a human/rodent somatic cell hybrid panel. With the use of Northern blot analysis, 5-HT3R transcripts were identified in human small intestine, colon, and brain regions including hippocampus, amygdala, and striatum. In human heart, 5-HT3R expression was not detectable even with reverse transcriptase-polymerase chain reaction analysis, although it was detectable in mouse heart. Transfection of COS-1 with human 5-HT3R cDNA induced specific binding of the 5-HT3R-selective radioligand [3H]YM060. Human 5-HT3R showed typical characteristics of the 5-HT3R, but its affinity for the 5-HT3R agonist m-chlorophenylbiguanide was much lower than that of rat 5-HT3R. When injected with human 5-HT3R cRNA, the oocytes responded to 5-HT3R agonists with a rapidly developing inward current. The potency of the agonists to induce inward current paralleled that to compete with the radioligand binding, and 2-methyl-5-hydroxytryptamine, a partial agonist for mouse 5-HT3R, was a full agonist for human 5-HT3R. Our data revealed that the 5-HT3R molecule has interspecies differences in both tissue distribution and functional profile.

Amino Acid Sequence↗

Cervical invasion of endometrial carcinoma--evaluation by parasagittal MR imaging.

Twenty-seven consecutive patients were examined by T2-(1,800/70 ms) and postcontrast T1-weighted (600/15) spin echo (SE) or dynamic (200/15) SE MR imaging to determine the usefulness of parasagittal MR imaging in assessing cervical invasion of endometrial carcinoma. The images were obtained in a direction parallel to the longitudinal axis of the uterus (parasagittal). The cervical epithelium, being hyperintense on the late phase dynamic and postcontrast T1-weighted SE images, had disappeared partially or totally in all 4 patients with cervical invasion. The enhanced cervical epithelium was completely seen in one patient with the tumor protruding into the cervical canal in a polyp-like form without cervical epithelial invasion. The same was also seen in the 22 patients with the tumor remaining in the corpus cavity. The enhanced parasagittal MR images facilitated the evaluation of the extent of the endometrial carcinoma.

Adenocarcinoma↗

Decidua is a possible source of serum mouse soluble interleukin-6 receptor (msIL-6R): gestational profile of serum msIL-6R concentration.

The serum concentration of msIL-6R of 10 weeks old virgin ICR mouse assessed by a RIA was 45.6 +/- 6.6 ng/ml. The serum msIL-6R concentration of the pregnant mouse mated at 10 weeks of age was 41.2 +/- 3.0 ng/ml on day 7 of pregnancy. The serum concentration gradually increased during gestation and reached peak on day 17 of pregnancy (149.1 +/- 13.7 ng/ml). On the third day of the puerperium, the serum msIL-6R concentration of the mother from which the pups had been removed on the day of delivery was decreased to the level of that on day 7 of pregnancy. IL-6R mRNA level in the decidua significantly increased during mid and late gestational stages. msIL-6R was detected in the medium of cultured decidual cells (8.03 +/- 0.28 ng/ml), but not placental cells. Western analysis for msIL-6R using the conditioned medium of the cultured decidual cells resulted in a single band at approximately 45 K. To find out a biological role of msIL-6R during gestation, placental cells were incubated with mIL-6 and msIL-6R, and mPL-I concentration in the medium was assessed by RIA. 2.5 nM mIL-6 did not affect the secretion of mPL-I in placental cells; however, addition of msIL-6R resulted in a significant stimulation of mPL-I secretion. These results suggest that serum msIL-6R, which is likely to be secreted from decidua, may play an important role during gestation.

Analysis of Variance↗

Implantation and growth of epidermal growth factor (EGF) receptor expressing human ovarian cancer xenografts in nude mice is dependent on EGF.

BACKGROUND: The importance of epidermal growth factor (EGF) receptor-dependent growth has not been clarified for in vivo growth of primary human ovarian cancers. METHODS: Seventeen primary human ovarian cancer tissue samples were examined for the presence of EGF receptors by a 125I-EGF-binding study. Three groups of mice were inoculated with EGF receptor expressing and not-expressing cancer tissues. The groups were as follows: control group, Sx group (mice that underwent sialoadenectomy; EGF depleted mice), and Sx+EGF (EGF-replaced) group. The ability of the inoculated tissues to implant and grow then was studied. RESULTS: Of the 17 primary ovarian cancers, 12 expressed EGF receptors and 5 did not. Eight of 12 EGF-receptor expressing cancer tissues implanted and formed growing tumors in control animals. None implanted in the Sx animals. Epidermal growth factor receptor-expressing cancers implanted in Sx animals that received EGF administration. Two of five EGF receptor-negative ovarian cancers implanted and grew in both control and Sx animals. CONCLUSION: Growth of EGF receptor-expressing primary human ovarian cancers may be dependent on EGF in vivo.

Animals↗

Transforming growth factor-beta 1 post-transcriptionally inhibits mouse growth hormone releasing factor secretion in placenta.

The aim of this study was to investigate whether TGF-beta 1 regulates mouse GHRF secretion by primary cultures of placental cells from day 12 of pregnancy. Ten ng/ml TGF-beta 1 significantly inhibited mouse GHRF secretion by the third day of culture. The lowest concentration of TGF-beta 1 that significantly inhibited mouse GHRF secretion was 1 ng/ml. The inhibitory effect of TGF-beta 1 on mouse GHRF secretion was completely eliminated by addition of an anti-TGF-beta 1 antibody. Steady-state levels of mouse GHRF mRNA as assessed by Northern analysis was not reduced by incubation of placental cells from day 12 of pregnancy with 10 ng/ml TGF-beta 1 for 5 days. Both placenta and decidua expressed TGF-beta 1 mRNA, and the level of TGF-beta 1 mRNA in decidua increased during gestation while the level of TGF-beta 1 mRNA was constant throughout gestation. These findings suggest that TGF-beta 1 is one of the potent regulators of mouse GHRF secretion and that TGF-beta 1 regulates mouse GHRF secretion in an autocrine or paracrine manner in the mouse placenta in vivo.

Animals↗

Characterization of activin A-, activin AB- and activin B-responding cells by their responses to hypothalamic releasing hormones.

Activin A-, AB- and B-responding cells were characterized by their responsiveness in cytosolic free calcium ([Ca2+])i) to four hypothalamic releasing hormones, CRH, GHRH, TRH and GnRH. First, rat pituitary cells responding to activin A, AB and B in [Ca2+]i were determined in a mixed population of pituitary cells. The populations of the activin A-, AB-, and B-responding cells were 13.9%, 9.3% and 13.2%, respectively. Overlapping of response among each population of activin- responding cells was present in some of the cells. The cells responding to activin A, AB, and B were then characterized by their responses to CRH, GHRH, TRH and GnRH. Most of the cells responding to activin A, AB, and B also responded to GHRH or TRH. These results reveal that there are distinct differences among each population of activin A-, AB- and B-responding cells and that there is still functional overlapping of responsiveness among these populations. The characterization of activin-responding cells suggests involvement of somatotropes and lactotropes in activin-induced biological events in the pituitary.

Activins↗

Tumor necrosis factor-alpha (TNF-alpha) inhibits expression of mouse placental lactogen-II through TNF-alpha type-I but not type-II receptor.

The aim of this study was to determine whether TNF-alpha inhibits mPL-II secretion through TNF-RI or TNF-RII, and to investigate the gestational profile of TNF-RI and TNF-RII gene expression. The mouse trophoblast cells from day 12 pregnancy were cultured with or without agonistic polyclonal antibodies directed against the individual TNF-alpha receptors, and mPL-II secretion in the medium was assessed by RIA. Anti-TNF-RI antibody significantly inhibited the mPL-II secretion in a dose- and time-dependent manner, but anti-TNF-RII antibody did not. Moreover, the TNF-RII antibody did not influence the inhibitory effect of the TNF-RI antibody on mPL-II secretion. TNF-RI antibody inhibited the mPL-II gene expression by Northern blot analysis. Amount of the gene expression of TNF-RI in the second half of pregnancy was higher than those in the first half of pregnancy both in vitro and in vivo, although the gene expression of TNF-RII was not detectable by Northern blot analysis using poly(A)+RNA. These results suggest that TNF-RI is a main receptor for TNF-alpha in mouse placenta and that TNF-alpha may have an important role in regulating mPL-II secretion after midpregnancy.

Animals↗

In vitro effects of CINC/gro, a member of the interleukin-8 family, on hormone secretion by rat anterior pituitary cells.

We investigated the effects of CINC/gro on hormone secretion using normal rat anterior pituitary cells. In normal anterior pituitary cells, 10-100 ng/ml of CINC/gro significantly increased the secretion of PRL within 3 h of incubation, and two-fold enhancement of PRL secretion was induced by 100 ng/ml of CINC/gro within 24-h incubation, while the response of GH and ACTH secretions to CINC/gro was weak. On the other hand, CINC/gro suppressed basal LH and FSH secretions in a concentration-dependent manner. The percent inhibition of basal secretion by CINC/gro (50 ng/ml) within 24-h incubation was 70% for LH and 43% for FSH. Twenty-four-hour incubation with 100 ng/ml of IAP completely blocked the CINC/gro-stimulated PRL and GH secretions and CINC/gro's suppression of both basal LH and FSH secretions. These data demonstrate a new biological activity for CINC/gro and provide evidence for immune system regulation of anterior pituitary hormone secretion.

Analysis of Variance↗

The production of CINC/gro, a member of the interleukin-8 family, in rat anterior pituitary gland.

We investigated the possibility of detection of CINC/gro, which is a IL-8-like neutrophil chemoattractant, immunoreactivity in rat normal anterior pituitary gland by immunohistochemistry, western blot analysis and an ELISA. We first ascertained the possibility of detection of CINC/gro immunoreactivity in the anterior pituitary gland by immunocytochemistry. In the anterior lobe of the pituitary gland, a few CINC/gro-like immunoreactive cells were observed (1-3% of all cells in the anterior pituitary). The positive cells were middle or large in size and looked angular in shape. Intense immunoreactivity was observed in the cytoplasm but not in the nucleus. Analysis by immunoblotting with anti-CINC/gro antiserum gave a characteristic single CINC/gro band with a molecular weight of 6.3 kDa. CINC/gro immunoreactivity was also detected in 3-h conditioned medium of normal anterior pituitary cells by an ELISA, and that immunoreactivity increased significantly in a time-dependent manner during 24-h incubation. This immunoreactivity could be induced by TNF-alpha in a dose-dependent manner. These findings indicate that CINC/gro is produced in pituitary gland and also suggests the possibility that CINC/gro may play some role asa modulator of anterior pituitary function, especially in the cross-talk mechanism between the immune and neuroendocrine systems.

Analysis of Variance↗

Dopamine inhibits TRH-induced MAP kinase activation in dispersed rat anterior pituitary cells.

We recently reported the existence of two separate pathways for thyrotropin-releasing hormone (TRH)-induced mitogen-activated protein (MAP) kinase activation in GH3 pituitary tumor cells. To test the role of MAP kinase in TRH action, we examined the effect of dopamine (DA) on TRH-induced MAP kinase in primary cultures of rat anterior pituitary cells. 1 microM of DA attenuated 1 microM TRH-induced MAP kinase activity and phosphorylation. 100 ng/ml of islet-activating protein (IAP) blocked these inhibitory effects of DA. These results suggest that crosstalk exists between the DA signaling pathway and the TRH-stimulated MAP kinase activating pathway in rat anterior pituitary cells.

Adenosine Triphosphate↗

In vitro effects of CINC/gro, a member of the interleukin-8 family, on interleukin-6 secretion by rat posterior pituitary cells.

We investigated the effects of CINC/gro on IL-6 secretion by rat posterior pituitary cells. CINC/gro immunoreactivity was already detected in 1-h conditioned medium of normal posterior pituitary cells, and it increased significantly in a time-dependent manner during the first 24 h of culture. This immunoreactivity could be induced by TNF-alpha in a dose-dependent manner. On the other hand, CINC/gro stimulated IL-6 secretion by posterior pituitary monolayer cultures in a concentration dependent manner. Thus, CINC/gro significantly (P < 0.01) increased the secretion of IL-6 within 13 h of incubation, and this effect continued throughout 24 h of incubation. The stimulatory effect of 100 ng/ml CINC/gro on IL-6 secretion was completely blocked by 24-h incubation with 100 ng/ml IAP. These data demonstrate a new biological activity for CINC/gro in the posterior pituitary system.

Animals↗