Decrease In Inositol Research Articles

Inositol 1,4,5-trisphosphate receptor type 2 is associated with the bone–vessel axis in chronic kidney disease–mineral bone disorder

Objective The pathogenesis of renal osteopathy and cardiovascular disease suggests the disordered bone–vessel axis in chronic kidney disease–mineral bone disorder (CKD–MBD). However, the mechanism of the bone–vessel axis in CKD–MBD remains unclear. Methods We established a CKD–MBD rat model to observe the pathophysiological phenotype of the bone–vessel axis and performed RNA sequencing of aortas to identify novel targets of the bone–vessel axis in CKD–MBD. Results The microarchitecture of the femoral trabecular bone deteriorated and alveolar bone loss was aggravated in CKD–MBD rats. The intact parathyroid hormone and alkaline phosphatase levels increased, 1,25-dihydroxyvitamin D3 levels decreased, and intact fibroblast growth factor-23 levels did not increase in CKD–MBD rats at 16 weeks; other bone metabolic parameters in the serum demonstrated dynamic characteristics. With calcium deposition in the abdominal aortas of CKD–MBD rats, RNA sequencing of the aortas revealed a significant decrease in inositol 1,4,5-trisphosphate receptor type 2 (ITPR2) gene levels in CKD–MBD rats. A similar trend was observed in rat aortic smooth muscle cells. As a secretory protein, ITPR2 serum levels decreased at 4 weeks and slightly increased without statistical differences at 16 weeks in CKD–MBD rats. ITPR2 serum levels were significantly increased in patients with vascular calcification, negatively correlated with blood urea nitrogen levels, and positively correlated with serum tartrate-resistant acid phosphatase 5b levels. Conclusion These findings provide preliminary insights into the role of ITPR2 in the bone–vessel axis in CKD–MBD. Thus, ITPR2 may be a potential target of the bone–vessel axis in CKD–MBD.

Neuroprotective Effects of Lithium in Patients with Bipolar Disorder

Background: Lithium has been considered the "gold standard" in the management of bipolar disorder, being the agent compared with "new mood stabilizer". It has also shown effect in the reduction of suicidal thoughts. Among the known mechanisms of action of lithium it has been described the inhibition in inositol monophosphatase (IMP) as resulting in non-competitive inhibition of IMP, resulting in a decrease of inositol in the central nervous system leading to a decrease in the synthesis of inositol triphosphate, affecting the transmission and translation of signals. Added to this, lithium inhibits kinase glycogen synthase enzyme (GSK) 3-β; it’s regulated by different signaling pathways and its dysfunction has been related to the pathophysiology of mood disorders, schizophrenia, diabetes and Alzheimer's disease. The neuroprotective effects have been shown of lithium in preclinical studies. The chronic administration of lithium enhances the proliferation of progenitor cells and inhibits cell loss induced by glutamate and glucocorticoids. In addition to a neurotrophic and neuroprotective role, lithium can also exert beneficial effect through its ability to improve mitochondrial function. Objective: Our aim was to explore the current information on the findings relating to lithium as a substance with possible neuroprotector effect in patients with diagnosis of bipolar disorder, as well as other conditions. Method: We performed a literature review regarding the use of Lithium and its potential neuroprotective effect in patients Conclusion: There is enough evidence that lithium may slow the progression of neurocognitive impairment in long term treatment patients, suggesting a potential use for the treatment of neurodegenerative diseases. Keywords: Bipolar disorder, mood disorders, lithium, neuroprotection, neurodegenerative, mood stabilizer.

Down-regulation of the Inositol 1,4,5-Trisphosphate Receptor in Mouse Eggs Following Fertilization or Parthenogenetic Activation

Fertilization in mammalian eggs is characterized by the presence of intracellular calcium ([Ca2+]i) oscillations. In mouse eggs, these oscillations cease after a variable period of time and this is accompanied by a decrease in inositol 1,4,5-trisphosphate receptor (IP3R) responsiveness and down-regulation of the IP3R type 1 (IP3R-1). To investigate the signaling pathway responsible for inducing IP3R-1 down-regulation during fertilization, mouse eggs were exposed to or injected with several Ca2+-releasing agonists and the amounts of IP3R-1 immunoreactivity evaluated by Western blotting. Exposure to ethanol or ionomycin, which induce a single [Ca2+]i rise, failed to signal down-regulation of IP3R-1. However, [Ca2+]i oscillations induced by injection of boar sperm fractions (SF), which presumably stimulate production of IP3, or adenophostin A, an IP3R agonist, both induced down-regulation of IP3R-1 of a magnitude similar to or greater than that observed after fertilization. Exposure to thimerosal, an oxidizing agent that modifies the IP3R without stimulating production of IP3, also initiated down-regulation of IP3R-1, although oscillations initiated by SrCl2 failed to evoke down-regulation of IP3R-1. The degradation of IP3R-1 in mouse eggs appears to be mediated by the proteasome pathway because it was inhibited by preincubation with lactacystin, a very specific proteasome inhibitor. We therefore suggest that persistent stimulation of the phosphoinositide pathway in mouse eggs by the sperm during fertilization or by injection of SF leads to down-regulation of the IP3R-1.

Ketamine Inhibits Inositol 1,4,5-Trisphosphate Production Depending on the Extracellular Ca2+ Concentration in Neonatal Rat Cardiomyocytes

We investigated the effect of ketamine on inositol 1,4,5-trisphosphate (IP3) formation in rat cardiomyocytes. After the addition of 1 μmol/L ketamine, IP3 production in the presence of 0.5, 1, 5, 10, and 30 mmol/L Ca2+ significantly decreased from 537.1 ± 8.3, 590.7 ± 12.9, 690.6 ± 7.9, 754.8 ± 12.5, and 823.7 ± 15.2 pmol/mg protein to 467.0 ± 8.3, 483.8 ± 11.0, 512.6 ± 21.3, 612.1 ± 16.9, and 652.6 ± 17.3 pmol/mg protein, respectively. When exposed to TMB-8 (a intracellular calcium inhibitor), IP3 production decreased significantly from 347.2 ± 27.3 to 283.8 ± 20.4 pmol/mg protein in the presence of 1 μmol/L ketamine, but A23187, which increases intracellular calcium, did not affect the inhibition of IP3 production by ketamine. These results demonstrate that ketamine decreases IP3 formation through inhibition of the calcium ion-sensing receptor and that IP3 formation reduced by ketamine is not affected by the alteration of intracellular calcium. Implications Ketamine has a negative inotropic effect in isolated cardiomyocytes. The negative inotropic effect was associated with a decrease in inositol 1,4,5-trisphosphate production, and the inhibitory action was enhanced depending on the concentration of extracellular Ca2+.

Ketamine inhibits inositol 1,4,5-trisphosphate production depending on the extracellular Ca2+ concentration in neonatal rat cardiomyocytes.

We investigated the effect of ketamine on inositol 1,4,5-trisphosphate (IP3) formation in rat cardiomyocytes. After the addition of 1 micromol/L ketamine, IP3 production in the presence of 0.5, 1, 5, 10, and 30 mmol/L Ca2+ significantly decreased from 537.1+/-8.3, 590.7+/-12.9, 690.6+/-7.9, 754.8+/-12.5, and 823.7+/-15.2 pmol/mg protein to 467.0+/-8.3, 483.8+/-11.0, 512.6+/-21.3, 612.1+/-16.9, and 652.6+/-17.3 pmol/mg protein, respectively. When exposed to TMB-8 (a intracellular calcium inhibitor), IP3 production decreased significantly from 347.2+/-27.3 to 283.8+/-20.4 pmol/mg protein in the presence of 1 micromol/L ketamine, but A23187, which increases intracellular calcium, did not affect the inhibition of IP3 production by ketamine. These results demonstrate that ketamine decreases IP3 formation through inhibition of the calcium ion-sensing receptor and that IP3 formation reduced by ketamine is not affected by the alteration of intracellular calcium. Ketamine has a negative inotropic effect in isolated cardiomyocytes. The negative inotropic effect was associated with a decrease in inositol 1,4,5-trisphosphate production, and the inhibitory action was enhanced depending on the concentration of extracellular Ca2+.

Poly-phosphoinositide-mediated messengers in focal cerebral ischemia and reperfusion

The receptor-mediated poly-phosphoinositide (PI) signalling pathway is known to play an important role in maintaining intracellular calcium homeostasis, which in turn, is critical for mediating neuronal function. In this study, we examined the effects of focal cerebral ischemia induced in rats by temporary occlusion of the middle cerebral artery (MCA) and both common carotid arteries (CCAs) on this signal transduction pathway. Results indicate that several parts of the pathway are altered, both during the early phase of focal cerebral ischemic insult and after recirculation. Cerebral ischemia induced a decrease in levels of phosphatidylinositol 4,5-biphosphate (PIP 2) in the ischemic MCA cortex, due partly to stimulated poly-PI hydrolysis and partly to the depletion of ATP required for resynthesis of this substrate. ATP depletion during ischemia was also attributed to a sustained decrease in inositol 1,4,5-triphosphate (IP 3) levels. On the other hand, the decline in IP 3 3-kinase activity after 30 min of ischemic insult was not related to ATP depletion. During reperfusion upon prolonged ischemic insult, neither IP 3 level nor IP 3 3-kinase activity were able to show recovery after reperfusion, despite that ATP levels recovered by 80%. In situ hybridization studies indicated a decrease in mRNA expression of IP 3 receptor but not IP 3 3-kinase during the initial 4 h of reperfusion after a 45 min ischemic insult. Under this same condition, insulted cortical neurons started to show morphological changes between 4 and 8 h after reperfusion and extensive cell death could be observed by 16 h. Taken together, these results demonstrated early and delayed changes in the poly-PI signalling pathway due to focal cerebral ischemia. These effects are likely to cause impairment in neuronal function and underline the process of cerebral ischemic damage.

Direct inhibitory mechanisms of halothane on human platelet aggregation.

Although halothane directly inhibits platelet aggregation, the mechanisms of this effect are still unknown. The current study aimed to clarify the inhibitory mechanisms of halothane on thrombin-induced human platelet aggregation by measuring (1) platelet-surface glycoprotein Ib expression, (2) the concentration of intracellular free Ca2+ ([Ca2+]i) measured simultaneously with aggregation, (3) the concentration of intracellular inositol 1,4,5-triphosphate, and (4) the concentration of intracellular cyclic 3',5'-adenosine monophosphate ([cAMP]i). Washed platelet suspensions, obtained from healthy volunteers, were preincubated with halothane (0-2 mM) for 2 min and then exposed to 0.02 units/ml thrombin for 3 min. The glycoprotein Ib bound to fluorescein-labeled antibody was measured by fluorescence flow cytometry. [Ca2+]i was measured, simultaneously with aggregation, in Fura-2 (Ca2+ indicator)-loaded platelets by use of a fluorometer. Inositol 1,4,5-triphosphate and [cAMP]i were measured by radioimmunoassay. Halothane had no effect on glycoprotein Ib expression with or without thrombin. Halothane decreased the thrombin stimulated [Ca2+]i transient and inhibited platelet aggregation in a dose-dependent manner, both in the presence and in the absence of external Ca2+. Isoflurane had no apparent effect on either platelet aggregation or [Ca2+]i in the absence of external Ca2+. Halothane inhibited the increase in inositol 1,4,5-triphosphate induced by thrombin. Halothane moderately but significantly increased [cAMP]i, but the adenylate cyclase activator forskolin (which has the same inhibitory ability on aggregation as halothane) increased [cAMP]i to a much greater extent than did halothane. Halothane inhibits thrombin-induced human platelet aggregation by decreasing [Ca2+]i without inhibiting agonist-receptor binding; the inhibitory effect of halothane on [Ca2+]i might be mediated by a decrease in inositol 1,4,5 triphosphate and in part by an increase in [cAMP]i.

Age-associated decrease in inositol 1,4,5-triphosphate and diacylglycerol generation by 1,25(OH) 2-vitamin D 3 in rat intestine

The hormonal form of vitamin D 3, 1,25(OH) 2-vitamin D 3(1,25[OH] 2D 3), stimulates the breakdown of membrane phosphoinositides, generating inositol-1,4,5-trisphosphate (IP 3) and diacyglycerol (DAG) in a variety of cell systems. Several studies suggest that alterations in the receptor-mediated phosphoinositide cascade are involved in the pathophysiology of aging. Therefore, the formation of IP 3 and DAG were determined under basal conditions and after stimulation with physiological concentrations of 1,25(OH) 2D 3 in duodenum from young (3-mo-old) and aged (24-mo-old) rats. The hormone induced a transient and biphasic formation of IP 3 and DAG. Values obtained in young rats peaking at 15 s (51% and 42% above basal levels for IP 3 and DAG, respectively) and at 3 min (90% and 74% above basal levels for IP 3 and DAG, respectively) were significantly decreased in duodenum from senescent animals (IP 3: + 20% and DAG: 18% above basal level at 15 s; and IP 3: + 18% and DAG: + 29% above basal level at 3 min). The 1,25(OH) 2D 3-induced generation of DAG in both young and aged duodenum was effectively inhibited in the presence of neomycin, a phospholipase C (PLC) inhibitor, and was dependent on extracellular Ca 2+. After the biphasic response, the levels of DAG generated by the hormone (10 min stimulation) remained elevated; the elevation occurred in the absence of IP 3 production; and the elevated levels were not abolished by neomycin, implying that phospholipids other than phosphoinositides are the source of DAG. This 1,25(OH) 2D 3-dependent late phase of DAG generation was also diminished in aged animals. The precise molecular basis and the physiological significance of decreased liberation of IP 3 and DAG by 1,25(OH) 2D 3 in the aged rat duodenum remains to be determined.

Identification of galactitol 2-phosphate and galactitol 3-phosphate in the lens of galactose-fed rats

Production of unusual phosphorylated metabolites in the lens is one of several changes caused by hyperglycemia. Sorbitol 3-phosphate (Sor-3P) and fructose 3-phosphate (Fru-3P) are two such compounds identified in the diabetic lens, and galactitol 2-phosphate (Gal-2P) and galactitol 3-phosphate (Gal-3P) are identified here in the galactosemic lens. These new compounds are the first example of galactitol metabolism in mammalian tissue other than liver. Sor-3P and Fru-3P are also present in the galactosemic lens, apparently synthesized directly from their precursors, sorbitol and fructose, which are elevated in the lens due to increased flux of glucose through the aldose reductase (AR) pathway. The NADPH necessary to support this increased flux is derived from activation of the hexose monophosphate shunt (HMPS), which is clearly demonstrated by a large increase in the concentration of sedoheptulose 7-phosphate (Sed-7P), a HMPS-specific metabolite. Additionally, during 3 weeks of galactose feeding there is a dramatic increase in lenticular concentrations of galactitol, sorbitol, galactose, and fructose and a sharp decrease in inositol. Glucose remains unchanged. A precipitous loss of both phosphorylated and nonphosphorylated metabolites occurs after 3 weeks, possibly due to lens rupture.

The effects of FMRFamide, 5-hydroxytryptamine and phorbol esters on the heart of the mussel Geukensia demissa.

The ventricle of the mussel Geukensia demissa is inhibited by 5-hydroxytryptamine and excited by the molluscan neuropeptide FMRFamide. Supra-threshold doses of amide result in marked positive chronotropy and inotropy within 5-15 s. 5-Hydroxytryptamine at 10(-8) M produces diastolic arrest within 10 s. A 1-min exposure to FMRFamide (5 x 10(-8) M) results in a small increase in the cytoplasmic levels of adenosine 3',5'-cyclic monophosphate; shorter or longer exposures have no effect. The cAMP content of ventricles incubated in 5 x 10(-8) M 5-hydroxytryptamine for 1 min decreases by 2.3 pmol/mg protein; longer or shorter incubations have no effect. Treatment with forskolin results in 3- or 4-fold increases in adenosine 3',5'-cyclic monophosphate, but forskolin has no effect on the mechanical activity of the ventricle. The levels of inositol monophosphate, inositol 1,4-diphosphate, and inositol 1,4,5-triphosphate in tissues exposed to 5-hydroxytryptamine are not different from levels in control tissues. FMRFamide decreases the levels of these phosphoinositides by 50% or more. Lower concentrations of phorbol 12,13-diacetate (10(-8) to 10(-7) M) and phorbol 12-myristate,13-acetate (10(-6) M) cause positive chronotropy in the isolated ventricle; higher concentrations induce systolic arrest. These results suggest that the effects of 5HT on the ventricle are not mediated by adenosine 3',5'-cyclic monophosphate or inositol 1,4,5-triphosphate. The effects of FMRFamide may involve a decrease in inositol 1,4,5-triphosphate. The effects of amide may involve a decrease in inositol 1,4,5-triphosphate. The response of the ventricles to phorbol esters suggest that protein kinase C may be involved in the regulation of cardiac contractility.

Second messenger imbalance hypothesis of schizophrenia

Based on the results of studies on intracellular signaling in platelets of schizophrenics, an imbalance of the second messenger system is proposed: Diacylglycerol (DG), which activates protein kinase C (PKC), was increased, while adenylate cyclase (AC)-cAMP function was decreased. It is proposed that the increased DG PKC function may entail a decrease in inositol 1,4,5-trisphosphate (IP 3) Ca 2+ function and lowering of phosphoinositide turnover. If such a pathological intracellular signaling takes place in the brain, it may cause a distorted balance of protein activation via phosphorylation in neurons, resulting in some of the deficits of schizophrenia. Neuroleptics have been reported to antagonize the above-mentioned pathological processes of intracellular signaling. The imbalance hypothesis of the DG PKC pathway and AC-cAMP pathway is not inconsistent with the dopamine hypothesis of schizophrenia, because dopamine receptor stimulation is indirectly related to reduction in IP 3 Ca 2+ function and lowering of phosphoinositide metabolism.

Phospholipase C-mediated hydrolysis of phosphatidylcholine is activated by cis-diamminedichloroplatinum(II).

We have investigated the effect of cis-diamminedichloroplatinum(II) (CDDP) on signal transduction pathways. CDDP treatment did not cause any change in the binding of [3H]-phorbol dibutyrate to PC-9 (human lung adenocarcinoma cell line) cells, a measure of protein kinase C activation. However, 2-h CDDP treatment (20 micrograms/ml) caused approximately 200% increase in 1,2-sn-diacylglycerol (DAG) production and approximately 50% decrease in inositol 1,4,5-triphosphate production. To explore the different source of DAG, we analyzed phospholipids labeled with [14C]choline by TLC and revealed that [14C]choline-labeled phosphatidylcholine (PC) was decreased to 50% by CDDP treatment. This suggested that PC turnover was increased by CDDP-treatment. PC-specific phospholipase C (PC-PLC) activity was increased to 2.5-fold (2.58 +/- 0.28 nmol/mg protein per min) by 2 h CDDP (20 micrograms/ml) treatment compared with control (1.05 +/- 0.24 nmol/mg protein per min). Treatment of CDDP also stimulated PC-PLC in the crude membrane extract from PC-9 cells. CDDP had no effect on the activities of phospholipase A2 and D. Trans-DDP, which has far less cytotoxicity than its stereoisomer, CDDP, did not cause any change in PC-PLC activity. A significant inhibition of DNA synthesis (less than 80%) occurred 4 h after 2 h CDDP (20 micrograms/ml) treatment. These results demonstrated that CDDP-induced PC-PLC activation was an early event in CDDP-induced cytotoxicity and suggested that the effects of CDDP on signal transduction pathways had an important role in CDDP-induced cytotoxicity.

?-interferon, retinoic acid, and cytosine arabinoside induce neuroblastoma differentiation by different mechanisms

1. The effects of gamma-interferon (gamma-IFN), retinoic acid (RA), and cytosine arabinoside (ARA-C) on the growth, morphology, and phenotype of the human neuroblastoma (NB) cell lines, LAN-1 and GI-ME-N, have been extensively tested. 2. RA, gamma-IFN, and ARA-C induced a dose-dependent morphological differentiation and growth inhibition, without affecting cell viability. Cells exposed to 10(-6) M RA or 1000 U/ml gamma-IFN significantly decreased their growth rate within the first 24 and 48 hr of culture, respectively. Cells became smaller and polygonal and sprouted long cellular processes with varicosities along their courses. In contrast, ARA-C-differentiated cells were larger and flattened, with few elongated dendritic processes. 3. Analysis of membrane and cytoskeletal markers by immunofluorescence and Western blot showed several changes in NB-specific antigen expression after 5 days of treatment with all inducing agents. Analysis of labeled phosphatidylinositol metabolites from prelabeled cells showed, within 1 min of treatment with RA, a rapid decrease in inositol 1,4,5-trisphosphate and of 1,2-diacylglycerol levels. No changes in inositol phospholipid metabolism were observed in gamma-IFN- or ARA-C-treated cells. 4. We conclude that RA-induced decrease in phosphatidylinositol (PI) hydrolysis is not likely to be a consequence of the acquisition of a different phenotype, as its changes precede the acquisition of neuronal markers. In addition, gamma-IFN and ARA-C, both inducing a mature phenotype, did not affect PI hydrolysis. 5. Decreased PI hydrolysis seems to be sufficient, although not necessary, to commit NB cells to neuronal differentiation. Analysis of molecular mechanisms associated with NB cell differentiation may be helpful to clarify the potential of various biological agents in affecting the development of the neural cell.

Second messenger imbalance hypothesis of schizophrenia

Abstract Based on the results of studies on intracellular signaling in platelets of schizophrenics, an imbalance of the second messenger system is proposed: Diacylglycerol (DG), which activates protein kinase C (PKC), was increased, while adenylate cyclase (AC)-cAMP function was decreased. It is proposed that the increased DG PKC function may entail a decrease in inositol 1,4,5-trisphosphate (IP 3 ) Ca 2+ function and lowering of phosphoinositide turnover. If such a pathological intracellular signaling takes place in the brain, it may cause a distorted balance of protein activation via phosphorylation in neurons, resulting in some of the deficits of schizophrenia. Neuroleptics have been reported to antagonize the above-mentioned pathological processes of intracellular signaling. The imbalance hypothesis of the DG PKC pathway and AC-cAMP pathway is not inconsistent with the dopamine hypothesis of schizophrenia, because dopamine receptor stimulation is indirectly related to reduction in IP 3 Ca 2+ function and lowering of phosphoinositide metabolism.

Decapitation-induced changes in inositol phosphates in rat brain

Decapitation resulted in a time-dependent production of inositol phosphates in rat brain. This production was analyzed by measuring both the radioactivity and the concentrations of inositol phosphates generated from [3H]inositol-labeled phospholipids. Both measurements produced the same time-dependent changes, including a rapid decrease in inositol 1,4,5-trisphosphate within 1.5 min, a 6-fold increase in inositol 1,4-bisphosphate to a maximum at 1.5 min, a 5-fold rise in inositol 4-monophosphate to a maximum at 2.5 min, and little change in inositol 1-monophosphate. The temporal changes in the mass and radioactivity of these compounds, together with the decrease in labeling of phosphatidylinositol 4,5-bisphosphates, support the idea that the inositol phosphates originate from the hydrolysis of phosphatidylinositol 4,5-bisphosphates and not from either the direct hydrolysis of phosphatidylinositol 4-phosphates or phosphatidylinositols.

Brain Cortical Fatty Acids and Phospholipids During and Following Complete and Severe Incomplete Ischemia

Abstract: To explore the possibility that peroxtdative degradation of brain tissue lipid constituents is an important mechanism of irreversible ischemic damage, we measured cortical fatty acids and phospholipids during reversible brain ischemia in the rat. Neither complete nor severe incomplete ischemia (5 and 30 min) caused any measurable breakdown of total or individual fatty acids or phospholipids. Except for a small (and reversible) decrease of inositol plus serine phosphoglycerides in the early postischemic period following 30 min of incomplete ischemia, there were no significant losses of fatty acids or phospholipids during recirculation. Since peroxidation, induced in brain cortical tissue in vitro, characteristically involves degradation of polyenoic fatty acids (arachidonic and docosahexaenoic acids) and of ethanolamine phosphoglycerides, the present in vivo results fail to support the hypothesis that peroxidation of membrane lipids is of primary importance for ischemic brain cell damage. Both complete and severe incomplete ischemia caused a similar increase in the tissue content of free fatty acids (FFA). Thus the FFA pool increased by about 10 times during a 30‐min ischemic period, to constitute 1 ‐ 2% of the total fatty acid pool. Since there was a relatively larger increase in polyenoic FFA (especially in arachidonic acid) than in saturated FFA, the release of FFA may be the result of activation of a phospholipase A2 unbalanced by reesterification. Increased levels of FFA persisted during the initial recirculation period, but a gradual normalization occurred and the ischemic changes were essentially reversed at 30 min after restoration of circulation. The pathophysiological implications of the changes in FFA are discussed with respect to mitochondrial dysfunction, formation of cellular edema and prostaglandin‐mediated deterioration of postischemic circulation.