37 results on '"Pannaccione, Anna"'
Search Results
2. Additional file 4 of Na+/Ca2+ exchanger isoform 1 takes part to the Ca2+-related prosurvival pathway of SOD1 in primary motor neurons exposed to beta-methylamino-l-alanine
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Petrozziello, Tiziana, Boscia, Francesca, Tedeschi, Valentina, Pannaccione, Anna, de Rosa, Valeria, Corvino, Angela, Severino, Beatrice, Annunziato, Lucio, and Secondo, Agnese
- Abstract
Additional file 3. Bar graph depicting the effect of L-BMAA (0.01-1 mM) on cell survival of differentiated NSC-34 cells. Data are expressed as mean��S.E. of three different experimental sessions. *p
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- 2022
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3. Additional file 2 of Na+/Ca2+ exchanger isoform 1 takes part to the Ca2+-related prosurvival pathway of SOD1 in primary motor neurons exposed to beta-methylamino-l-alanine
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Petrozziello, Tiziana, Boscia, Francesca, Tedeschi, Valentina, Pannaccione, Anna, de Rosa, Valeria, Corvino, Angela, Severino, Beatrice, Annunziato, Lucio, and Secondo, Agnese
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Additional file 1. (A) Immunolocalization of NCX1 (a,d) and MAP2 (b,e) within a motor-neuron enriched culture under control conditions. Nuclei were stained with nuclear DNA stain 4, 6-diamino-2-phenylinndole (DAPI). Arrows indicate MAP2-positive cells with higher level of NCX1 expression. (B) Immunolocalization of NCX1 and NCX3 in differentiated NSC-34 cells. (C) Quantification of SOD1-induced [Ca2+]i in presence of CNQX (20 ��M), MK801 (10 ��M), or CB-DMB (1 ��M) in motor neurons expressed as ���% of increase. All the experiments were repeated at least three times on at least 35 cells for each group; *p < 0.001 vs control (basal values of [Ca2+]i) .
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- 2022
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4. 7-nitro-5-phenyl-1-(pyrrolidin-1-ylmethyl)-1H-benzo[E][1,4]diazepin-2(3H)-one and other benzodiazepine derivatives
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PIGNATARO, GIUSEPPE, ANNUNZIATO, LUCIO, MOLINARO, PASQUALE, SCORZIELLO, ANTONELLA, SECONDO, AGNESE, PANNACCIONE, ANNA, CUOMO, ORNELLA, CANTILE, MARIA, DI RENZO, GIANFRANCO MARIA LUIGI, CALIENDO, GIUSEPPE, SANTAGADA, VINCENZO, SEVERINO, BEATRICE, FIORINO, FERDINANDO, Pignataro, Giuseppe, Annunziato, Lucio, Molinaro, Pasquale, Scorziello, Antonella, Secondo, Agnese, Pannaccione, Anna, Cuomo, Ornella, Cantile, Maria, DI RENZO, GIANFRANCO MARIA LUIGI, Caliendo, Giuseppe, Santagada, Vincenzo, Severino, Beatrice, and Fiorino, Ferdinando
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sodium and calcium homeostasis ,Benzodiazepine ,ischemia - Abstract
The present invention describes compounds of formula (I), and in particular the 7- nitro-5-phenyl-1 -(pyrrolidin-1 - ylmethyl)-1 H-benzo[e][1,4]diazepin-2(3H)-one compound and their pharmaceuticall acceptable salts. The compounds of the invention are useful for medical use, particularly as active ingredient potentially useful for the treatment of cerebral ischemia and other disease characterized by deregulation of sodium and calcium homeostasis
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- 2012
5. 7-nitro-5-fenil-1(pirrolidin-1-ilmetil)-1H-benzo[e][1,4] diazepin-2(3H)-one ed altri composti derivati delle benzodiazepine
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DI RENZO, GIANFRANCO MARIA LUIGI, ANNUNZIATO, LUCIO, PIGNATARO, GIUSEPPE, SCORZIELLO, ANTONELLA, MOLINARO, PASQUALE, CUOMO, ORNELLA, SECONDO, AGNESE, PANNACCIONE, ANNA, CANTILE, MARIA, CALIENDO, GIUSEPPE, SANTAGADA, VINCENZO, SEVERINO, BEATRICE, FIORINO, FERDINANDO, DI RENZO, GIANFRANCO MARIA LUIGI, Annunziato, Lucio, Pignataro, Giuseppe, Scorziello, Antonella, Molinaro, Pasquale, Cuomo, Ornella, Secondo, Agnese, Pannaccione, Anna, Cantile, Maria, Caliendo, Giuseppe, Santagada, Vincenzo, Severino, Beatrice, and Fiorino, Ferdinando
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Benzodiazepine ,ischemia cerebrale ,degenerazione neuronale - Abstract
Il brevetto riguarda composti di formula (I), ed in particolare 7-nitro-5-fenil-1(pirrolidin-1-ilmetil)-1H-benzo[e][1,4] diazepin-2(3H)-one, e loro sali farmaceuticamente accettabili, per l’uso medico ed in particolare come principio farmacologicamente attivo per il trattamento dell’ischemia cerebrale e di ogni altra patologia caratterizzata dalla perdita del controllo dell’omeostasi degli ioni sodio e calcio.
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- 2010
6. Cardiovascular side-effects of second-generation H1 receptor antagonists
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TAGLIALATELA, MAURIZIO, GENOVESE, ARTURO, MARONE, GIANNI, ANNUNZIATO, LUCIO, PANNACCIONE, ANNA, Castaldo P., Giorgio G., Taglialatela, Maurizio, Pannaccione, Anna, Castaldo, P., Giorgio, G., Genovese, Arturo, Marone, Gianni, and Annunziato, Lucio
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- 1999
7. Modulation of the K+ channels encoded by the Human ether-a-gogo-related gene 1 (hERG1) by nitric oxide
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TAGLIALATELA M., PANNACCIONE, ANNA, CASTALDO P, IOSSA S., L. ANNUNZIATO, ANNUNZIATO, LUCIO, Taglialatela, M., Pannaccione, Anna, Castaldo, P, Iossa, S., Annunziato, Lucio, and L., Annunziato
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- 1999
8. Cardiac ion channels and antihistamines: possible mechanisms of cardiotoxicity
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TAGLIALATELA M, CASTALDO P, PANNACCIONE, ANNA, GIORGIO G, ANNUNZIATO L., TAGLIALATELA, MAURIZIO, GENOVESE, ARTURO, MARONE, GIANNI, Taglialatela, M, Castaldo, P, Pannaccione, Anna, Giorgio, G, Genovese, Arturo, Marone, Gianni, Annunziato, L., and Taglialatela, Maurizio
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- 1999
9. Human ether-a-gogo related gene (HERG) K+ channels as pharmacological targets: present and future implications
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TAGLIALATELA, MAURIZIO, CASTALDO, PASQUALINA, PANNACCIONE, ANNA, ANNUNZIATO, LUCIO, Giorgio G, Taglialatela, M, Castaldo, P, Pannaccione, Anna, Giorgio, G, Annunziato, L., Taglialatela, Maurizio, Castaldo, Pasqualina, and Annunziato, Lucio
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Central Nervous System ,ERG1 Potassium Channel ,Potassium Channels ,oxygen free radical ,Myocardium ,Heart ,second-generation antihistamine ,Ether-A-Go-Go Potassium Channels ,DNA-Binding Proteins ,antiarrhythmic ,Oxidative Stress ,human arrhythmia ,Transcriptional Regulator ERG ,Potassium Channels, Voltage-Gated ,long QT syndrome ,Histamine H1 Antagonists ,Trans-Activators ,human ether-a-gogo related gene (HERG) K+ channels ,Animals ,Humans ,Reactive Oxygen Species ,Anti-Arrhythmia Agents ,Cation Transport Proteins - Abstract
Electrophysiological and molecular biology techniques have widely expanded our knowledge of the diverse functions where K+ channels are implicated as potential and proven pharmacological targets. The aim of the present commentary is to review the recent progress in the understanding of the functional role of the K+ channels encoded by the human ether-a-gogo related gene (HERG), with particular emphasis on their direct pharmacological modulation by drugs, or on their regulation by pharmacologically relevant phenomena. About 3 years have passed since the cloning, expression, and description of the pathophysiological role of HERG K+ channels in human cardiac repolarization. Despite this short lapse of time, these K+ channels have already gained considerable attention as pharmacological targets. In fact, interference with HERG K+ channels seems to be the main mechanism explaining both the therapeutic actions of the class III antiarrhythmics and the potential cardiotoxicity of second-generation H1 receptor antagonists such as terfenadine and astemizole, as well as of psychotropic drugs such as some antidepressants and neuroleptics. It seems possible to anticipate that the main tasks for future investigation will be, on the one side, the better understanding of the intimate mechanism of action of HERG K+ channel-blocking drugs in order to elucidate the conditions regulating the delicate balance between antiarrhythmic and proarrhythmic potential and, on the other, to unravel the pathophysiological role of this K+ channel in the function of the brain and of other excitable tissues.
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- 1998
10. Regulation of the human ether-a-gogo-related gene (hERG)K+ channels by reactive oxygen species
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TAGLIALATELA M., CASTALDO P., IOSSA S., PANNACCIONE, ANNA, ANNUNZIATO L., ANNUNZIATO, LUCIO, Taglialatela, M., Castaldo, P., Iossa, S., Pannaccione, Anna, Annunziato, Lucio, and Annunziato, L.
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- 1997
11. Size-Based Effects of Anthropogenic Ultrafine Particles on Lysosomal TRPML1 Channel and Autophagy in Motoneuron-like Cells
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Silvia Sapienza, Valentina Tedeschi, Barbara Apicella, Francesco Palestra, Carmela Russo, Ilaria Piccialli, Anna Pannaccione, Stefania Loffredo, Agnese Secondo, Sapienza, Silvia, Tedeschi, Valentina, Apicella, Barbara, Palestra, Francesco, Russo, Carmela, Piccialli, Ilaria, Pannaccione, Anna, Loffredo, Stefania, and Secondo, Agnese
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Motor Neurons ,Organic Chemistry ,air pollution ,ROS ,General Medicine ,ultrafine particle ,Motor Neuron ,Lysosome ,Catalysis ,Antioxidants ,Computer Science Applications ,Inorganic Chemistry ,ultrafine particles ,TRPML1 ,autophagy ,motor neurons ,neurotoxicity ,lysosome ,mitochondrial dysfunction ,Transient Receptor Potential Channels ,Autophagy ,Particulate Matter ,Physical and Theoretical Chemistry ,Antioxidant ,Lysosomes ,Molecular Biology ,Spectroscopy - Abstract
Background: An emerging body of evidence indicates an association between anthropogenic particulate matter (PM) and neurodegeneration. Although the historical focus of PM toxicity has been on the cardiopulmonary system, ultrafine PM particles can also exert detrimental effects in the brain. However, only a few studies are available on the harmful interaction between PM and CNS and on the putative pathomechanisms. Methods: Ultrafine PM particles with a diameter < 0.1 μm (PM0.1) and nanoparticles < 20 nm (NP20) were sampled in a lab-scale combustion system. Their effect on cell tracking in the space was studied by time-lapse and high-content microscopy in NSC-34 motor neurons while pHrodo™ Green conjugates were used to detect PM endocytosis. Western blotting analysis was used to quantify protein expression of lysosomal channels (i.e., TRPML1 and TPC2) and autophagy markers. Current-clamp electrophysiology and Fura2-video imaging techniques were used to measure membrane potential, intracellular Ca2+ homeostasis and TRPML1 activity in NSC-34 cells exposed to PM0.1 and NP20. Results: NP20, but not PM0.1, reduced NSC-34 motor neuron movement in the space. Furthermore, NP20 was able to shift membrane potential of motor neurons toward more depolarizing values. PM0.1 and NP20 were able to enter into the cells by endocytosis and exerted mitochondrial toxicity with the consequent stimulation of ROS production. This latter event was sufficient to determine the hyperactivation of the lysosomal channel TRPML1. Consequently, both LC3-II and p62 protein expression increased after 48 h of exposure together with AMPK activation, suggesting an engulfment of autophagy. The antioxidant molecule Trolox restored TRPML1 function and autophagy. Conclusions: Restoring TRPML1 function by an antioxidant agent may be considered a protective mechanism able to reestablish autophagy flux in motor neurons exposed to nanoparticles.
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- 2022
12. Genetic Up-Regulation or Pharmacological Activation of the Na+/Ca2+ Exchanger 1 (NCX1) Enhances Hippocampal-Dependent Contextual and Spatial Learning and Memory
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Angelo Serani, Agnese Secondo, Roselia Ciccone, Ferdinando Fiorino, Gianfranco Di Renzo, Anna Pannaccione, Beatrice Severino, Lucia D’Esposito, Lucrezia Calabrese, Serenella Anzilotti, Francesco Frecentese, Silvia Natale, Tiziana Petrozziello, A.G. Sadile, Antonio Vinciguerra, Pasquale Molinaro, Simona Cabib, Ornella Cuomo, Lucio Annunziato, Natale, S., Anzilotti, S., Petrozziello, T., Ciccone, R., Serani, A., Calabrese, L., Severino, B., Frecentese, F., Secondo, A., Pannaccione, Anna, Fiorino, F., Cuomo, O., Vinciguerra, A., D'Esposito, L., Sadile, A. G., Cabib, S., Di Renzo, G., Annunziato, L., and Molinaro, P.
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0301 basic medicine ,Neuroscience (miscellaneous) ,Hippocampus ,Anxiety ,Hippocampal formation ,Amygdala ,Long-term memory ,Synaptic plasticity ,03 medical and health sciences ,Cellular and Molecular Neuroscience ,0302 clinical medicine ,medicine ,Fear conditioning ,anxiety ,drug discovery ,long-term memory ,synaptic plasticity ,Drug discovery ,Chemistry ,Barnes maze ,030104 developmental biology ,medicine.anatomical_structure ,Neurology ,cardiovascular system ,Memory consolidation ,Neuroscience ,030217 neurology & neurosurgery - Abstract
The Na+/Ca2+ exchanger 1 (NCX1) participates in the maintenance of neuronal Na+ and Ca2+ homeostasis, and it is highly expressed at synapse level of some brain areas involved in learning and memory processes, including the hippocampus, cortex, and amygdala. Furthermore, NCX1 increases Akt1 phosphorylation and enhances glutamate-mediated Ca2+ influx during depolarization in hippocampal and cortical neurons, two processes involved in learning and memory mechanisms. We investigated whether the modulation of NCX1 expression/activity might influence learning and memory processes. To this aim, we used a knock-in mouse overexpressing NCX1 in hippocampal, cortical, and amygdala neurons (ncx1.4over) and a newly synthesized selective NCX1 stimulating compound, named CN-PYB2. Both ncx1.4over and CN-PYB2-treated mice showed an amelioration in spatial learning performance in Barnes maze task, and in context-dependent memory consolidation after trace fear conditioning. On the other hand, these mice showed no improvement in novel object recognition task which is mainly dependent on non-spatial memory and displayed an increase in the active phosphorylated CaMKIIα levels in the hippocampus. Interestingly, both of these mice showed an increased level of context-dependent anxiety. Altogether, these results demonstrate that neuronal NCX1 participates in spatial-dependent hippocampal learning and memory processes.
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- 2020
13. ORAI1/STIM1 Interaction Intervenes in Stroke and in Neuroprotection Induced by Ischemic Preconditioning Through Store-Operated Calcium Entry
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Roselia Ciccone, Giuseppe Pignataro, Antonio Vinciguerra, Anna Pannaccione, Tiziana Petrozziello, Agnese Secondo, Valentina Tedeschi, Lucio Annunziato, Pasquale Molinaro, Francesca Boscia, Secondo, A., Petrozziello, T., Tedeschi, V., Boscia, F., Vinciguerra, A., Ciccone, R., Pannaccione, Anna, Molinaro, P., Pignataro, G., and Annunziato, L.
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ORAI1 Protein ,Glucose-regulated protein ,calcium homeostasi ,Neuroprotection ,Brain ischemia ,03 medical and health sciences ,0302 clinical medicine ,Animals ,Medicine ,rat ,Stromal Interaction Molecule 1 ,Rats, Wistar ,Cells, Cultured ,030304 developmental biology ,Cerebral Cortex ,Advanced and Specialized Nursing ,0303 health sciences ,biology ,ORAI1 ,business.industry ,Calcium channel ,Calcium-Binding Proteins ,Membrane Proteins ,STIM1 ,primary cortical neuron ,medicine.disease ,stroke ,Store-operated calcium entry ,Rats ,Cell biology ,ischemic preconditioning ,biology.protein ,Ischemic preconditioning ,Neurology (clinical) ,Cardiology and Cardiovascular Medicine ,business ,030217 neurology & neurosurgery - Abstract
Background and Purpose— Disturbance of endoplasmic reticulum (ER) Ca 2+ homeostasis causes neuronal cell injury in stroke. By contrast, ischemic preconditioning (IPC)—a brief sublethal ischemic episode affording tolerance to a subsequent ischemic insult—restores ER Ca 2+ homeostasis. Under physiological conditions, ER calcium content is continuously refilled by the interaction between the ER-located Ca 2+ sensor STIM (stromal interacting molecule) 1 and the plasma membrane channel ORAI1 (a structural component of the CRAC calcium channel)—2 key mediators of the store-operated calcium entry (SOCE) mechanism. However, the role played by ORAI1 and STIM1 in stroke and in IPC-induced neuroprotection during stroke remains unknown. Therefore, we explored whether ORAI1 and STIM1 might be involved in stroke pathogenesis and in IPC-induced neuroprotection. Methods— Primary cortical neurons were subjected to oxygen and glucose deprivation+reoxygenation to reproduce in vitro brain ischemia. Focal brain ischemia and IPC were induced in rats by transient middle cerebral artery occlusion. Expression of ORAI1 and STIM1 transcripts and proteins and their immunosignals were detected by qRT-PCR, Western blot, and immunocytochemistry, respectively. SOCE and Ca 2+ release–activated Ca 2+ currents (I CRAC ) were measured by Fura-2 AM video imaging and patch-clamp electrophysiology in whole-cell configuration, respectively. Results— STIM1 and ORAI1 protein expression and immunosignals decreased in the ipsilesional temporoparietal cortex of rats subjected to transient middle cerebral artery occlusion followed by reperfusion. Analogously, in primary hypoxic cortical neurons, STIM1 and ORAI1 transcript and protein levels decreased concurrently with SOCE and Ca 2+ release–activated Ca 2+ currents. By contrast, IPC induced SOCE and Ca 2+ release–activated Ca 2+ current upregulation, thereby preventing STIM1 and ORAI1 downregulation induced by oxygen and glucose deprivation+reoxygenation. Silencing of STIM1 or ORAI1 prevented IPC-induced tolerance and caused ER stress, as measured by GRP78 (78-kDa glucose regulated protein) and caspase-3 upregulation. Conclusions— ORAI1 and STIM1, which participate in SOCE, take part in stroke pathophysiology and play an important role in IPC-induced neuroprotection.
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- 2019
14. The expression and activity of K V 3.4 channel subunits are precociously upregulated in astrocytes exposed to Aβ oligomers and in astrocytes of Alzheimer's disease Tg2576 mice
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Francesca Boscia, Antonella Casamassa, Valeria de Rosa, Cristina Franco, Roselia Ciccone, Antonio Vinciguerra, Ilaria Piccialli, Lucio Annunziato, Gianfranco Di Renzo, Anna Pannaccione, Boscia, Francesca, Pannaccione, Anna, Ciccone, Roselia, Casamassa, Antonella, Franco, Cristina, Piccialli, Ilaria, DE ROSA, Valeria, Vinciguerra, Antonio, DI RENZO, GIANFRANCO MARIA LUIGI, and Annunziato, Lucio
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0301 basic medicine ,Aging ,Protein subunit ,Biology ,Hippocampal formation ,Tg2576 ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Aβ-oligomer ,Gene knockdown ,Glial fibrillary acidic protein ,GFAP ,General Neuroscience ,Voltage-gated potassium channel ,Alzheimer's disease ,Potassium channel ,Aβ(1-42) ,Cell biology ,030104 developmental biology ,medicine.anatomical_structure ,Cerebral cortex ,biology.protein ,K(V)3.4 potassium channel ,Neurology (clinical) ,Geriatrics and Gerontology ,Astrocyte ,Neuroscience ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Astrocyte dysfunction emerges early in Alzheimer's disease (AD) and may contribute to its pathology and progression. Recently, the voltage gated potassium channel KV3.4 subunit, which underlies the fast-inactivating K(+) currents, has been recognized to be relevant for AD pathogenesis and is emerging as a new target candidate for AD. In the present study, we investigated both in in vitro and in vivo models of AD the expression and functional activity of KV3.4 potassium channel subunits in astrocytes. In primary astrocytes our biochemical, immunohistochemical, and electrophysiological studies demonstrated a time-dependent upregulation of KV3.4 expression and functional activity after exposure to amyloid-β (Aβ) oligomers. Consistently, astrocytic KV3.4 expression was upregulated in the cerebral cortex, hippocampus, and cerebellum of 6-month-old Tg2576 mice. Further, confocal triple labeling studies revealed that in 6-month-old Tg2576 mice, KV3.4 was intensely coexpressed with Aβ in nonplaque associated astrocytes. Interestingly, in the cortical and hippocampal regions of 12-month-old Tg2576 mice, plaque-associated astrocytes much more intensely expressed KV3.4 subunits, but not Aβ. More important, we evidenced that the selective knockdown of KV3.4 expression significantly downregulated both glial fibrillary acidic protein levels and Aβ trimers in the brain of 6-month-old Tg2576 mice. Collectively, our results demonstrate that the expression and function of KV3.4 channel subunits are precociously upregulated in cultured astrocytes exposed to Aβ oligomers and in reactive astrocytes of AD Tg2576 mice.
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- 2017
15. Lavender and coriander essential oils and their main component linalool exert a protective effect against amyloid-β neurotoxicity
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Lucia Caputo, Antonio Massa, Vincenzo De Feo, Roselia Ciccone, Ilaria Piccialli, Anna Pannaccione, Paolo De Caprariis, Caputo, L., Piccialli, I., Ciccone, R., de Caprariis, P., Massa, A., De Feo, V., and Pannaccione, Anna
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Amyloid β ,linalool ,Lavender ,coriander essential oil ,Acyclic Monoterpenes ,amyloid-β oligomer ,Coriandrum ,Pharmacology ,Neuroprotection ,PC12 Cells ,03 medical and health sciences ,chemistry.chemical_compound ,0302 clinical medicine ,Linalool ,In vivo ,Alzheimer Disease ,lavender essential oil ,medicine ,Oils, Volatile ,Animals ,Plant Oils ,Cognitive Dysfunction ,Cognitive decline ,0303 health sciences ,Amyloid beta-Peptides ,030302 biochemistry & molecular biology ,Neurodegeneration ,Neurotoxicity ,Alzheimer's disease ,medicine.disease ,amyloid-β oligomers ,Peptide Fragments ,Rats ,Lavandula ,Neuroprotective Agents ,chemistry ,030220 oncology & carcinogenesis ,neuroprotection ,Cognition Disorders ,Reactive Oxygen Species - Abstract
Alzheimer's disease (AD) is a neurodegenerative disorder leading to cognitive deficits and cognitive decline. Since no cure or preventing therapy is currently available to counteract AD, natural-derived compounds are investigated to find new potential neuroprotective agents for its treatment. In the present study, we tested the neuroprotective effect of lavender and coriander essential oils (EOs) and their main active constituent linalool, against the neurotoxicity elicited by Aβ1-42 oligomers, a key molecular factor in the neurodegeneration of AD. Importantly, our findings on neuronally differentiated PC12 cells exposed to Aβ1-42 oligomers are in accordance with previous in vivo studies reporting the neuroprotective potential of lavender and coriander EOs and linalool. We found that lavender and coriander EOs at the concentration of 10 μg/mL as well as linalool at the same concentration were able to improve viability and to reduce nuclear morphological abnormalities in cells treated with Aβ1-42 oligomers for 24 hours. Lavender and coriander EOs and linalool also showed to counteract the increase of intracellular reactive oxygen species production and the activation of the pro-apoptotic enzyme caspase-3 induced by Aβ1-42 oligomers. Our findings provide further evidence that these EOs and their main constituent linalool could be natural agents of therapeutic interest against Aβ1-42 -induced neurotoxicity.
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- 2019
16. Synthesis and Pharmacological Evaluation of a Novel Peptide Based on Anemonia sulcata BDS-I Toxin as a New KV3.4 Inhibitor Exerting a Neuroprotective Effect Against Amyloid-β Peptide
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Anna Pannaccione, Ilaria Piccialli, Paolo Grieco, Lucio Annunziato, Francesco Merlino, Roselia Ciccone, Ciccone, Roselia, Piccialli, Ilaria, Grieco, Paolo, Merlino, Francesco, Annunziato, Lucio, and Pannaccione, Anna
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Programmed cell death ,Amyloid beta ,Peptide ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,lcsh:Chemistry ,chemistry.chemical_compound ,BDS-I ,Aβ peptide ,Peptide synthesis ,Patch clamp ,chemistry.chemical_classification ,voltage gated potassium channel ,biology ,Chinese hamster ovary cell ,General Chemistry ,Voltage-gated potassium channel ,Alzheimer's disease ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,Cell biology ,lcsh:QD1-999 ,KV3.4 ,chemistry ,biology.protein ,0210 nano-technology ,Marine toxin - Abstract
There is increasing evidence that the fast-inactivating potassium current IA, encoded by KV3. 4 channels, plays an important role in Alzheimer's Disease (AD), since the neurotoxic β-amyloid peptide1-42 (Aβ1−42) increases the IA current triggering apoptotic processes. The specific inhibition of KV3.4 by the marine toxin extracted from Anemonia sulcata, named blood depressing substance-I (BDS-I), reverts the Aβ peptide-induced cell death. The aim of the present study was to identify the smallest fragments of BDS-I, obtained by peptide synthesis, able to inhibit KV3.4 currents. For this purpose, whole-cell patch clamp technique was used to evaluate the effects of BDS-I fragments on KV3.4 currents in CHO cells heterologously expressing KV3.4. We found that BDS-I[1-8] fragment, containing the N-terminal octapeptide sequence of full length BDS-I, was able to inhibit KV3.4 currents in a concentration dependent manner, whereas the scrambled sequence of BDS-I[1-8] and all the other fragments obtained from BDS-I full length were ineffective. As we demonstrated in a previous study, BDS-I full length is able to counteract Aβ1−42-induced enhancement of KV3.4 activity, preventing Aβ1−42-induced caspase-3 activation and the abnormal nuclear morphology in NGF-differentiated PC-12 cells. Similarly to BDS-I, we found that BDS-I[1-8] blocking KV3.4 currents prevented Aβ1−42-induced caspase-3 activation and apoptotic processes. Collectively, these results suggest that BDS-I[1-8] could represent a lead compound to be developed as a new drug targeting KV3.4 channels.
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- 2019
17. The Na+/Ca2+exchanger in Alzheimer’s disease
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Roselia Ciccone, Agnese Secondo, Ilaria Piccialli, Anna Pannaccione, Pasquale Molinaro, Francesca Boscia, Lucio Annunziato, Pannaccione, Anna, Piccialli, I., Secondo, A., Ciccone, R., Molinaro, P., Boscia, F., and Annunziato, L.
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0301 basic medicine ,Gene isoform ,Physiology ,2+ ,Hippocampal formation ,+ ,exchanger ,Neuroprotection ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Ionic deregulation ,Na ,Molecular Biology ,Ca ,Chemistry ,Endoplasmic reticulum ,Neurodegeneration ,Cell Biology ,Alzheimer's disease ,medicine.disease ,NCX3 ,030104 developmental biology ,Apoptosis ,Neuroscience ,030217 neurology & neurosurgery ,Homeostasis ,Intracellular - Abstract
As a pivotal player in regulating sodium (Na+) and calcium (Ca2+) homeostasis and signalling in excitable cells, the Na+/Ca2+ exchanger (NCX) is involved in many neurodegenerative disorders in which an imbalance of intracellular Ca2+ and/or Na+ concentrations occurs, including Alzheimer's disease (AD). Although NCX has been mainly implicated in neuroprotective mechanisms counteracting Ca2+ dysregulation, several studies highlighted its role in the neuronal responses to intracellular Na+ elevation occurring in several pathophysiological conditions. Since the alteration of Na+ and Ca2+ homeostasis significantly contributes to synaptic dysfunction and neuronal loss in AD, it is of crucial importance to analyze the contribution of NCX isoforms in the homeostatic responses at neuronal and synaptic levels. Some studies found that an increase of NCX activity in brains of AD patients was correlated with neuronal survival, while other research groups found that protein levels of two NCX subtypes, NCX2 and NCX3, were modulated in parietal cortex of late stage AD brains. In particular, NCX2 positive synaptic terminals were increased in AD cohort while the number of NCX3 positive terminals were reduced. In addition, NCX1, NCX2 and NCX3 isoforms were up-regulated in those synaptic terminals accumulating amyloid-beta (Aβ), the neurotoxic peptide responsible for AD neurodegeneration. More recently, the hyperfunction of a specific NCX subtype, NCX3, has been shown to delay endoplasmic reticulum stress and apoptotic neuronal death in hippocampal neurons exposed to Aβ insult. Despite some issues about the functional role of NCX in synaptic failure and neuronal loss require further studies, these findings highlight the putative neuroprotective role of NCX in AD and open new strategies to develop new druggable targets for AD therapy.
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- 2020
18. Nuclear localization of NCX: Role in Ca2+ handling and pathophysiological implications
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Anna Pannaccione, Tiziana Petrozziello, Francesca Boscia, Agnese Secondo, Valentina Tedeschi, Lucio Annunziato, Pasquale Molinaro, Secondo, A., Petrozziello, T., Tedeschi, V., Boscia, F., Pannaccione, Anna, Molinaro, P., and Annunziato, L.
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0301 basic medicine ,Physiology ,2+ ,Nuclear Na ,+ ,exchanger ,03 medical and health sciences ,0302 clinical medicine ,Inner membrane ,Nuclear pore ,Nuclear Ca ,Molecular Biology ,Ion channel ,Ca ,Nucleoplasm ,Chemistry ,Endoplasmic reticulum ,homeostasi ,Cell Biology ,Cytosol ,030104 developmental biology ,Neuronal differentiation ,Cytoplasm ,Biophysics ,Akt pathway ,030217 neurology & neurosurgery ,Nuclear localization sequence - Abstract
Numerous lines of evidence indicate that nuclear calcium concentration ([Ca2+]n) may be controlled independently from cytosolic events by a local machinery. In particular, the perinuclear space between the inner nuclear membrane (INM) and the outer nuclear membrane (ONM) of the nuclear envelope (NE) likely serves as an intracellular store for Ca2+ ions. Since ONM is contiguous with the endoplasmic reticulum (ER), the perinuclear space is adjacent to the lumen of ER thus allowing a direct exchange of ions and factors between the two organelles. Moreover, INM and ONM are fused at the nuclear pore complex (NPC), which provides the only direct passageway between the nucleoplasm and cytoplasm. However, due to the presence of ion channels, exchangers and transporters, it has been generally accepted that nuclear ion fluxes may occur across ONM and INM. Within the INM, the Na+/Ca2+ exchanger (NCX) isoform 1 seems to play an important role in handling Ca2+ through the different nuclear compartments. Particularly, nuclear NCX preferentially allows local Ca2+ flowing from nucleoplasm into NE lumen thanks to the Na+ gradient created by the juxtaposed Na+/K+-ATPase. Such transfer reduces abnormal elevation of [Ca2+]n within the nucleoplasm thus modulating specific transductional pathways and providing a protective mechanism against cell death. Despite very few studies on this issue, here we discuss those making major contribution to the field, also addressing the pathophysiological implication of nuclear NCX malfunction.
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- 2020
19. Synthesis and Biological Evaluation of a New Structural Simplified Analogue of cADPR, a Calcium-Mobilizing Secondary Messenger Firstly Isolated from Sea Urchin Eggs
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Bruno Catalanotti, Nicola Borbone, Ilaria Piccialli, Giorgia Oliviero, Tiziana Petrozziello, Luciano Mayol, Agnese Secondo, Anna Pannaccione, Gennaro Piccialli, Stefano D'Errico, Valeria Costantino, D'Errico, Stefano, Borbone, Nicola, Catalanotti, Bruno, Secondo, Agnese, Petrozziello, Tiziana, Piccialli, Ilaria, Pannaccione, Anna, Costantino, Valeria, Mayol, Luciano, Piccialli, Gennaro, and Oliviero, Giorgia
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cyclization ,PC12 neuronal cells ,Stereochemistry ,Pharmaceutical Science ,chemistry.chemical_element ,Calcium ,010402 general chemistry ,01 natural sciences ,Pyrophosphate ,PC12 Cells ,Article ,chemistry.chemical_compound ,Structure-Activity Relationship ,biology.animal ,Cell Line, Tumor ,Drug Discovery ,Ribose ,Moiety ,Animals ,Nucleotide ,Pharmacology, Toxicology and Pharmaceutics (miscellaneous) ,Sea urchin ,lcsh:QH301-705.5 ,Ovum ,chemistry.chemical_classification ,Neurons ,cADPR ,Cyclic ADP-Ribose ,biology ,010405 organic chemistry ,PC12 neuronal cell ,nucleotides ,0104 chemical sciences ,Rats ,chemistry ,lcsh:Biology (General) ,calcium mobilization ,Sea Urchins ,Second messenger system ,macrocycle conformational sampling ,Intracellular ,Signal Transduction - Abstract
Herein, we reported on the synthesis of cpIPP, which is a new structurally-reduced analogue of cyclic ADP-ribose (cADPR), a potent Ca2+-releasing secondary messenger that was firstly isolated from sea urchin eggs extracts. To obtain cpIPP the “northern” ribose of cADPR was replaced by a pentyl chain and the pyrophosphate moiety by a phophono-phosphate anhydride. The effect of the presence of the new phosphono-phosphate bridge on the intracellular Ca2+ release induced by cpIPP was assessed in PC12 neuronal cells in comparison with the effect of the pyrophosphate bridge of the structurally related cyclic N1-butylinosine diphosphate analogue (cbIDP), which was previously synthesized in our laboratories, and with that of the linear precursor of cpIPP, which, unexpectedly, revealed to be the only one provided with Ca2+ release properties.
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- 2018
20. Pharmacological Characterization of the Newly Synthesized 5-Amino-N-butyl-2-(4-ethoxyphenoxy)-benzamide Hydrochloride (BED) as a Potent NCX3 Inhibitor That Worsens Anoxic Injury in Cortical Neurons, Organotypic Hippocampal Cultures, and Ischemic Brain
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Giuseppe Caliendo, Maria Cantile, Lucio Annunziato, Serenella Anzilotti, Giuseppe Pignataro, Gianfranco Di Renzo, Agnese Secondo, Maria Josè Sisalli, Ornella Cuomo, Natascia Guida, Alba Esposito, Francesca Boscia, Antonella Scorziello, Anna Pannaccione, Beatrice Severino, Vincenzo Santagada, Pasquale Molinaro, Paolo Ambrosino, Ferdinando Fiorino, Secondo, Agnese, Pignataro, Giuseppe, Ambrosino, Paolo, Pannaccione, Anna, Molinaro, Pasquale, Boscia, Francesca, Cantile, Maria, Cuomo, Ornella, Esposito, Alba, Sisalli, MARIA JOSE', Scorziello, Antonella, Guida, Natascia, Anzilotti, Serenella, Fiorino, Ferdinando, Severino, Beatrice, Santagada, Vincenzo, Caliendo, Giuseppe, DI RENZO, GIANFRANCO MARIA LUIGI, and Annunziato, Lucio
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Gene isoform ,Programmed cell death ,Sodium calcium exchanger ,NCX isoform ,Physiology ,Cognitive Neuroscience ,Drug Evaluation, Preclinical ,Hippocampal formation ,Pharmacology ,Biochemistry ,Sodium-Calcium Exchanger ,cerebral ischemia ,Brain Ischemia ,Cell Line ,Tissue Culture Techniques ,Brain ischemia ,chemistry.chemical_compound ,Dogs ,Cricetinae ,medicine ,Animals ,Protein Isoforms ,Benzamide ,NCX3 inhibitor ,Neurons ,Cell Death ,Dose-Response Relationship, Drug ,Sodium-calcium exchanger ,OGD ,Brain ,Infarction, Middle Cerebral Artery ,Cell Biology ,General Medicine ,medicine.disease ,Cell Hypoxia ,Rats ,Mice, Inbred C57BL ,Disease Models, Animal ,Glucose ,chemistry ,Benzamides ,Mutation ,Calcium ,Homeostasis ,Intracellular ,Central Nervous System Agents - Abstract
The Na(+)/Ca(2+) exchanger (NCX), a 10-transmembrane domain protein mainly involved in the regulation of intracellular Ca(2+) homeostasis, plays a crucial role in cerebral ischemia. In the present paper, we characterized the effect of the newly synthesized compound 5-amino-N-butyl-2-(4-ethoxyphenoxy)-benzamide hydrochloride (BED) on the activity of the three NCX isoforms and on the evolution of cerebral ischemia. BED inhibited NCX isoform 3 (NCX3) activity (IC50 = 1.9 nM) recorded with the help of single-cell microflorimetry, (45)Ca(2+) radiotracer fluxes, and patch-clamp in whole-cell configuration. Furthermore, this drug displayed negligible effect on NCX2, the other isoform expressed within the CNS, and it failed to modulate the ubiquitously expressed NCX1 isoform. Concerning the molecular site of action, the use of chimera strategy and deletion mutagenesis showed that α1 and α2 repeats of NCX3 represented relevant molecular determinants for BED inhibitory action, whereas the intracellular regulatory f-loop was not involved. At 10 nM, BED worsened the damage induced by oxygen/glucose deprivation (OGD) followed by reoxygenation in cortical neurons through a dysregulation of [Ca(2+)]i. Furthermore, at the same concentration, BED significantly enhanced cell death in CA3 subregion of hippocampal organotypic slices exposed to OGD and aggravated infarct injury after transient middle cerebral artery occlusion in mice. These results showed that the newly synthesized 5-amino-N-butyl-2-(4-ethoxyphenoxy)-benzamide hydrochloride is one of the most potent inhibitor of NCX3 so far identified, representing an useful tool to dissect the role played by NCX3 in the control of Ca(2+) homeostasis under physiological and pathological conditions.
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- 2015
21. Involvement of the Na+/Ca2+ exchanger isoform 1 (NCX1) in Neuronal Growth Factor (NGF)-induced Neuronal Differentiation through Ca2+-dependent Akt Phosphorylation
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Anna Pannaccione, Maria Cantile, Agnese Secondo, Francesca Boscia, Gianfranco Di Renzo, Roselia Ciccone, Maria Josè Sisalli, Alba Esposito, Antonella Scorziello, Rossana Sirabella, Pasquale Molinaro, Lucio Annunziato, Secondo, Agnese, Esposito, Alba, Sirabella, Rossana, Boscia, Francesca, Pannaccione, Anna, Molinaro, Pasquale, Cantile, Maria, Ciccone, Roselia, Sisalli, MARIA JOSE', Scorziello, Antonella, DI RENZO, GIANFRANCO MARIA LUIGI, and Annunziato, Lucio
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Patch-Clamp Techniques ,Patch-Clamp Technique ,Cellular differentiation ,Endoplasmic Reticulum ,PC12 Cells ,PI3K ,Biochemistry ,Phosphatidylinositol 3-Kinases ,Neurobiology ,Nerve Growth Factor ,Homeostasis ,Phosphorylation ,RNA, Small Interfering ,Calcium Transport ,Neurons ,Brain ,Cell Differentiation ,Calcium Imaging ,Cell biology ,cardiovascular system ,Signal transduction ,Signal Transduction ,Neurite ,Biology ,Sodium-Calcium Exchanger ,Neurite Outgrowth ,Homeostasi ,Neurites ,Animals ,Rats, Wistar ,Molecular Biology ,Protein kinase B ,PI3K/AKT/mTOR pathway ,Sodium-calcium exchanger ,Animal ,Akt ,Sodium ,Cell Biology ,Neuron ,Molecular biology ,PC12 Cell ,Rats ,Enzyme Activation ,Nerve growth factor ,nervous system ,Mutation ,Rat ,Calcium ,Phosphatidylinositol 3-Kinase - Abstract
NGF induces neuronal differentiation by modulating [Ca(2+)]i. However, the role of the three isoforms of the main Ca(2+)-extruding system, the Na(+)/Ca(2+) exchanger (NCX), in NGF-induced differentiation remains unexplored. We investigated whether NCX1, NCX2, and NCX3 isoforms could play a relevant role in neuronal differentiation through the modulation of [Ca(2+)]i and the Akt pathway. NGF caused progressive neurite elongation; a significant increase of the well known marker of growth cones, GAP-43; and an enhancement of endoplasmic reticulum (ER) Ca(2+) content and of Akt phosphorylation through an early activation of ERK1/2. Interestingly, during NGF-induced differentiation, the NCX1 protein level increased, NCX3 decreased, and NCX2 remained unaffected. At the same time, NCX total activity increased. Moreover, NCX1 colocalized and coimmunoprecipitated with GAP-43, and NCX1 silencing prevented NGF-induced effects on GAP-43 expression, Akt phosphorylation, and neurite outgrowth. On the other hand, the overexpression of its neuronal splicing isoform, NCX1.4, even in the absence of NGF, induced an increase in Akt phosphorylation and GAP-43 protein expression. Interestingly, tetrodotoxin-sensitive Na(+) currents and 1,3-benzenedicarboxylic acid, 4,4'-[1,4,10-trioxa-7,13-diazacyclopentadecane-7,13-diylbis(5-methoxy-6,12-benzofurandiyl)]bis-, tetrakis[(acetyloxy)methyl] ester-detected [Na(+)]i significantly increased in cells overexpressing NCX1.4 as well as ER Ca(2+) content. This latter effect was prevented by tetrodotoxin. Furthermore, either the [Ca(2+)]i chelator(1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid) (BAPTA-AM) or the PI3K inhibitor LY 294002 prevented Akt phosphorylation and GAP-43 protein expression rise in NCX1.4 overexpressing cells. Moreover, in primary cortical neurons, NCX1 silencing prevented Akt phosphorylation, GAP-43 and MAP2 overexpression, and neurite elongation. Collectively, these data show that NCX1 participates in neuronal differentiation through the modulation of ER Ca(2+) content and PI3K signaling.
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- 2015
22. Anoxia-Induced NF-kB-Dependent Upregulation of NCX1 Contributes to Ca 2+ Refilling Into Endoplasmic Reticulum in Cortical Neurons
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Leonilda Bilo, Anna Pannaccione, Valeria Valsecchi, Gianfranco Di Renzo, Agnese Secondo, Rossana Sirabella, Lucio Annunziato, Antonella Scorziello, Annagrazia Adornetto, Sirabella, Rossana, Secondo, Agnese, Pannaccione, Anna, Scorziello, Antonella, Valsecchi, V, Adornetto, A, Bilo, Leonilda, DI RENZO, GIANFRANCO MARIA LUIGI, and Annunziato, Lucio
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Gene isoform ,medicine.medical_specialty ,Blotting, Western ,Endoplasmic Reticulum ,Neuroprotection ,Sodium-Calcium Exchanger ,Western blot ,Downregulation and upregulation ,Pregnancy ,[Ca2+]i homeostasi ,Internal medicine ,medicine ,Animals ,Calcium Signaling ,RNA, Small Interfering ,Rats, Wistar ,Caspase 12 ,Fluorescent Dyes ,Cerebral Cortex ,Neurons ,Advanced and Specialized Nursing ,Cell Death ,medicine.diagnostic_test ,Reverse Transcriptase Polymerase Chain Reaction ,business.industry ,Endoplasmic reticulum ,NF-kappa B ,Transcription Factor RelA ,OGD ,Cell Hypoxia ,Rats ,Up-Regulation ,Cell biology ,Enzyme Activation ,Cytosol ,Glucose ,Endocrinology ,Na+-Ca2+ exchanger ,Unfolded protein response ,ER stre ,Female ,RNA Interference ,neuroprotection ,Neurology (clinical) ,Fura-2 ,Cardiology and Cardiovascular Medicine ,business ,Homeostasis - Abstract
Background and Purpose— The 3 gene products of the Na + /Ca 2+ exchanger (NCX), viz, NCX1, NCX2, and NCX3, may play a pivotal role in the pathophysiology of brain ischemia. The aim of this study was to investigate the transductional and posttranslational mechanisms involved in the expression of these isoforms during oxygen and glucose deprivation and their role in endoplasmic reticulum Ca 2+ refilling in cortical neurons. Methods— NCX1, NCX2, and NCX3 transcript and protein expression was evaluated in primary cortical neurons by reverse transcriptase–polymerase chain reaction and Western blot. NCX currents (I NCX ) and cytosolic Ca 2+ concentrations ([Ca 2+ ] i ) were monitored by means of patch-clamp in whole-cell configuration and Fura-2AM single-cell video imaging, respectively. Results— Exposure of cortical neurons to 3 hours of oxygen and glucose deprivation yielded dissimilar effects on the 3 isoforms. First, it induced an upregulation in NCX1 transcript and protein expression. This change was exerted at the transcriptional level because the inhibition of nuclear factor kappa B translocation by small interfering RNA against p65 and SN-50 prevented oxygen and glucose deprivation-induced NCX1 upregulation. Second, it elicited a downregulation of NCX3 protein expression. This change, unlike NCX1, was exerted at the posttranscriptional level because it was prevented by the proteasome inhibitor MG-132. Finally, we found that it significantly increased I NCX both in the forward and reverse modes of operation and promoted an increase in ER Ca 2+ accumulation. Interestingly, such accumulation was prevented by the silencing of NCX1 and the NCX inhibitor CB-DMB that triggered caspase-12 activation. Conclusions— These results suggest that nuclear factor kappa B-dependent NCX1 upregulation may play a fundamental role in Ca 2+ refilling in the endoplasmic reticulum, thus helping neurons to prevent endoplasmic reticulum stress during oxygen and glucose deprivation.
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- 2009
23. A Critical Role for the Potassium-Dependent Sodium–Calcium Exchanger NCKX2 in Protection against Focal Ischemic Brain Damage
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Gianfranco Di Renzo, Rosaria Gala, Francesca Boscia, Lucio Annunziato, Jonathan Lytton, Xiao Fang Li, Pasquale Molinaro, Anna Pannaccione, Davide Viggiano, Antonella Scorziello, Agnese Secondo, Ornella Cuomo, Giuseppe Pignataro, Annagrazia Adornetto, Cuomo, Ornella, Gala, Rosaria, Pignataro, Giuseppe, Boscia, Francesca, Secondo, Agnese, Scorziello, Antonella, Pannaccione, Anna, Viggiano, Davide, Adornetto, Annagrazia, Molinaro, Pasquale, Li, Xf, Lytton, J, DI RENZO, GIANFRANCO MARIA LUIGI, and Annunziato, Lucio
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Male ,Gene isoform ,antisense strategy ,medicine.medical_specialty ,Patch-Clamp Techniques ,sodium–calcium exchanger ,Ischemia ,Phosphorothioate Oligonucleotides ,Blood Pressure ,In Vitro Techniques ,cerebral ischemia ,Sodium-Calcium Exchanger ,Brain Ischemia ,Rats, Sprague-Dawley ,Brain ischemia ,Mice ,Ischemic brain ,Internal medicine ,medicine ,Animals ,NCKX ,knock-out ,Cerebral Cortex ,Mice, Knockout ,Neurons ,Analysis of Variance ,Messenger RNA ,Sodium-calcium exchanger ,biology ,Chemistry ,General Neuroscience ,Articles ,Infusion Pumps, Implantable ,medicine.disease ,Potassium-dependent sodium-calcium exchanger ,Rats ,Mice, Inbred C57BL ,Disease Models, Animal ,Endocrinology ,Cerebrovascular Circulation ,Phosphopyruvate Hydratase ,biology.protein ,Autoradiography ,Calcium ,MCAO ,Blood Gas Analysis ,Neuroscience ,Homeostasis - Abstract
The superfamily of cation/Ca2+plasma–membrane exchangers contains two branches, the K+-independent Na+–Ca2+exchangers (NCXs) and the K+-dependent Na+–Ca2+exchangers (NCKXs), widely expressed in mammals. NCKX2 is the major neuronally expressed isoform among NCKX members. Despite its importance in maintaining Na+, Ca2+, and K+homeostasis in the CNS, the role of NCKX2 during cerebral ischemia, a condition characterized by an alteration of ionic concentrations, has not yet been investigated. The present study examines NCKX2 role in the development of ischemic brain damage in permanent middle cerebral artery occlusion (pMCAO) and transient middle cerebral artery occlusion. Furthermore, to evaluate the effect of nckx2 ablation on neuronal survival,nckx2−/− primary cortical neurons were subjected to oxygen glucose deprivation plus reoxygenation. NCKX2 mRNA and protein expression was evaluated in the ischemic core and surrounding ipsilesional areas, at different time points after pMCAO in rats. In ischemic core and in periinfarctual area, NCKX2 mRNA and protein expression were downregulated. In addition, NCKX2 knock-down by antisense oligodeoxynucleotide and NCKX2 knock-out by genetic disruption dramatically increased infarct volume. Accordingly,nckx2−/− primary cortical neurons displayed a higher vulnerability and a greater [Ca2+]iincrease under hypoxic conditions, compared withnckx2+/+ neurons. In addition, NCKX currents both in the forward and reverse mode of operation were significantly reduced innckx2−/− neurons compared withnckx2+/+ cells. Overall, these results indicate that NCKX2 is involved in brain ischemia, and it may represent a new potential target to be investigated in the study of the molecular mechanisms involved in cerebral ischemia.
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- 2008
24. Nitric oxide induces [Ca2+]ioscillations in pituitary GH3cells: involvement ofIDRand ERG K+currents
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Rossana Sirabella, Agnese Secondo, Lucio Annunziato, Luigi Formisano, Anna Pannaccione, Mauro Cataldi, Gianfranco Di Renzo, Secondo, Agnese, Pannaccione, Anna, Cataldi, Mauro, Sirabella, Rossana, Formisano, Luigi, DI RENZO, GIANFRANCO MARIA LUIGI, and Annunziato, Lucio
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Nitroprusside ,medicine.medical_specialty ,Pituitary gland ,Arginine ,Physiology ,S-Nitroso-N-Acetylpenicillamine ,Nitric Oxide ,Cell Line ,Nitric oxide ,chemistry.chemical_compound ,Software Design ,Internal medicine ,medicine ,Animals ,Nitric Oxide Donors ,Calcium Signaling ,Enzyme Inhibitors ,voltage-gated potassium channel ,biology ,fast-inactivating outward currents ,Cell Biology ,Voltage-gated potassium channel ,Ether-A-Go-Go Potassium Channels ,Rats ,Nitric oxide synthase ,NG-Nitroarginine Methyl Ester ,Endocrinology ,medicine.anatomical_structure ,chemistry ,Pituitary Gland ,Potassium ,Biophysics ,biology.protein ,ether-à-go-go-related gene potassium channel ,slow-inactivating outward current ,Nitric Oxide Synthase ,S-Nitroso-N-acetylpenicillamine ,Erg ,Algorithms ,Intracellular ,Nitroso Compounds - Abstract
The role of nitric oxide (NO) in the occurrence of intracellular Ca2+concentration ([Ca2+]i) oscillations in pituitary GH3cells was evaluated by studying the effect of increasing or decreasing endogenous NO synthesis with l-arginine and nitro-l-arginine methyl ester (l-NAME), respectively. When NO synthesis was blocked with l-NAME (1 mM) [Ca2+]i, oscillations disappeared in 68% of spontaneously active cells, whereas 41% of the quiescent cells showed [Ca2+]ioscillations in response to the NO synthase (NOS) substrate l-arginine (10 mM). This effect was reproduced by the NO donors NOC-18 and S-nitroso- N-acetylpenicillamine (SNAP). NOC-18 was ineffective in the presence of the L-type voltage-dependent Ca2+channels (VDCC) blocker nimodipine (1 μM) or in Ca2+-free medium. Conversely, its effect was preserved when Ca2+release from intracellular Ca2+stores was inhibited either with the ryanodine-receptor blocker ryanodine (500 μM) or with the inositol 1,4,5-trisphosphate receptor blocker xestospongin C (3 μM). These results suggest that NO induces the appearance of [Ca2+]ioscillations by determining Ca2+influx. Patch-clamp experiments excluded that NO acted directly on VDCC but suggested that NO determined membrane depolarization because of the inhibition of voltage-gated K+channels. NOC-18 and SNAP caused a decrease in the amplitude of slow-inactivating ( IDR) and ether-à-go-go-related gene ( ERG) hyperpolarization-evoked, deactivating K+currents. Similar results were obtained when GH3cells were treated with l-arginine. The present study suggests that in GH3cells, endogenous NO plays a permissive role for the occurrence of spontaneous [Ca2+]ioscillations through an inhibitory effect on IDRand on IERG.
- Published
- 2006
25. First- and second-generation H1 antihistamines: from the molecular basis of their interaction with HERG K+ channels to physiological and pathophysiological implication
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Agnese Secondo, Pasqualina Castaldo, Anna Pannaccione, Lucio Annunziato, Mauro Cataldi, Maurizio Taglialatela, Francesca Boscia, Taglialatela, Maurizio, Castaldo, P., Pannaccione, Anna, Secondo, Agnese, Cataldi, Mauro, Boscia, Francesca, and Annunziato, Lucio
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biology ,HERG CHANNELS ,Chemistry ,ARRHYTHMIA ,Immunology ,hERG ,Cardiac action potential ,Pharmacology ,Potassium channel ,biology.protein ,Immunology and Allergy ,Neuroscience ,ANTIHISTAMINES ,K channels - Abstract
Summary The present article reports on the recent findings addressing the molecular basis for some rare but serious cardiovascular side-effects exerted by some non-sedating H1-blocking antihistamines. These latest developments, which have allowed fundamental insights into the role played by a specific class of potassium channels, the so-called human ether-a-gogo-related gene (HERG) channels, in the regulation of cardiac action potential duration and rhythm regulation, have also opened new areas of investigation into its participation in neuronal and endocrine (adenohypophyseal, chromaffin, and pancreatic) cells functioning.
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- 2004
26. Histidines 578 and 587 in the S5-S6Linker of the Human Ether-a-gogo Related Gene-1K+ Channels Confer Sensitivity to Reactive Oxygen Species
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Maurizio Taglialatela, Eckhard Ficker, Anna Pannaccione, Pasqualina Castaldo, Lucio Annunziato, Pannaccione, Anna, Castaldo, Pasqualina, Ficker, E, Annunziato, Lucio, and Taglialatela, Maurizio
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ERG1 Potassium Channel ,Potassium Channels ,Stereochemistry ,Molecular Sequence Data ,Xenopus ,Ether ,Deferoxamine ,medicine.disease_cause ,Biochemistry ,Histidines 578 and 587 ,Structure-Activity Relationship ,Xenopus laevis ,chemistry.chemical_compound ,medicine ,Animals ,Humans ,Histidine ,Amino Acid Sequence ,Molecular Biology ,Gene ,chemistry.chemical_classification ,Mutation ,Reactive oxygen species ,biology ,Chemistry ,Cell Biology ,biology.organism_classification ,Ether-A-Go-Go Potassium Channels ,Enzyme ,Potassium Channels, Voltage-Gated ,Catalase ,biology.protein ,Biophysics ,Reactive Oxygen specie ,K+ channels ,Reactive Oxygen Species ,Linker ,Phenanthrolines - Abstract
The K(+) channels encoded by the human Ether-a-gogo Related Gene-1 (hERG1) are crucially involved in controlling heart and brain excitability and are selectively influenced by reactive oxygen species (ROS). To localize the molecular regions involved in ROS-induced modulation of hERG1, segmental exchanges between the ROS-sensitive hERG1 and the ROS-insensitive bovine ether-a-gogo gene (bEAG) K(+) channels were generated, and the sensitivity of these chimeric channels to ROS was studied with the two-microelectrode voltage-clamp technique upon their expression in Xenopus oocytes. Substitution of the S(5)-S(6) linker of hERG1 with the corresponding bEAG region removed channel sensitivity to ROS, whereas the reverse chimeric exchange introduced ROS sensitivity into bEAG. Mutation of each of the two hERG1 histidines at positions 578 and 587 within the S(5)-S(6) linker generated K(+) channels insensitive to modulation by ROS. In addition, the two iron chelators desferrioxamine (1 mm) and o-phenanthroline (0.2 mm) significantly inhibited hERG1 outward K(+) currents and prevented hERG1 inhibition induced by the ROS-scavenging enzyme catalase (1000 units/ml). Finally, the hERG1-inhibitory effect exerted by the iron chelators was prevented by the hERG1 H578D/H587Y double mutation. Collectively, the results obtained suggest that histidines at positions 578 and 587 in the S(5)-S(6) linker region of hERG1 K(+) channels are crucial players in ROS-induced modulation of hERG1 K(+) channels.
- Published
- 2002
27. Genetically Modified Mice as a Strategy to Unravel the Role Played by the Na+/Ca2+ Exchanger in Brain Ischemia and in Spatial Learning and Memory Deficits
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Giuseppe Pignataro, Ornella Cuomo, Francesca Boscia, Agnese Secondo, Mauro Cataldi, André Herchuelz, Rossana Sirabella, Antonella Scorziello, Anna Pannaccione, Lucio Annunziato, Sophie Sokolow, Pasquale Molinaro, Davide Viggiano, Gianfranco Di Renzo, Molinaro, Pasquale, Cataldi, Mauro, Cuomo, Ornella, Viggiano, D, Pignataro, Giuseppe, Sirabella, Rossana, Secondo, Agnese, Boscia, Francesca, Pannaccione, Anna, Scorziello, Antonella, Sokolow, S, Herchuelz, A, DI RENZO, GIANFRANCO MARIA LUIGI, and Annunziato, Lucio
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NCX1 ,Pathology ,medicine.medical_specialty ,Sodium-calcium exchanger ,Antiporter ,Ischemia ,Biology ,medicine.disease ,NCX2 ,NCX3 ,Learning and memory ,Brain ischemia ,Conditional gene knockout ,Knockout mouse ,cardiovascular system ,medicine ,Memory consolidation ,Neuroscience ,Homeostasis ,Knockout mice - Abstract
Because no isoform-specific blocker of NCX has ever been synthesized, a more selective strategy to identify the role of each antiporter isoform in the brain was represented by the generation of knockout and knockin mice for the different isoforms of the antiporter.Experiments performed in NCX2 and NCX3 knockout mice provided evidence that these two isoforms participate in spatial learning and memory consolidation, although in an opposite manner. These new data from ncx2-/- and ncx3-/- mice may open new experimental avenues for the development of effective therapeutic compounds that, by selectively inhibiting or activating these molecular targets, could treat patients affected by cognitive impairment including Alzheimer's, Parkinson's, Huntington's diseases, and infarct dementia.More importantly, knockout and knockin mice also provided new relevant information on the role played by NCX in maintaining the intracellular Na(+) and Ca(2+) homeostasis and in protecting neurons during brain ischemia. In particular, both ncx2-/- and ncx3-/- mice showed an increased neuronal vulnerability after the ischemic insult induced by transient middle cerebral artery occlusion.As the ubiquitous deletion of NCX1 brings about to an early death of embryos because of a lack of heartbeat, this strategy could not be successfully pursued. However, information on the role of NCX1 in normal and ischemic brain could be obtained by developing conditional knockout mice lacking NCX1 in the brain. Preliminarily results obtained in these conditional mice suggest that also NCX1 protects neurons from ischemic cell death.Overall, the use of genetic-modified mice for NCX1, NCX2, and NCX3 represents a fruitful strategy to characterize the physiological role exerted by NCX in CNS and to identify the isoforms of the antiporter as potential molecular targets for therapeutic intervention in cerebral ischemia.
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- 2012
28. NCX1 expression and functional activity increase in microglia invading the infarct core
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Agnese Secondo, Gianfranco Di Renzo, Francesca Boscia, Anna Pannaccione, Lucio Annunziato, Rosaria Gala, Antonella Scorziello, Boscia, Francesca, Gala, Rosaria, Pannaccione, Anna, Secondo, Agnese, Scorziello, Antonella, DI RENZO, GIANFRANCO MARIA LUIGI, and Annunziato, Lucio
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Male ,Pathology ,medicine.medical_specialty ,cerebral ischemia ,Sodium-Calcium Exchanger ,Brain ischemia ,Rats, Sprague-Dawley ,Downregulation and upregulation ,Cell Movement ,Medicine ,Animals ,Cells, Cultured ,Advanced and Specialized Nursing ,Na+/Ca2+ exchanger ,NCX1 ,Microglia ,Sodium-calcium exchanger ,business.industry ,Cerebral infarction ,Cerebral Infarction ,medicine.disease ,Rats ,Up-Regulation ,Blot ,medicine.anatomical_structure ,Gene Expression Regulation ,IB4 ,cardiovascular system ,pMCAO ,Immunohistochemistry ,Neurology (clinical) ,Cardiology and Cardiovascular Medicine ,business ,Ex vivo - Abstract
Background and Purpose— The sodium–calcium exchanger NCX1 represents a key mediator for maintaining [Na + ] i and [Ca 2+ ] i in anoxic conditions. To date, no information is available on NCX1 protein expression and activity in microglial cells under ischemic conditions. Methods— By means of Western blotting, patch-clamp electrophysiology, single-cell Fura-2 acetoxymethyl-ester microfluorometry, immunohistochemistry, and confocal microscopy, we investigated the regional and temporal changes of NCX1 protein in microglial cells of the peri-infarct and core regions after permanent middle cerebral artery occlusion. The exchanger expression and activity were measured in primary microglia isolated ex vivo from the core region of adult rat brains 7 days after permanent middle cerebral artery occlusion and in cultured microglia under in vitro hypoxia. Results— One day after permanent middle cerebral artery occlusion, NCX1 protein expression was detected in some microglial cells adjacent to the soma of neurons in the infarct core. More interestingly, 3 and 7 days after permanent middle cerebral artery occlusion, NCX1 signal strongly increased in the round-shaped microglia invading the infarct core. Cultured microglial cells obtained from the core also displayed increased NCX1 expression as compared with contralateral cells and showed enhanced NCX activity in the reverse mode of operation. Similarly, NCX activity and NCX1 protein expression were significantly enhanced in BV2 microglia exposed to oxygen and glucose deprivation, whereas NCX2 and NCX3 were downregulated. Interestingly, in NCX1-silenced cells, [Ca 2+ ] i increase induced by hypoxia was completely prevented. Conclusion– The upregulation of NCX1 expression and activity observed in microglia after permanent middle cerebral artery occlusion suggests a relevant role of NCX1 in modulating microglia functions in the postischemic brain.
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- 2009
29. Targeted disruption of Na+/Ca2+ exchanger 3 (NCX3) gene leads to a worsening of ischemic brain damage
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Francesca Boscia, Rosaria Gala, Annagrazia Adornetto, Sophie Sokolow, Gianfranco Di Renzo, Rossana Sirabella, André Herchuelz, Agnese Secondo, Antonella Scorziello, Lucio Annunziato, Davide Viggiano, Anna Pannaccione, Ornella Cuomo, Giuseppe Pignataro, Stéphane Schurmans, Pasquale Molinaro, Molinaro, Pasquale, Cuomo, Ornella, Pignataro, Giuseppe, Boscia, Francesca, Sirabella, Rossana, Pannaccione, Anna, Secondo, Agnese, Scorziello, Antonella, Adornetto, A, Gala, Rosaria, Viggiano, D, Sokolow, S, Herchuelz, A, Schurmans, S, DI RENZO, GIANFRANCO MARIA LUIGI, and Annunziato, Lucio
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Gene isoform ,medicine.medical_specialty ,Cell Survival ,Ischemia ,Brain damage ,Hippocampal formation ,Biology ,Hippocampus ,cerebral ischemia ,Sodium-Calcium Exchanger ,Brain Ischemia ,Mice ,Mice, Congenic ,Organ Culture Techniques ,Internal medicine ,medicine ,Animals ,Homeostasis ,Hypoxia, Brain ,Mice, Knockout ,Neurons ,Sodium-calcium exchanger ,Cell Death ,General Neuroscience ,OGD ,Membrane Transport Proteins ,Articles ,Hypoxia (medical) ,medicine.disease ,sodium calcium exchanger ,Mice, Inbred C57BL ,Endocrinology ,organotypic hippocampal cultures ,Gene Targeting ,Disease Progression ,medicine.symptom ,MCAO ,Neuroscience ,Intracellular ,NCX ,Signal Transduction - Abstract
Na+/Ca2+exchanger 3 (NCX3), one of the three isoforms of the NCX family, is highly expressed in the brain and is involved in the maintenance of intracellular Na+and Ca2+homeostasis. Interestingly, whereas the function of NCX3 under physiological conditions has been determined, its role under anoxia is still unknown. To assess NCX3 role in cerebral ischemia, we exposed ncx3−/− mice to transient middle cerebral artery occlusion followed by reperfusion. In addition, to evaluate the effect of ncx3 ablation on neuronal survival, organotypic hippocampal cultures and primary cortical neurons from ncx3−/− mice were subjected to oxygen glucose deprivation (OGD) plus reoxygenation. Here we report that ncx3 gene suppression leads to a worsening of brain damage after focal ischemia and to a massive neuronal death in all the hippocampal fields of organotypic cultures as well as in cortical neurons from ncx3−/− mice exposed to OGD plus reoxygenation. In addition, in ncx3−/− cortical neurons exposed to hypoxia, NCX currents, recorded in the reverse mode of operation, were significantly lower than those detected in ncx3+/+. From these results, NCX3 protein emerges as a new molecular target that may have a potential therapeutic value in modulating cerebral ischemia.
- Published
- 2008
30. Up-regulation and increased activity of KV3.4 channels and their accessory subunit MinK-related peptide 2 induced by amyloid peptide are involved in apoptotic neuronal death
- Author
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Maurizio Taglialatela, G.F. Di Renzo, Anna Pannaccione, Pasqualina Castaldo, Lucio Annunziato, Francesca Boscia, Rossana Sirabella, Annagrazia Adornetto, Antonella Scorziello, Pannaccione, Anna, Boscia, Francesca, Scorziello, Antonella, Adornetto, Annagrazia, Castaldo, P, Sirabella, Rossana, Taglialatela, Maurizio, DI RENZO, GIANFRANCO MARIA LUIGI, and Annunziato, Lucio
- Subjects
Programmed cell death ,Patch-Clamp Techniques ,Protein subunit ,Peptide ,Apoptosis ,Biology ,Hippocampus ,PC12 Cells ,Cnidarian Venoms ,medicine ,Animals ,Patch clamp ,RNA, Messenger ,Rats, Wistar ,Cells, Cultured ,Pharmacology ,chemistry.chemical_classification ,Neurons ,Amyloid beta-Peptides ,Cell Death ,Neurodegeneration ,P3 peptide ,NF-kappa B ,NFKB1 ,medicine.disease ,Molecular biology ,Peptide Fragments ,Rats ,Up-Regulation ,Electrophysiology ,Nerve growth factor ,Sea Anemones ,chemistry ,Shaw Potassium Channels ,Molecular Medicine ,Peptides - Abstract
The aim of the present study was to investigate whether K(V)3.4 channel subunits are involved in neuronal death induced by neurotoxic beta-amyloid peptides (Abeta). In particular, to test this hypothesis, three main questions were addressed: 1) whether the Abeta peptide can up-regulate both the transcription/translation and activity of K(V)3.4 channel subunit and its accessory subunit, MinK-related peptide 2 (MIRP2); 2) whether the increase in K(V)3.4 expression and activity can be mediated by the nuclear factor-kappaB (NF-kappaB) family of transcriptional factors; and 3) whether the specific inhibition of K(V)3.4 channel subunit reverts the Abeta peptide-induced neurodegeneration in hippocampal neurons and nerve growth factor (NGF)-differentiated PC-12 cells. We found that Abeta(1-42) treatment induced an increase in K(V)3.4 and MIRP2 transcripts and proteins, detected by reverse transcription-polymerase chain reaction and Western blot analysis, respectively, in NGF-differentiated PC-12 cells and hippocampal neurons. Patch-clamp experiments performed in whole-cell configuration revealed that the Abeta peptide caused an increase in I(A) current amplitude carried by K(V)3.4 channel subunits, as revealed by their specific blockade with blood depressing substance-I (BDS-I) in both hippocampal neurons and NGF-differentiated PC-12 cells. The inhibition of NF-kappaB nuclear translocation with the cell membrane-permeable peptide SN-50 prevented the increase in K(V)3.4 protein and transcript expression. In addition, the SN-50 peptide was able to block Abeta(1-42)-induced increase in K(V)3.4 K(+) currents and to prevent cell death caused by Abeta(1-42) exposure. Finally, BDS-I produced a similar neuroprotective effect by inhibiting the increase in K(V)3.4 expression. As a whole, our data indicate that K(V)3.4 channels could be a novel target for Alzheimer's disease pharmacological therapy.
- Published
- 2007
31. GABA and glutamate receptors are involved in modulating pacemaker activity in hydra
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G Kass-Simon, P Pierobon, A Pannaccione, KASS SIMON, G, Pannaccione, Anna, and Pierobon, P.
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Agonist ,medicine.medical_specialty ,Physiology ,medicine.drug_class ,Hydra ,Glutamic Acid ,glutamate ,Kainate receptor ,AMPA receptor ,Biology ,Biochemistry ,GABA Antagonists ,GABA ,chemistry.chemical_compound ,Receptors, GABA ,Biological Clocks ,Internal medicine ,Culture Techniques ,Quinoxalines ,medicine ,Excitatory Amino Acid Agonists ,Animals ,GABA-A Receptor Agonists ,GABA-A Receptor Antagonists ,Molecular Biology ,GABA Agonists ,alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid ,gamma-Aminobutyric Acid ,Kainic Acid ,Glutamate receptor ,LGIC ,Bicuculline ,GABA receptor antagonist ,Endocrinology ,nervous system ,chemistry ,Receptors, Glutamate ,pacemaker systems ,embryonic structures ,NBQX ,Excitatory Amino Acid Antagonists ,medicine.drug ,Ionotropic effect - Abstract
The effects of gamma-amino butyric acid (GABA) and glutamate, their ionotropic agonists and antagonists on hydra’s ectodermal and endodermal pacemaker systems were studied. GABA decreased ectodermal body contraction bursts (CBs) and the number of pulses in a burst (PyCB) and endodermal rhythmic potentials (RPs); tentacle pulses (TPs) were not affected. The GABA agonist, muscimol, and the benzodiazepine receptor agonist, diazepam, mimicked the A effects of GABA on the endodermal system. The GABA antagonist bicuculline counteracted GABA’s effects. Low A concentrations of glutamate increased CBs and RPs. Higher concentrations required concanavalin A (Con A) to produce the same effect on CBs and PyCB. TPs were increased by high concentrations of glutamate and kainate. The ionotropic glutamate agonist, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) also required Con A to increase CBs and RPs. The effects of AMPA were antagonized by 6-nitro-7-sulfamoylbenzowfxquinoxaline-2,3-dione (NBQX), which, per se, decreased CBs. The results indicate that GABA and glutamate, acting on their ionotropic receptors, modify the impulses of hydra’s pacemaker systems. On the whole GABA decreased the outputs of both ectodermal and endodermal impulse generating systems, while glutamate increased them. 2003 Elsevier Science Inc. All rights reserved.
- Published
- 2003
32. Apoptosis induced in neuronal cells by oxidative stress: role played by caspases and intracellular calcium ions
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Giuseppe Pignataro, Lucio Annunziato, Luigi Sibaud, Rossana Sirabella, Mauro Cataldi, Angela D'Alessio, Salvatore Amoroso, G.F. Di Renzo, Agnese Secondo, Anna Pannaccione, Annunziato, L, Amoroso, S, Pannaccione, Anna, Cataldi, Mauro, Pignataro, Giuseppe, D'Alessio, A, Sirabella, Rossana, Secondo, Agnese, Sibaud, L, DI RENZO, GIANFRANCO MARIA LUIGI, Annunziato, L., Amoroso, S., D'Alessio, A., Sibaud, L., Annunziato, Lucio, Amoroso, Salvatore, and D'Alessio, Angela
- Subjects
Programmed cell death ,Free Radicals ,Caspase 2 ,Caspase 3 ,Apoptosis ,Biology ,Toxicology ,medicine.disease_cause ,Calcium in biology ,medicine ,Animals ,Humans ,Calcium ions ,chemistry.chemical_classification ,Neurons ,Reactive oxygen species ,General Medicine ,Caspase ,Cell biology ,Neuronal apoptosi ,Enzyme Activation ,Oxidative Stress ,chemistry ,Caspases ,biology.protein ,Oxidative stre ,Calcium ,Oxidative stress ,Intracellular - Abstract
Reactive oxygen species (ROS) have been implicated in the pathophysiology of many neurologic disorders and brain dysfunction. In the same pathological settings evidence has been provided in favour of a participation of intracellular Ca(2+) concentration altered homeostasis in the chain of events leading to neuronal apoptosis. In the present review literature reports and experimental data on the relationship between caspase activation and alteration of intracellular calcium concentrations in the mechanisms triggering neuronal apoptosis are discussed. The data gathered support the conclusion that during oxidative stress in neuronal cells the production of ROS triggers a mechanism that, through the release of cytochrome c from mitochondria and caspase-3 activation, leads to apoptosis; the concomitant ROS-mediated elevation of intracellular Ca(2+) concentration triggers caspase-2 activation but both events do not seem to be involved in cell death.
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- 2003
33. Modulation of ion channels by reactive oxygen and nitrogen species: a pathophysiological role in brain aging?
- Author
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Maurizio Taglialatela, Mauro Cataldi, Pasqualina Castaldo, Gianfranco Di Renzo, Anna Pannaccione, Agnese Secondo, Lucio Annunziato, Annunziato, Lucio, Pannaccione, Anna, Cataldi, Mauro, Secondo, Agnese, Castaldo, Pasqualina, DI RENZO, GIANFRANCO MARIA LUIGI, Taglialatela, Maurizio, Annunziato, L, Castaldo, P, and Taglialatela, M.
- Subjects
Voltage-gated calcium channel ,Aging ,Neuronal death ,Ion Channels ,chemistry.chemical_compound ,Voltage-gated sodium channel ,Humans ,Voltage-gated potassium channel ,Ion channel ,Reactive nitrogen species ,Human ether-a-gogo related gene-1 (hERG1 or KCNH2) ,Aged ,Voltage-dependent calcium channel ,Chemistry ,General Neuroscience ,Sodium channel ,Reactive oxygen species (ROS) ,Apoptosi ,Brain ,Reactive Nitrogen Species ,Electrophysiology ,Ion homeostasis ,Biochemistry ,Reactive nitrogen species (RNS) ,Biophysics ,Neurology (clinical) ,Geriatrics and Gerontology ,Reactive Oxygen Species ,Intracellular ,Developmental Biology - Abstract
An ever increasing number of reports shows the involvement of free radicals in the functional and structural changes occurring in the brain as a part of the "normal" aging process. Given that plasma membrane and intracellular ion channels play a critical role in maintaining intracellular ion homeostasis, which is crucial for neuronal cell survival, in the present review we have attempted to elaborate on the idea that functional changes in ion channel activity induced by reactive oxygen species (ROS) and reactive nitrogen species (RNS) might occur during the aging process. To this aim, we have reviewed the available literature and the data obtained in our laboratory on the ability of ROS and RNS to modify the activity of several plasma membrane and intracellular ion channels and transporters, in an attempt to correlate such changes with those occurring with the aging process. Particular emphasis is given to voltage-gated Na(+), Ca(2+), and K(+) channels, although second messenger-activated channels like Ca(2+)- and ATP-dependent K(+) channels, and intracellular channels controlling intracellular Ca(2+) storage and release will also be discussed. On the basis of the available data it is not yet possible to establish a strict correlation between the changes in neuronal electrophysiological properties induced by oxidative modification at the level of ion channels and the neurodegenerative process accompanying brain aging. However, an increasing amount of information suggests that the modulatory effects exerted by ROS and RNS on ion channel proteins might have a relevant role for neuronal cell survival or death. Obviously, more work is needed to establish the possible involvement of ion channels and of their modulation by ROS and RNS as important mechanisms for the aging process. Only when a more complete molecular picture of the aging process will be available, it will be possible to test the fascinating hypothesis that aging might be pharmacologically delayed by modulating ROS and RNS action on ion channels or the biochemical pathways involved in their modulation.
- Published
- 2002
34. 'Janus face' of nitric oxide action on plasma membrane and intracellular ionic channels
- Author
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Agnese Secondo, Anna Pannaccione, G.F. Di Renzo, Maurizio Taglialatela, Pasqualina Castaldo, Mauro Cataldi, S. Iossa, L. Annunziato, Taglialatela, Maurizio, Pannaccione, Anna, Cataldi, Mauro, Castaldo, P, Secondo, Agnese, Iossa, S, DI RENZO, GIANFRANCO MARIA LUIGI, Annunziato, L., Taglialatela, M, M., Taglialatela, P., Castaldo, S., Iossa, and Annunziato, Lucio
- Subjects
Aging ,chemistry.chemical_compound ,Health (social science) ,Membrane ,Chemistry ,Biophysics ,Plasma ,Janus ,Geriatrics and Gerontology ,Gerontology ,Ionic Channels ,Intracellular ,Nitric oxide - Published
- 2001
35. Inhibition of depolarization-induced [3H]noradrenaline release from SH-SY5Y human neuroblastoma cells by some second-generation H(1) receptor antagonists through blockade of store-operated Ca(2+) channels (SOCs)
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Taglialatela, Maurizio, Secondo, A, Fresi, A, Rosati, B, Pannaccione, A, Castaldo, P, Giorgio, G, Wanke, E, Annunziato, L., Taglialatela, Maurizio, Secondo, Agnese, Fresi, A, Rosati, B, Pannaccione, Anna, Castaldo, P, Giorgio, G, Wanke, E, Annunziato, L., Taglialatela, M, and Annunziato, Lucio
- Subjects
ERG1 Potassium Channel ,Potassium Channels ,Depolarization-induced norepinephrine release ,Tritium ,Neuroblastoma ,Norepinephrine ,Transcriptional Regulator ERG ,Tumor Cells, Cultured ,Humans ,Receptors, Histamine H1 ,Cation Transport Proteins ,Second-generation antihistamine ,Ca2+ channels activated by Cai2+ store depletion ,hERG K+ channel ,Astemizole ,Loratadine ,Calcium Channel Blockers ,Cetirizine ,Ether-A-Go-Go Potassium Channels ,DNA-Binding Proteins ,Potassium Channels, Voltage-Gated ,Hydroxyzine ,Histamine H1 Antagonists ,Trans-Activators ,SH-SY5Y human neuroblastoma cell ,Calcium ,Calcium Channels ,Terfenadine ,Long QT syndrome - Abstract
In the present study, the effect of the blockade of membrane calcium channels activated by intracellular Ca(2+) store depletion on basal and depolarization-induced [3H]norepinephrine ([3H]NE) release from SH-SY5Y human neuroblastoma cells was examined. The second-generation H(1) receptor blockers astemizole, terfenadine, and loratadine, as well as the first-generation compound hydroxyzine, inhibited [3H]NE release induced by high extracellular K(+) concentration ([K(+)](e)) depolarization in a concentration-dependent manner (the IC(50)s were 2.3, 1.7, 4.8, and 9.4 microM, respectively). In contrast, the more hydrophilic second-generation H(1) receptor blocker cetirizine was completely ineffective (0.1-30 microM). The inhibition of high [K(+)](e)-induced [3H]NE release by H(1) receptor blockers seems to be related to their ability to inhibit Ca(2+) channels activated by Ca(i)(2+) store depletion (SOCs). In fact, astemizole, terfenadine, loratadine, and hydroxyzine, but not cetirizine, displayed a dose-dependent inhibitory action on the increase in intracellular Ca(2+) concentrations ([Ca(2+)](i)) obtained with extracellular Ca(2+) reintroduction after Ca(i)(2+) store depletion with thapsigargin (1 microM), an inhibitor of the sarcoplasmic-endoplasmic reticulum calcium ATPase (SERCA) pump. The rank order of potency for SOC inhibition by these compounds closely correlated with their inhibitory properties on depolarization-induced [3H]NE release from SH-SY5Y human neuroblastoma cells. Nimodipine (1 microM) plus omega-conotoxin (100 nM) did not interfere with the present model for SOC activation. In addition, the inhibition of depolarization-induced [3H]NE release does not seem to be attributable to the blockade of the K(+) currents carried by the K(+) channels encoded by the human Ether-a-Gogo Related Gene (I(HERG)) by these antihistamines. In fact, whole-cell voltage-clamp experiments revealed that the IC(50) for astemizole-induced hERG blockade is about 300-fold lower than that for the inhibition of high K(+)-induced [3H]NE release. Furthermore, current-clamp experiments in SH-SY5Y cells showed that concentrations of astemizole (3 microM) which were effective in preventing depolarization-induced [3H]NE release were unable to interfere with the cell membrane potential under depolarizing conditions (100 mM [K(+)](e)), suggesting that hERG K(+) channels do not contribute to membrane potential control during exposure to elevated [K(+)](e). Collectively, the results of the present study suggest that, in SH-SY5Y human neuroblastoma cells, the inhibition of SOCs by some second-generation antihistamines can prevent depolarization-induced neurotransmitter release.
- Published
- 2001
36. Inhibition of HERG1 K+ channels by the novel second-generation antihistamine mizolastine
- Author
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Taglialatela, M., Anna PANNACCIONE, Castaldo, P., Giorgio, G., Annunziato, L., Taglialatela, Maurizio, Pannaccione, Anna, Castaldo, P, Giorgio, G, Annunziato, Lucio, and Annunziato, L.
- Subjects
drug cardiotoxicity ,ERG1 Potassium Channel ,Potassium Channels ,Xenopus ,Astemizole ,antihistamine ,Ether-A-Go-Go Potassium Channels ,Cell Line ,DNA-Binding Proteins ,Transcriptional Regulator ERG ,Potassium Channels, Voltage-Gated ,Papers ,Histamine H1 Antagonists ,Trans-Activators ,mizolastine ,Animals ,Humans ,Benzimidazoles ,Female ,Potassium channel ,Cation Transport Proteins ,arrhythmias - Abstract
1. Ventricular arrhythmias are rare but life-threatening side effects of therapy with the second-generation H(1) receptor antagonists terfenadine and astemizole. Blockade of the K(+) channels encoded by the Human Ether-à-go-go-Related Gene 1 (HERG1) K(+) channels, which is the molecular basis of the cardiac repolarizing current I(Kr), by prolonging cardiac repolarization, has been recognized as the mechanism underlying the cardiac toxicity of these compounds. 2. In the present study, the potential blocking ability of the novel second-generation H(1) receptor antagonist mizolastine of the HERG1 K(+) channels heterologously expressed in XENOPUS: oocytes and in HEK 293 cells or constitutively present in SH-SY5Y human neuroblastoma cells has been examined and compared to that of astemizole. 3. Mizolastine blocked HERG1 K(+) channels expressed in XENOPUS: oocytes with an estimated IC(50) of 3.4 microM. Mizolastine blockade was characterized by a fast dissociation rate when compared to that of astemizole; when fitted to a monoexponential function, the time constants for drug dissociation from the K(+) channel were 72.4+/-11.9 s for 3 microM mizolastine, and 1361+/-306 s for 1 microM astemizole. 4. In human embryonic kidney 293 cells (HEK 293 cells) stably transfected with HERG1 cDNA, extracellular application of mizolastine exerted a dose-related inhibitory action on I(HERG1), with an IC(50) of 350+/-76 nM. Furthermore, mizolastine dose-dependently inhibited HERG1 K(+) channels constitutively expressed in SH-SY5Y human neuroblastoma clonal cells. 5. The results of the present study suggest that the novel second-generation H(1) receptor antagonist mizolastine, in concentrations higher than those achieved in vivo during standard therapy, is able to block in some degree both constitutively and heterologously expressed HERG1 K(+) channels, and confirm the heterogeneity of molecules belonging to this therapeutical class with respect to their HERG1-inhibitory action.
- Published
- 2000
37. Modulation of the K+ channels encoded by the human ethera-gogo-related gene-1 (hERG1) by nitric oxide
- Author
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Taglialatela, M., Pannaccione, A., Iossa, S., Castaldo, P., LUCIO ANNUNZIATO, Taglialatela, M, Pannaccione, Anna, Iossa, S, Castaldo, P, Annunziato, Lucio, and Taglialatela, Maurizio
- Subjects
K(+) channels. human ether-a-gogo-related gene-1 (hERG1) ,nitric oxide - Abstract
The inhibition of nitric oxide synthase by N-nitro-L-arginine methyl ester (0.03-3 mM) dose-dependently reduced nitric oxide (NO(*)) levels and enhanced the outward currents carried by human ether-a-gogo-related gene-1 (hERG1) K(+) channels expressed in Xenopus laevis oocytes, whereas the increase in NO(*) levels achieved by exposure to L-arginine (0.03-10 mM) inhibited these currents. Furthermore, four NO(*) donors belonging to such different chemical classes as sodium nitroprusside (1-1000 microM), 3-morpholino-sydnonimine (100-1000 microM), (Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1- ium-1, 2-diolate (NOC-18; 1-300 microM), and S-nitroso N-acetylpenicillamine (1-300 microM) dose-dependently inhibited hERG1 outward K(+) currents. By contrast, the NO(*) donor NOC-18 (0.3 mM) did not affect other cloned K(+) channels such as rat neuroblastoma-glioma K(+) channel 2, rat delayed rectifier K(+) channel 1, bovine ether-a-gogo gene, rat ether-a-gogo-related gene-2, and rat ether-a-gogo-related gene-3. The inhibitory effect of NO(*) donors on hERG1 K(+) channels was prevented by the NO(*) scavengers 2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide and hemoglobin. The membrane permeable analog of cGMP, 8-bromo-cGMP (1 mM), failed to reproduce the inhibitory action of NO(*) donors on hERG1 outward currents; furthermore, the specific inhibitor of the NO(*)-dependent guanylyl cyclase, 1H-[1,2,4]oxadiazolo[4, 3-a]quinoxalin-1-one (50 microM), neither interfered with outward hERG1 K(+) currents nor prevented their inhibition by 0.3 mM NOC-18. Both L-arginine (10 mM) and NOC-18 (0.3 mM) counteracted the stimulatory effect on hERG1 outward currents induced by the radical oxygen species-generating system FeSO(4) (25 microM)/ascorbic acid (50 microM; Fe/Asc). Finally, L-arginine (10 mM) and NOC-18 (0.3 mM) inhibited both basal and Fe/Asc (0.1 mM/0.2 mM)-stimulated lipid peroxidation in X. laevis oocytes. Collectively, the present results suggest that NO(*), both endogenously produced and pharmacologically delivered, may exert in a cGMP-independent way an inhibitory effect on hERG1 outward K(+) currents via an interaction with radical oxygen species either generated under resting conditions or triggered by Fe/Asc.
- Published
- 1999
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