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dottorato internazionale in neurofarmacologia rivista trimestrale on-line anno vi numero 2 aprile giugno 2011 il prometeo direttore prof filippo drago pubblicazioni del dottorato internazionale di neurofarmacologia 2009 2011 neuropsychopharmacology 2009 feb;343 593-606 epub 2008 jun 25 anxiolytic effects in mice of a dual blocker of fatty acid amide hydrolase and transient receptor potential vanilloid type-1 channels micale v cristino l tamburella a petrosino s leggio gm drago f di marzo v department of experimental and clinical pharmacology university of catania medical school catania italy dottorato internazionale di neurofarmacologia the endocannabinoid-inactivating enzyme fatty acid amide hydrolase faah and the transient receptor potential vanilloid type-1 trpv1 channel are new targets for the development of anxiolytic drugs we studied the effect on anxiety-like behavior in the elevated plus maze of a dual faah/trpv1 blocker n-arachidonoyl-serotonin aa-5-ht in male c57bl/6j mice acute intraperitoneal administration of aa-5-ht 0.1-2.5 mg kg increased both the time spent and the number of entries in the open arm while being inactive at the highest dose tested 5 mg/kg aa-5-ht was more potent than selective blockers of faah or trpv1 urb597 and sb366791 respectively in male swiss mice aa-5-ht had to be administered chronically to observe an anxiolytic effect at an intermediate dose 2.5 mg/kg the highest dose 5 mg/kg being anxiogenic and 1 mg kg being ineffective in both strains the anxiolytic effects of aa-5-ht were paralleled by elevation of brain endocannabinoid levels and were reversed by per se inactive doses of the cannabinoid receptors of type-1 cb1 receptor antagonist am251 or the trpv1 agonist olvanil immunohistochemical localization of cb1 and trpv1 receptors was observed in mouse prefrontal cortex nucleus accumbens amygdala and hippocampus simultaneous `indirect activation of cb1 receptors following faah inhibition and antagonism at trpv1 receptors might represent a new therapeutic strategy against anxiety br j ophthalmol 2009 feb;932 254-7 epub 2008 oct 31 università degli studi di catania dipartimento di biomedicina clinica e molecolare viale andrea doria 6 catania tel +39 095 7384237 tel +39 095.7384238 il_prometeo@email.it http www.unict.it/dfsc a water-soluble carbon monoxide-releasing molecule corm-3 lowers intraocular pressure in rabbits sezione di farmacologia e biochimica stagni e privitera mg bucolo c leggio gm motterlini r drago f department of experimental and clinical pharmacology university of catania medical school viale a doria 6 catania italy background:carbon monoxide-releasing molecules corms are a novel group of substances that are capable of modulating physiological functions via the liberation of co aims:this study was undertaken to investigate the effects of corm-3 a water-soluble co-releasing agent on two rabbit models of ocular hypertension methods:ocular hypertension was induced by injecting alpha-chymotrypsin in the rabbit eye the dose-response effect of corm-3 on iop was assessed by topical administration of the drug 0.001 0.01 0.1 and 1 ocular hypertension was also obtained by weekly subconjunctival injection of betamethasone and animals were treated topically with corm-3 a group of animals in both models was treated with the inactive form of the drug icorm-3 results:corm-3 induced a dose-dependent reduction in iop in rabbits treated with alpha-chymotrypsin a similar reduction in iop was observed in rabbits with betamethasone-induced ocular hypertension treated with the drug treatment with the icorm-3 had no effect on iop in both models conclusions:treatment with corm-3 is associated with a reduction in iop in two different rabbit models of ocular hypertension these results support previous findings on the effect of haem oxygenase-derived co on iop and suggest a direct involvement of co system in the regulation of ocular pressure probably through the modulation of aqueous humour dynamics.

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neurochem res 2009 feb;342 327-32 epub 2008 jul 9 parkin expression profile in dopamine d3 receptor knock-out mice brains d agata v tiralongo a castorina a leggio gm micale v carnazza ml drago f department of anatomy diagnostic pathology legal medicine hygiene and public health university of catania medical school via s sofia 87 95123 catania italy mory enhancement by activation of ppar-alpha either directly by administering a ppar-alpha agonist or indirectly by administering a faah inhibitor j neurochem 2009 jun;1096 1733-44 epub 2009 apr 4 prenatal stress alters glutamatergic system responsiveness in adult rat prefrontal cortex fumagalli f pasini m frasca a drago f racagni g riva ma center of neuropharmacology department of pharmacological sciences university of milan milan italy patients affected by autosomic recessive juvenile parkinsonism arjp exhibit parkin gene mutations with brain decrease in dopamine d2/d3 binding sites to date there are no data indicating whether the reduction in dopamine d3 receptors drd3 may be associated with the expression of specific parkin variants in the present study we investigated parkin expression profile in drd3 knock-out mice brains rt-pcr analysis was performed to assess qualitative changes in parkin isoforms distribution pattern and in exons expression both in wild type controls and dopamine d3 receptor s knock-out mice real-time pcr was performed to quantify single exons mrna results demonstrated that exons 1 2 4 6 7 8 were more expressed in wild type compared to dopamine d3 receptor ko mice brains while some other 3 9 10 were lower expressed the expression levels of exons 5 11 and 12 did not change in both animal groups our analysis was confirmed by western blot which showed that parkin protein levels were influenced by the absence of drd3 neurobiol dis 2009 mar;333 415-21 epub 2008 dec 11 enhanced expression of eralpha in astrocytes modifies the response of cortical neurons to betaamyloid toxicity carbonaro v caraci f giuffrida ml merlo s canonico pl drago f copani a sortino ma department of experimental and clinical pharmacology university of catania viale andrea doria 6 95125 catania italy estrogen receptor alpha eralpha is over-expressed in reactive glia under conditions of neuronal damage to elucidate the functional significance of eralpha overexpression an in vitro model of reactive astrocytes with enhanced expression of eralpha was obtained by growth in g5 culture supplement exposure of cortical neurons to beta-amyloid in the presence of either conditioned medium from reactive astrocytes previously treated with 17betaestradiol 17betae2 or transferring of 17betae2-pretreated astrocytes caused a greater neuroprotective effect compared to the respective control conditions although reactive glia resulted being per se neuroprotective blockade of eralpha overexpression by the er antagonist ici182,780 was not successful as ici182,780 behaved as an agonist however complete prevention of 17betae2 effect by ici182,780 produced an increased sensitivity of neurons to beta-amyloid toxicity a similar effect was observed when eralpha knock-down was induced by sirna it is suggested that increased eralpha expression in reactive glia may have a role in limiting neuronal damage learn mem 2009 apr 29;165 332-7 print 2009 may exposure to stress during gestation alters brain development resulting in permanent alterations that may increase susceptibility to subsequent cognitive or neuropsychiatric disorders in this manuscript we examined the effects of prenatal stress on critical determinants of the glutamatergic synapse under basal conditions as well as in response to acute stress the main finding of this work is that gestational stress altered the responsiveness of the glutamatergic system following a challenge at adulthood in fact while in control animals acute swim stress enhanced the phosphorylation levels of the nmda receptor subunits nr-1ser896 and nr-2bser1303 as well as the phosphorylation levels of alpha calcium/calmodulin-dependent protein kinase ii thr286 a crucial sensor of calcium fluctuations prenatal stress prevented or attenuated such activation this dynamic modulation is restricted to prefrontal cortex since no changes were observed in the hippocampus in line with the different maturational profile of these brain regions changes were also observed in the phosphorylation of the alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate subunit glur-1ser831 which however relied on the acute stress exposure and were independent of gestational stress these effects point to a unique interference of chronic prenatal stress with the responsiveness of specific determinants of the glutamatergic synapse at adulthood in a region specific manner the inability to mount an homeostatic glutamatergic response to subsequent stress at adulthood may impair the normal responses of the cell to challenging situations prog neuropsychopharmacol biol psychiatry 2009 oct 1;337 1205-10 epub 2009 jul 19 antidepressant properties of the 5-ht4 receptor partial agonist sl65.0155 behavioral and neurochemical studies in rats tamburella a micale v navarria a drago f department of experimental and clinical pharmacology faculty of medicine university of catania 95125 catania italy fatty acid amide hydrolase faah inhibition enhances memory acquisition through activation of ppar-alpha nuclear receptors mazzola c medalie j scherma m panlilio lv solinas m tanda g drago f cadet jl goldberg sr yasar s preclinical pharmacology section behavioral neuroscience research branch intramural research program nida nih dhhs baltimore maryland 21224 usa inhibitors of fatty acid amide hydrolase faah increase endogenous levels of anandamide a cannabinoid cb1 receptor ligand and oleoylethanolamide and palmitoylethanolamide oea and pea ligands for alpha-type peroxisome proliferator-activated nuclear receptors ppar-alpha when and where they are naturally released in the brain using a passive-avoidance task in rats we found that memory acquisition was enhanced by the faah inhibitor urb597 or by the ppar-alpha agonist wy14643 and these enhancements were blocked by the ppar-alpha antagonist mk886 these findings demonstrate novel mechanisms for me this study was undertaken to investigate the potential antidepressant-like properties of sl65.0155 a serotonin 5-ht4 receptor partial agonist in male rats of the wistar strain tested in the forced swim test fst an experimental model widely used to assess antidepressant-like activity the expression of hippocampal neurotrophic factors such as the brain-derived neurotrophic factor bdnf the phosphorilated camp response element-binding protein p-creb the b cell lymphoma-2 bcl-2 the bax and the vascular endothelium growth factor vegf were also evaluated by western blot analysis different groups of rats received intraperitoneally i.p injections of sl65.0155 0.1 0.5 and 1 mg/kg clomipramine 50 mg/kg citalopram 15 mg/kg or vehicle respectively 24 5 and 1 h prior to the fst compared to the control group sl65.0155 0.5 and 1 mg/kg clomipramine or citalopram injected animals showed an increased swimming and climbing behavior and reduced immobility time in the fst interestingly this effect was not due to changes in the locomotor activity since all treated groups failed to show any change in motor ability as assessed in the open field test western blot analysis of hippocampal homogenates showed an enhancement of p-creb bdnf bcl-2 and vegf protein levels in sl65.0155 treated groups but not in citalopram or clomipramine treated groups used here as positive control no change was found in bax expression in any treated group these findings give further support to the hypothesis that the stimulation of serotonin 5-ht4 receptors may be a therapeutic target for depression.

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neurobiol dis 2009 oct;361 70-80 epub 2009 jul 8 altered responses of dopamine d3 receptor null mice to excitotoxic or anxiogenic stimuli possible involvement of the endocannabinoid and endovanilloid systems micale v cristino l tamburella a petrosino s leggio gm drago f di marzo v department of experimental and clinical pharmacology university of catania medical school catania italy dopamine and the endocannabinoids anandamide and 2-arachidonoylglycerol interact at several levels in the brain with the involvement of both cannabinoid cb1 receptors and transient receptor potential vanilloid type-1 trpv1 channels which are alternative anandamide receptors using pharmacological immunohistochemical and analytical approaches we investigated the response of dopamine d3 receptor null d3r mice in models of epilepsy and anxiety in relation to their brain endocannabinoid and endovanilloid tone compared to wild-type mice d3r mice exhibited a delayed onset of clonic seizures enhanced survival time reduced mortality rate and more sensitivity to anticonvulsant effects of diazepam after intraperitoneal administration of picrotoxin 7 mg/kg and a less anxious-like behaviour in the elevated plus maze test d3r mice also exhibited different endocannabinoid and trpv1 but not cb1 levels in the hippocampus nucleus accumbens amygdala and striatum given the role played by cb1 and trpv1 in neuroprotection and anxiety and based on data obtained here with pharmacological tools we suggest that the alterations of endocannabinoid and endovanilloid tone found in d3r mice might account for part of their altered responses to excitotoxic and anxiogenic stimuli cns neurosci ther 2009 nov 19 [epub ahead of print female volunteers were treated with prulifloxacin or amoxicillin clavulanate in this open randomized parallel-group repeateddose study vaginal signs and symptoms were assessed at the first doctor s visit 0 3 weeks prior to the start of the study and subsequent examinations 1 3 5 6 7 and 8 followup some volunteers treated with amoxicillin-clavulanate showed increased ph values and 73.3 of them had lower lactobacillus flora at visit 3 this reduction was still present in 66.7 3 days after the last dose and in 26.7 of subjects at the follow-up about 7 13 days after the last dose the situation was completely normalized at the second follow-up about one month after treatment stop on the contrary the repeated administration of prulifloxacin 600 mg tablets affected neither the ph nor the lactobacillus component of the vaginal flora in healthy fertile women the oral administration of prulifloxacin may have advantages over some other antimicrobial agents because it preserves the normal vaginal microbiota in healthy women pharmacology 2010;863 182-8 epub 2010 aug 18 modulation of c6 glioma cell proliferation by ureido-calix[8]arenes viola s merlo s consoli gm drago f geraci c sortino ma department of experimental and clinical pharmacology university of catania italy tgf-beta1 pathway as a new target for neuroprotection in alzheimer s disease caraci f battaglia g bruno v bosco p carbonaro v giuffrida ml drago f sortino ma nicoletti f copani a department of pharmaceutical sciences university of catania 95125 catania italy alzheimer s disease ad is a neurodegenerative disorder that affects more than 37 million people worldwide current drugs for ad are only symptomatic but do not interfere with the underlying pathogenic mechanisms of the disease ad is characterized by the presence of ss-amyloid abeta plaques neurofibrillary tangles and neuronal loss the identification of the molecular determinants underlying ad pathogenesis is a fundamental step to design new disease-modifying drugs recently a specific impairment of transforming-growth-factor-beta1 tgf-beta1 signaling pathway has been demonstrated in ad brain the deficiency of tgf-beta1 signaling has been shown to increase both abeta accumulation and abeta-induced neurodegeneration in ad models the loss of function of tgf-ss1 pathway seems also to contribute to tau pathology and neurofibrillary tangle formation growing evidence suggests a neuroprotective role for tgf-beta1 against abeta toxicity both in vitro and in vivo models of ad different drugs such as lithium or group ii mglu receptor agonists are able to increase tgf-beta1 levels in the central nervous system cns and might be considered as new neuroprotective tools against abeta-induced neurodegeneration in the present review we examine the evidence for a neuroprotective role of tgf-beta1 in ad and discuss the tgf-beta1 signaling pathway as a new pharmacological target for the treatment of ad j chemother 2009 dec;216 646-50 calixarenes are synthetic macrocyclic compounds that may serve as scaffolds for biologically active molecules and have been proposed as potential anticancer agents we synthesized a ureido-calix[8]arene carrying n-acetyl-d-glucosamine residue compound 1 and had previously demonstrated that it inhibits c6 glioma cell migration and proliferation with divergent mechanisms in the present work we explored in more detail the antiproliferative effect of compound 1 comparing it to related compounds lacking either the sugar moieties compound 2 the multiple ureido groups compound 3 or both compound 4 the results show that the action of compound 1 is independent of the n-acetyl-d-glucosamine residues requires the presence of multiple ureido groups and does not seem to involve focal adhesion kinase signaling inhibition of proliferation is reduced by preincubation with epidermal growth factor egf and vascular endothelial growth factor 20 ng/ml with compound 1 and extracellular-related kinase phosphorylation is reduced by treatment with compound 1 in both basal and egf-stimulated conditions suggesting that the observed effect depends on a direct interference with growth factor signaling j ocul pharmacol ther 2010 feb;261 31-5 morphine-induced ocular hypotension is modulated by nitric oxide and carbon monoxide role of mu3 receptors stagni e bucolo c motterlini r drago f department of experimental and clinical pharmacology school of medicine university of catania catania italy the impact of prulifloxacin on vaginal lactobacillus microflora an in vivo study tempera g furneri pm cianci a incognito t marano mr drago f department of microbiological and gynaecological sciences university of catania catania italy the aim of this study was to evaluate the in vivo effect of a repeated-dose regimen with prulifloxacin in comparison to amoxicillin/clavulanate on vaginal lactobacillus microflora thirty healthy purpose:recent findings generated from our laboratory have demonstrated the involvement of nitric oxide no in morphineinduced reduction of intraocular pressure iop the present study was designed to investigate the possible involvement of carbon monoxide co in morphine-induced reduction of iop and the role of mu3 opioid receptors methods:new zealand rabbits were used in this study they were pretreated with the nitric oxide synthase inhibitor nomeganitro-l-arginine methyl ester l-name 1 30 microl or an inhibitor of heme oxygenase ho zinc protoporphyrin-ix znpp-ix 0.1 mg/kg i.v the same animals were then treated with morphine 100 microg/30 microl with or without no or co donors administration sodium nitroprusside snp and tricarbonylchloroglycinatorutheniumii corm-3 respectively a separate set of animals were pretreated with the nonselective opioid receptor antagonist naloxone 100 microg/30 microl or the micro3 opioid receptor inhibitor l-glutathione gsh 1 30 microl in the presence of snp or corm-3 followed by morphine administration iop measurements were taken at different times after monolateral instillation of morphine results:morphine induced a significant decrease in iop and pretreatment with znpp-ix or l-name significantly prevented

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this effect whereas administration of no or co donors amplified morphine-induced decrease in iop this effect was partially abrogated both by pretreatment with znpp-ix or l-name and by pretreatment with naloxone and gsh suggesting that the decrease in iop relies on exogenous no and co liberated from snp and corm-3 respectively conclusions:we conclude that the endogenous no/co system and micro3 receptors contribute to morphine-induced ocular hypotension and that the reduction of iop elicited by morphine can be augmented by exogenous no and co neuropeptides 2010 feb;441 45-51 pacap and vip affect nf1 expression in rat malignant peripheral nerve sheath tumor mpnst cells giunta s castorina a adorno a mazzone v carnazza ml d agata v department of anatomy diagnostic pathology legal medicine hygiene and public health university of catania catania italy in our previous study we have identified pacap vip and their receptors in rat malignant peripheral nerve sheath tumor mpnst cells thus showing anti-apoptotic roles recently it has been shown that the tumor suppressor neurofibromin encoded by the neurofibromatosis type i nf1 gene promotes mpnst cells sensitivity to apoptosis after serum withdrawal in the present study we investigated whether pacap or vip negatively regulate nf1 expression under normal or serum-dependent pro-apoptotic culture conditions results indicated that serum itself significantly influenced gene and protein levels in fact the low nf1 levels of cells cultured in normal serum-containing medium were remarkably increased in cells switched to low or no-serum after 24h and 48 h treatment with 100 nm pacap or vip did not affect nf1 expression when using normal amounts of serum whereas it significantly inhibited transcript and protein levels both in low or no-serum cultured cells in particular pacap reduced nf1 levels already after 24h in low-serum cultured cells while vip showed a similar effect only after serum deprivation however both pacap and vip downregulated gene and protein levels within 48 h either in low-dose and serum-starved cells results were confirmed by fluorescence microscopy showing that 100 nm pacap or vip attenuated neurofibromin cytoplasmic localization only in low or no-serum cultured cells the present study provides a comprehensive analysis of both neuropeptides effect on nf1 expression in normal low or serum-starved mpnst cells ameliorating the hypothesis that resistance to apoptosis in serum-deprived cells might be correlated to pacap vip-induced nf1 inhibition trends pharmacol sci 2010 apr;314 153-60 epub 2010 jan 11 dopamine d3 receptors drsd3 seem to have a pivotal role in mood disorders using the elevated plus maze epm and the novelty-induced grooming test ngt we assessed the responses of drd3-deficient d3 mice to the acute treatment different testing time with the anxiolytic drug diazepam d3 mice treated with diazepam 0.1 or 0.5mg/kg exhibited a better behavioral response in the epm than their wild type wt furthermore in d3 mice but not in wt 1mg/kg diazepam induced anxiolytic effects at all testing times the contribution of drsd3 in the anxiolytic effects of diazepam was confirmed by similar results obtained in epm by using the selective drd3 antagonist u99194a 10mg/kg in combination with diazepam in wt animals d3 mice treated with diazepam all doses also showed a decrease in grooming behavior however the 3 h]flunitrazepam autoradiographic analysis revealed no significant changes in d3 mice compared to wt suggesting that if aminobutyric acid receptor gabaa changes are involved they do not occur at the level of binding to benzodiazepine site these data suggest that d3 mice exhibit low baseline anxiety levels and provide the evidence that the drd3 is involved in the modulation of benzodiazepine anxiolytic effects pharmacol res 2010 jun;616 531-6 epub 2010 feb 10 enhanced cognitive performance of dopamine d3 receptor knock-out mice in the step-through passive-avoidance test assessing the role of the endocannabinoid/endovanilloid systems micale v cristino l tamburella a petrosino s leggio gm di marzo v drago f department of experimental and clinical pharmacology university of catania medical school v.le a doria 6 95125 catania italy transcriptional regulation of type-2 metabotropic glutamate receptors an epigenetic path to novel treatments for chronic pain chiechio s copani a zammataro m battaglia g gereau rw 4th nicoletti f department of pharmaceutical sciences university of catania catania italy activation of metabotropic glutamate 2 mglu2 receptors inhibits pain transmission at the synapses between primary afferent fibers and neurons in the dorsal horn of the spinal cord in addition mglu2 receptors are found in peripheral nociceptors and in pain-regulatory centers of the brain stem and forebrain mglu2 receptor agonists produce analgesia in models of inflammatory and neuropathic pain but their use is limited by the development of tolerance a new therapeutic strategy could be based on the transcriptional regulation of mglu2 receptors via the acetylationpromoted activation of the p65/rela transcription factor epigenetic drugs that increase mglu2 receptor expression including l-acetylcarnitine and inhibitors of histone deacetylases have a different analgesic profile with no tolerance to the therapeutic effect after repeated dosing eur neuropsychopharmacol 2011 apr;214 325-32 epub 2010 jun 8 increasing evidence suggests a pivotal role of the d3 receptor d3r in cognitive processes and the involvement of endocannabinoid/endovanilloid signaling in the pathophysiology of neurodegenerative disorders such as alzheimer s disease this study was undertaken to investigate both the basal and beta-amyloid peptide 1-42 bap 1-42 impaired cognitive performance of d3r mice and the role therein of endocannabinoids/endovanilloids d3r mice were either untreated or injected i.c.v with 400 pmol bap 1-42 or vehicle to be tested 14 days later in a step-through passive-avoidance paradigm the cb1 receptor antagonist rimonabant 1mg/kg or the transient receptor potential vanilloid-type 1 channel trpv1 antagonist sb366791 were injected intraperitoneally for 11 or 7 days the retention test was performed 1 7 and 14 days after the learning trial wild-type wt mice were subjected to the same procedures d3r mice exhibited a better basal cognitive performance as compared to wt mice p<0.001 which was reversed by trpv1 antagonism bap 1-42 induced a pronounced worsening of the passive-avoidance response in all tests and in both genotypes p<0.001 rimonabant treatment never affected the cognitive performance of healthy mice but fully counteracted bap 1-42-induced amnesic effects in both d3r and wt mice only when administered for 11 days whereas when administered for 7 days only transiently affected wt mice and caused more prolonged cognitive ameliorations in d3r mice these results support the involvement of d3r and trpv1 in cognitive processes and the concept that a beta peptides inhibit memory retention in mice through the involvement of endocannabinoids drug news perspect 2010 sep;237 430-7 therapeutic potential of nitric oxide modulation in ocular diseases drago f bucolo c department of experimental and clinical pharmacology medical school university of catania catania italy dopamine d3 receptor knock-out mice exhibit increased behavioral sensitivity to the anxiolytic drug diazepam leggio gm micale v le foll b mazzola c nobrega jn drago f department of experimental and clinical pharmacology university of catania v.le a doria 6 95125 catania italy nitric oxide no is an organic gas ubiquitously synthesized in mammalian tissues by no synthase nos over the past 20 years remarkable progress has been made in explaining the mechanism/s of no and its functions in different biological systems no is produced as metabolic endproduct in specific cell life phases and may act as a atypical neuronal messenger no is an important regulator of homeostatic processes in the eye and

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changes in its synthesis could lead to a variety of eye diseases such as glaucoma retinal degeneration and uveitis both overexpression and underexpression of no could contribute to pathological conditions in the eye many works have highlighted the role of no in a wide range of ocular diseases and recent studies from our laboratory and others have shown that a suppressive action of inducible nosderived no production lowers the intraocular pressure indeed from a clinical perspective a precise regulation of no may lead to new therapeutic options likely safer and more efficacious than currently available treatments for various sight-threatening eye diseases eur neuropsychopharmacol 2010 oct;2010 704-13 epub 2010 may 26 the upregulation of nf1 transcripts and protein levels induced by serum withdrawal were remarkably attenuated by 10 6 and 10 5 m agonist treatment within 24 h p<0.01 and p<0.001 respectively whereas similar effects were observed already at a lower concentration 10 7 m after 48 h treatment p<0.001 in conclusion these results suggest that d3r might mediate the protective response to serum deprivation in mpnst cells through the inhibition of nf1 gene expression further underlying a subtle role of these receptors in mpnst development peptides 2010 dec;3112 2276-83 epub 2010 aug 26 the beta3 adrenoceptor agonist amibegron sr58611a counteracts stress-induced behavioral and neurochemical changes tamburella a micale v leggio gm drago f department of experimental and clinical pharmacology university of catania medical school viale a doria 6 95125 catania italy effects of pacap and vip on hyperglycemia-induced proliferation in murine microvascular endothelial cells castorina a giunta s mazzone v cardile v d agata v department of anatomy diagnostic pathology legal medicine hygiene and public health university of catania catania italy these experiments were made to study the mechanisms underlying the antidepressant-like effects of the beta3 adrenoceptor agonist amibegron sr58611a to this purpose the expression levels of the hippocampal cyclic adenosine monophosphate camp response element binding protein creb brain-derived neurotrophic factor bdnf b-cell lymphoma-2 bcl-2 and bax proteins were assessed by using western blot analysis in rats tested in the forced swim test fst under basal conditions no previous exposure to stressors different groups of male wistar rats received acutely or repeatedly once/day for 7days intraperitoneal i.p injections of amibegron 1 5 and 10mg/kg the tricyclic antidepressant tca clomipramine 50mg/kg the selective serotonin reuptake inhibitor ssri citalopram 15mg/kg or their vehicles the influence of stress-related conditions was studied in rats subjected to acute 4h or repeated 4h/day for 7days restraint stress applied prior to the fst procedure compared to the control groups both stressor procedures increased the immobility time in the fst and reduced hippocampal bdnf and bcl-2/bax ratio proteins expression which were counteracted by amibegron 5 and 10mg/kg treatment opposite effects were found in the creb expression since it was lower after acute and higher after repeated stress procedure respectively again these effects were reversed by amibegron treatment different results were obtained in animals treated with clomipramine or citalopram hence it is likely that the observed behavioral effects of amibegron could be due at least in part to its action on hippocampal expression of neurotrophic and/or anti-apoptotic factors supporting the hypothesis that beta3 adrenoceptors may be a therapeutic target for the treatment of stress-related disorders int j oncol 2010 oct;374 927-34 hyperglycemia is implicated both in micro and macro-vascular complications in diabetes mellitus pituitary adenylate cyclase-activating polypeptide pacap and vasoactive intestinal polypeptide vip are two known nonclassic regulators of angiogenesis although their biological role on endothelial cell proliferation remains poorly defined in the present study we hypothesized that either peptides might play an inhibitory role on hyperglycemia-induced cell growth to this end we investigated the effect of both pacap and vip on cell proliferation in murine microvascular endothelial cells h5v cultured both under euglycemic and hyperglycemic conditions 5 and 25 mm glucose respectively for 24 48 h 7 and 15 days results demonstrated that high glucose treatment induced a time-dependent increase in cell viability after 48 h p<0.05 which was much more evident after 7 and 15 days p<0.001 similar effects were observed in cell proliferation although significant changes were obtained after prolonged exposures to high glucose 7 and 15 days p<0.001 the proliferative response to the glucose-enriched environment was correlated to changes in the expression of pac1 and to a minor extent to vpac2 but not vpac1 receptors as measured by quantitative real-time pcr these results were further confirmed by western blot and immunofluorescence analyses interestingly 10?7 m pacap or vip treatment significantly attenuated hyperglycemiainduced increase in cell viability and proliferation after 7 and 15 days taken together our findings demonstrate that both pacap and vip peptides exert an inhibitory activity on hyperglycemia-induced endothelial cell proliferation thus suggesting that the effect might be mediated by pac1 and vpac2 receptors neurochem res 2011 mar;363 426-34 epub 2010 dec 19 protective effect of the dopamine d3 receptor agonist 7-oh-pipat against apoptosis in malignant peripheral nerve sheath tumor mpnst cells neurofibromin and amyloid precursor protein expression in dopamine d3 receptor knock-out mice brains castorina a leggio gm giunta s magro g scapagnini g drago f d agata v department of anatomy diagnostic pathology legal medicine hygiene and public health university of catania via s sofia 87 95123 catania italy castorina a giunta s d agata v department of anatomy diagnostic pathology legal medicine hygiene and public health 95123 catania italy emerging evidence indicates that the dopamine d3 receptor d3 r mediates protective roles both in neuronal and non-neuronal cell lines in a previous study we proposed that neurofibromin a large tumor suppressor protein encoded by the neurofibromatosis type 1 gene nf1 may increase susceptibility to apoptosis after serum deprivation in malignant peripheral nerve sheath tumor mpnst cells thus acting as a proapoptotic gene in addition it has been observed that d3rs are functionally correlated to neurofibromin in this study we examined whether 7-oh-pipat a potent dopamine d3 r agonist exerts an antiapoptotic role under the same culture conditions and then correlated this effect to changes in nf1 expression results showed that serum deprivation caused a significant reduction of cell viability mtt assay both after 24 and 48 h p<0.001 treatment with increasing concentrations of 7-oh-pipat 10 910 5 m induced a progressive increase in cell viability both after 24 and 48 h as compared to vehicle-treated cells with significant changes at the highest concentrations tested 10 6 and 10 5 m consistently at the latter two concentrations a significant reduction in oligonucleosomes formation was observed thus suggesting an antiapoptotic role of 7-oh-pipat these results were confirmed by hoechst 33254 nuclear staining to investigate whether these effects were correlated to changes in nf1 transcript and protein expression quantitative real-time pcr western blot and immunofluorescence analyses were performed results demonstrated that recently it has been proposed that neurofibromin nf1 forms a binding complex with amyloid precursor protein app that interacts with the dopamine d3 receptor d3r in the present study we investigated whether the absence of the d3r is correlated to modifications in the expression of both nf1 and app quantitative real-time pcr analyses of both transcripts showed that nf1 mrna levels were significantly reduced whereas app levels were strikingly increased in d3r knock-out d3r ko as compared to wild type wt mice brains western blot analyses using mice whole brains produced comparable results with those obtained by mrna measurements moreover immunohistochemical analyses revealed a similar brain regional distribution of app protein in the hippocampus in the cerebral and cerebellar cortex of d3r ko mice conversely hippocampal nf1 immunoreactivity did not seem to be affected by the absence of d3rs further analyses confirmed that regional nf1 protein expression in the hippocampus was not affected by the absence of the d3r whereas app levels were still increased in this specific brain region in conclusion these results show the existence of a correlation among the d3r nf1 and app in mice brains and thus show the regional-specific regulation of nf1 in brains of d3r ko which may contribute to gain insights into the comprehension of novel underlying mechanisms that regulate brain function.

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