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Y Oiso

Publications and source records attributed to Y Oiso.

At least 19 recordsLinked to original sources

Analysis of the vasopressin system and water regulation in genetically polydipsic mice.

Polydipsic mice, STR/N, which show extreme polydipsia and polyuria, were discovered in 1958. In the STR/N, urine outputs are much higher than in control mice. The possibility of an abnormal regulation of the arginine vasopressin (AVP) system, or an abnormality in the renal susceptibility to AVP, should be considered. In this study we investigated the AVP system and water regulation in STR/N. We sequenced the AVP and the AVP V(2)-receptor genes of the STR/N by direct sequencing. No mutation was found in either of them. AVP gene expression examined by in situ hybridization and plasma sodium in 8-wk-old STR/N was significantly lower than in control mice, whereas it was significantly higher at 20 wk. Renal sensitivity to injected AVP was attenuated in 20-wk-old STR/N. The suppression of AVP synthesis due to excessive water retention in 8-wk-old STR/N suggests that polydipsia may be the primary cause in this strain. The 20-wk-old STR/N became dehydrated with the acceleration of AVP synthesis, which might have resulted from secondary desensitization to AVP.

Amino Acid Sequence

Administration of oxytocin affects vasopressin V2 receptor and aquaporin-2 gene expression in the rat.

Oxytocin (OT) binds to the vasopressin V2 receptor (V2R) because of its structural similarity to arginine vasopressin (AVP). Though the affinity of OT for V2R is low, it is known that OT causes antidiuresis. To clarify the effect of OT as an agonist of V2R, we investigated the influence of acute elevation of plasma OT levels on the rat mRNA expression of V2R and aquaporin-2 (AQP2), the water channel regulated by V2R. The plasma OT level increased from 11.1+/-1.6 pg/ml to 331.0+/-67.9 pg/ml by 1 h after subcutaneousinjection of 20 microg OT. V2R mRNA expression decreased to 68.3+/-4.1% of the control at 3 h, and AQP2 mRNA expression increased to 239.3+/-26.8% of the control at 6 h. The plasma AVP level did not change significantly during the experiment. The influence of a subcutaneous injection of 20 microg OT on V2R and AQP2 mRNA expression is comparable to that of 10 microg AVP that we documented in the previous study. In conclusion, OT can downregulate V2R mRNA expression and upregulate AQP2 mRNA expression in the collecting duct as an agonist of the V2R like AVP.

Animals

Rapid and sensitive vasopressin heteronuclear RNA responses to changes in plasma osmolality.

The plasma concentration of arginine vasopressin (AVP) is closely regulated by plasma osmolality. In this study, we used intronic in-situ hybridization to investigate the transcriptional activity of the AVP gene in the supraoptic nucleus (SON) and paraventricular nucleus (PVN) after intraperitoneal (i.p.) injection of hypertonic saline inducing small changes in plasma osmolality in rats. After ip injection of 900 mOsm/kg saline (2% BW), plasma [Na] reached the highest level at 10 min (900 mOsm/kg, 146+/-1 mEq/l; 290 mOsm/kg, 139+/-1 mEq/l, P<0.01) and maintained that level until 30 min. The expression of AVP heteronuclear (hn) RNA in the SON and PVN increased significantly as early as 10 min and peaked 30 min after ip injection of the hypertonic saline. AVP hnRNA expression showed a significant increase even after a 2 mEq/L elevation in plasma [Na] concentration, and increased dose-dependently in response to the concentration of saline injection (290-900 mOsm/kg). These results demonstrate that, similar to AVP secretion, AVP gene transcriptional activity in the SON and PVN is closely regulated by plasma osmolality.

Animals

Molecular mechanisms of the negative effect of insulin-like growth factor-I on growth hormone gene expression in MtT/S somatotroph cells.

Although insulin-like growth factor-I (IGF-I) is shown to have a suppressive effect on GH gene expression at the pituitary level, its molecular mechanism has not yet been clarified. To study the issue, we established a new in vitro system using MtT/S, a recently established rat somatotroph tumor cell line that retains the basic characteristics of somatotroph function. Plasmids containing the GH 5' promoter (approximately 1.75 kb or shorter)-luciferase fusion gene were transfected stably or transiently into the cells, and the effect of IGF-I on the GH promoter activity was estimated by a luciferase assay. The results showed that IGF-I inhibited GH promotor activity (more than 50% suppression) in a time- and dose-related manner. IGF-I also inhibited GH secretion. A study using deletion mutants of the GH promoter revealed that the negative effect was maintained in the shortest construct (-80 to +6), suggesting that IGF-I-related factor is acting at the region very close to the minimal promoter. Interestingly, the negative effect was completely eliminated by a PI3 kinase inhibitor wortmannin (1 microM), whereas a MAP kinase inhibitor PD98059 (20 microM) or S6 kinase inhibitor rapamycin (10 nM) did not influence the effect. Our results suggest that IGF-I suppresses GH gene expression at the transcriptional level and that the PI3 kinase-mediated signaling pathway plays a major role in the negative effect of IGF-I. We believe that our system using MtT/S cells is an excellent experimental model system for studying the cellular and molecular mechanisms of the transcriptional regulation of GH in vitro.

Androstadienes

Central administration of urocortin inhibits vasopressin release in conscious rats.

Urocortin (UCN) is a new mammalian member of the corticotropin releasing factor (CRF) family and supposed to be an endogenous ligand for type 2 CRF receptors. Previous studies have revealed that UCN mRNA exists in the supraoptic nucleus (SON), and that water deprivation increases UCN immunoreactivity in SON. In this study, we examined the effect of centrally-administered UCN on arginine vasopressin (AVP) release in conscious rats. Intracerebroventricular (i.c.v.) injection of UCN (5.0 microg/rat) significantly attenuated AVP release induced by hyperosmolality at 30 min after the injection. In contrast, CRF (5.0 microg/rat) injected i.c.v. had no significant effect on AVP release. These results suggest that central UCN play an inhibitory role in osmoregulation of AVP release.

Animals

Age-associated decrease in response of rat aquaporin-2 gene expression to dehydration.

It is well known that urine-concentrating ability decreases with aging and that this decreasing ability results from a reduced sensitivity of the renal collecting duct to arginine vasopressin (AVP). AVP regulates the water channel (aquaporin-2:AQP2) through V2 receptors and increases the water permeability of the collecting duct. To elucidate the mechanism of change with aging in urine-concentrating ability, we investigated the change of V2 receptor and AQP2 mRNA expression in young (8-week-old) and older (7-month-old) rats after dehydration for 2 days. After dehydration, plasma AVP levels in older rats were higher than young rats, and urinary osmolality in older rats was lower than young rats. By Northern blot analysis, there was no significant difference between young and older rats in both V2 receptor and AQP2 mRNA expression before dehydration. After dehydration, V2 receptor mRNA expression in young and older rats decreased in the same degree, suggesting the downregulation of V2 receptors may occur in the mRNA level. Northern blot analysis and in situ hybridization histochemistry showed that AQP2 mRNA expression increased and the increased expression in older rats was less than in young rats. The present study suggests the reduced response of AQP2 mRNA expression to dehydration, observed in 7-month-old rats, might be partially responsible for the decreasing urine-concentrating ability with aging.

Aging

Antiserum against neuropeptide FF augments vasopressin release in conscious rats.

We previously reported that centrally administered neuropeptide FF (NPFF) inhibited arginine vasopressin (AVP) release. In this study, immunoneutralization of central NPFF was performed to evaluate the role of endogenous NPFF in the regulation of AVP release. Intracerebroventricular (ICV) injection of antiserum against NPFF (Anti-NPFF) significantly augmented the plasma AVP increase induced by hyperosmolality [intraperitoneal injection of hypertonic saline (600 mOsm/kg, 2% BW)] at 60 min after ICV injection compared with normal rabbit serum (NRS) (NRS: 4.20+/-0.30 pg/ml, Anti-NPFF: 5.83+/-0.46 pg/ml, p < 0.01). Anti-NPFF did not cause significant change in plasma osmolality, plasma volume or arterial blood pressure. This evidence indicates that endogenous NPFF might be physiologically involved in osmoregulation of the plasma AVP level through its inhibitory action.

Amino Acid Sequence

Anorectic effect of pituitary adenylate cyclase activating polypeptide (PACAP) in rats: lack of evidence for involvement of hypothalamic neuropeptide gene expression.

We investigated the effect of centrally administered pituitary adenylate cyclase activating polypeptide (PACAP) on feeding in rats, and the involvement of hypothalamic neuropeptide gene expression using in situ hybridization. Intracerebroventricular injection of PACAP (1000 pmol/rat) significantly decreased food intake in a dose-dependent manner. In PACAP-treated rats, neuropeptide Y (NPY) mRNA levels in the arcuate nucleus and galanin mRNA levels in the paraventricular nucleus increased, and corticotropin-releasing hormone (CRH) mRNA levels in the paraventricular nucleus decreased. In rats fasted for 72 h, NPY mRNA levels increased, and CRH mRNA levels decreased, but galanin mRNA levels were unchanged. These results indicate that the anorectic function of PACAP is not mediated by NPY or CRH, and that PACAP increases galanin synthesis.

Animals

Regulation of vasopressin synthesis and release by area postrema in rats.

There is evidence indicating that the area postrema (AP), the most caudal circumventricular organ located on the dorsal surface of the medulla, is involved in several physiological regulations. In this study, we investigated the role of AP in the regulation of arginine vasopressin (AVP) synthesis and release, using rats of which the AP was lesioned 6 weeks previously. The level of plasma AVP in the AP lesioned (APX) group was significantly lower than in the sham operated (Sham) group in the basal state. AVP release induced by either hyperosmolality or hypovolemia was significantly attenuated by APX. To clarify the role of AP in AVP synthesis in the hypothalamus, we examined the AVP gene expression using in situ hybridization. AVP messenger RNA levels in paraventricular (PVN) and supraoptic nuclei (SON) in the APX group were significantly lower than in the Sham group in the basal state. Moreover, the AVP messenger RNA levels in PVN and SON in the APX group were also significantly lower than in the Sham group after water deprivation for 3 days. These results suggest that AVP synthesis and release are tonically stimulated by AP in the basal state and that AVP synthesis and release in stimulated states are also regulated, at least partially, by AP.

Animals

Cytokine regulation of the rat proopiomelanocortin gene expression in AtT-20 cells.

Although cytokines are known to be involved in the regulation of ACTH secretion, their effects, along with the molecular mechanisms, on POMC gene expression are not thoroughly characterized. In this study we examined the effects of representative cytokines on transcription of the POMC gene in corticotrophs in vitro using AtT20PL, a clone of the AtT20 cell line stably transfected with approximately 0.7 kilobase of the rat POMC 5'-promoter-luciferase fusion gene. In each experiment, cells were incubated with the cytokine tested, and the changes in POMC 5'-promoter activity were determined by a luciferase assay. The results showed that interleukin-1beta (IL-1beta) stimulated promoter activity in a biphasic manner [weak short term effects (2-3 h) followed by potent long term effects (>12-16 h)]. Tumor necrosis factor-alpha had similar effects, but much less potency. IL-6 showed a profound stimulatory, but only a long term (>20 h), effect. IL-2 did not influence POMC expression. In contrast, interferon-alpha (IFN alpha) and IFN-gamma showed acute stimulatory effects (approximately 4 h) followed by marked inhibitory effects (>8 h). Although the acute effects of IL-1beta, IL-6, and tumor necrosis factor-alpha alone were minimal, they significantly potentiated the stimulatory effect of CRH on POMC expression. Finally, pretreatment of the cells with a broad spectrum tyrosine kinase inhibitor, genistein, abolished or significantly diminished the effects of all cytokines except IFNs. Our results suggest that 1) each cytokine tested has a distinct effect on POMC gene expression; 2) there are positive cross-talk effects between CRH and cytokines at the corticotroph level; and 3) tyrosine phosphorylation cascades are involved in the intracellular signaling mechanisms of some cytokines.

Adrenocorticotropic Hormone

Influence of acute elevation of plasma AVP level on rat vasopressin V2 receptor and aquaporin-2 mRNA expression.

It is known that vasopressin V2 receptor (V2R) mRNA is downregulated by elevated plasma arginine vasopressin (AVP) following chronic osmotic stimulation. To elucidate the response in V2R mRNA expression to acute elevation of the plasma AVP level, we investigated the time-course change of rat V2R mRNA expression after subcutaneous injection of AVP (10 microg/body). Plasma AVP levels increased from 1.4+/-0.3 pg/ml to 56.8+/-10.7 pg/ml by an hour after injection,and returned to the control level at 6 h. By Northern blotanalysis, V2R mRNA expression decreased to 52.7+/-3.7% of the control level at 2 h, and then returned to the control level by 6 h. Furthermore we investigated the time-course change of aquaporin-2 (AQP2) mRNA expression. AQP2 mRNA expression increased gradually after injection and reached 240.3+/-7.5% of the control level at 6 h. Then it returned to the control level. This study showed that the downregulation of V2R mRNA occurred rapidly after acute elevation of the plasma AVP level, and AQP2 mRNA expression was upregulated despite the downregulation of V2R mRNA.

Animals

Non-genomic mechanisms of glucocorticoid inhibition of adrenocorticotropin secretion: possible involvement of GTP-binding protein.

We investigated non-genomic mechanisms of glucocorticoid negative feedback regulation on pituitary corticotroph cells using the AtT20 mouse corticotroph tumor cell line. A synthetic glucocorticoid dexamethasone (100 nM) potently suppressed forskolin-induced cAMP generation, adrenocorticotropin (ACTH) secretion, and proopiomelanocortin gene expression. When de novo gene expression was inhibited by actinomycin D (1 microM), dexamethasone still suppressed cAMP efflux and ACTH release, although less potently. Interestingly, under the same conditions, pretreatment of the cells with pertussis toxin (50 ng/ml) completely abolished the suppressive effect of dexamethasone on both parameters. These results suggest that non-genomic and genomic mechanisms are involved in the glucocorticoid negative regulation of ACTH expression, and a pertussis toxin-sensitive GTP-binding protein might, at least partly, participate in the non-genomic effect.

8-Bromo Cyclic Adenosine Monophosphate