Metal complexes driven from Schiff bases and semicarbazones for biomedical and allied applications: a review.

Schiff bases are versatile organic compounds which are widely used and synthesized by condensation reaction of different amino compound with aldehydes or ketones known as imine. Schiff base ligands are considered as privileged ligands as they are simply synthesized by condensation. They show broad range of application in medicine, pharmacy, coordination chemistry, biological activities, industries, food packages, dyes, and polymer and also used as an O2 detector. Semicarbazone is an imine derivative which is derived from condensation of semicarbazide and suitable aldehyde and ketone. Imine ligand-containing transition metal complexes such as copper, zinc, and cadmium have shown to be excellent precursors for synthesis of metal or metal chalcogenide nanoparticles. In recent years, the researchers have attracted enormous attention toward Schiff bases, semicarbazones, thiosemicarbazones, and their metal complexes owing to numerous applications in pharmacology such as antiviral, antifungal, antimicrobial, antimalarial, antituberculosis, anticancer, anti-HIV, catalytic application in oxidation of organic compounds, and nanotechnology. In this review, we summarize the synthesis, structural, biological, and catalytic application of Schiff bases as well as their metal complexes.

Chitosan: A macromolecule as green corrosion inhibitor for mild steel in sulfamic acid useful for sugar industry.

The present investigation aims at investigation of low cost nontoxic carbohydrate biopolymer chitosan as corrosion inhibitor alone and in combination with KI for mild steel in 1M sulfamic acid medium using gravimetric, electrochemical and surface analysis techniques. It is found that chitosan alone exhibits inhibition efficiency of 73.8% at 200ppm concentration. However, in combination with KI (5ppm), it gave more than 90% inhibition efficiency. The significant increase in the inhibition performance of chitosan has been explained by the synergistic mechanism. The results of Potentiodynamic polarization study shows that chitosan and its blend with KI decreases both anodic and cathodic reactions occurring at mild steel surface in 1M sulfamic acid medium by blocking active sites of the metal and acts as mixed type inhibitor. EIS study reveals that the polarization resistance increases with increase in the concentration of inhibitors which increases charge transfer resistance across the metal/solution interface. The adsorption of chitosan followed the Langmuir adsorption isotherm. The formation of inhibitor film on metal surface was supported by scanning electron microscopy (SEM) and atomic force microscopy (AFM) surface studies.

Activation of Src/kinase/phospholipase C/mitogen-activated protein kinase and induction of neurite expression by ATP, independent of nerve growth factor.

Extracellular ATP has been reported to potentiate the neurite outgrowth induced by nerve growth factor. In the present study the neurotrophic effect of ATP and other nucleotides was examined in mouse neuroblastoma neuro2a cells which lack nerve growth factor receptor. Exposure of neuro2a cells to ATP resulted in a dramatic increase in neurite bearing cells as compared with untreated control cells. Experiments performed with purinergic receptor agonists and antagonists suggest that the ATP stimulates neurite outgrowth via P2 receptors. Neurite outgrowth was completely blocked by P2 receptor antagonist suramin whereas the P1 receptor antagonist CGS15943 was ineffective. P1 receptor agonist 5'-(N-ethylcarboxamido)adenosine failed to induce neurite outgrowth. The potency order of different P2 receptor agonists was ATP=ATPgammaS>ADP>>2Me-S-ATP. It was insensitive to UTP and antagonist pyridoxal phosphate-6-azo (benzene-2,4-disulfonic acid) suggesting the involvement of P2Y11 receptor in the observed neuritogenic effect. The signaling pathway leading to ATP-induced neuritogenesis was investigated. The neuritogenic effect of ATP is independent of rise in intracellular Ca(2+) as pharmacological profile of neuritogenic P2Y receptor does not match with that of P2Y2 receptor associated with [Ca(2+)](i) signaling cascade. Exposure of cells to ATP caused activation of Src kinase, phospholipase Cgamma and extracellular signal-regulated kinases ERK1/2. Mitogen-activated protein kinase (MAPK) inhibitor U0126 drastically reduced the number of neurite bearing cells in ATP-treated cultures implying that the neurotrophic effect of ATP is mediated by MAPK. Our results demonstrate that ATP can stimulate neurite outgrowth independent of other neurotrophic factors and can be an effective trophic agent.

Alterations in plasma membrane of glioblastoma cells by photodynamic action of merocyanine 540.

Photodynamic action of merocyanine 540 (MC540) on the plasma membrane of human glioblastoma(U-87MG) cells has been investigated. Plasma membrane was labeled with lipid specific probe 1,(4-trimethylammonium),6-diphenyl-1,3,5-hexatriene. Steady-state anisotropy, decay time and time-dependent anisotropy of TMA-DPH in U-87MG cells have been measured as a function of light dose. A decrease in the steady-state anisotropy and decay time of TMA-DPH in MC540-treated cells was observed upon light irradiation. The time-dependent anisotropy measurements showed a decrease in the limiting anisotropy (r infinity) and an increase in the rotational relaxation time (phi) of the probe upon photosensitization of cells. Analysis of these data using wobbling in cone model for probe rotation in the membrane indicated an increase in the cone angle (theta c) and a decrease in the order parameter (S). Protein specific probe N-(1-pyrene)-maleimide was used to study the effect of photosensitization on the plasma membrane proteins. An increase in the rotational relaxation time and a decrease in the ratio of excimer to monomer fluorescence intensity of PM was observed on photosensitization. Photodynamic action of MC540 also caused an inhibition of protein SH groups and Na(+)-K(+)-ATPase activity of plasma membrane. Our results demonstrate that the photodynamic action of MC540 decreases the order of the lipid bilayer and reduces the mobility of the proteins in the plasma membrane of cells.

Binding of monomeric and oligomeric porphyrins to human glioblastoma (U-87MG) cells and their photosensitivity.

The binding of monomeric (Hp) and oligomeric (PHE) forms of porphyrin to glioblastoma (U-87MG) cells and the photosensitization of these cells have been studied. Upon binding to U-87MG cells, Hp and PHE exhibited fluorescence bands at 615 and 636 nm, respectively. The fluorescence and absorption spectra of Hp, HpD and PHE, measured in different solvents, suggest that the 615 nm band may arise due to the binding of monomeric as well as aggregated forms of porphyrin to the hydrophilic sites in the cells whereas the 636 nm band may be due to the binding of an aggregated form of porphyrin to the hydrophobic sites. The photosensitivity of cells and photo-induced lipid peroxidation were measured as a function of light dose. Cells were found most photosensitive to PHE followed by HpD and Hp. The photosensitivity of cells correlates well with the fluorescence intensity of cell bound dye at 636 nm. These results suggest that the binding of the oligomeric component of HpD to hydrophobic sites in the cells is responsible for the enhancement in the photosensitivity.

Galactocerebroside mediates Ca2+ signaling in cultured glioma cells.

Expression of galactocerebroside (GalC) was detected in human glioma cell line (U-87 MG). Exposure of cells to antibody against GalC and fluoresceinated second antibody showed intense fluorescence on the plasma membrane. Possible involvement of GalC in receptor-mediated transmembrane signaling was explored in this cell line. Antibodies raised against GalC were used as ligands. Binding of anti-GalC to these cells caused a transient increase in intracellular free calcium ([Ca2+]i). The response was observed both in the presence and absence of extracellular calcium demonstrating that the rise in [Ca2+]i induced by anti-GalC was due to an influx of Ca2+ through plasma membrane as well as the release of Ca2+ from intracellular pools. Ca2+ influx was blocked by verapamil, indicating that influx is mediated by voltage-sensitive channels. Our results suggest that GalC can play a role in transmembrane signaling by modulation of voltage-sensitive Ca2+ channels.

Gangliosides asymmetrically alter the membrane order in cultured PC-12 cells.

Exogenous gangliosides readily associate with the cell membranes and produce marked effects on cell growth and differentiation. We have studied the effect of bovine brain gangliosides (BBG) on the membrane dynamics of intact cells. The structural and dynamic changes in the cell membrane were monitored by the fluorescence probes DPH, TMA-DPH and laurdan. Incorporation of BBG into the cell membrane decreased the fluorescence intensity, lifetime and the steady state anisotropy of TMA-DPH. Analysis of the time resolved anisotropy decay by wobbling in the cone model revealed that BBG decreased the order parameter, and increased the cone angle without altering the rotational relaxation rate. The fluorescence intensity and lifetime of DPH were unaffected by BBG incorporation, however, a modest increase was observed in the steady state anisotropy. BBG incorporation reduced the total fluorescence intensity of laurdan with pronounced quenching of the 440-nm band. The wavelength sensitivity of generalized polarization of laurdan manifested an ordered liquid crystalline environment of the probe in the cell membrane. BBG incorporation reduced the GP values and augmented the liquid crystalline behavior of the cell membrane. BBG incorporation also influenced the permeability of cell membranes to cations. An influx of Na+ and Ca2+ and an efflux of K+ was observed. The data demonstrate that incorporation of gangliosides into the cell membrane substantially enhances the disorder and hydration of the lipid bilayer region near the exoplasmic surface. The inner core region near the center of the bilayer becomes slightly more ordered and remains highly hydrophobic. Such changes in the structure and dynamics of the membrane could play an important role in modulation of transmembrane signaling events by the gangliosides.

Binding of hematoporphyrin derivative to brain tumor cells--a fluorescence spectroscopic study.

The binding of hematoporphyrin derivative (HpD) to brain tumor cells and their photosensitivity was studied as a function of HpD concentration, time of incubation and growth phase of cells. Upon binding to cells, HpD showed three fluorescence bands at 616, 636 and 678 nm. In plateau phase cells a fluorescence band at 636 nm was predominant, which was further enhanced by increasing HpD concentration and/or increasing incubation time. In exponential phase cells the maximum fluorescence was exhibited at 616 nm. After 1 h incubation of exponential phase cells with increasing HpD concentration an overall intensity enhancement occurred with no change in the distribution of bands, whereas longer incubation time caused an increase in relative intensity of the 636 nm band similar to that observed in plateau phase cells. After 1 h incubation with HpD plateau phase cells were more photosensitive than exponential phase cells, although cell bound HpD was much less in the former case. Incubation of cells for 24 h drastically enhanced the photosensitivity irrespective of the growth phase. Our results suggest a relationship between the fluorescence emission band of HpD at 636 nm and photosensitivity of cells.

Ca2+ influx induced by photodynamic action in human cerebral glioma (U-87 MG) cells: possible involvement of a calcium channel.

The plasma membrane has been implicated as a critical target of photodynamic action on cells. We have observed that the photosensitization of human cerebral glioma (U-87 MG) cells by hematoporphyrin derivative (HpD) causes a large increase in intracellular calcium [Ca2+]i. This increase in [Ca2+]i was solely due to the influx of extracellular Ca2+ through the plasma membrane and showed a dependence on HpD concentration, light dose and concentration of calcium in the extracellular medium. The magnitude of the Ca2+ influx decreased with increasing postirradiation time, which suggests that the cell membrane partially recovers from the photodynamic injury. The photoinduced Ca2+ influx was inhibited by the Ca2+ channel blocker diltiazem and the reducing agent dithioerythritol. These findings are discussed in terms of possible activation of a Ca2+ channel as a result of photosensitization.

A novel type of Ca2+ channel in U-87 MG cells activated by anti-galactocerebroside.

Antibody to galactocerebroside (GalC) evokes a Ca2+ response in cultured glioma U-87 MG cells. The rise in intracellular calcium [Ca2+]i occurs largely due to the influx of Ca2+ through a plasma membrane channel, though the release of Ca2+ from intracellular stores also contributes. We characterized the channel activated by anti-GalC. The channel activity was transient and the inactivation appeared to be Ca2+ dependent. The channel was impermeant to monovalent ions Na+ and K+ and also to Mn2+. Ni2+ and Co2+ neither permeate through the channel nor inhibit the Ca2+ influx. In contrast Cd2+ the most potent inorganic blocker of Ca2+ channels permeated through this channel. The Ca2+ influx was inhibited by verapamil with IC50 of 65 +/- 8 microM. The Ca2+ influx as well as the intracellular release were markedly inhibited by neomycin sulfate and phorbol dibutyrate, suggesting that the Ca2+ influx may be mediated by IP3 (1). Depletion of intracellular Ca2+ stores by thapsigargin was followed by Ca2+ influx. This represents the capacitative Ca2+ entry pathway and is distinct from the channel activated by anti -GalC.

Differential effects of tricyclic antidepressant drugs on membrane dynamics--a fluorescence spectroscopic study.

The effect of tricyclic antidepressant drugs amitriptyline, nortriptyline, imipramine and desipramine on synaptosomal membrane and lipid bilayer was studied using steady state and time dependent fluorescence spectroscopy of lipid specific fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH). The synaptosomal membrane was prepared from rat brain while liposomes were prepared from dimyristoyl phosphatidyl choline (DMPC) alone and a mixture of DMPC and cholesterol. Upon treatment with amitriptyline and nortriptyline a decrease was observed in the steady state anisotropy of DPH in DMPC liposomes as well as in rat brain synaptosomes. On the other hand, imipramine and desipramine did not cause any significant change. Amitriptyline and nortriptyline also decreased the steady state anisotropy of DPH in liposomes prepared from a mixture of DMPC and cholesterol. Fluorescence decay time and time dependent anisotropy of DPH in both the membranes were measured and the decay of anisotropy was analyzed using wobbling in cone model. Amitriptyline and nortriptyline treatment decreased the limiting anisotropy and order parameter, while the cone angle increased. Imipramine and desipramine did not cause significant change in these parameters. In addition to structural alterations, these drugs inhibited the activity of Na+-K+-ATPase in synaptosomal membrane, however, the decrease was more in case of amitriptyline and nortriptyline as compared to imipramine and desipramine. Our results suggest that the perturbation in membrane order caused by antidepressant drugs could depend on the net charge on the drug molecule.

MC540 induced photosensitization of glioma & neuroblastoma cells.

Binding and photodynamic action of merocyanine 540 (MC540) has been studied in glioma (U-87MG) and neuroblastoma (Neuro 2A) cells as a function of dye concentration, incubation time of cells with MC540 and growth phase of cells. In the plateau phase, U-87MG cells accumulated more MC540 as compared to exponentially growing cells, whereas in Neuro 2A cells the opposite effect was observed. Exponentially growing U-87MG cells were more photosensitive than plateau phase cells. However, the photosensitivity of Neuro 2A cells was not dependent on the growth phase. Thus, MC540 mediated photosensitization may be useful for photodynamic therapy of brain tumours.

Effect of hematoporphyrin derivative and light on sulfhydryl groups in brain tumour cells.

Effect of hematoporphyrin derivative (HpD) and light on sulfhydryl (SH) groups in brain tumor cells was studied. Sulfhydryl groups were measured by 5,5'-dithiobis-(2-nitrobenzoic acid) (DTNB) and a fluorescent probe 3,4-maleimidylphenyl-4-methyl-7-diethylaminocoumarin (CPM). Incubation of cells with HpD in dark resulted in the loss of DTNB as well as CPM reactive SH groups. After 2 hr of incubation DTNB reactive SH groups showed a negligible change while a continuous decrease was observed in CPM reactive SH groups. Cells treated with HpD showed a further degradation of SH groups upon light irradiation. A comparison of cytotoxicity and SH groups under identical conditions showed that blockage of SH groups by HpD binding is not leathal to the cells where as photoinduced cell death was observed on photodegradation of SH groups.

Hematoporphyrin derivative binding and photosensitization in human glioblastoma cells: comparison of exponential and plateau phase cells.

Plateau phase glioblastoma (U 87MG) cells were found more photosensitive than the exponentially growing cells. In both phases of growth, the photosensitivity showed further enhancement on incubating the cells with HpD for longer duration. Plateau phase cells accumulated more HpD than exponential phase cells for shorter duration of incubation with HpD, however, for longer duration of incubation, the amount of drug uptake was almost the same in both phases of growth. Fluorescence spectra of cell bound HpD showed a difference in spectral intensity distribution in exponential and plateau phase cells. In exponential phase cells, the fluorescence maximum of cell bound HpD was at 615 nm whereas in plateau phase cells the same was at 636 nm.

Depolarization of synaptosomal membranes: a study of mechanism by which rhodamine 6G measures membrane potential.

The fluorescence intensity of Rhodamine 6G in synaptosomal suspensions has been measured to monitor the membrane potential changes in pre-synaptic nerve terminals. The fluorescence response of the dye was seen to be a function of potential-dependent partitioning of dye molecules between the synaptosomes and the extracellular medium. Binding of dye molecules to the hydrophobic regions of membranes results in the quenching of fluorescence. Upon depolarization of the synaptosomal membrane, the dye molecules are released from the cells. The effect of changing extracellular ionic composition was also studied. The membrane potential increased linearly with log of [K]0. The resting membrane potential in buffer containing 5 mM K+ was calculated to be -60 mV. Raising the extracellular Ca2+ and Mg2+ from 1.2 mM to 10 mM did not change the membrane potential. Ca2+ ionophore A23187, in the presence of Ca2+ was found to depolarize the membranes.

Mechanism of photosensitization of glioblastoma and neuroblastoma cells by merocyanine 540: a lipid peroxidation study.

Mechanism of merocyanine 540 (MC540) mediated photosensitization in glioblastoma (U-87MG) and neuroblastoma (Neuro 2a) cells was investigated. Photoinduced lipid peroxidation was measured in the presence of mechanistic probes-deuterium oxide (D2O), sodium azide, superoxide dismutase (SOD), mannitol and sodium benzoate. In both the types of cells, the photoinduced lipid peroxidation was enhanced in D2O whereas it showed inhibition in the presence of sodium azide. SOD also inhibited the lipid peroxidation while sodium benzoate and mannitol had no effect. These results suggest that photosensitization of U-87MG and Neuro 2a cells by MC 540 involves both type I (free radical mediated) and type II (singlet oxygen mediated) mechanisms.

Structural alterations induced by photodynamic action of hematoporphyrin derivative (HpD) in plasma membrane of glioblastoma (U-87MG) cells: time dependent fluorescence spectroscopic study.

Photodynamic action of hematoporphyrin derivative (HpD) on the plasma membrane of human glioblastoma U-87MG cells was investigated using lipid and protein specific fluorescent probes trimethylammonium-1,6-diphenyl 1,3,5-hexatriene (TMA-DPH) and N-(1-pyrene)-maleimide (PM) respectively. Steady state anisotropy, decay time and time dependent anisotropy of these probes in U-87MG cells were measured. Light irradiation caused an increase in the steady state anisotropy of TMA-DPH in cells treated with HpD; however, no change in decay time was observed. Time dependent anisotropy measurements were performed and the data were analyzed using wobbling in cone model. A decrease in the rotational relaxation time (phi) as well as the cone angle (theta(c)) and an increase in the order parameter (S) of TMA-DPH were observed on photosensitization of cells. A decrease in the order parameter (S) of TMA-DPH were observed on photosensitization of cells. A decrease in the steady rate anisotropy and the rotational relaxation time (phi) of PM and enhancement in the lipid peroxidation were also observed. Our results show that the photodynamic action of HpD increases the order in the lipid bilayer and the mobility of the proteins in the plasma membrane of cells.

Regulation of transmembrane signaling by ganglioside GM1: interaction of anti-GM1 with Neuro2a cells.

Interaction of antibodies to ganglioside GM1 with Neuro2a cells was studied to investigate the role of GM1 in cell signaling. Binding of anti-GM1 to Neuro2a cells induced the formation of 3H-inositol phosphates (3H-IPs) and elevated the intracellular Ca2+ concentration [Ca2+]i. The rise in [Ca2+]i was due to the influx of Ca2+ from the extracellular medium and release from intracellular Ca2+ pools. The Ca2+ influx pathway did not allow the permeation of Na+ or K+. The influx was inhibited by amiloride, a specific blocker of T-type Ca2+ channels, whereas nifedipine and diltiazem, blockers of L-type Ca2+ channels, did not have any effect. Thus, anti-GM1 appears to activate a T-type Ca2+ channel in Neuro2a cells. The intracellular Ca2+ release was inhibited by pretreatment of cells with neomycin sulfate, phorbol dibutyrate, and pertussis toxin (PTx), which also inhibited the 3H-IP formation in Neuro2a cells. Addition of caffeine neither elevated the [Ca2+]i nor affected the anti-GM1-induced [Ca2+]i rise. The data reveal that the binding of anti-GM1 to Neuro2a cells activates phospholipase C via a PTx-sensitive G protein, which leads to formation of IPs and release of Ca2+ from inositol trisphosphate-sensitive pool of endoplasmic reticulum. Anti-GM1 also arrested the differentiation of Neuro2a cells in culture and significantly stimulated their proliferation. This stimulatory effect of anti-GM1 on cell proliferation was blocked by amiloride but not by PTx, suggesting that the influx of Ca2+ was essentially required for cell proliferation. Our data suggest a role for GM1 in the regulation of transmembrane signaling events and cell growth.

Calcium sequestering ability of mitochondria modulates influx of calcium through glutamate receptor channel.

The excitotoxicity of glutamate is believed to be mediated by sustained increase in the cytosolic Ca2+ concentration. Mitochondria play a vital role in buffering the cytosolic calcium overload in stimulated neurons. Here we have studied the glutamate induced Ca2+ signals in cortical brain slices under physiological conditions and the conditions that modify the mitochondrial functions. Exposure of slices to glutamate caused a rapid increase in [Ca2+]i followed by a slow and persistently rising phase. The rapid increase in [Ca2+]i was mainly due to influx of Ca2+ through the N-methyl-D-aspartate (NMDA) receptor channels. Glutamate stimulation in the absence of Ca2+ in the extracellular medium elicited a small transient rise in [Ca2+]i which can be attributed to the mobilization of Ca2+ from IP3 sensitive endoplasmic reticulum pools consequent to activation of metabotropic glutamate receptors. The glutamate induced Ca2+ influx was accompanied by depolarization of the mitochondrial membrane, which was inhibited by ruthenium red, the blocker of mitochondrial Ca2+ uniporter. These results imply that mitochondria sequester the Ca2+ loaded into the cytosol by glutamate stimulation. Persistent depolarization of mitochondrial membrane observed in presence of extracellular Ca2+ caused permeability transition and released the sequestered Ca2+ which is manifested as slow rise in [Ca2+]i. Protonophore carbonyl cyanide m-chlorophenyl-hydrazone (CCCP) depolarized the mitochondrial membrane and enhanced the glutamate induced [Ca2+]i response. Contrary to this, treatment of slices with mitochondrial inhibitor oligomycin or ruthenium red markedly reduced the [Ca2+]i response. Combined treatment with oligomycin and rotenone further diminished the [Ca2+]i response and also abolished the CCCP mediated rise in [Ca2+]i. However, rotenone alone had no effect on glutamate induced [Ca2+]i response. These changes in glutamate-induced [Ca2+]i response could not be explained on the basis of deficient mitochondrial Ca2+ sequestration or ATP dependent Ca2+ buffering. The mitochondrial inhibitors reduced the cellular ATP/ADP ratio, however, this would have restrained the ATP dependent Ca2+ buffering processes leading to elevation of [Ca2+]i. In contrast our results showed repression of Ca2+ signal except in case of CCCP which drastically reduced the ATP/ADP ratio. It was inferred that, under the conditions that hamper the Ca2+ sequestering ability of mitochondria, the glutamate induced Ca2+ influx could be impeded. To validate this, influx of Mn2+ through ionotropic glutamate receptor channel was monitored by measuring the quenching of Fura-2 fluorescence. Treatment of slices with oligomycin and rotenone prior to glutamate exposure conspicuously reduced the rate of glutamate induced fluorescence quenching as compared to untreated slices. Thus our data establish that the functional status of mitochondria can modify the activity of ionotropic glutamate receptor and suggest that blockade of mitochondrial Ca2+ sequestration may desensitize the NMDA receptor operated channel.

High concentrations of tricyclic antidepressants increase intracellular Ca2+ in cultured neural cells.

We examined the effect of tricyclic antidepressants on intracellular Ca2+ signalling in cultured cells of neuronal and glial origin. High concentrations of amitriptyline and desipramine increased the intracellular Ca2+ in PC-12 and U-87 MG cells. In PC-12 cells amitriptyline induced a biphasic rise in intracellular Ca2+. A rapid and transient increase due to release of Ca2+ from intracellular pools was followed by sustained elevation of [Ca2+]i due to influx from the extracellular medium. Desipramine evoked the Ca2+ release from intracellular pools but the influx of Ca2+ was not elicited. In U-87 MG cells both the drugs induced Ca2+ release from intracellular pools, however amitriptyline also induced a transient influx of Ca2+. To delineate the mechanisms involved in mobilization of Ca2+ by the drugs pharmacological agents that inhibit IP3 formation in cells and Ca2+ channel blockers were used and changes in [Ca2+]i and membrane potential were monitored. The results show that both the drugs release Ca2+ from IP3 sensitive pools by activation of phospholipase C and amitriptyline in addition activates a non specific cation channel in the plasma membrane of cells. Paradoxically at relatively lower concentrations (< 50 microM) amitriptyline and desipramine inhibited the Ca2+ signal induced by adenosine triphosphate in both the cell types. Our data demonstrate that tricyclic antidepressants at different doses may have inhibitory or stimulatory effects on cellular Ca2+ signalling.