Adaptive Modifications in the Calpain/Calpastatin System in Brain Cells after Persistent Alteration in Ca2+ Homeostasis

Stifanese, Roberto; Averna, Monica; Tullio, Roberta De; Pedrazzi, Marco; Beccaria, Francesco; Salamino, Franca; Milanese, Marco; Bonanno, Giambattista; Pontremoli, S.; Melloni, Edon

doi:10.1074/jbc.m109.031674

Cited by 42 publications

(34 citation statements)

References 57 publications

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“…3f). Since complex protein conformations may display a higher electrophoretic mobility compared to unbound structures in native gels [86], we performed calpain digestion of recombinant α-syn to analyze migration pattern of truncated and full-length α-syn. Under these conditions the full-length, monomeric α-syn migrated at ~60 kDa.…”

Section: Resultsmentioning

confidence: 99%

“…In order to provide additional experimental evidence in support of calpain implicated in the detected OB synucleinopathy, we measured protein expression levels of calpain and calpastatin. Calpastatin is a selective inhibitor and substrate of calpain, and its formation into complexes is indicative for increased calpain activity [22, 86]. When compared to A30P+D mice, A30P and paraquat-treated mice showed increased expression levels of calpain (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…We therefore utilized native PAGE, enabling the detection of free and calpain-associated complex calpastatin (CAST) formations by its shift in electrophoretic mobility. CAST complex formations are thought to correlate with calpain activation [86]. The calculated ratio between free CAST and complex CAST revealed an increase in presence of the complex formation in A30P+P mice compared to A30P mice and Ntg controls (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Environmental neurotoxic challenge of conditional alpha-synuclein transgenic mice predicts a dopaminergic olfactory-striatal interplay in early PD

et al. 2014

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The olfactory bulb (OB) is one of the first brain regions in Parkinson’s disease (PD) to contain alpha-synuclein (α-syn) inclusions, possibly associated with nonmotor symptoms. Mechanisms underlying olfactory synucleinopathy, its contribution to progressive aggregation pathology and nigrostriatal dopaminergic loss observed at later stages, remain unclear. A second hit, such as environmental toxins, is suggestive for α-syn aggregation in olfactory neurons, potentially triggering disease progression. To address the possible pathogenic role of olfactory α-syn accumulation in early PD, we exposed mice with site-specific and inducible overexpression of familial PD-linked mutant α-syn in OB neurons to a low dose of the herbicide paraquat. Here, we found that olfactory α-syn per se elicited structural and behavioral abnormalities, characteristic of an early time point in models with widespread α-syn expression, including hyperactivity and increased striatal dopaminergic marker. Suppression of α-syn reversed the dopaminergic phenotype. In contrast, paraquat treatment synergistically induced degeneration of olfactory dopaminergic cells and opposed the higher reactive phenotype. Neither neurodegeneration nor behavioral abnormalities were detected in paraquat-treated mice with suppressed α-syn expression. By increasing calpain activity, paraquat induced a pathological cascade leading to inhibition of autophagy clearance and accumulation of calpain-cleaved truncated and insoluble α-syn, recapitulating biochemical and structural changes in human PD. Thus our results underscore the primary role of proteolytic failure in aggregation pathology. In addition, we provide novel evidence that olfactory dopaminergic neurons display an increased vulnerability toward neurotoxins in dependence to presence of human α-syn, possibly mediating an olfactory-striatal dopaminergic network dysfunction in mouse models and early PD.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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Environmental neurotoxic challenge of conditional alpha-synuclein transgenic mice predicts a dopaminergic olfactory-striatal interplay in early PD

et al. 2014

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“…There was a marked upregulation of calpain without a change in CAST in AR2 mice ( Supplementary Fig. S4), in contrast to the decreased expression levels of calpains and an increase in CAST and cleaved forms of CAST in SOD1 Tg mice [50][51][52] . The difference between these two ALS model lines suggests a mechanistic difference between SOD1-associated ALS and sporadic ALS.…”

Section: )mentioning

confidence: 98%

A role for calpain-dependent cleavage of TDP-43 in amyotrophic lateral sclerosis pathology

et al. 2012

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Both mislocalization of TDP-43 and downregulation of RNA-editing enzyme ADAR2 colocalize in the motor neurons of amyotrophic lateral sclerosis patients, but how they are linked is not clear. Here we demonstrate that activation of calpain, a Ca 2 þ -dependent cysteine protease, by upregulation of Ca 2 þ -permeable AMPA receptors generates carboxyterminal-cleaved TDP-43 fragments and causes mislocalization of TDP-43 in the motor neurons expressing glutamine/arginine site-unedited GluA2 of conditional ADAR2 knockout (AR2) mice that mimic the amyotrophic lateral sclerosis pathology. These abnormalities are inhibited in the AR2res mice that express Ca 2 þ -impermeable AMPA receptors in the absence of ADAR2 and in the calpastatin transgenic mice, but are exaggerated in the calpastatin knockout mice. Additional demonstration of calpain-dependent TDP43 fragments in the spinal cord and brain of amyotrophic lateral sclerosis patients, and high vulnerability of amyotrophic lateral sclerosis-linked mutant TDP43 to cleavage by calpain support the crucial role of the calpain-dependent cleavage of TDP43 in the amyotrophic lateral sclerosis pathology.

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“…Cleavage to a 76KD (p76) active form is triggered by increased intracellular calcium which reduces binding to calpastatin [20,25]. Given the ability of exogenous calpain inhibitor to attenuate eosinophil apoptosis [Figure 1A,1C], we evaluated the kinetics of calpain cleavage after the addition of IL-5, TGF-β1, neither or both cytokines by immunoblot analysis.…”

Section: Resultsmentioning

confidence: 99%

Transforming Growth Factor-β1 Antagonizes Interleukin-5 Pro-Survival Signaling by Activating Calpain-1 in Primary Human Eosinophils

Xie

Shen²,

Oh³

et al. 2011

J Clin Cell Immunol

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Background Eosinophils rapidly undergo apoptosis unless exposed to prosurvival cytokines such as interleukin 5 (IL-5) or granulocyte-macrophage colony stimulating factor (GM-CSF). In vivo, eosinophils are exposed to TGF-β 1 which can induce apoptosis suggesting it may function to counteract the effects of IL-5 or GM-CSF and limit, in vivo tissue eosinophilia. Objective The objective of this study was to investigate the proapoptotic effects of TGF-β alone and in combination with IL-5 on eosinophils. Methods Peripheral blood eosinophil (PBEos) viability was assessed using flow cytometry after exposure to TGF-β1 and IL-5. Calpain-1 activation was determined in cell extracts by western blot analysis of endogenous substrates and with a fluorogenic α-spectrin substrate. Molecular interactions between calpain1 and calpastatin were assessed by immunoprecipitation and western blotting. Results Physiologic concentrations of TGF-β1 significantly antagonized the prosurvival effects of IL-5. TGF-β1-induced apoptosis was suppressed by inhibitors of calpain, or its downstream target, caspase 3. TGF-β1 signaling through Smad3 was unaffected by IL-5 and was required for the pro-apoptotic effects of TGF-β1. However, IL-5 induced Akt phosphorylation was inhibited by TGF-β1 and was associated with accelerated calpain cleavage and eosinophil death. Conclusion TGF-β1 induces calpain-1 activation through antagonism of Akt which induces caspase activation and eosinophil apoptosis.

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Adaptive Modifications in the Calpain/Calpastatin System in Brain Cells after Persistent Alteration in Ca2+ Homeostasis

Cited by 42 publications

References 57 publications

Environmental neurotoxic challenge of conditional alpha-synuclein transgenic mice predicts a dopaminergic olfactory-striatal interplay in early PD

Environmental neurotoxic challenge of conditional alpha-synuclein transgenic mice predicts a dopaminergic olfactory-striatal interplay in early PD

A role for calpain-dependent cleavage of TDP-43 in amyotrophic lateral sclerosis pathology

Transforming Growth Factor-β1 Antagonizes Interleukin-5 Pro-Survival Signaling by Activating Calpain-1 in Primary Human Eosinophils

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