Comparison of inositol 1,4,5-trisphosphate and inositol 1,3,4,5-tetrakisphosphate binding sites in cerebellum.
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Animals↗
Biomedical subjects
R A Challiss
Publications and source records attributed to R A Challiss.
Depolarization and agonist-stimulated changes in inositol 1,4,5-trisphosphate and inositol 1,3,4,5-tetrakisphosphate mass accumulation in rat cerebral cortex.
Muscarinic receptor stimulation or depolarization with elevated extracellular K+ induced rapid and sustained increases in mass accumulations of myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and myo-inositol 1,3,4,5-tetrakisphosphate [Ins(1,3,4,5)P4] in cerebral cortex slices. Synergistic but transient responses of both inositol polyphosphate second messengers were observed when slices were stimulated with carbachol under depolarizing conditions; this synergy was observed as an increase in the maximal responsiveness, with no significant change in EC50 values for carbachol. Omission of buffer Ca2+ ([Ca2+]e 10-20 microM) reduced basal Ins(1,4,5)P3 and Ins(1,3,4,5)P4 concentrations; the relative stimulatory effects of muscarinic receptor stimulation were maintained, but the effects of depolarization were markedly attenuated under these conditions. A component of the response to depolarization appeared to be indirectly mediated by the release of acetylcholine, because the K(+)-evoked increase in Ins(1,3,4,5)P4 was enhanced by the cholinesterase inhibitor physostigmine, and was partially attenuated by atropine. An additive suppression by nitrendipine suggests that entry of Ca2+ through L-type Ca2+ channels may serve to accelerate phosphorylation of Ins(1,4,5)P3 by 3-kinase. Norepinephrine did not significantly increase Ins(1,4,5)P3 or Ins(1,3,4,5)P4 accumulation; however, in the presence of depolarizing K+, norepinephrine caused a dramatic increase in Ins(1,3,4,5)P4 mass accumulation. In contrast, the excitatory amino acid quisqualate caused significant increases in the mass accumulations of both inositol polyphosphates measured, with no further increase being observed under depolarizing conditions. The results are discussed with respect to the interactive effects of agonist and depolarization stimuli on inositol polyphosphate accumulation which might more accurately reflect the conditions pertaining in vivo.
Animals↗
Lack of effect of zaprinast on methacholine-induced contraction and inositol 1,4,5-trisphosphate accumulation in bovine tracheal smooth muscle.
1. The effects of zaprinast (M&B 22948), a selective guanosine 3':5'-cyclic monophosphate (cyclic GMP) phosphodiesterase inhibitor, and sodium nitroprusside on cyclic GMP content, phosphoinositide hydrolysis and airway smooth muscle tone were examined in flurbiprofen pretreated bovine tracheal smooth muscle (BTSM). 2. Anion-exchange chromatography of the soluble fraction of BTSM homogenates resolved three peaks of Ca2+/calmodulin-independent phosphodiesterase (PDE) activity that corresponded to type Ia (cyclic GMP-specific, zaprinast-inhibitable), type II (cyclic GMP-stimulated) and type IV (Ro 20 1724-inhibitable) PDE isoenzymes. Zaprinast caused a selective inhibition of the type Ia PDE isoenzyme (IC50 0.94 microM) with respect to the type II and IV (IC50 s 93 microM and 197 microM respectively) isoenzymes. 3. Pretreatment of BTSM strips with zaprinast (10 microM) for 20 min affected neither the initial rate of force development, nor the resultant magnitude of contraction induced by methacholine (10 microM). In addition, zaprinast (10 microM; 20 min) did not affect the cumulative concentration-response relationship induced by methacholine. In contrast, sodium nitroprusside (300 microM) either alone, or in combination with zaprinast (10 microM), significantly attenuated tone induced by low, but not high concentrations of methacholine. This resulted in a non-parallel, rightwards shift of the methacholine concentration-response curves (nitroprusside: 4.0 fold; nitroprusside/zaprinast: 4.8 fold at the EC50 values), without a reduction in the maximum tone generated. 4. In BTSM slices, zaprinast (10 or 100 microM) did not influence basal or methacholine (10 microM)-stimulated cyclic GMP accumulation or inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) mass accumulation over a 60s incubation period, although it did significantly increase cyclic GMP content over longer (30 min) stimulation periods. 5. In [3H]-inositol prelabelled BTSM slices, stimulated in the presence of 5mM LiCl, methacholine (10 microM) caused a marked increase in total [3H]-inositol phosphate accumulation. This effect was not inhibited by zaprinast (10 microM), sodium nitroprusside (300 microM), or a combination of these drugs despite these agents markedly increasing tissue cyclic GMP content. 6. These findings demonstrate that despite zaprinast being a potent and selective inhibitor of the type Ia PDE isoenzyme in a cell-free system, this drug only increases cyclic GMP content in BTSM following prolonged agonist-stimulation. This may explain its lack of inhibitory effect on methacholine-induced tone. The inability of drugs which increase tissue cyclic GMP content and exhibit anti-spasmogenic activity to inhibit methacholine-stimulated Ins(1,4,5)P3 formation suggests that, unlike vascular smooth muscle, cyclic GMP-dependent mechanisms do not regulate receptor-mediated phosphoinositide hydrolysis in BTSM.
3',5'-Cyclic-GMP Phosphodiesterases↗
Inositol 1,4,5-triphosphate-stimulated calcium release from permeabilized cerebellar granule cells.
1. Muscarinic cholinoceptor stimulation of phosphoinositide hydrolysis in rat cultured cerebellar granule cells results in a rapid, transient accumulation of inositol 1,4,5-trisphosphate (Ins(1,4,5)P3), which has been implicated in the release of non-mitochondrial intracellular Ca2+ stores. In the present study, the release of Ca2+ from intracellular stores and the Ins(1,4,5)P3 receptor responsible for this process have been investigated. 2. Monolayers of saponin-permeabilized granule cells accumulate 45Ca2+ in an ATP-dependent manner and the sequestered 45Ca2+ can be concentration-dependently released by Ins(1,4,5)P3 by a stereospecific and heparin-sensitive mechanism. The EC50 for Ins(1,4,5)P3-stimulated 45Ca2+ release was 80 +/- 3 nM. 3. Radioligand binding studies performed on a crude granule cell membrane fraction indicated the presence of an apparently homogeneous population of stereo-specific Ins(1,4,5)P3 receptors (KD 54.7 +/- 2.0 nM; Bmax 1.37 +/- 0.29 pmol mg-1 protein). 4. This study provides evidence for Ins(1,4,5)P3-sensitive intracellular Ca2+ stores in primary cultures of cerebellar granule cells and suggest that these cells provide an excellent model neuronal system in which to study the relative functional roles of Ca2+ release from intracellular stores and Ca(2+)-entry in neuronal Ca2+ homeostasis.
Animals↗
Role for ionotropic and metabotropic receptors in quisqualate-stimulated inositol polyphosphate accumulation in rat cerebral cortex.
The actions of the excitatory amino acid quisqualate (QA) on inositol polyphosphate accumulation in cerebral cortex slices have been assessed using both [3H]inositol prelabeling and mass measurements over relatively short incubation periods. QA stimulated accumulation of all the inositol polyphosphates, with similar EC50 values (2.8 +/- 0.7 microM). High performance liquid chromatography analysis of isomeric forms of inositol polyphosphates and specific mass assays revealed that both phosphorylation and dephosphorylation products of inositol-1,4,5-trisphosphate accumulate. A large component of the QA-stimulated inositol polyphosphate accumulation was inhibited by the ionotropic antagonist 6,7-dinitroquinoxaline-2,3-dione in a competitive manner. This implied that the QA response may be due to entry of Ca2+ via voltage-sensitive calcium channels as a consequence of an ionotropic receptor-induced depolarization. In support of this mechanism, the QA-induced response was dependent on the presence of extracellular calcium, whereas the well characterized muscarinic receptor agonist response to carbachol showed only a slight reduction under the same conditions. The concentration-dependent (EC50 8.8 +/- 3 microM) response to the selective ionotropic agonist amino-3-hydroxy-5-methyl-4-isoazolepropionic acid (AMPA) differed from that to QA or carbachol, in that accumulation of only [3H]inositol mono- and bisphosphates was stimulated, with no increase in the [3H]inositol tris- or tetrakisphosphates. Use of the metabotropic agonist (trans)-(+/-)-1-aminocyclopentyl-1,3-dicarboxylate (ACPD), however, produced concentration-dependent increases in all [3H]inositol polyphosphates. Although both AMPA and ACPD responses alone were smaller in magnitude than that to QA, when present together AMPA and ACPD produced additive responses on [3H]inositol mono- and bisphosphate and a marked synergistic increase in [3H]inositol tetrakisphosphate accumulation, resulting in a response similar to that seen for QA. These data suggest that QA-evoked responses in rat cortex slices are the result of a complex interaction mediated through both ionotropic and metabotropic receptors, in which Ca2+ entry may stimulate accumulation of inositol mono- and bisphosphate directly and divert the metabolism of inositol-1,4,5-trisphosphate to inositol-1,3,4,5-tetrakisphosphate.
Animals↗
Coupling of muscarinic receptors to the mobilization of intracellular Ca2+ stores in permeabilized SH-SY5Y human neuroblastoma cells.
Intracellular stores of Ca2+ were mobilized transiently by carbachol in suspensions of electrically permeabilized SH-SY5Y cells. The kinetics and the dose-dependence of this mobilization paralleled carbachol-induced increases in inositol 1,4,5-trisphosphate (InsP3) mass [for both parameters EC50 (concn. giving half-maximal response) = 60-70 microM]. Guanosine 5'-[gamma-thio]triphosphate enhanced the maximal effect and the potency of carbachol on Ca2+ mobilization and InsP3 mass, but caused separation of the dose-response curves (EC50 = 0.6 microM and 5.6 microM respectively). These data show that functional coupling of muscarinic receptors to Ca2+ mobilization can be maintained after permeabilization, reveal major effects of guanine nucleotides on agonist-induced Ca2+ mobilization and provide a basis for explanation of discrepancies between agonist potency on InsP3 concentration and Ca2+ mobilization in intact cells.
Calcium↗
Characteristics of inositol 1,4,5-trisphosphate binding to rat cerebellar and bovine adrenal cortical membranes: evidence for the heterogeneity of binding sites.
The equilibrium and kinetic binding characteristics of D-inositol 1,4,5-trisphosphate (Ins(1,4,5)P3) have been studied in membrane preparations of rat cerebellum and bovine adrenal cortex. Saturation analysis of isotopic dilution binding data demonstrated apparent KD values for Ins(1,4,5)P3 binding of 23 +/- 5 nM and 3.0 +/- 1.3 nM for cerebellar and adrenal cortical preparations, respectively, with approximately 20-fold greater receptor density present in the cerebellar preparation (Bmax: 10.2 +/- 2.5 pmol/mg protein). Kinetic analysis confirmed the equilibrium binding-derived KD value for cerebellum (KD: 39.9 nM), but revealed a second, very high affinity site (KD: 0.06 nM) to be present in adrenal cortex. The affinity differences between the investigated preparations was also observed with respect to the IC50 values obtained for inhibition of specific [3H]Ins(1,4,5)P3 binding by a number of inositol polyphosphate analogues including D-inositol 2,4,5-trisphosphate, DL-inositol 1,4,5-trisphosphorothioate and L-Ins(1,4,5)P3. In contrast, the Ins(1,4,5)P3-receptor antagonist heparin displayed greater potency for the cerebellar (IC50: 16.5 +/- 6.2 micrograms . ml-1) compared to the adrenal cortical preparation (IC50: 51.0 +/- 6.1 micrograms . ml-1). The apparent differences between the Ins(1,4,5)P3 receptors characterized in the two tissue preparations are discussed.
Adrenal Cortex↗
Spatially resolved changes in diabetic rat skeletal muscle metabolism in vivo studied by 31P-n.m.r. spectroscopy.
Phase-modulated rotating-frame imaging (p.m.r.f.i.), a localization technique for 31P-n.m.r. spectroscopy, has been applied to obtain information on the heterogeneity of phosphorus-containing metabolites and pH in the skeletal muscle of control and streptozotocin-diabetic rats. Using this method, the metabolic changes in four spatially resolved longitudinal slices (where slice I is superficial and slice IV is deep muscle) through the ankle flexor muscles have been investigated at rest and during steady-state isometric twitch-contraction at 2 Hz. At rest, intracellular pH was lower, and phosphocreatine (PCr)/ATP was higher, throughout the muscle mass in diabetic compared with control animals. The change in PCr/ATP in diabetic muscle correlated with a decrease in the chemically determined ATP concentration. During the muscle stimulation period, the decrease in pH observed in diabetic muscle at rest was maintained, but not exacerbated, by the contractile stimulus. Stimulation of muscle contraction caused more marked changes in PCr/(PCr + Pi), PCr/ATP and Pi/ATP in the diabetic group. These changes were most evident in slice III, which contains the greatest proportion of fast glycolytic-oxidative (type IIa) fibres, in which statistically significant differences were observed for all metabolite ratios. The results presented suggest that some degree of heterogeneity occurs in diabetic skeletal muscle in vivo with respect to the extent of metabolic dysfunction caused by the diabetic insult and that regions of the muscle containing high proportions of type IIa fibres appear to be most severely affected.
Adenosine Triphosphate↗
Reduced inositol polyphosphate accumulation and inositol supply induced by lithium in stimulated cerebral cortex slices.
The ability of lithium to interfere with phosphoinositide metabolism in rat cerebral cortex slices has been examined by monitoring the accumulation of CMP-phosphatidate (CMP-PtdOH) and the reduction in Ins(1,4,5)P3 and Ins(1,3,4,5)P4 levels. A small accumulation of [14C]CMP-PtdOH was seen in slices prelabelled with [14C]cytidine and stimulated with carbachol (1 mM) or Li+ (1 mM). However, simultaneous addition of both agents for 30 min produced a 22-fold accumulation, with Li+ producing a half-maximal effect at a concentration of 0.61 +/- 0.19 mM. Kinetic studies revealed that the effects of carbachol and Li+ on CMP-PtdOH accumulation occurred with no initial lag apparent under these conditions and that preincubation with myo-inositol (10 or 30 mM) dramatically attenuated CMP-PtdOH accumulation. myo-Inositol could also attenuate the rate of accumulation of CMP-PtdOH when added 20 min after carbachol and Li+; these effects were not observed when equimolar concentrations of scyllo-inositol were added. Use of specific radioreceptor assays allowed the mass accumulations of Ins(1,4,5)P3 and Ins(1,3,4,5)P4 to be monitored. Following a lag of 5-10 min, Li+ resulted in a marked reduction in the accumulation of both inositol polyphosphates resulting from muscarinic-cholinergic stimulation. Preincubation of cerebral cortex slices with myo- (but not scyllo-) inositol delayed, but did not prevent, the reduction in the accumulation of Ins(1,4,5)P3 or Ins(1,3,4,5)P4. The results suggest that cerebral cortex, at least in vitro, is very sensitive to myo-inositol depletion under conditions of muscarinic receptor stimulation. The relationship of such depletion to the generation of inositol polyphosphate second messengers is discussed.
Animals↗
Heterogeneity of [3H]inositol 1,4,5-trisphosphate binding sites in adrenal-cortical membranes. Characterization and validation of a radioreceptor assay.
1. The characterization of a radioreceptor assay for determining Ins(1,4,5)P3 concentration in tissue extracts is described which utilizes the binding of [3H]Ins(1,4,5)P3 to an adrenal-cortex membrane fraction. 2. Analysis of [3H]Ins(1,4,5)P3 binding by isotope dilution demonstrated an apparent single population of binding sites (KD 3.65 +/- 0.18 nM, Bmax. 872 +/- 70 fmol/mg of protein). Specific binding of [3H]Ins(1,4,5)P3 was enhanced at alkaline pH values (maximum at pH 8.5), with complete loss of specific binding at pH less than 6. These binding sites displayed strict stereo- and positional specificity for Ins(1,4,5)P3, with L-Ins(1,4,5)P3, Ins(1,3,4)P3 and DL-Ins(1,3,4,5)P4 causing 50% displacement of specific [3H]Ins(1,4,5)P3 binding (IC50 values) at concentrations of 14 +/- 3 microM, 3.0 +/- 0.3 microM and 0.53 +/- 0.03 microM respectively. 3. Kinetic analysis of binding data, however, revealed a high-affinity [3H]Ins(1,4,5)P3 binding site (KD 0.052 nM) in addition to the lower-affinity site (KD 2.53 nM) already demonstrated in displacement studies. 4. It is shown that the presence of the high-affinity site can be exploited to increase the sensitivity of the [3H]Ins(1,4,5)P3 radioreceptor assay, allowing accurate detection of 20 fmol of Ins(1,4,5)P3 in 300 microliters of tissue extract. 5. Further validation of the specificity of the above assay for Ins(1,4,5)P3 was provided by incubating tissue extracts with either a 5-phosphatase or 3-kinase preparation. It was shown that identical loss occurred of both Ins(1,4,5)P3 mass and [3H]Ins(1,4,5)P3, added to parallel incubations. 6. The ability of the assay to measure basal and agonist-stimulated increases in Ins(1,4,5)P3 concentration has been demonstrated with rat cerebral cortex and bovine tracheal smooth-muscle slices and a range of cultured and isolated cell preparations.
Adrenal Cortex↗
Neurotransmitter and depolarization-stimulated accumulation of inositol 1,3,4,5-tetrakisphosphate mass in rat cerebral cortex slices.
[32P]Inositol 1,3,4,5-tetrakisphosphate ([32P]Ins(1,3,4,5)P4) binds to a rat cerebellar membrane site with high affinity (KD = 2.8 +/- 0.6 nM) and low capacity (Bmax = 176 +/- 34 fmol/mg of protein). Evidence for a low-affinity site (KD = 164 +/- 48 nM) was also apparent. The high-affinity site displayed marked specificity for the Ins(1,3,4,5)P4 isomer, compared with several other inositol polyphosphates, and has been used as the basis of a radioreceptor assay for Ins(1,3,4,5)P4 in extracts of rat cerebral cortex slices. The resting Ins(1,3,4,5)P4 concentration (1.89 +/- 0.11 pmol/mg of protein) in the slices was rapidly and dramatically increased by carbachol and quisqualate. K+ depolarization of cerebral cortex slices also stimulated Ins(1,3,4,5)P4 accumulation, with at least 50% of the response being sensitive to atropine, a result indicating that muscarinic receptor stimulation by released acetylcholine contributes significantly to the K+ effect.
Animals↗
Effects of selective phosphodiesterase inhibition on cyclic AMP hydrolysis in rat cerebral cortical slices.
1. The effects of selective inhibition of phosphodiesterase activities on the concentration and rate of hydrolysis of adenosine 3':5' cyclic-monophosphate (cyclic AMP) in rat cerebral cortical slices has been studied. 2. Isoprenaline caused a rapid, concentration-dependent increase in cyclic AMP concentration to new steady-state levels (basal: 7.1 +/- 0.7; 10 microM isoprenaline: 14.3 +/- 1.4 pmol mg-1 protein). Addition of a beta-adrenoceptor antagonist to isoprenaline-stimulated cerebral cortical slices caused a rapid decrease in cyclic AMP concentration to basal levels (t1/2: 58 +/- 18 s). 3. Preincubation of slices for 30 min with the phosphodiesterase inhibitors 1-methyl-3-isobutylxanthine, denbufylline, rolipram or Ro20,1724 caused concentration-dependent increases in basal and isoprenaline-stimulated cyclic AMP concentrations and decreased the rate of cyclic AMP hydrolysis measured after addition of a beta-adrenoceptor antagonist. However, SKF 94120 and zaprinast had none of these effects. 4. The results are discussed with respect to previous studies of phosphodiesterase isozymic activities isolated from cerebrum and it is suggested that the Ca2+/calmodulin-independent, low Km cyclic AMP phosphodiesterase isozyme, which is selectively inhibited by denbufylline, rolipram and Ro20,1724, and is present in cerebrum is of critical importance to the regulation of cyclic AMP concentration in this tissue.
Animals↗
Characterisation of stereospecific binding sites for inositol 1,4,5-trisphosphate in airway smooth muscle.
1. A 'P2' membrane fraction of bovine tracheal smooth muscle displays high affinity (KD 3.8 +/- 0.2 nM), saturable (Bmax 1003 +/- 170 fmol mg-1 protein) and reversible binding of D-myo[3H]-inositol 1,4,5-trisphosphate ([3H]-Ins(1,4,5)P3). 2. This binding site shows strict stereo- and positional specificity for the D-Ins(1,4,5)P3 isomer with L-Ins(1,4,5)P3, DL-Ins(1,3,4,5)P4 and D-Ins(1,3,4)P3 displacing [3H]-Ins(1,4,5)P3 with Ki values of 20 microM, 0.35 microM and 2.4 microM, respectively. 3. Specific binding of [3H]-Ins(1,4,5)P3 is enhanced at alkaline pH values (maximal at pH 7.75) and, in distinct contrast to [3H]-Ins(1,4,5)P3 binding in rat cerebellum membranes, is not inhibited by Ca2+ (5-500 microM). 4. Heparin displaces [3H]-Ins(1,4,5)P3 specific binding with an IC50 of 7.6 +/- 1.0 micrograms ml-1. 5. Comparative studies demonstrated specific [3H]-Ins(1,4,5)P3 binding in bovine cardiac atrial preparations (Bmax 75 +/- 5 fmol mg-1 protein) and very low specific [3H]-Ins(1,4,5)P3 binding in bovine cardiac ventricle and skeletal muscle membranes (less than or equal to 25 fmol mg-1 protein).
Animals↗
The role of prostaglandins as modulators of insulin-stimulated glucose metabolism in skeletal muscle.
Prostaglandins of the E series are known to increase the sensitivity of glucose uptake to insulin in isolated, incubated skeletal muscle of the rat. In the present study the relationships between changes in both insulin sensitivity and prostaglandins E2 metabolism were investigated. Chronic treatment of rats with the synthetic glucocorticoid dexamethasone causes an insulin resistant state in vivo. Also, the sensitivity of glucose metabolism to insulin from normal rats varies over a 24-h period. In both these rat models the sensitivity of glucose metabolism to insulin in isolated skeletal muscle was greatest when the content or rates of release of prostaglandin E2 were highest. These results strongly suggest that changes in PGE2 metabolism can influence the sensitivity of glucose metabolism to insulin in skeletal muscle.
Animals↗
Mass changes of inositol(1,4,5)trisphosphate in trachealis muscle following agonist stimulation.
Agonist-stimulated changes in D-Ins(1,4,5)P3 has been examined in tracheal smooth muscle using a specific radioreceptor assay utilizing the binding of [3H] or [32P]Ins(1,4,5)P3 to a bovine adrenal cortex preparation. Carbachol produced a maximal increase in Ins(1,4,5)P3 concentration after 5 s (basal: 12.9 +/- 0.8; stimulated: 27 +/- 1.5 pmol/mg protein) with an EC50 of 6.7 +/- 0.8 microM. With longer stimulation periods Ins(1,4,5)P3 rapidly returned to basal values by 30 s. This is the first report of mass measurements of Ins(1,4,5)P3 in smooth muscle.
Animals↗
31P NMR magnetization-transfer measurements of ATP turnover during steady-state isometric muscle contraction in the rat hind limb in vivo.
31P NMR magnetization-transfer measurements have been used to measure the flux between ATP and inorganic phosphate during steady-state isometric muscle contraction in the rat hind limb in vivo. Steady-state contraction was obtained by supramaximal sciatic nerve stimulation. Increasing the stimulation pulse width from 10 to 90 ms, at a pulse frequency of 1 Hz, or increasing the frequency of a 10-ms pulse from 0.5 to 2 Hz resulted in an increase in the flux which was an approximately linear function of the increase in the tension-time integral. The flux showed an approximately linear dependence on the calculated free cytosolic ADP concentration up to an ADP concentration of about 90 microM. The data are consistent with control of mitochondrial ATP synthesis by the cytosolic ADP concentration and indicate that the apparent Km of the mitochondria for ADP is at least 30 microM.
Adenosine Triphosphate↗
A gated 31P-n.m.r. study of bioenergetic recovery in rat skeletal muscle after tetanic contraction.
1. Gated 31P-n.m.r. spectra were obtained from the ankle flexor muscles of the rat at various times after 3 s isometric tetanic contraction. This allowed the time course of changes in phosphocreatine (PCr), Pi and free ADP concentrations and intracellular pH to be monitored in skeletal muscle in vivo with 1 s time resolution. 2. ATP concentration did not change significantly, either during the recovery from a 3 s tetanus or during the overall protocol. 3. The calculated rate of recovery of ADP towards pre-stimulation levels was very rapid (t1/2 less than 5 s). The rate of Pi disappearance (t1/2 = 14 s) was more rapid than the rate of PCr synthesis (t1/2 = 24 s), resulting in a significant transient decrease in n.m.r.-visible PCr + Pi between 25 and 45 s after tetanic contraction. 4. The rates of PCr, Pi and ADP recovery are higher than those previously reported for recovery from steady-state exercise in humans or twitch isometric contraction in animals.
Adenosine Diphosphate↗
Lithium reduces the accumulation of inositol polyphosphate second messengers following cholinergic stimulation of cerebral cortex slices.
The ability of lithium to interfere with the metabolism of inositol phosphates in brain may underlie its therapeutic action in manic-depressive illness. In these experiments, lithium, at therapeutic concentrations, enhanced the accumulation of [3H]inositol monophosphate but suppressed the accumulation of the putative second messengers [3H]inositol 1,4,5-trisphosphate [( 3H]Ins(1,4,5)P3) and [3H]inositol 1,3,4,5-tetrakisphosphate following stimulation of cerebral cortex slices with carbachol. Mass measurements of Ins(1,4,5)P3 showed similar inhibitory effects, which could be prevented by preincubation with myo-inositol. These data may reveal the mechanism by which lithium can reduce polyphosphoinositide-mediated neurotransmission in brain.
Animals↗