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Biomedical subjects

S Vijayaraghavan

Publications and source records attributed to S Vijayaraghavan.

At least 19 recordsLinked to original sources
PubMed1999

Effect of acute exposure of the organophosphate insecticide Rogor on some biochemical aspects of Clarias batrachus (Linnaeus).

Indian catfish (Clarias batrachus), a common fish of commercial importance, was exposed to a sublethal concentration (1/3 of 96 h LC50 value) of Rogor for periods ranging from 24 to 192 h. The changes in glycogen, lactate, lactate dehydrogenase, and glycogen phosphorylase (a, ab) contents of muscle tissue were studied. A gradual decrease in muscle glycogen and an increase in lactate contents were observed. The activity level of lactate dehydrogenase showed a sharp rise initially for 48 h followed by a decline after 96 and 192 h. The glycogen phosphorylase a and ab in muscle tissue enhanced in response to Rogor exposure.

Animals↗
PubMed1999

Self-injurious behavior is decreased by cyproterone acetate in adult male rhesus (Macaca mulatta).

Self-injurious behavior (SIB) presents a serious problem in laboratory macaques that cannot be socially housed for scientific reasons and among institutionalized children and adults where it is often associated with different forms of brain dysfunction. We have experienced limited success in reducing SIB in macaques by enhancing their environment with enrichment devices. Psychotropic drugs also help, but problems are associated with their use. Because sexual and aggressive behavioral problems in men have been treated with progestational drugs, we tested the efficacy of cyproterone acetate (CA, 5-10 mg/kg/week) on reducing SIB in 8 singly housed, adult male rhesus macaques. The main findings were: (1) SIB and other atypical behaviors were significantly reduced during CA treatment; (2) serum testosterone was significantly reduced during CA treatment; (3) cerebral spinal fluid (CSF) levels of 5HIAA and HVA, metabolites of serotonin and dopamine, respectively, declined significantly during CA treatment; (4) the duration of SIB positively correlated with levels of 5HIAA in CSF; but (5) sperm counts were not reduced during treatment. Thus, CA was a partially effective treatment (3 months) for adult male macaques whose behavioral problems include SIB. In summary, CA reduced SIB, overall aggression, serum testosterone, CSF 5HIAA, and CSF HVA. We hypothesized that the progestin activity of CA represses the hypothalamic gonadal axis and decreases testosterone, which in turn decreases SIB. In addition, we speculate that the decrease in 5HIAA and HVA in CSF may have been caused by progestins decreasing the activity of MAO. Therefore, the reduction of SIB may also be related to an increase in the availability of active monoamines in the CNS.

Age Factors↗
PubMed1998

Nicotinic receptor-induced apoptotic cell death of hippocampal progenitor cells.

Nicotine has many effects on CNS functions, presumably through its action on neuronal nicotinic acetylcholine receptors (AChRs). One subclass of AChRs that binds the snake venom toxin alpha-bungarotoxin (alpha-Bgt-AChRs) has been shown to modulate neurotransmission in the brain. We now show that alpha-Bgt-AChR activation by low doses of nicotine results in apoptotic cell death of both primary and immortalized hippocampal progenitor cells. In HC2S2-immortalized hippocampal progenitors, nicotine is cytotoxic to undifferentiated cells, whereas it spares the same cells once differentiation has been induced. The activation of alpha-Bgt-AChRs by nicotine results in the induction of the tumor suppressor protein p53 and the cdk inhibitor p21. The cytotoxic effect of nicotine is dependent on extracellular calcium levels and is probably attributable to the poor ability of undifferentiated progenitors to buffer calcium loads. The major calcium buffer in these cells, calbindin D28K, is present only after differentiation has been induced. Furthermore transfection of undifferentiated cells with calbindin results in dramatic protection against the cytotoxic effects of nicotine. These results show that nicotine abuse could have significant effects on the survival of progenitor populations in the developing and adult brain and also suggest an endogenous role for alpha-Bgt-AChRs in neuronal development and differentiation.

Animals↗
PubMed1997

Protein kinase A-anchoring inhibitor peptides arrest mammalian sperm motility.

Cyclic AMP-dependent protein kinase (PKA) is anchored at specific subcellular sites through the interaction of the regulatory subunit (R) with protein kinase A-anchoring proteins (AKAPs) via an amphipathic helix binding motif. Synthetic peptides containing this amphipathic helix domain competitively disrupt PKA binding to AKAPs and cause a loss of PKA modulation of cellular responses. In this report we use S-Ht31, a cell-permeant anchoring inhibitor peptide, to study the role of PKA anchoring in sperm. Our analysis of three species of mammalian sperm detected three isoforms of PKA (RIIalpha, RIIbeta, and RIbeta) and one 110-kDa AKAP. The addition of S-Ht31 to bovine caudal epididymal sperm inhibits motility in a time- and concentration-dependent manner. A control peptide, S-Ht31-P, identical to S-Ht31 except for a proline for isoleucine substitution to prevent amphipathic helix formation, had no effect on motility. The inhibition of motility by S-Ht31 is reversible but only if calcium is present in the suspension buffer, suggesting a role for PKA anchoring in regulating cellular calcium homeostasis. Surprisingly, inhibition of PKA catalytic activity had little effect on basal motility or motility stimulated by agents previously thought to work via PKA activation. These data suggest that the interaction of the regulatory subunit of PKA with sperm AKAPs, independent of PKA catalytic activity, is a key regulator of sperm motility and that disruption of this interaction using cell-permeable anchoring inhibitor peptides may form the basis of a sperm-targeted contraceptive.

Animals↗
PubMed1997

A tyrosine-phosphorylated 55-kilodalton motility-associated bovine sperm protein is regulated by cyclic adenosine 3',5'-monophosphates and calcium.

Sperm motility is regulated by protein phosphorylation. We have recently shown that a serine/threonine phosphatase system is involved in motility regulation. Two of the components of the phosphatase system, GSK-3 and PP1gamma2, are regulated by tyrosine phosphorylation. During our investigation of sperm tyrosine-phosphorylated proteins we discovered a 55-kDa protein whose tyrosine phosphorylation correlates closely to the motility state of sperm. This protein is tyrosine phosphorylated to a much higher degree in motile caudal than in immotile caput epididymal sperm. Motility inhibition of caudal epididymal sperm by protein kinase A (PKA) anchoring inhibition or by ionomycin-induced calcium overload led to the virtual disappearance of tyrosine phosphorylation of the 55-kDa protein. Conversely, treatment of sperm with motility activators, isobutylmethylxanthine or 8-bromo-cAMP, resulted in increased tyrosine phosphorylation of the protein. The protein was present in the soluble 100 000 x g supernatants of sperm extracts and was heat labile. Chromatography through diethylaminoethyl-cellulose and Western blot analysis showed that this 55-kDa protein is not a regulatory subunit of PKA or alpha-tubulin. Our results represent the identification of a soluble protein whose tyrosine phosphorylation varies directly with motility and suggest that motility regulation may involve cross talk between PKA, calcium, and tyrosine kinase pathways.

Amino Acid Sequence↗
PubMed1997

Subcellular localization of the regulatory subunits of cyclic adenosine 3',5'-monophosphate-dependent protein kinase in bovine spermatozoa.

Cyclic AMP (cAMP) is a regulator of sperm flagellar activity. The action of this cyclic nucleotide is presumably mediated by cAMP-dependent protein kinase (PKA). PKA is localized or targeted to specific subcellular sites through the interaction of PKA regulatory subunits with A-kinase anchoring proteins (AKAPs). We have recently shown that the addition of PKA anchoring inhibitor peptides to spermatozoa leads to the complete arrest of motility. A knowledge of the subcellular localization of PKA and AKAPs is essential for an understanding of how cAMP acts in spermatozoa. In this report, monospecific, affinity-purified, antipeptide antibodies were used to determine the distribution of the regulatory (R) subunit isoforms. Immunocytochemistry staining revealed that RIalpha and RIbeta subunits are both localized predominantly in the acrosomal segment of the head, although they have distinct staining patterns within this region. In addition to the head, RIbeta was observed in the midpiece of the tail while RIalpha was detected in the connecting piece. RIIalpha is prominent in the axonemal region of the flagellum but was not observed in the head region. These data suggest distinct roles for each of these isoforms in sperm functions such as motility and the acrosome reaction.

Acrosome↗
PubMed1997

Hypercalcaemia.

Explore the source record for details and available documents.

Female↗
PubMed1996

Post-translational regulation of neuronal acetylcholine receptors stably expressed in a mouse fibroblast cell line.

Second messenger regulation of neuronal acetylcholine receptors (AChRs) was investigated in a mouse fibroblast cell line, M10, stably transfected with chicken alpha 4 and beta 2 cDNAs. Both forskolin and 8-bromo-cyclic adenosine 3',5'-monophosphate (cAMP) induced large increases in the numbers of AChRs. The increases were due in part to increased transcription and translation of the alpha 4 and beta 2 genes. Blockade of protein synthesis with cycloheximide, however, revealed that forskolin also exerts a post-translational effect, increasing the number of surface receptors by twofold. Immunoblot analysis of sucrose gradient fractions confirmed that the cells had a large fraction of unassembled subunits potentially available for receptor assembly. The post-translational effect of forskolin was blocked by H-89, an inhibitor of cAMP-dependent protein kinase, and by okadaic acid, an inhibitor of phosphatases 1 and 2A. Nicotine also acted post-translationally to induce a twofold increase in the number of surface receptors, but the mechanism differed from that utilized by forskolin, since the effects of the two agents were additive and were differentially affected by okadaic acid. The results suggest that protein phosphorylation-dephosphorylation mechanisms act post-translationally to increase the number of neuronal AChRs maintained on the cell surface. This could be achieved by increasing the efficiency of receptor assembly, transport, or stabilization on the cell surface.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1996

Elevation of intracellular calcium levels in neurons by nicotinic acetylcholine receptors.

The recognition that intracellular free calcium serves as a ubiquitous intracellular signal has motivated efforts to elucidate mechanisms by which cells regulate calcium influx. One route of entry that may offer both spatial and temporal fine resolution for altering calcium levels is that provided by cation-permeable, ligand-gated ion channels. Biophysical measurements as well as calcium imaging techniques demonstrate that neuronal nicotinic acetylcholine receptors as a class have a high relative permeability to calcium; some subtypes equal or exceed all other known receptors in this respect. Activation of nicotinic receptors on neurons can produce substantial increases in intracellular calcium levels by direct passage of calcium through the receptor channel. When multiple classes of nicotinic receptors are expressed by the same neuron, each appears capable of increasing calcium in the cell but may differ with respect to location, temporal response, agonist sensitivity, or regulation in achieving it. As a result, nicotinic receptors must be considered strong candidates for signaling molecules through which neurons regulate a diverse array of cellular events.

Acetylcholine↗
PubMed1996

Sperm motility development in the epididymis is associated with decreased glycogen synthase kinase-3 and protein phosphatase 1 activity.

Immotile bovine caput epididymal sperm contain levels of protein phosphatase activity twofold higher than do mature motile caudal sperm. Comparison of the inhibition profiles of endogenous phosphatase activities detected by okadaic acid (OA) and calyculin A (CA) revealed a pattern consistent with the predominance of a type 1 protein phosphatase (PP1). Immunoblot analysis identified PP1 gamma 2 (the testis-specific isoform of PP1) as the only PP1 isoform in sperm and showed little protein phosphatase 2A (PP2A). In addition, of the known PP1 inhibitors, i.e., DARPP-32, inhibitor 1 (I1), and inhibitor 2 (I2), only I2-like activity was detected in sperm. Inhibition of PP1 by the heat-stable I2-like activity purified from sperm could be reversed with purified glycogen synthase kinase-3 (GSK-3). Furthermore, sperm extracts contain an inactive complex of PP1 and I2 (termed PP1I) that could also be activated by purified GSK-3. The presence of GSK-3 in sperm was demonstrated by activation of purified PP1I, and quantitation revealed that immotile caput sperm contained sixfold higher GSK-3 activity than motile caudal sperm. Immunoblot analysis confirmed the expression of GSK-3 in sperm and revealed the occurrence of both the alpha and beta isoforms. Our findings suggest that the higher PP1 activity measured in immotile sperm, presumably due to higher GSK-3 activity, is responsible for holding motility in check. This conclusion was supported by the observation that the phosphatase inhibitors OA and CA, at micromolar and nanomolar levels, respectively, were able to induce motility in completely immotile bovine caput epididymal sperm and to stimulate the kinetic activity of mature caudal sperm. The intrasperm levels of cAMP, pH, and calcium were unaltered by treatment with these inhibitors. The results suggest a biochemical basis for the development and regulation of sperm motility and a possible physiological role for the PP1/I2/GSK-3 system.

Amino Acid Sequence↗
PubMed1996

Primate sperm contain protein phosphatase 1, a biochemical mediator of motility.

Sperm motility initiation, capacitation, and hyperactivation are modulated by an interplay of intracellular Ca2+, cAMP, and pH. Mechanisms by which these mediators alter sperm function have not been elucidated but may involve reversible alterations in regulatory protein phosphorylation. Studies were designed 1) to investigate the influence of the protein phosphatase (PP) inhibitor calyculin A (CA) on human sperm motility and 2) to characterize the CA-sensitive PP and its endogenous regulators in human and rhesus monkey sperm. CA (50 nM) treatment of human sperm resulted in an increase in percentage motility and an acceleration in mean path velocity. Inhibition of either protein phosphatase-1 (PP1) or protein phosphatase-2A (PP2A) could be responsible for this motility stimulation, since both of these phosphatases are sensitive to nanomolar quantities of CA. PP activity in human (n = 4) and rhesus monkey (n = 4) sperm sonicates was measured using [32P]-phosphorylase-a, the preferred substrate for PP1 and PP2A, in the absence of divalent cations. Human (6.2 +/- 4.5 x 10(-2) mU/10(6) sperm) and monkey (4.3 +/- 0.8 x 10(-2) mU/10(6) sperm) sonicates contained activity tentatively identified as PP1 on the basis of inhibition profiles in okadaic acid (OA) and CA. Western blot analysis with antibodies against various isoforms of PP1 subsequently documented the presence of PP1 gamma 2 in human and monkey sperm. PP1 activity in most tissues is regulated by the heat-stable inhibitors I1 or I2. Sperm sonicates contained inhibitor activity similar to I2 as well as glycogen synthase kinase-3 (GSK-3) activity, which is involved in the activation of the PP1-I2 complex. These results indicate, for the first time, that human and rhesus monkey sperm contain PP1 and regulators of PP1 and that inhibition of PP1 activity by CA can enhance motility.

Adult↗
PubMed1995

Acetylcholine differentially affects intracellular calcium via nicotinic and muscarinic receptors on the same population of neurons.

Multiple receptor subtypes activated by the same ligand but coupled to different second messengers can produce divergent signaling in a cell, while receptors activated by different ligands but sharing the same second messenger can produce convergent signaling. We show here that chick ciliary ganglion neurons have three classes of receptors activated by the same neurotransmitter, acetylcholine, and that all three regulate the same second messenger, intracellular free calcium. Activation of muscarinic receptors on the neurons stimulates phosphatidylinositol turnover and induces calcium oscillations that are initiated and maintained by calcium release from caffeine/ryanodine-insensitive intracellular stores. Extracellular calcium is required to sustain the oscillations, while cadmium abolishes them. Activation of either of two classes of nicotinic receptors, distinguished both by location on the neurons and by subunit composition, induces a single, rapid elevation in intracellular calcium without inducing phosphatidylinositol turnover. The nicotinic responses are entirely dependent on extracellular calcium, show no dependence on release from internal stores, and do not display oscillations. Low concentrations of the native agonist, acetylcholine, induce repetitive calcium spikes in the neurons characteristic of muscarinic receptors, while higher concentrations induce nonoscillating increases in intracellular calcium that include contributions from nicotinic receptors. The three classes of receptors also differ in the acetylcholine concentration required to elicit a response. These differences, together with differences in receptor location and sources of calcium mobilized, may enable the receptor subtypes to target different sets of calcium-dependent processes for regulation.

Acetylcholine↗
PubMed1995

Carbohydrate metabolism in hepatic tissue of freshwater catfish Clarias batrachus L. during dimethoate exposure.

Clarias batrachus L., a freshwater catfish of commercial importance, was exposed to a sublethal concentration of dimethoate (Rogor). The acute effect of dimethoate on some aspects of carbohydrate metabolism of hepatic tissue was studied after 1, 2, 4 and 8 days. The glycogen content was depleted, whereas the lactate level was elevated throughout the experimental period and an increase in lactate dehydrogenase was observed. The activity levels of glycogen phosphorylase a and ab in the hepatic tissue were increased during dimethoate stress. The results suggest that carbohydrate metabolism was adversely affected in the hepatic tissue by the organophosphate insecticide dimethoate.

Animals↗
PubMed1995

Arachidonic acid as a possible negative feedback inhibitor of nicotinic acetylcholine receptors on neurons.

Neuronal acetylcholine receptors, being highly permeable to calcium, are likely to regulate calcium-dependent events in neurons. Arachidonic acid is a membrane-permeant second messenger that can be released from membrane phospholipids by phospholipases in a calcium-dependent manner. We show here that activation of neuronal acetylcholine receptors triggers release of 3H-arachidonic acid in a calcium-dependent manner from neurons preloaded with the fatty acid. Moreover, low concentrations of arachidonic acid reversibly inhibit the receptors and act most efficiently on receptors likely to have the highest permeability to calcium, namely receptors containing alpha 7 subunits. Low concentrations of arachidonic acid also reversibly inhibit alpha 7-containing receptors expressed in Xenopus oocytes following injection of alpha 7 cRNA. The oocyte results indicate following injection of alpha 7 cRNA. The oocyte results indicate that the inhibition is a feature of the receptors rather than a consequence of neuron-specific machinery. The inhibition is not mediated by specific metabolites of arachidonic acid because the effects can be mimicked by other fatty acids; their effectiveness correlates with their content of double bonds. In contrast to arachidonic effects on calcium currents, inhibition of neuronal nicotinic receptors by the fatty acid cannot be prevented by blocking production of free radicals or by inhibiting protein kinase C. An alternative mechanism is that arachidonic acid binds directly to the receptors or perturbs the local environment in such a manner as to constrain receptor function.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Science for art's sake.

Hollywood's absent-minded professor, always a cliché, is now more unrealistic than ever before. Just how can biological research move forward in the modern era?

Art↗
PubMed1994

Evidence against a functional ATP-dependent calcium extrusion mechanism in bovine epididymal sperm.

Bovine epididymal sperm resuspended in ionic buffers take up relatively large amounts of calcium. This uptake, which is almost entirely mitochondrial, apparently bypasses the sperm cytosol. The direct mitochondrial loading is an unusual aspect of sperm calcium uptake, which suggests that the plasma membrane region surrounding the mitochondria should be highly permeable to calcium, whereas the membrane domains surrounding the head and tail regions of sperm should be impermeable. This study was undertaken to determine the role of a plasma membrane calcium ATPase in sperm calcium homeostasis. Kinetics of calcium (45Ca2+) uptake into intact and permeabilized caudal epididymal sperm confirmed that mitochondrial calcium uptake occurs with virtually no resistance from the surrounding plasma membrane. Cytoplasmic calcium accumulation by sperm depleted of intracellular ATP, measured in the presence of mitochondrial calcium uptake inhibitors, showed no increase upon energy depletion as would be expected if an ATP-dependent calcium extrusion mechanism were present. Furthermore, lowering the incubation temperature to further reduce the activity of the calcium ATPase in these energy-depleted sperm was also without effect on calcium accumulation. The calcium ATPase inhibitor vanadate, even at high concentrations, failed to increase intracellular 45Ca2+ accumulation. However, vanadate was effective in inhibiting motility showing that the compound was accumulated into sperm to inhibit flagellar dyenin ATPase. Therefore, the lack of effect of vanadate on 45Ca2+ accumulation was not due to its inability to enter sperm. Other calcium ATPase inhibitors such as quercetin, thapsigargin, and cyclopiazonic acid, which readily demonstrate ATP-dependent calcium extrusion in other somatic cells, were also without effect on sperm calcium accumulation.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗