62 results on '"Borsello, T."'
Search Results
2. Contribution of serine racemase/d-serine pathway to neuronal apoptosis
- Author
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Esposito, S, Pristerà, A, Maresca, G, Cavallaro, S, Felsani, A, Florenzano, F, Manni, L, Ciotti, M, Pollegioni, L, Borsello, T, and Canu, N
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Proteasome Endopeptidase Complex ,Racemases and Epimerases ,Apoptosis ,serine.racemase ,In Vitro Techniques ,Settore BIO/09 ,Receptors, N-Methyl-D-Aspartate ,neuronal death, proteasome, c.Jun N-terminal kinase, NMDA receptor, serine.racemase, cerebellar granule neurons, apoptosis ,Cerebellum ,Serine ,Animals ,RNA, Messenger ,cerebellar granule neurons ,RNA, Small Interfering ,Rats, Wistar ,Cellular Senescence ,Neurons ,JNK Mitogen-Activated Protein Kinases ,Stereoisomerism ,NMDA receptor ,neuronal death ,Rats ,proteasome ,nervous system ,c.Jun N-terminal kinase ,Signal Transduction - Abstract
Recent data indicate that age-related N-methyl-d-aspartate receptor (NMDAR) transmission impairment is correlated with the reduction in serine racemase (SR) expression and d-serine content. As apoptosis is associated with several diseases and conditions that generally occur with age, we investigated the modulation of SR/d-serine pathway during neuronal apoptosis and its impact on survival. We found that in cerebellar granule neurons (CGNs), undergoing apoptosis SR/d-serine pathway is crucially regulated. In the early phase of apoptosis, the expression of SR is reduced, both at the protein and RNA level through pathways, upstream of caspase activation, involving ubiquitin proteasome system (UPS) and c-Jun N-terminal kinases (JNKs). Forced expression of SR, together with treatment with NMDA and d-serine, blocks neuronal death, whereas pharmacological inhibition and Sh-RNA-mediated suppression of endogenous SR exacerbate neuronal death. In the late phase of apoptosis, the increased expression of SR contribute to the last, NMDAR-mediated, wave of cell death. These findings are relevant to our understanding of neuronal apoptosis and NMDAR activity regulation, raising further questions as to the role of SR/d-serine in those neuro-pathophysiological processes, such as aging and neurodegenerative diseases characterized by a convergence of apoptotic mechanisms and NMDAR dysfunction.
- Published
- 2012
3. Exploring the role of MKK7 in excitotoxicity and cerebral ischemia: a novel pharmacological strategy against brain injury
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Vercelli, A, primary, Biggi, S, additional, Sclip, A, additional, Repetto, I E, additional, Cimini, S, additional, Falleroni, F, additional, Tomasi, S, additional, Monti, R, additional, Tonna, N, additional, Morelli, F, additional, Grande, V, additional, Stravalaci, M, additional, Biasini, E, additional, Marin, O, additional, Bianco, F, additional, di Marino, D, additional, and Borsello, T, additional
- Published
- 2015
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4. c-Jun N-terminal kinase has a key role in Alzheimer disease synaptic dysfunction in vivo
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Sclip, A, primary, Tozzi, A, additional, Abaza, A, additional, Cardinetti, D, additional, Colombo, I, additional, Calabresi, P, additional, Salmona, M, additional, Welker, E, additional, and Borsello, T, additional
- Published
- 2014
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5. c-Jun N-terminal kinase pathway activation in human and experimental traumatic brain injury: neuroprotective effects of its inhibition
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Ortolano, F, primary, Zanier, ER, additional, Colombo, A, additional, Sclip, A, additional, Longhi, L, additional, Perego, C, additional, Borsello, T, additional, Stocchetti, N, additional, and De Simoni, MG, additional
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- 2008
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6. The TAT-JNK inhibitor peptide interferes with beta amyloid protein stability
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Colombo, A, primary, Repici, M, additional, Pesaresi, M, additional, Santambrogio, S, additional, Forloni, G, additional, and Borsello, T, additional
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- 2007
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7. C-JUN NH2-TERMINAL KINASE MEDIATES THE HEPATIC STRESS RESPONSE AFTER RAT HEMORRHAGE/RESUSCITATION
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Relja, B., primary, Schwestka, B., additional, Sun-Young Lee, V., additional, Henrich, D., additional, Czerny, C., additional, Borsello, T., additional, Lehnert, M., additional, and Marzi, I., additional
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- 2006
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8. Role of the JNK pathway in NMDA-mediated excitotoxicity of cortical neurons
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Centeno, C, primary, Repici, M, additional, Chatton, J-Y, additional, Riederer, B M, additional, Bonny, C, additional, Nicod, P, additional, Price, M, additional, Clarke, P G H, additional, Papa, S, additional, Franzoso, G, additional, and Borsello, T, additional
- Published
- 2006
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9. Blocking ADAM10 synaptic trafficking generates a model of sporadic Alzheimer's disease.
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Epis R, Marcello E, Gardoni F, Vastagh C, Malinverno M, Balducci C, Colombo A, Borroni B, Vara H, Dell'agli M, Cattabeni F, Giustetto M, Borsello T, Forloni G, Padovani A, Di Luca M, Epis, Roberta, Marcello, Elena, Gardoni, Fabrizio, and Vastagh, Csaba
- Abstract
We describe here an innovative, non-transgenic animal model of Alzheimer's disease. This model mimics early stages of sporadic disease, which represents the vast majority of cases. The model was obtained by interfering with the complex between a disintegrin and metalloproteinase domain containing protein 10 (ADAM10), the main α-secretase candidate, and its partner, synapse-associated protein 97, a protein of the postsynaptic density-membrane associated guanylate kinase family. Association of ADAM10 with synapse-associated protein 97 governs enzyme trafficking and activity at synapses. Interfering with the ADAM10/synapse-associated protein 97 complex for 2 weeks by means of a cell-permeable peptide strategy is sufficient to shift the metabolism of the amyloid precursor protein towards amyloidogenesis and allows the reproduction of initial phases of sporadic Alzheimer's disease. After 2 weeks of treatment, we detected progressive Alzheimer's disease-like neuropathology, with an increase of β-amyloid aggregate production and of tau hyperphosphorylation, and a selective alteration of N-methyl-d-aspartic acid receptor subunit composition in the postsynaptic compartment of mouse brain. Behavioural and electrophysiological deficits were also induced by peptide treatment. [ABSTRACT FROM AUTHOR]
- Published
- 2010
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10. Role of the JNK pathway in NMDA-mediated excitotoxicity of cortical neurons.
- Author
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Centeno, C., Repici, M., Chatton, J.-Y., Riederer, B. M., Bonny, C., Nicod, P., Price, M., Clarke, P. G. H., Papa, S., Franzoso, G., and Borsello, T.
- Subjects
JNK mitogen-activated protein kinases ,NEURODEGENERATION ,METHYL aspartate ,PHOSPHORYLATION ,CEREBRAL ischemia ,PEPTIDES ,EXCITATORY amino acids ,CELLULAR control mechanisms - Abstract
Excitotoxic insults induce c-Jun N-terminal kinase (JNK) activation, which leads to neuronal death and contributes to many neurological conditions such as cerebral ischemia and neurodegenerative disorders. The action of JNK can be inhibited by the D-retro-inverso form of JNK inhibitor peptide (D-JNKI1), which totally prevents death induced by N-methyl-D-aspartate (NMDA) in vitro and strongly protects against different in vivo paradigms of excitotoxicity. To obtain optimal neuroprotection, it is imperative to elucidate the prosurvival action of D-JNKI1 and the death pathways that it inhibits. In cortical neuronal cultures, we first investigate the pathways by which NMDA induces JNK activation and show a rapid and selective phosphorylation of mitogen-activated protein kinase kinase 7 (MKK7), whereas the only other known JNK activator, mitogen-activated protein kinase kinase 4 (MKK4), was unaffected. We then analyze the action of D-JNKI1 on four JNK targets containing a JNK-binding domain: MAPK-activating death domain-containing protein/differentially expressed in normal and neoplastic cells (MADD/DENN), MKK7, MKK4 and JNK-interacting protein-1 (IB1/JIP-1).Cell Death and Differentiation (2007) 14, 240–253. doi:10.1038/sj.cdd.4401988; published online 23 June 2006 [ABSTRACT FROM AUTHOR]
- Published
- 2007
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11. C-JUN NH2-TERMINAL KINASE MEDIATES THE HEPATIC STRESS RESPONSE AFTER RAT HEMORRHAGE/RESUSCITATION.
- Author
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Relja, B., Schwestka, B., Sun-Young Lee, V., Henrich, D., Czerny, C., Borsello, T., Lehnert, M., and Marzi, I.
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- 2006
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12. Super-resolution study of PIAS SUMO E3-ligases in hippocampal and cortical neurons
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Tiziana Borsello, Luca Colnaghi, Clara Alice Musi, Luca Russo, Andrea Conz, Conz, A., Musi, C. A., Russo, L., Borsello, T., and Colnaghi, L.
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Gene isoform ,Histology ,hippocampus ,QH301-705.5 ,Ubiquitin-Protein Ligases ,hipppocampus ,Biophysics ,SUMO protein ,Regulator ,neurons ,Hippocampal formation ,Biology ,Hippocampus ,Article ,Synapse ,Mice ,synapse ,Animals ,Premovement neuronal activity ,Protein inhibitor of activated STAT ,Biology (General) ,Cerebral Cortex ,Neurons ,Microscopy ,Sumoylation ,Cell Biology ,Protein Inhibitors of Activated STAT ,Cell biology ,cortex ,SUMO ,Synaptic plasticity ,Cortex - Abstract
The SUMOylation machinery is a regulator of neuronal activity and synaptic plasticity. It is composed of SUMO isoforms and specialized enzymes named E1, E2 and E3 SUMO ligases. Recent studies have highlighted how SUMO isoforms and E2 enzymes localize with synaptic markers to support previous functional studies but less information is available on E3 ligases. PIAS proteins - belonging to the protein inhibitor of activated STAT (PIAS) SUMO E3-ligase family - are the best-characterized SUMO E3-ligases and have been linked to the formation of spatial memory in rodents. Whether however they exert their function co-localizing with synaptic markers is still unclear. In this study, we applied for the first time structured illumination microscopy (SIM) to PIAS ligases to investigate the co-localization of PIAS1 and PIAS3 with synaptic markers in hippocampal and cortical murine neurons. The results indicate partial co-localization of PIAS1 and PIAS3 with synaptic markers in hippocampal neurons and much rarer occurrence in cortical neurons. This is in line with previous super-resolution reports describing the co-localization with synaptic markers of other components of the SUMOylation machinery.
- Published
- 2021
13. Neuronal Localization of SENP Proteins with Super Resolution Microscopy
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Luana Fioriti, Luca Colnaghi, Luca Russo, Mario Salmona, Andrea Conz, Tiziana Borsello, Clara Alice Musi, Colnaghi, L, Conz, A, Russo, L, Musi, Ca, Fioriti, L, Borsello, T, and Salmona
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Protein sumoylation ,SENP1 ,medicine.medical_treatment ,SENP6 ,SUMO protein ,neurons ,Article ,lcsh:RC321-571 ,03 medical and health sciences ,0302 clinical medicine ,SENPs ,medicine ,lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry ,030304 developmental biology ,0303 health sciences ,Protease ,biology ,Chemistry ,General Neuroscience ,Colocalization ,Cysteine protease ,Cell biology ,SUMO ,super resolution microscopy ,Synaptophysin ,biology.protein ,030217 neurology & neurosurgery - Abstract
SUMOylation of proteins plays a key role in modulating neuronal function. For this reason, the balance between protein SUMOylation and deSUMOylation requires fine regulation to guarantee the homeostasis of neural tissue. While extensive research has been carried out on the localization and function of small ubiquitin-related modifier (SUMO) variants in neurons, less attention has been paid to the SUMO-specific isopeptidases that constitute the human SUMO-specific isopeptidase (SENP)/Ubiquitin-Specific Protease (ULP) cysteine protease family (SENP1-3 and SENP5-7). Here, for the first time, we studied the localization of SENP1, SENP6, and SENP7 in cultured hippocampal primary neurons at a super resolution detail level, with structured illumination microscopy (SIM). We found that the deSUMOylases partially colocalize with pre- and post-synaptic markers such as synaptophysin and drebrin. Thus, further confirming the presence with synaptic markers of the negative regulators of the SUMOylation machinery.
- Published
- 2020
14. The cell-permeable Aβ1-6A2VTAT(D) peptide reverts synaptopathy induced by Aβ1-42wt
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Mario Salmona, Giuseppe Di Fede, Fabrizio Tagliavini, Sara Cimini, Laura Colombo, Alfredo Cagnotto, Simona Mancini, Alessandra Sclip, Tiziana Borsello, Massimo Messa, Cimini, S, Sclip, A, Mancini, S, Colombo, L, Messa, M, Cagnotto, A, Di Fede, G, Tagliavini, F, Salmona, M, and Borsello, T
- Subjects
0301 basic medicine ,Cell Membrane Permeability ,Aβ oligomers ,Synaptic injury ,Aβ oligomer ,Dendritic Spines ,Peptide ,Mice, Transgenic ,Biology ,medicine.disease_cause ,Neuroprotection ,Hippocampus ,lcsh:RC321-571 ,03 medical and health sciences ,Mice ,In vivo ,Alzheimer Disease ,Brainbow hippocampal neurons ,medicine ,Animals ,lcsh:Neurosciences. Biological psychiatry. Neuropsychiatry ,chemistry.chemical_classification ,Neurons ,Mutation ,Cell-permeable peptide ,Amyloid beta-Peptides ,Wild type ,Long-term potentiation ,Alzheimer's disease ,medicine.disease ,Peptide Fragments ,Cell biology ,Disease Models, Animal ,030104 developmental biology ,Biochemistry ,chemistry ,Brainbow hippocampal neuron ,Neurology ,Synapses ,Synaptopathy ,A673V mutation - Abstract
Alzheimer disease (AD) is the most prevalent form of dementia. Loss of hippocampal synapses is the first neurodegenerative event in AD. Synaptic loss has been associated with the accumulation in the brain parenchyma of soluble oligomeric forms of amyloid β peptide (Aβ1-42wt). Clinical observations have shown that a mutation in the APP protein (A673V) causes an early onset AD-type dementia in homozygous carriers while heterozygous carriers are unaffected. This mutation leads to the formation of mutated Aβ peptides (Aβ1-42A2V) in homozygous patients, while in heterozygous subjects both Aβ1-42wt and Aβ1-42A2V are present. To better understand the impact of the A673V mutation in AD, we analyzed the synaptotoxic effect of oligomers formed by aggregation of different Aβ peptides (Aβ1-42wt or Aβ1-42A2V) and the combination of the two Aβ1-42MIX (Aβ1-42wt and Aβ1-42A2V) in an in vitro model of synaptic injury. We showed that Aβ1-42A2V oligomers are more toxic than Aβ1-42wt oligomers in hippocampal neurons, confirming the results previously obtained in cell lines. Furthermore, we reported that oligomers obtained by the combination of both wild type and mutated peptides (Aβ1-42MIX) did not exert synaptic toxicity. We concluded that the combination of Aβ1-42wt and Aβ1-42A2V peptides hinders the toxicity of Aβ1-42A2V and counteracts the manifestation of synaptopathy in vitro. Finally we took advantage of this finding to generate a cell-permeable peptide for clinical application, by fusing the first six residues of the Aβ1-42A2V to the TAT cargo sequence (Aβ1-6A2VTAT(D)). Noteworthy, the treatment with Aβ1-6A2VTAT(D) confers neuroprotection against both in vitro and in vivo synaptopathy models. Therefore Aβ1-6A2VTAT(D) may represent an innovative therapeutic tool to prevent synaptic degeneration in AD.
- Published
- 2015
15. A New Fluorogenic Peptide Determines Proteasome Activity in Single Cells
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Rodolfo Gonella Diaza, Mario Salmona, Simone Cenci, Enrico Monzani, Paolo Cascio, Roberto Sitia, Tiziana Borsello, Enrico Davoli, Pietro Veglianese, Niccolò Pengo, Ada De Luigi, Eugenio Erba, Andrea Carrà, Silvana Anna Maria Urru, Elena Fumagalli, Gianluigi Forloni, URRU S. A., M, Veglianese, P, DE LUIGI, A, Fumagalli, E, Erba, E, GONELLA DIAZA, R, Carra, A, Davoli, E, Borsello, T, Forloni, G, Pengo, N, Monzani, E, Cascio, P, Cenci, S, Sitia, R, and Salmona, M
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Models, Molecular ,Proteasome Endopeptidase Complex ,Cell ,Proteasome ,Protein degradation ,Proteasomal activity ,TAT ,Peptide ,Hippocampus ,Flow cytometry ,Mice ,chemistry.chemical_compound ,In vivo ,Cell Line, Tumor ,Drug Discovery ,medicine ,Animals ,Humans ,Fluorescent Dyes ,Neurons ,chemistry.chemical_classification ,medicine.diagnostic_test ,Chemistry ,Flow Cytometry ,In vitro ,Cell biology ,Protein Subunits ,medicine.anatomical_structure ,Microscopy, Fluorescence ,Biochemistry ,EDANS ,Molecular Medicine ,Peptides - Abstract
The ubiquitin−proteasome system plays a critical role in many diseases, making it an attractive biomarker and therapeutic target. However, the impact of results obtained in vitro using purified proteasome particles or whole cell extracts is limited by the lack of efficient methods to assess proteasome activity in living cells. We have engineered an internally quenched fluorogenic peptide with a proteasome-specific cleavage motif fused to TAT and linked to the fluorophores DABCYL and EDANS. This peptide penetrates cell membranes and is rapidly cleaved by the proteasomal chymotrypsin-like activity, generating a quantitative fluorescent reporter of in vivo proteasome activity as assessed by time-lapse or flow cytometry fluorescence analysis. This reporter is an innovative tool for monitoring proteasomal proteolytic activities in physiological and pathological conditions.
- Published
- 2010
16. Soluble Aβ oligomer-induced synaptopathy: c-Jun N-terminal kinase's role
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Laura Colombo, Egbert Welker, Federica Morelli, Simona Mancini, Andrea Arnaboldi, Pietro Veglianese, Isabella Colombo, Tiziana Borsello, Mario Salmona, Massimo Messa, Alessandra Sclip, Xanthi Antoniou, Sara Cimini, Sclip, A, Arnaboldi, A, Colombo, I, Veglianese, P, Colombo, L, Messa, M, Mancini, S, Cimini, S, Morelli, F, Antoniou, X, Welker, E, Salmona, M, and Borsello, T
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Dendritic spine ,Dendritic Spines ,Caspase 3 ,Alzhreimer's disease ,Mitogen-activated protein kinase kinase ,Models, Biological ,Receptors, N-Methyl-D-Aspartate ,Oligomer ,Mice ,chemistry.chemical_compound ,Alzheimer Disease ,Genetics ,medicine ,Animals ,Humans ,Receptors, AMPA ,Molecular Biology ,Amyloid beta-Peptides ,Chemistry ,Kinase ,c-jun ,JNK Mitogen-Activated Protein Kinases ,Cell Biology ,General Medicine ,medicine.disease ,c-Jun N-terminal kinase, JNK, Amyloid-beta oligomers ,Cell biology ,Synaptopathy ,Signal transduction ,Signal Transduction - Published
- 2013
17. JNK Activation Correlates with Cognitive Impairment and Alteration of the Post-Synaptic Element in the 5xFAD AD Mouse Model.
- Author
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Priori EC, Musi CA, Giani A, Colnaghi L, Milic I, Devitt A, Borsello T, and Repici M
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- Animals, Mice, MAP Kinase Signaling System, Phosphorylation, Disease Models, Animal, Alzheimer Disease metabolism, Cognitive Dysfunction metabolism, JNK Mitogen-Activated Protein Kinases metabolism
- Abstract
The c-Jun N-terminal kinases (JNKs) are a family of proteins that, once activated by stress stimuli, can alter neuronal functions and survival. The JNK cascade plays a crucial role in the post-synaptic neuronal compartment by altering its structural organization and leading, at worst, to an overall impairment of neuronal communication. Increasing evidence suggests that synaptic impairment is the first neurodegenerative event in Alzheimer's disease (AD). To better elucidate this mechanism, we longitudinally studied 5xFAD mice at three selected time points representative of human AD symptom progression. We tested the mice cognitive performance by using the radial arm water maze (RAWM) in parallel with biochemical evaluations of post-synaptic enriched protein fraction and total cortical parenchyma. We found that 5xFAD mice presented a strong JNK activation at 3.5 months of age in the post-synaptic enriched protein fraction. This JNK activation correlates with a structural alteration of the post-synaptic density area and with memory impairment at this early stage of the disease that progressively declines to cause cell death. These findings pave the way for future studies on JNK as a key player in early neurodegeneration and as an important therapeutic target for the development of new compounds able to tackle synaptic impairment in the early phase of AD pathology.
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- 2023
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18. Synaptic alterations as a common phase in neurological and neurodevelopmental diseases: JNK is a key mediator in synaptic changes.
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Musi CA, Bonadonna C, and Borsello T
- Abstract
Competing Interests: None
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- 2023
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19. Salivary microRNA profiling dysregulation in autism spectrum disorder: A pilot study.
- Author
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Kalemaj Z, Marino MM, Santini AC, Tomaselli G, Auti A, Cagetti MG, Borsello T, Costantino A, Inchingolo F, Boccellino M, Di Domenico M, and Tartaglia GM
- Abstract
Introduction: Autism spectrum disorders (ASD) are the most prevalent neurobiological disorders in children. The etiology comprises genetic, epigenetic, and environmental factors such as dysfunction of the immune system. Epigenetic mechanisms are mainly represented by DNA methylation, histone modifications, and microRNAs (miRNA). The major explored epigenetic mechanism is mediated by miRNAs which target genes known to be involved in ASD pathogenesis. Salivary poly-omic RNA measurements have been associated with ASD and are helpful to differentiate ASD endophenotypes. This study aims to comprehensively examine miRNA expression in children with ASD and to reveal potential biomarkers and possible disease mechanisms so that they can be used to improve faction between individuals by promoting more personalized therapeutic approaches., Materials and Methods: Saliva samples were collected from 10 subjects: 5 samples of children with ASD and 5 from healthy controls. miRNAs were analyzed using an Illumina Next-Generation-Sequencing (NGS) system., Results: Preliminary data highlighted the presence of 365 differentially expressed miRNAs. Pathway analysis, molecular function, biological processes, and target genes of 41 dysregulated miRNAs were assessed, of which 20 were upregulated, and 21 were downregulated in children with ASD compared to healthy controls., Conclusion: The results of this study represent preliminary but promising data, as the identified miRNA pathways could represent useful biomarkers for the early non-invasive diagnosis of ASD., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Kalemaj, Marino, Santini, Tomaselli, Auti, Cagetti, Borsello, Costantino, Inchingolo, Boccellino, Di Domenico and Tartaglia.)
- Published
- 2022
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20. Effect of 3D Synthetic Microscaffold Nichoid on the Morphology of Cultured Hippocampal Neurons and Astrocytes.
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Musi CA, Colnaghi L, Giani A, Priori EC, Marchini G, Tironi M, Conci C, Cerullo G, Osellame R, Raimondi MT, Remuzzi A, and Borsello T
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- Coculture Techniques, Hippocampus, Humans, Neurons metabolism, Astrocytes, Brain Diseases metabolism
- Abstract
The human brain is the most complex organ in biology. This complexity is due to the number and the intricate connections of brain cells and has so far limited the development of in vitro models for basic and applied brain research. We decided to create a new, reliable, and cost-effective in vitro system based on the Nichoid, a 3D microscaffold microfabricated by two-photon laser polymerization technology. We investigated whether these 3D microscaffold devices can create an environment allowing the manipulation, monitoring, and functional assessment of a mixed population of brain cells in vitro. With this aim, we set up a new model of hippocampal neurons and astrocytes co-cultured in the Nichoid microscaffold to generate brain micro-tissues of 30 μm thickness. After 21 days in culture, we morphologically characterized the 3D spatial organization of the hippocampal astrocytes and neurons within the microscaffold, and we compared our observations to those made using the classical 2D co-culture system. We found that the co-cultured cells colonized the entire volume of the 3D devices. Using confocal microscopy, we observed that within this period the different cell types had become well-differentiated. This was further elaborated with the use of drebrin, PSD-95, and synaptophysin antibodies that labeled the majority of neurons, both in the 2D as well as in the 3D co-cultures. Using scanning electron microscopy, we found that neurons in the 3D co-culture displayed a significantly larger amount of dendritic protrusions compared to neurons in the 2D co-culture. This latter observation indicates that neurons growing in a 3D environment may be more prone to form connections than those co-cultured in a 2D condition. Our results show that the Nichoid can be used as a 3D device to investigate the structure and morphology of neurons and astrocytes in vitro. In the future, this model can be used as a tool to study brain cell interactions in the discovery of important mechanisms governing neuronal plasticity and to determine the factors that form the basis of different human brain diseases. This system may potentially be further used for drug screening in the context of various brain diseases.
- Published
- 2022
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21. Colocalization and Interaction Study of Neuronal JNK3, JIP1, and β-Arrestin2 Together with PSD95.
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Musi CA, Marchini G, Giani A, Tomaselli G, Priori EC, Colnaghi L, and Borsello T
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- Disks Large Homolog 4 Protein metabolism, Humans, JNK Mitogen-Activated Protein Kinases metabolism, Male, Neurons metabolism, Phosphorylation, beta-Arrestin 1, Mitogen-Activated Protein Kinase 10 metabolism, Mitogen-Activated Protein Kinases metabolism
- Abstract
c-Jun N-terminal kinases (JNKs) are stress-activated serine/threonine protein kinases belonging to the mitogen-activated protein kinase (MAPK) family. Among them, JNK3 is selectively expressed in the central nervous system, cardiac smooth muscle, and testis. In addition, it is the most responsive JNK isoform to stress stimuli in the brain, and it is involved in synaptic dysfunction, an essential step in neurodegenerative processes. JNK3 pathway is organized in a cascade of amplification in which signal transduction occurs by stepwise, highly controlled phosphorylation. Since different MAPKs share common upstream activators, pathway specificity is guaranteed by scaffold proteins such as JIP1 and β-arrestin2. To better elucidate the physiological mechanisms regulating JNK3 in neurons, and how these interactions may be involved in synaptic (dys)function, we used (i) super-resolution microscopy to demonstrate the colocalization among JNK3-PSD95-JIP1 and JNK3-PSD95-β-arrestin2 in cultured hippocampal neurons, and (ii) co-immunoprecipitation techniques to show that the two scaffold proteins and JNK3 can be found interacting together with PSD95. The protein-protein interactions that govern the formation of these two complexes, JNK3-PSD95-JIP1 and JNK3-PSD95-β-arrestin2, may be used as targets to interfere with their downstream synaptic events.
- Published
- 2022
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22. JNK signaling provides a novel therapeutic target for Rett syndrome.
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Musi CA, Castaldo AM, Valsecchi AE, Cimini S, Morello N, Pizzo R, Renieri A, Meloni I, Bonati M, Giustetto M, and Borsello T
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- Animals, Disease Models, Animal, MAP Kinase Signaling System, Mice, Neurons metabolism, Synapses metabolism, Rett Syndrome genetics, Rett Syndrome metabolism, Rett Syndrome therapy
- Abstract
Background: Rett syndrome (RTT) is a monogenic X-linked neurodevelopmental disorder characterized by loss-of-function mutations in the MECP2 gene, which lead to structural and functional changes in synapse communication, and impairments of neural activity at the basis of cognitive deficits that progress from an early age. While the restoration of MECP2 in animal models has been shown to rescue some RTT symptoms, gene therapy intervention presents potential side effects, and with gene- and RNA-editing approaches still far from clinical application, strategies focusing on signaling pathways downstream of MeCP2 may provide alternatives for the development of more effective therapies in vivo. Here, we investigate the role of the c-Jun N-terminal kinase (JNK) stress pathway in the pathogenesis of RTT using different animal and cell models and evaluate JNK inhibition as a potential therapeutic approach., Results: We discovered that the c-Jun N-terminal kinase (JNK) stress pathway is activated in Mecp2-knockout, Mecp2-heterozygous mice, and in human MECP2-mutated iPSC neurons. The specific JNK inhibitor, D-JNKI1, promotes recovery of body weight and locomotor impairments in two mouse models of RTT and rescues their dendritic spine alterations. Mecp2-knockout presents intermittent crises of apnea/hypopnea, one of the most invalidating RTT pathological symptoms, and D-JNKI1 powerfully reduces this breathing dysfunction. Importantly, we discovered that also neurons derived from hiPSC-MECP2 mut show JNK activation, high-phosphorylated c-Jun levels, and cell death, which is not observed in the isogenic control wt allele hiPSCs. Treatment with D-JNKI1 inhibits neuronal death induced by MECP2 mutation in hiPSCs mut neurons., Conclusions: As a summary, we found altered JNK signaling in models of RTT and suggest that D-JNKI1 treatment prevents clinical symptoms, with coherent results at the cellular, molecular, and functional levels. This is the first proof of concept that JNK plays a key role in RTT and its specific inhibition offers a new and potential therapeutic tool to tackle RTT., (© 2021. The Author(s).)
- Published
- 2021
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23. Super-resolution study of PIAS SUMO E3-ligases in hippocampal and cortical neurons.
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Conz A, Musi CA, Russo L, Borsello T, and Colnaghi L
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- Animals, Cerebral Cortex ultrastructure, Hippocampus ultrastructure, Mice, Neurons ultrastructure, Cerebral Cortex enzymology, Hippocampus enzymology, Microscopy methods, Neurons enzymology, Protein Inhibitors of Activated STAT metabolism, Sumoylation, Ubiquitin-Protein Ligases metabolism
- Abstract
The SUMOylation machinery is a regulator of neuronal activity and synaptic plasticity. It is composed of SUMO isoforms and specialized enzymes named E1, E2 and E3 SUMO ligases. Recent studies have highlighted how SUMO isoforms and E2 enzymes localize with synaptic markers to support previous functional studies but less information is available on E3 ligases. PIAS proteins - belonging to the protein inhibitor of activated STAT (PIAS) SUMO E3-ligase family - are the best-characterized SUMO E3-ligases and have been linked to the formation of spatial memory in rodents. Whether however they exert their function co-localizing with synaptic markers is still unclear. In this study, we applied for the first time structured illumination microscopy (SIM) to PIAS ligases to investigate the co-localization of PIAS1 and PIAS3 with synaptic markers in hippocampal and cortical murine neurons. The results indicate partial co-localization of PIAS1 and PIAS3 with synaptic markers in hippocampal neurons and much rarer occurrence in cortical neurons. This is in line with previous super-resolution reports describing the co-localization with synaptic markers of other components of the SUMOylation machinery.
- Published
- 2021
- Full Text
- View/download PDF
24. c-Jun N-terminal kinase 1 (JNK1) modulates oligodendrocyte progenitor cell architecture, proliferation and myelination.
- Author
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Lorenzati M, Boda E, Parolisi R, Bonato M, Borsello T, Herdegen T, Buffo A, and Vercelli A
- Subjects
- Animals, Mice, Mice, Knockout, Mitogen-Activated Protein Kinase 8 genetics, Myelin Sheath genetics, Cell Proliferation, MAP Kinase Signaling System, Mitogen-Activated Protein Kinase 8 metabolism, Myelin Sheath metabolism, Oligodendrocyte Precursor Cells enzymology, Oligodendroglia enzymology
- Abstract
During Central Nervous System ontogenesis, myelinating oligodendrocytes (OLs) arise from highly ramified and proliferative precursors called oligodendrocyte progenitor cells (OPCs). OPC architecture, proliferation and oligodendro-/myelino-genesis are finely regulated by the interplay of cell-intrinsic and extrinsic factors. A variety of extrinsic cues converge on the extracellular signal-regulated kinase/mitogen activated protein kinase (ERK/MAPK) pathway. Here we found that the germinal ablation of the MAPK c-Jun N-Terminal Kinase isoform 1 (JNK1) results in a significant reduction of myelin in the cerebral cortex and corpus callosum at both postnatal and adult stages. Myelin alterations are accompanied by higher OPC density and proliferation during the first weeks of life, consistent with a transient alteration of mechanisms regulating OPC self-renewal and differentiation. JNK1 KO OPCs also show smaller occupancy territories and a less complex branching architecture in vivo. Notably, these latter phenotypes are recapitulated in pure cultures of JNK1 KO OPCs and of WT OPCs treated with the JNK inhibitor D-JNKI-1. Moreover, JNK1 KO and WT D-JNKI-1 treated OLs, while not showing overt alterations of differentiation in vitro, display a reduced surface compared to controls. Our results unveil a novel player in the complex regulation of OPC biology, on the one hand showing that JNK1 ablation cell-autonomously determines alterations of OPC proliferation and branching architecture and, on the other hand, suggesting that JNK1 signaling in OLs participates in myelination in vivo.
- Published
- 2021
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25. Neuronal Localization of SENP Proteins with Super Resolution Microscopy.
- Author
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Colnaghi L, Conz A, Russo L, Musi CA, Fioriti L, Borsello T, and Salmona M
- Abstract
SUMOylation of proteins plays a key role in modulating neuronal function. For this reason, the balance between protein SUMOylation and deSUMOylation requires fine regulation to guarantee the homeostasis of neural tissue. While extensive research has been carried out on the localization and function of small ubiquitin-related modifier (SUMO) variants in neurons, less attention has been paid to the SUMO-specific isopeptidases that constitute the human SUMO-specific isopeptidase (SENP)/Ubiquitin-Specific Protease (ULP) cysteine protease family (SENP1-3 and SENP5-7). Here, for the first time, we studied the localization of SENP1, SENP6, and SENP7 in cultured hippocampal primary neurons at a super resolution detail level, with structured illumination microscopy (SIM). We found that the deSUMOylases partially colocalize with pre- and post-synaptic markers such as synaptophysin and drebrin. Thus, further confirming the presence with synaptic markers of the negative regulators of the SUMOylation machinery.
- Published
- 2020
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26. JNK3 as Therapeutic Target and Biomarker in Neurodegenerative and Neurodevelopmental Brain Diseases.
- Author
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Musi CA, Agrò G, Santarella F, Iervasi E, and Borsello T
- Subjects
- Biomarkers metabolism, Brain pathology, Humans, MAP Kinase Signaling System genetics, Molecular Targeted Therapy, Neurodegenerative Diseases metabolism, Neurodegenerative Diseases pathology, Neurodevelopmental Disorders metabolism, Neurodevelopmental Disorders pathology, Phosphorylation, Brain metabolism, Mitogen-Activated Protein Kinase 10 genetics, Neurodegenerative Diseases genetics, Neurodevelopmental Disorders genetics
- Abstract
The c-Jun N -terminal kinase 3 (JNK3) is the JNK isoform mainly expressed in the brain. It is the most responsive to many stress stimuli in the central nervous system from ischemia to Aβ oligomers toxicity. JNK3 activity is spatial and temporal organized by its scaffold protein, in particular JIP-1 and β-arrestin-2, which play a crucial role in regulating different cellular functions in different cellular districts. Extensive evidence has highlighted the possibility of exploiting these adaptors to interfere with JNK3 signaling in order to block its action. JNK plays a key role in the first neurodegenerative event, the perturbation of physiological synapse structure and function, known as synaptic dysfunction. Importantly, this is a common mechanism in many different brain pathologies. Synaptic dysfunction and spine loss have been reported to be pharmacologically reversible, opening new therapeutic directions in brain diseases. Being JNK3-detectable at the peripheral level, it could be used as a disease biomarker with the ultimate aim of allowing an early diagnosis of neurodegenerative and neurodevelopment diseases in a still prodromal phase.
- Published
- 2020
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27. JNK signaling activation in the Ube3a maternal deficient mouse model: its specific inhibition prevents post-synaptic protein-enriched fraction alterations and cognitive deficits in Angelman Syndrome model.
- Author
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Musi CA, Agrò G, Buccarello L, Camuso S, and Borsello T
- Subjects
- Animals, Cell Nucleus metabolism, Disease Models, Animal, Female, Hippocampus metabolism, Male, Mice, Neurons metabolism, Angelman Syndrome metabolism, Cognitive Dysfunction metabolism, MAP Kinase Signaling System, Synapses metabolism, Ubiquitin-Protein Ligases metabolism
- Abstract
Deficiency of the E3 ubiquitin ligase UBE3A leads to the neurodevelopmental disorder Angelman syndrome (AS), while higher levels are linked to autism spectrum disorder. The mechanisms underlying the downstream effects of UBE3A loss or gain of function in these disorders are still not well understood, and treatments are still lacking. Here, using the Ube3a maternal loss (Ube3a
m-/p+ ) mouse model, we report an important JNK signaling activation in the hippocampus, cortex and cerebellum correlating with the onset of behavioral defects and biochemical marker alterations in the post-synaptic element, suggesting important spine pathology. JNK activation occurs at 7 and persists up till 23 weeks in Ube3am-/p+ mice in two different cellular compartments: the nucleus and the post-synaptic protein-enriched fraction. To study JNK's role in Ube3am-/p+ pathology we treated mice with the specific JNK inhibitor peptide, D-JNKI1, from 7 to 23 weeks of age. Preventing JNK action in vivo restores the post-synaptic protein-enriched fraction defects and the cognitive impairment in these mice. Our results imply a critical role of UBE3A-JNK signaling in the pathogenesis of UBE3A-related disorders. In particular, it was clear that JNK is a key player in regulating AS synaptic alterations and the correlated cognitive impairments, in fact, its specific inhibition tackles Ube3am-/p+ pathology. This study sheds new light on the neuronal functions of UBE3A and offers new prospects for understanding the pathogenesis of UBE3A-related disorders., Competing Interests: Declaration of Competing Interest The authors declare no actual or potential conflicts of interest., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)- Published
- 2020
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28. Pharmacological c-Jun NH 2 -Terminal Kinase (JNK) Pathway Inhibition Reduces Severity of Spinal Muscular Atrophy Disease in Mice.
- Author
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Schellino R, Boido M, Borsello T, and Vercelli A
- Abstract
Spinal muscular atrophy (SMA) is a severe neurodegenerative disorder that occurs in early childhood. The disease is caused by the deletion/mutation of the survival motor neuron 1 (SMN1) gene resulting in progressive skeletal muscle atrophy and paralysis, due to the degeneration of spinal motor neurons (MNs). Currently, the cellular and molecular mechanisms underlying MN death are only partly known, although recently it has been shown that the c-Jun NH
2 -terminal kinase (JNK)-signaling pathway might be involved in the SMA pathogenesis. After confirming the activation of JNK in our SMA mouse model (SMN2+/+; SMNΔ7+/+; Smn-/-), we tested a specific JNK-inhibitor peptide (D-JNKI1) on these mice, by chronic administration from postnatal day 1 to 10, and histologically analyzed the spinal cord and quadriceps muscle at age P12. We observed that D-JNKI1 administration delayed MN death and decreased inflammation in spinal cord. Moreover, the inhibition of JNK pathway improved the trophism of SMA muscular fibers and the size of the neuromuscular junctions (NMJs), leading to an ameliorated innervation of the muscles that resulted in improved motor performances and hind-limb muscular tone. Finally, D-JNKI1 treatment slightly, but significantly increased lifespan in SMA mice. Thus, our results identify JNK as a promising target to reduce MN cell death and progressive skeletal muscle atrophy, providing insight into the role of JNK-pathway for developing alternative pharmacological strategies for the treatment of SMA.- Published
- 2018
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29. Axonal transport in a peripheral diabetic neuropathy model: sex-dimorphic features.
- Author
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Pesaresi M, Giatti S, Spezzano R, Romano S, Diviccaro S, Borsello T, Mitro N, Caruso D, Garcia-Segura LM, and Melcangi RC
- Subjects
- Animals, Female, Ganglia, Spinal metabolism, Gonadal Steroid Hormones metabolism, Kinesins genetics, Male, Myosin Type V genetics, Myosin Type V metabolism, Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha genetics, Rats, Sprague-Dawley, Sciatic Nerve metabolism, Axonal Transport, Diabetes Mellitus, Experimental genetics, Diabetes Mellitus, Experimental metabolism, Diabetic Neuropathies genetics, Diabetic Neuropathies metabolism, Sex Characteristics
- Abstract
Background: Disruption of axonal transport plays a pivotal role in diabetic neuropathy. A sex-dimorphism exists in the incidence and symptomatology of diabetic neuropathy; however, no studies so far have addressed sex differences in axonal motor proteins expression in early diabetes as well as the possible involvement of neuroactive steroids. Interestingly, recent data point to a role for mitochondria in the sexual dimorphism of neurodegenerative diseases. Mitochondria have a fundamental role in axonal transport by producing the motors' energy source, ATP. Moreover, neuroactive steroids can also regulate mitochondrial function., Methods: Here, we investigated the impact of short-term diabetes in the peripheral nervous system of male and female rats on key motor proteins important for axonal transport, mitochondrial function, and neuroactive steroids levels., Results: We show that short-term diabetes alters mRNA levels and axoplasm protein contents of kinesin family member KIF1A, KIF5B, KIF5A and Myosin Va in male but not in female rats. Similarly, the expression of peroxisome proliferator-activated receptor γ co-activator-1α, a subunit of the respiratory chain complex IV, ATP levels and the key regulators of mitochondrial dynamics were affected in males but not in females. Concomitant analysis of neuroactive steroid levels in sciatic nerve showed an alteration of testosterone, dihydrotestosterone, and allopregnanolone in diabetic males, whereas no changes were observed in female rats., Conclusions: These findings suggest that sex-specific decrease in neuroactive steroid levels in male diabetic animals may cause an alteration in their mitochondrial function that in turn might impact in axonal transport, contributing to the sex difference observed in diabetic neuropathy.
- Published
- 2018
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30. The c-jun N-terminal kinase plays a key role in ocular degenerative changes in a mouse model of Alzheimer disease suggesting a correlation between ocular and brain pathologies.
- Author
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Buccarello L, Sclip A, Sacchi M, Castaldo AM, Bertani I, ReCecconi A, Maestroni S, Zerbini G, Nucci P, and Borsello T
- Abstract
Recently a range of ocular manifestations such as retinal and lens amyloid-beta accumulation and retinal nerve fiber layer loss have been proposed as potential biomarkers in Alzheimer disease (AD). The TgCRND8 mouse model of AD exhibits age-dependent amyloid β (Aβ) oligomers accumulation and cognitive defects, amyloid plaques and hyperphosphorylated Tau deposition and inflammation. We proved the correlation between ocular pathologies and AD, observing increased levels of p-APP and p-Tau, accumulation of Aβ oligomers in the retina, eye, and optic nerve. The accumulation of amyloid markers was significantly stronger in the retinal ganglion cell (RGC) layer, suggesting that RGC might be more susceptible to degeneration. We detected a thinning of the RGC layer as well as RGC death in the retina of TgCRND8 mice, by using a combination of Optical Coherence Tomography (OCT), immunofluorescence, immunohistochemistry and Western blotting techniques. We proved for the first time the key role of C-Jun N-terminal Kinase (JNK) in the ocular degeneration. In support of this, the administration of the JNK inhibitor, D-JNKI1, was able to counteract the Aβ and p-Tau accumulation in the retina of TgCRND8 mice, and consequently reduce RGCs loss. These results confirm that degenerative changes in the retina/eye of AD mouse model mirrors the events observed in the brain parenchyma. Ocular changes can be detected by non-invasive imaging techniques, such as OCT, to study and test different therapeutic strategies against degenerative events associated to AD., Competing Interests: CONFLICTS OF INTEREST The authors declare no actual or potential conflicts of interest.
- Published
- 2017
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31. The Tat-Aβ1-6A2V(D) peptide against AD synaptopathy.
- Author
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Buccarello L and Borsello T
- Subjects
- Animals, Brain metabolism, Cognitive Dysfunction metabolism, Disks Large Homolog 4 Protein metabolism, Humans, Mutation, Peptide Fragments, Phosphorylation, Protein Denaturation, Protein Folding, Receptors, AMPA metabolism, Receptors, N-Methyl-D-Aspartate metabolism, Synucleins metabolism, tau Proteins chemistry, Alzheimer Disease metabolism, Amyloid beta-Peptides chemistry, Synapses metabolism
- Published
- 2017
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- View/download PDF
32. Evidence of Presynaptic Localization and Function of the c-Jun N-Terminal Kinase.
- Author
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Biggi S, Buccarello L, Sclip A, Lippiello P, Tonna N, Rumio C, Di Marino D, Miniaci MC, and Borsello T
- Subjects
- Animals, Cerebral Cortex physiology, Excitatory Amino Acid Agonists pharmacology, Excitatory Postsynaptic Potentials, Female, Glycine pharmacology, Male, Mice, Mitogen-Activated Protein Kinase 10 metabolism, Mitogen-Activated Protein Kinase 9 metabolism, N-Methylaspartate pharmacology, Synaptosomes metabolism, Cerebral Cortex metabolism, JNK Mitogen-Activated Protein Kinases metabolism, Presynaptic Terminals enzymology, Receptors, N-Methyl-D-Aspartate metabolism, SNARE Proteins metabolism
- Abstract
The c-Jun N-terminal kinase (JNK) is part of a stress signalling pathway strongly activated by NMDA-stimulation and involved in synaptic plasticity. Many studies have been focused on the post-synaptic mechanism of JNK action, and less is known about JNK presynaptic localization and its physiological role at this site. Here we examined whether JNK is present at the presynaptic site and its activity after presynaptic NMDA receptors stimulation. By using N-SIM Structured Super Resolution Microscopy as well as biochemical approaches, we demonstrated that presynaptic fractions contained significant amount of JNK protein and its activated form. By means of modelling design, we found that JNK, via the JBD domain, acts as a physiological effector on T-SNARE proteins; then using biochemical approaches we demonstrated the interaction between Syntaxin-1-JNK, Syntaxin-2-JNK, and Snap25-JNK. In addition, taking advance of the specific JNK inhibitor peptide, D-JNKI1, we defined JNK action on the SNARE complex formation. Finally, electrophysiological recordings confirmed the role of JNK in the presynaptic modulation of vesicle release. These data suggest that JNK-dependent phosphorylation of T-SNARE proteins may have an important functional role in synaptic plasticity., Competing Interests: There is no actual or potential conflict of interests.
- Published
- 2017
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33. Molecular and Cellular Mechanisms of Synaptopathies.
- Author
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Ardiles AO, Grabrucker AM, Scholl FG, Rudenko G, and Borsello T
- Subjects
- Brain physiopathology, Synapses physiology
- Published
- 2017
- Full Text
- View/download PDF
34. Epidermal RAF prevents allergic skin disease.
- Author
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Raguz J, Jeric I, Niault T, Nowacka JD, Kuzet SE, Rupp C, Fischer I, Biggi S, Borsello T, and Baccarini M
- Subjects
- Animals, Mice, Proto-Oncogene Proteins B-raf genetics, Proto-Oncogene Proteins c-raf genetics, Dermatitis, Atopic pathology, Dermatitis, Atopic prevention & control, Keratinocytes physiology, Proto-Oncogene Proteins B-raf metabolism, Proto-Oncogene Proteins c-raf metabolism
- Abstract
The RAS pathway is central to epidermal homeostasis, and its activation in tumors or in Rasopathies correlates with hyperproliferation. Downstream of RAS, RAF kinases are actionable targets regulating keratinocyte turnover; however, chemical RAF inhibitors paradoxically activate the pathway, promoting epidermal proliferation. We generated mice with compound epidermis-restricted BRAF/RAF1 ablation. In these animals, transient barrier defects and production of chemokines and Th2-type cytokines by keratinocytes cause a disease akin to human atopic dermatitis, characterized by IgE responses and local and systemic inflammation. Mechanistically, BRAF and RAF1 operate independently to balance MAPK signaling: BRAF promotes ERK activation, while RAF1 dims stress kinase activation. In vivo, JNK inhibition prevents disease onset, while MEK/ERK inhibition in mice lacking epidermal RAF1 phenocopies it. These results support a primary role of keratinocytes in the pathogenesis of atopic dermatitis, and the animals lacking BRAF and RAF1 in the epidermis represent a useful model for this disease., Competing Interests: The authors declare that no competing interests exist.
- Published
- 2016
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35. A cationic tetrapyrrole inhibits toxic activities of the cellular prion protein.
- Author
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Massignan T, Cimini S, Stincardini C, Cerovic M, Vanni I, Elezgarai SR, Moreno J, Stravalaci M, Negro A, Sangiovanni V, Restelli E, Riccardi G, Gobbi M, Castilla J, Borsello T, Nonno R, and Biasini E
- Subjects
- Amyloid beta-Peptides metabolism, Animals, Binding Sites, Cell Line, Tumor, HEK293 Cells, Humans, Metalloporphyrins pharmacology, Mice, Inbred C57BL, Mice, Knockout, Mutation, Porphyrins, PrPC Proteins chemistry, PrPC Proteins genetics, Prion Proteins chemistry, Protein Binding, Recombinant Proteins metabolism, Tetrapyrroles pharmacology, Metalloporphyrins chemistry, PrPC Proteins metabolism, Prion Proteins antagonists & inhibitors, Tetrapyrroles chemistry
- Abstract
Prion diseases are rare neurodegenerative conditions associated with the conformational conversion of the cellular prion protein (PrP(C)) into PrP(Sc), a self-replicating isoform (prion) that accumulates in the central nervous system of affected individuals. The structure of PrP(Sc) is poorly defined, and likely to be heterogeneous, as suggested by the existence of different prion strains. The latter represents a relevant problem for therapy in prion diseases, as some potent anti-prion compounds have shown strain-specificity. Designing therapeutics that target PrP(C) may provide an opportunity to overcome these problems. PrP(C) ligands may theoretically inhibit the replication of multiple prion strains, by acting on the common substrate of any prion replication reaction. Here, we characterized the properties of a cationic tetrapyrrole [Fe(III)-TMPyP], which was previously shown to bind PrP(C), and inhibit the replication of a mouse prion strain. We report that the compound is active against multiple prion strains in vitro and in cells. Interestingly, we also find that Fe(III)-TMPyP inhibits several PrP(C)-related toxic activities, including the channel-forming ability of a PrP mutant, and the PrP(C)-dependent synaptotoxicity of amyloid-β (Aβ) oligomers, which are associated with Alzheimer's Disease. These results demonstrate that molecules binding to PrP(C) may produce a dual effect of blocking prion replication and inhibiting PrP(C)-mediated toxicity.
- Published
- 2016
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- View/download PDF
36. Differential Relevance of NF-κB and JNK in the Pathophysiology of Hemorrhage/Resususcitation-Induced Liver Injury after Chronic Ethanol Feeding.
- Author
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Relja B, Weber R, Maraslioglu M, Wagner N, Borsello T, Jobin C, Marzi I, and Lehnert M
- Subjects
- Alcoholism complications, Alcoholism physiopathology, Animals, Disease Models, Animal, Ethanol toxicity, Gene Expression Regulation, Interleukin-6 blood, Liver Diseases etiology, Liver Diseases metabolism, MAP Kinase Kinase 4 antagonists & inhibitors, Male, Mice, Inbred C57BL, NF-kappa B genetics, Peptides pharmacology, Phosphorylation, Resuscitation methods, Shock, Hemorrhagic metabolism, Tumor Necrosis Factor-alpha metabolism, Alcoholism metabolism, Liver Diseases physiopathology, MAP Kinase Kinase 4 metabolism, NF-kappa B metabolism, Resuscitation adverse effects, Shock, Hemorrhagic physiopathology
- Abstract
Background: Chronic ethanol (EtOH) abuse worsens pathophysiological derangements after hemorrhagic shock and resuscitation (H/R) that induce hepatic injury and strong inflammatory changes via JNK and NF-κB activation. Inhibiting JNK with a cell-penetrating, protease-resistant peptide D-JNKI-1 after H/R in mice with healthy livers ameliorated these effects. Here, we studied if JNK inhibition by D-JNKI-1 in chronically EtOH-fed mice after hemorrhagic shock prior to the onset of resuscitation also confers protection., Methods: Male mice were fed a Lieber-DeCarli diet containing EtOH or an isocaloric control (ctrl) diet for 4 weeks. Animals were hemorrhaged for 90 min (32 ± 2 mm Hg) and randomly received either D-JNKI-1 (11 mg/kg, intraperitoneally, i. p.) or sterile saline as vehicle (veh) immediately before the onset of resuscitation. Sham animals underwent surgical procedures without H/R and were either D-JNKI-1 or veh treated. Two hours after resuscitation, blood samples and liver tissue were harvested., Results: H/R induced hepatic injury with increased systemic interleukin (IL)-6 levels, and enhanced local gene expression of NF-κB-controlled genes such as intercellular adhesion molecule (ICAM)-1 and matrix metallopeptidase (MMP)9. c-Jun and NF-κB phosphorylation were increased after H/R. These effects were further increased in EtOH-fed mice after H/R. D-JNKI-1 application inhibited the proinflammatory changes and reduced significantly hepatic injury after H/R in ctrl-fed mice. Moreover, D-JNKI-1 reduces in ctrl-fed mice the H/R-induced c-Jun and NF-κB phosphorylation. However, in chronically EtOH-fed mice, JNK inhibition did not prevent the H/R-induced hepatic damage and proinflammatory changes nor c-Jun and NF-κB phosphorylation after H/R., Conclusions: These results indicate, that JNK inhibition is protective only in not pre-harmed liver after H/R. In contrast, the pronounced H/R-induced liver damage in mice being chronically fed with ethanol cannot be prevented by JNK inhibition after H/R and seems to be under the control of NF-κB.
- Published
- 2015
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37. Determination of tissue levels of a neuroprotectant drug: the cell permeable JNK inhibitor peptide.
- Author
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Davoli E, Sclip A, Cecchi M, Cimini S, Carrà A, Salmona M, and Borsello T
- Subjects
- Animals, Animals, Newborn, Blood-Brain Barrier metabolism, Brain metabolism, Cell Membrane drug effects, Cell Membrane metabolism, Kidney metabolism, Liver metabolism, Mice, Neurons drug effects, Peptides antagonists & inhibitors, Cell Membrane Permeability drug effects, Neuroprotective Agents metabolism, Neuroprotective Agents pharmacology, Peptides metabolism, Peptides pharmacology
- Abstract
Introduction: Cell permeable peptides (CPPs) represent a novel tool for the delivery of bioactive molecules into scarcely accessible organs, such as the brain. CPPs have been successfully used in pre-clinical studies for a variety of diseases, ranging from cancer to neurological disorders. However, the mechanisms by which CPPs cross biological membranes, as well as their pharmacokinetic properties, have been poorly explored due to the lack of specific and sensitive analytical methods., Methods: In this paper we describe a protocol to quantitatively determine the amount of CPPs in in vitro and in vivo experimental models. To this end we selected the peptide D-JNKI1 that was shown to prevent neurodegeneration in both acute and chronic degenerative disorders. This method allows an accurate quantitative analysis of D-JNKI1 in both neuronal lysates and tissue homogenates using mass spectrometry and stable isotope dilution approach., Results: We found that D-JNKI1 crosses cellular membranes with fast kinetics, through an active and passive mechanism. After acute intraperitoneal (ip) administration of D-JNKI1 in mice, the peptide was found in the main organs with particular regard to the liver and kidney. Interestingly, D-JNKI1 crosses the blood brain barrier (BBB) and reaches the brain, where it remains for one week., Discussion: The challenge lies in developing the clinical application of therapeutic cell permeable peptides. Discerning pharmacokinetic properties is a high priority to produce a powerful therapeutic strategy. Overall, our data shed light on the pharmacokinetic properties of D-JNKI1 and supports its powerful neuroprotective effect., (Copyright © 2014 Elsevier Inc. All rights reserved.)
- Published
- 2014
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38. Soluble Aβ oligomer-induced synaptopathy: c-Jun N-terminal kinase's role.
- Author
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Sclip A, Arnaboldi A, Colombo I, Veglianese P, Colombo L, Messa M, Mancini S, Cimini S, Morelli F, Antoniou X, Welker E, Salmona M, and Borsello T
- Subjects
- Alzheimer Disease, Amyloid beta-Peptides pharmacology, Animals, Caspase 3 metabolism, Caspase 3 physiology, Dendritic Spines drug effects, Dendritic Spines ultrastructure, Humans, JNK Mitogen-Activated Protein Kinases metabolism, Mice, Models, Biological, Receptors, AMPA metabolism, Receptors, N-Methyl-D-Aspartate metabolism, Signal Transduction, Amyloid beta-Peptides metabolism, Dendritic Spines metabolism, JNK Mitogen-Activated Protein Kinases physiology
- Published
- 2013
- Full Text
- View/download PDF
39. An N-terminal fragment of the prion protein binds to amyloid-β oligomers and inhibits their neurotoxicity in vivo.
- Author
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Fluharty BR, Biasini E, Stravalaci M, Sclip A, Diomede L, Balducci C, La Vitola P, Messa M, Colombo L, Forloni G, Borsello T, Gobbi M, and Harris DA
- Subjects
- Alzheimer Disease metabolism, Amyloidogenic Proteins chemistry, Animals, Benzothiazoles, Caenorhabditis elegans metabolism, Hippocampus metabolism, Mice, Mice, Inbred C57BL, Models, Biological, Neurodegenerative Diseases metabolism, Neurons metabolism, Peptides chemistry, Protein Binding, Protein Structure, Tertiary, Surface Plasmon Resonance, Synapses metabolism, Thiazoles chemistry, Amyloid beta-Peptides chemistry, Prions chemistry
- Abstract
A hallmark of Alzheimer disease (AD) is the accumulation of the amyloid-β (Aβ) peptide in the brain. Considerable evidence suggests that soluble Aβ oligomers are responsible for the synaptic dysfunction and cognitive deficit observed in AD. However, the mechanism by which these oligomers exert their neurotoxic effect remains unknown. Recently, it was reported that Aβ oligomers bind to the cellular prion protein with high affinity. Here, we show that N1, the main physiological cleavage fragment of the cellular prion protein, is necessary and sufficient for binding early oligomeric intermediates during Aβ polymerization into amyloid fibrils. The ability of N1 to bind Aβ oligomers is influenced by positively charged residues in two sites (positions 23-31 and 95-105) and is dependent on the length of the sequence between them. Importantly, we also show that N1 strongly suppresses Aβ oligomer toxicity in cultured murine hippocampal neurons, in a Caenorhabditis elegans-based assay, and in vivo in a mouse model of Aβ-induced memory dysfunction. These data suggest that N1, or small peptides derived from it, could be potent inhibitors of Aβ oligomer toxicity and represent an entirely new class of therapeutic agents for AD.
- Published
- 2013
- Full Text
- View/download PDF
40. The neurodegeneration in Alzheimer disease and the prion protein.
- Author
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Forloni G, Sclip A, Borsello T, and Balducci C
- Subjects
- Amyloid beta-Peptides analysis, Amyloid beta-Peptides toxicity, Animals, Brain metabolism, Humans, Prions analysis, Alzheimer Disease metabolism, Alzheimer Disease pathology, Amyloid beta-Peptides metabolism, Brain pathology, Prions metabolism
- Abstract
The concept of "prion-like" has been proposed to explain the pathogenic mechanism of the principal neurodegenerative disorders associated with protein misfolding, including Alzheimer disease (AD). Other evidence relates prion protein with AD: the cellular prion protein (PrP(C)) binds β amyloid oligomers, allegedly responsible for the neurodegeneration in AD, mediating their toxic effects. We and others have confirmed the high-affinity binding between β amyloid oligomers and PrP(C), but we were not able to assess the functional consequences of this interaction using behavioral investigations and in vitro tests. This discrepancy rather than being resolved with the classic explanations, differencies in methodological aspects, has been reinforced by new data from different sources. Here we present data obtained with PrP antibody that not interfere with the neurotoxic activity of β amyloid oligomers. Since the potential role of the PrP(C) in the neuronal dysfunction induced by β amyloid oligomers is an important issue, find reasonable explanation of the inconsistent results is needed. Even more important however is the relevance of this interaction in the context of the disease, so as to develop valid therapeutic strategies.
- Published
- 2013
- Full Text
- View/download PDF
41. Contribution of serine racemase/d-serine pathway to neuronal apoptosis.
- Author
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Esposito S, Pristerà A, Maresca G, Cavallaro S, Felsani A, Florenzano F, Manni L, Ciotti MT, Pollegioni L, Borsello T, and Canu N
- Subjects
- Animals, Apoptosis genetics, Cellular Senescence genetics, Cellular Senescence physiology, Cerebellum cytology, Cerebellum metabolism, In Vitro Techniques, JNK Mitogen-Activated Protein Kinases metabolism, Proteasome Endopeptidase Complex metabolism, RNA, Messenger genetics, RNA, Messenger metabolism, RNA, Small Interfering genetics, Racemases and Epimerases antagonists & inhibitors, Racemases and Epimerases genetics, Rats, Rats, Wistar, Receptors, N-Methyl-D-Aspartate metabolism, Serine chemistry, Signal Transduction, Stereoisomerism, Apoptosis physiology, Neurons cytology, Neurons metabolism, Racemases and Epimerases metabolism, Serine metabolism
- Abstract
Recent data indicate that age-related N-methyl-d-aspartate receptor (NMDAR) transmission impairment is correlated with the reduction in serine racemase (SR) expression and d-serine content. As apoptosis is associated with several diseases and conditions that generally occur with age, we investigated the modulation of SR/d-serine pathway during neuronal apoptosis and its impact on survival. We found that in cerebellar granule neurons (CGNs), undergoing apoptosis SR/d-serine pathway is crucially regulated. In the early phase of apoptosis, the expression of SR is reduced, both at the protein and RNA level through pathways, upstream of caspase activation, involving ubiquitin proteasome system (UPS) and c-Jun N-terminal kinases (JNKs). Forced expression of SR, together with treatment with NMDA and d-serine, blocks neuronal death, whereas pharmacological inhibition and Sh-RNA-mediated suppression of endogenous SR exacerbate neuronal death. In the late phase of apoptosis, the increased expression of SR contribute to the last, NMDAR-mediated, wave of cell death. These findings are relevant to our understanding of neuronal apoptosis and NMDAR activity regulation, raising further questions as to the role of SR/d-serine in those neuro-pathophysiological processes, such as aging and neurodegenerative diseases characterized by a convergence of apoptotic mechanisms and NMDAR dysfunction., (© 2012 The Authors. Aging Cell © 2012 Blackwell Publishing Ltd/Anatomical Society of Great Britain and Ireland.)
- Published
- 2012
- Full Text
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42. c-Jun N-terminal kinase regulates soluble Aβ oligomers and cognitive impairment in AD mouse model.
- Author
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Sclip A, Antoniou X, Colombo A, Camici GG, Pozzi L, Cardinetti D, Feligioni M, Veglianese P, Bahlmann FH, Cervo L, Balducci C, Costa C, Tozzi A, Calabresi P, Forloni G, and Borsello T
- Subjects
- Alzheimer Disease genetics, Animals, Cognition Disorders metabolism, Disease Models, Animal, Electrophysiology, Humans, Maze Learning, Memory Disorders genetics, Mice, Models, Biological, Peptides chemistry, Signal Transduction, Time Factors, Alzheimer Disease metabolism, Amyloid beta-Peptides chemistry, Gene Expression Regulation, Enzymologic, JNK Mitogen-Activated Protein Kinases metabolism
- Abstract
Alzheimer disease (AD) is characterized by cognitive impairment that starts with memory loss to end in dementia. Loss of synapses and synaptic dysfunction are closely associated with cognitive impairment in AD patients. Biochemical and pathological evidence suggests that soluble Aβ oligomers correlate with cognitive impairment. Here, we used the TgCRND8 AD mouse model to investigate the role of JNK in long term memory deficits. TgCRND8 mice were chronically treated with the cell-penetrating c-Jun N-terminal kinase inhibitor peptide (D-JNKI1). D-JNKI1, preventing JNK action, completely rescued memory impairments (behavioral studies) as well as the long term potentiation deficits of TgCRND8 mice. Moreover, D-JNKI1 inhibited APP phosphorylation in Thr-668 and reduced the amyloidogenic cleavage of APP and Aβ oligomers in brain parenchyma of treated mice. In conclusion, by regulating key pathogenic mechanisms of AD, JNK might hold promise as innovative therapeutic target.
- Published
- 2011
- Full Text
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43. Crosstalk between JNK and SUMO signaling pathways: deSUMOylation is protective against H2O2-induced cell injury.
- Author
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Feligioni M, Brambilla E, Camassa A, Sclip A, Arnaboldi A, Morelli F, Antoniou X, and Borsello T
- Subjects
- Cell Death, Cell Line, Tumor, Cell Survival, Enzyme Activation, Gene Expression Regulation, Enzymologic, Humans, Immunohistochemistry methods, Immunoprecipitation, Oxidative Stress, Plasmids metabolism, Proto-Oncogene Proteins c-jun metabolism, Signal Transduction, Tetrazolium Salts pharmacology, Thiazoles pharmacology, Time Factors, Hydrogen Peroxide metabolism, JNK Mitogen-Activated Protein Kinases metabolism, Small Ubiquitin-Related Modifier Proteins metabolism
- Abstract
Background: Oxidative stress is a key feature in the pathogenesis of several neurological disorders. Following oxidative stress stimuli a wide range of pathways are activated and contribute to cellular death. The mechanism that couples c-Jun N-terminal kinase (JNK) signaling, a key pathway in stress conditions, to the small ubiquitin-related modifier (SUMO), an emerging protein in the field, is largely unknown., Methodology/principal Findings: With this study we investigated if SUMOylation participates in the regulation of JNK activation as well as cellular death in a model of H(2)O(2) induced-oxidative stress. Our data show that H(2)O(2) modulates JNK activation and induces cellular death in neuroblastoma SH-SY5Y cells. Inhibition of JNK's action with the D-JNKI1 peptide rescued cells from death. Following H(2)O(2), SUMO-1 over-expression increased phosphorylation of JNK and exacerbated cell death, although only in conditions of mild oxidative stress. Furthermore inhibition of SUMOylation, following transfection with SENP1, interfered with JNK activation and rescued cells from H(2)O(2) induced death. Importantly, in our model, direct interaction between these proteins can occur., Conclusions/significance: Taken together our results show that SUMOylation may significantly contribute to modulation of JNK activation and contribute to cell death in oxidative stress conditions.
- Published
- 2011
- Full Text
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44. Cell Permeable Peptides: A Promising Tool to Deliver Neuroprotective Agents in the Brain.
- Author
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Antoniou X and Borsello T
- Abstract
The inability of most drugs to cross the blood-brain barrier and/or plasma membrane limits their use for biomedical applications in the brain. Cell Permeable Peptides (CPPs) overcome this problem and are effective in vivo, crossing the plasma membrane and the blood-brain barrier. CPPs deliver a wide variety of compounds intracellularly in an active form. In fact, many bioactive cargoes have neuroprotective properties, and due to their ability to block protein-protein interactions, offer exciting perspectives in the clinical setting. In this review we give an overview of the Cell Permeable Peptides strategy to deliver neuroprotectants against neurodegeneration in the CNS.
- Published
- 2010
- Full Text
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45. Synthetic amyloid-beta oligomers impair long-term memory independently of cellular prion protein.
- Author
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Balducci C, Beeg M, Stravalaci M, Bastone A, Sclip A, Biasini E, Tapella L, Colombo L, Manzoni C, Borsello T, Chiesa R, Gobbi M, Salmona M, and Forloni G
- Subjects
- Alzheimer Disease etiology, Amyloid beta-Peptides chemical synthesis, Amyloid beta-Peptides chemistry, Animals, Cognition Disorders etiology, Cognition Disorders metabolism, Humans, Injections, Intraventricular, Male, Memory physiology, Mice, Mice, Inbred C57BL, Mice, Knockout, Neuronal Plasticity drug effects, Neuronal Plasticity physiology, Peptide Fragments chemical synthesis, Peptide Fragments chemistry, Prion Proteins, Prions genetics, Prions metabolism, Protein Binding, Surface Plasmon Resonance, Amyloid beta-Peptides pharmacology, Memory drug effects, Peptide Fragments pharmacology, PrPC Proteins metabolism
- Abstract
Inability to form new memories is an early clinical sign of Alzheimer's disease (AD). There is ample evidence that the amyloid-beta (Abeta) peptide plays a key role in the pathogenesis of this disorder. Soluble, bio-derived oligomers of Abeta are proposed as the key mediators of synaptic and cognitive dysfunction, but more tractable models of Abeta-mediated cognitive impairment are needed. Here we report that, in mice, acute intracerebroventricular injections of synthetic Abeta(1-42) oligomers impaired consolidation of the long-term recognition memory, whereas mature Abeta(1-42) fibrils and freshly dissolved peptide did not. The deficit induced by oligomers was reversible and was prevented by an anti-Abeta antibody. It has been suggested that the cellular prion protein (PrP(C)) mediates the impairment of synaptic plasticity induced by Abeta. We confirmed that Abeta(1-42) oligomers interact with PrP(C), with nanomolar affinity. However, PrP-expressing and PrP knock-out mice were equally susceptible to this impairment. These data suggest that Abeta(1-42) oligomers are responsible for cognitive impairment in AD and that PrP(C) is not required.
- Published
- 2010
- Full Text
- View/download PDF
46. Role of Glycogen Synthase Kinase-3β in APP Hyperphosphorylation Induced by NMDA Stimulation in Cortical Neurons.
- Author
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Ploia C, Sclip A, Colombo A, Repici M, Gardoni F, Di Luca M, Forloni G, Antoniou X, and Borsello T
- Abstract
The phosphorylation of Amyloid Precursor Protein (APP) at Thr
668 plays a key role in APP metabolism that is highly relevant to AD. The c-Jun-N-terminal kinase (JNK), glycogen synthase kinase-3β (GSK-3β) and cyclin-dependent kinase 5 (Cdk5) can all be responsible for this phosphorylation. These kinases are activated by excitotoxic stimuli fundamental hallmarks of AD. The exposure of cortical neurons to a high dose of NMDA (100 μM) for 30'-45' led to an increase of P-APP Thr668 . During NMDA stimulation APP hyperphosphorylation has to be assigned to GSK-3β activity, since addition of L803-mts, a substrate competitive inhibitor of GSK-3β reduced APP phosphorylation induced by NMDA. On the contrary, inhibition of JNK and Cdk5 with D-JNKI1 and Roscovitine respectively did not prevent NMDA-induced P-APP increase. These data show a tight connection, in excitotoxic conditions, between APP metabolism and the GSK-3β signaling pathway.- Published
- 2010
- Full Text
- View/download PDF
47. JNK contributes to Hif-1alpha regulation in hypoxic neurons.
- Author
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Antoniou X, Sclip A, Ploia C, Colombo A, Moroy G, and Borsello T
- Subjects
- Aging pathology, Amino Acid Sequence, Animals, Cell Death, Cell Hypoxia, Humans, Hypoxia-Inducible Factor 1, alpha Subunit chemistry, MAP Kinase Signaling System, Molecular Sequence Data, Protein Binding, Protein Structure, Secondary, Protein Structure, Tertiary, Rats, Hypoxia-Inducible Factor 1, alpha Subunit metabolism, JNK Mitogen-Activated Protein Kinases metabolism, Neurons enzymology, Neurons pathology
- Abstract
Hypoxia is an established factor of neurodegeneration. Nowadays, attention is directed at understanding how alterations in the expression of stress-related signaling proteins contribute to age dependent neuronal vulnerability to injury. The purpose of this study was to investigate how Hif-1alpha, a major neuroprotective factor, and JNK signaling, a key pathway in neurodegeneration, relate to hypoxic injury in young (6DIV) and adult (12DIV) neurons. We could show that in young neurons as compared to mature ones, the protective factor Hif-1alpha is more induced while the stress protein phospho-JNK displays lower basal levels. Indeed, changes in the expression levels of these proteins correlated with increased vulnerability of adult neurons to hypoxic injury. Furthermore, we describe for the first time that treatment with the D-JNKI1, a JNK-inhibiting peptide, rescues adult hypoxic neurons from death and contributes to Hif-1alpha upregulation, probably via a direct interaction with the Hif-1alpha protein.
- Published
- 2009
- Full Text
- View/download PDF
48. Inhibition of c-Jun N-terminal kinase after hemorrhage but before resuscitation mitigates hepatic damage and inflammatory response in male rats.
- Author
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Relja B, Schwestka B, Lee VS, Henrich D, Czerny C, Borsello T, Marzi I, and Lehnert M
- Subjects
- Alanine Transaminase blood, Animals, Apoptosis drug effects, In Situ Nick-End Labeling, Inflammation pathology, Interleukin-6 blood, Interleukin-6 metabolism, Lactate Dehydrogenases blood, Liver metabolism, Male, Necrosis prevention & control, Random Allocation, Rats, Rats, Sprague-Dawley, Time Factors, JNK Mitogen-Activated Protein Kinases antagonists & inhibitors, Liver drug effects, Liver pathology, Peptides pharmacology, Resuscitation methods, Shock, Hemorrhagic
- Abstract
Inhibition of c-Jun N-terminal kinase (JNK) by a cell-penetrating, protease-resistant JNK peptide (D-JNKI-1) before hemorrhage and resuscitation (H/R) ameliorated the H/R-induced hepatic injury and blunted the proinflammatory changes. Here we tested the hypothesis if JNK inhibition at a later time point-after hemorrhagic shock but before the onset of resuscitation-in a rat model of H/R also confers protection. Twenty-four male Sprague-Dawley rats (250 - 350 g) were randomly divided into 4 groups: 2 groups of shock animals were hemorrhaged to a MAP of 32 to 37 mmHg for 60 min and randomly received either D-JNKI-1 (11 mg/kg i.p.) or sterile saline as vehicle immediately before the onset of resuscitation. Two groups of sham-operated animals underwent surgical procedures without H/R and were either D-JNKI-1 or vehicle treated. Rats were killed 2 h later. Serum activity of alanine aminotransferase and serum lactate dehydrogenase after H/R increased 3.5-fold in vehicle-treated rats as compared with D-JNKI-1-treated rats. Histopathological analysis revealed that hepatic necrosis and apoptosis (hematoxylin-eosin, TUNEL, and M30, respectively) were significantly inhibited in D-JNKI-1-treated rats after H/R. Hepatic oxidative (4-hydroxynonenal) and nitrosative (3-nitrotyrosine) stress as well as markers of inflammation (hepatic and serum IL-6 levels and hepatic infiltration with polymorphonuclear leukocytes) were also reduced in D-JNKI-1-treated rats. LPS-stimulated TNF-alpha release from whole blood from hemorrhaged and resuscitated animals was higher in vehicle-treated rats as compared with D-JNKI-1-treated rats. c-Jun N-terminal kinase inhibition after hemorrhage before resuscitation resulted in a reduced activation of c-Jun. Taken together, these results indicate that D-JNKI-1 application after hemorrhagic shock before resuscitation blunts hepatic damage and proinflammatory changes during resuscitation. Hence, JNK inhibition is even protective when initiated after blood loss before resuscitation. These experimental results indicate that the JNK pathway may be a possible treatment option for the harmful consequences of H/R.
- Published
- 2009
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49. A peptide inhibitor of C-jun N-terminal kinase modulates hepatic damage and the inflammatory response after hemorrhagic shock and resuscitation.
- Author
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Lehnert M, Relja B, Sun-Young Lee V, Schwestka B, Henrich D, Czerny C, Froh M, Borsello T, and Marzi I
- Subjects
- Animals, Blood Pressure physiology, Humans, Inflammation enzymology, Inflammation physiopathology, Inflammation prevention & control, Inflammation Mediators administration & dosage, JNK Mitogen-Activated Protein Kinases physiology, Liver blood supply, Liver enzymology, Male, Peptides administration & dosage, Random Allocation, Rats, Rats, Sprague-Dawley, Shock, Hemorrhagic pathology, Shock, Hemorrhagic physiopathology, Inflammation Mediators therapeutic use, JNK Mitogen-Activated Protein Kinases antagonists & inhibitors, Liver pathology, Peptides therapeutic use, Resuscitation methods, Shock, Hemorrhagic enzymology, Shock, Hemorrhagic prevention & control
- Abstract
Hemorrhage and resuscitation (H/R) leads to phosphorylation of mitogen-activated stress kinases, an event that is associated with organ damage. Recently, a specific, cell-penetrating, protease-resistant inhibitory peptide of the mitogen-activated protein kinase c-JUN N-terminal kinase (JNK) was developed (D-JNKI-1). Here, using this peptide, we tested if inhibition of JNK protects against organ damage after H/R. Male Sprague-Dawley rats were treated with D-JNKI-1 (11 mg/kg, i.p.) or vehicle. Thirty minutes later, rats were hemorrhaged for 1 h to a MAP of 30 to 35 mmHg and then resuscitated with 60% of the shed blood and twice the shed blood volume as Ringer lactate. Tissues were harvested 2 h later. ANOVA with Tukey post hoc analysis or Kruskal-Wallis ANOVA on ranks, P < 0.05, was considered significant. c-JUN N-terminal kinase inhibition decreased serum alanine aminotransferase activity as a marker of liver injury by 70%, serum creatine kinase activity by 67%, and serum lactate dehydrogenase activity by 60% as compared with vehicle treatment. The histological tissue damage observed was blunted after D-JNKI-1 pretreatment both for necrotic and apoptotic cell death. Hepatic leukocyte infiltration and serum IL-6 levels were largely diminished after D-JNKI-1 pretreatment. The extent of oxidative stress as evaluated by immunohistochemical detection of 4-hydroxynonenal was largely abrogated after JNK inhibition. After JNK inhibition, activation of cJUN after H/R was also reduced. Hemorrhage and resuscitation induces a systemic inflammatory response and leads to end-organ damage. These changes are mediated, at least in part, by JNK. Therefore, JNK inhibition deserves further evaluation as a potential treatment option in patients after resuscitated blood loss.
- Published
- 2008
- Full Text
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50. A peptide c-Jun N-terminal kinase (JNK) inhibitor blocks mechanical allodynia after spinal nerve ligation: respective roles of JNK activation in primary sensory neurons and spinal astrocytes for neuropathic pain development and maintenance.
- Author
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Zhuang ZY, Wen YR, Zhang DR, Borsello T, Bonny C, Strichartz GR, Decosterd I, and Ji RR
- Subjects
- Animals, Astrocytes drug effects, Enzyme Activation drug effects, Ganglia, Spinal drug effects, Hyperalgesia complications, Hyperalgesia prevention & control, MAP Kinase Kinase 4 antagonists & inhibitors, Male, Neuralgia complications, Neuralgia prevention & control, Neurons, Afferent drug effects, Rats, Rats, Sprague-Dawley, Spinal Nerves drug effects, Spinal Nerves injuries, Astrocytes enzymology, Ganglia, Spinal enzymology, Hyperalgesia enzymology, MAP Kinase Kinase 4 metabolism, Neuralgia enzymology, Neurons, Afferent enzymology, Peptides administration & dosage
- Abstract
Optimal management of neuropathic pain is a major clinical challenge. We investigated the involvement of c-Jun N-terminal kinase (JNK) in neuropathic pain produced by spinal nerve ligation (SNL) (L5). SNL induced a slow (>3 d) and persistent (>21 d) activation of JNK, in particular JNK1, in GFAP-expressing astrocytes in the spinal cord. In contrast, p38 mitogen-activated protein kinase activation was found in spinal microglia after SNL, which had fallen to near basal level by 21 d. Intrathecal infusion of a JNK peptide inhibitor, D-JNKI-1, did not affect normal pain responses but potently prevented and reversed SNL-induced mechanical allodynia, a major symptom of neuropathic pain. Intrathecal D-JNKI-1 also suppressed SNL-induced phosphorylation of the JNK substrate, c-Jun, in spinal astrocytes. However, SNL-induced upregulation of GFAP was not attenuated by spinal D-JNKI-1 infusion. Furthermore, SNL induced a rapid (<12 h) but transient activation of JNK in the L5 (injured) but not L4 (intact) DRG. JNK activation in the DRG was mainly found in small-sized C-fiber neurons. Infusion of D-JNKI-1 into the L5 DRG prevented but did not reverse SNL-induced mechanical allodynia. Finally, intrathecal administration of an astroglial toxin, l-alpha-aminoadipate, reversed mechanical allodynia. Our data suggest that JNK activation in the DRG and spinal cord play distinct roles in regulating the development and maintenance of neuropathic pain, respectively, and that spinal astrocytes contribute importantly to the persistence of mechanical allodynia. Targeting the JNK pathway in spinal astroglia may present a new and efficient way to treat neuropathic pain symptoms.
- Published
- 2006
- Full Text
- View/download PDF
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