Neuronal loss in primary long-term cortical culture involves neurodegeneration-like cell death via calpain and p35 processing, but not developmental apoptosis or aging

Kim, Min-Ju; Oh, Soo‐Jin; Park, Seong Hoon; Kang, Hongjun; Won, Moo Ho; Kang, Tae‐Cheon; Park, Jae-Bong; Kim, Jong‐Il; Kim, Jaebong; Lee, Jae Yong

doi:10.1038/emm.2007.3

Cited by 31 publications

(21 citation statements)

References 49 publications

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“…Furthermore, collapsin response mediator-protein 3 triggers neuronal death after cleavage by calpain (Hou et al, 2006). In a more chronic model, Kim et al (2007) could show that degeneration of cortical neurons in vitro depends on calpain cleavage of p35. If calpain is part of such degenerative cascades, then inhibition of calpain should attenuate neuronal decline.…”

Section: Discussionmentioning

confidence: 91%

Inhibition of Calpain Prevents N-Methyl-d-aspartate-Induced Degeneration of the Nucleus Basalis and Associated Behavioral Dysfunction

Nimmrich¹,

Szabo²,

Nyakas³

et al. 2008

J Pharmacol Exp Ther

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N-Methyl-D-aspartate (NMDA) receptor-mediated excitotoxicity is thought to underlie a variety of neurological disorders, and inhibition of either the NMDA receptor itself, or molecules of the intracellular cascade, may attenuate neurodegeneration in these diseases. Calpain, a calcium-dependent cysteine protease, has been identified as part of such an NMDA receptorinduced excitotoxic signaling pathway. The present study addressed the question of whether inhibition of calpain can prevent neuronal cell death and associated behavioral deficits in a disease-relevant animal model, which is based on excitotoxic lesions of the cholinergic nucleus basalis magnocellularis of Meynert. Excitotoxic lesions of the nucleus basalis with NMDA induced a markedly impaired performance in the novel object recognition test. Treatment with the calpain inhibitor,, dose-dependently prevented the behavioral deficit. Subsequent analysis of choline acetyltransferase in the cortical mantle of the lesioned animals revealed that application of A-705253 dose-dependently and significantly attenuated cholinergic neurodegeneration. Calpain inhibition also significantly diminished the accompanying gliosis, as determined by immunohistochemical analysis of microglia activation. Finally, inhibition of calpain by A-705253 and the peptidic calpain inhibitor N-acetyl-Leu-LeuNle-CHO did not impair long-term potentiation in hippocampal slices, indicating that calpain inhibition interrupts NMDA excitotoxicity pathways without interfering with NMDA receptormediated signaling involved in cognition. We conclude that inhibition of calpains may represent a valuable strategy for the prevention of excitotoxicity-induced neuronal decline without interfering with the physiological neuronal functions associated with learning and memory processes. Thus, calpain inhibition may be a promising and novel approach for the treatment of various neurodegenerative disorders.Calpains in the central nervous system are Ca 2ϩ -dependent cysteine proteases, which can modulate the conformation, localization, and activity of various proteins. Calpaininduced modifications have a profound affect on biological processes as different as cell cycle progression and vesicular trafficking. They are an integrative part of several signaling cascades, but they can also contribute to apoptotic and necrotic cell death in pathological conditions. For this reason, calpains have been discussed extensively as target for therapeutic interventions in a number of neurodegenerative disease that are associated with neuronal loss (for review, see Huang and Wang, 2001;Goll et al., 2003;Zatz and Starling, 2005).In that respect, it has recently been shown that inhibition of calpains prevents excitotoxic neuronal cell death in vitro (Caba et al., 2002;Ray et al., 2006) and in vivo (Chiu et al., 2005;Takano et al., 2005). Excitotoxicity is caused by an overstimulation of N-methyl-D-aspartate (NMDA) receptors, and there is now growing evidence that calpain cleaves sevArticle, publication date, and cita...

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Section: Discussionmentioning

confidence: 91%

Inhibition of Calpain Prevents N-Methyl-d-aspartate-Induced Degeneration of the Nucleus Basalis and Associated Behavioral Dysfunction

Nimmrich¹,

Szabo²,

Nyakas³

et al. 2008

J Pharmacol Exp Ther

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“…In fact, the mechanism of neurodegeneration in culture could be different from that in vivo, because in primary cultures, neurons died by an apoptotic process involving DNA breakage as indicated by TUNEL staining, which was not observed in vivo in brain of the p25 ON mice. A similar situation was recently observed in a different but related model system, 51 reiterating the question of the relevance of cellular models for in vivo phenomena.…”

Section: Microgliosismentioning

confidence: 85%

“…Similar observations were recently reported in a different but related model system. 51 Significantly more p25-expressing neurons were TUNELpositive compared with wild-type neurons, when co-cultured with astroglia ( Figure 7D, left histogram). Surprisingly, the fraction of TUNEL-positive p25 neurons was lower when co-cultured with N11 microglia than with astroglia and not significantly different from wild-type neurons ( Figure 7D, right histogram).…”

Section: Primary Hippocampal Neurons Activate Microglia In Culturementioning

confidence: 99%

Neurodegeneration and Neuroinflammation in cdk5/p25-Inducible Mice

Muyllaert

Terwel

Kremer

et al. 2008

The American Journal of Pathology

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The cyclin-dependent kinase cdk5 is atypically active in postmitotic neurons and enigmatic among the kinases proposed as molecular actors in neurodegeneration. We generated transgenic mice to express p25, the N-terminally truncated p35 activator of cdk5, in forebrain under tetracycline control (TET-off). Neuronal expression of p25 (p25 ON ) caused high mortality postnatally and early in life. Mortality was completely prevented by administration of doxycycline in the drinking water of pregnant dams and litters until P42, allowing us to study the action of p25 in adult mouse forebrain. Neuronal p25 triggered neurodegeneration and also microgliosis, rapidly and intensely in hippocampus and cortex. Progressive neurodegeneration was severe with marked neuron loss, causing brain atrophy (40% loss at age 5 months) with nearly complete elimination of the hippocampus. Neurodegeneration did not involve phosphorylation of protein tau or generation of amyloid peptide. Degenerating neurons did not stain for terminal deoxynucleotidyl transferase-mediated dUTP nickend labeling or activated caspase-3 but were marked by FluoroJadeB in early stages. Diseased neurons were always closely associated with activated microglia already very early in the disease process. Primary neurons derived from p25 embryos were more prone to apoptosis than wild-type neurons, and they activated microglial cells in co-culture. The inducible p25 mice present as a model for neurodegeneration in hippocampal sclerosis and neocortical degeneration, with important contributions of activated microglia.

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“…Primary cortical neuron cells were established by a modification of previously described procedure (Kim et al, 2007). The cortices from rat embryos of embryonic day 18 were dissected and freed of meninges.…”

Section: Methodsmentioning

confidence: 99%

Nitric Oxide Is an Essential Mediator for Neuronal Differentiation of Rat Primary Cortical Neuron Cells

Heo

Kho

et al. 2010

Exp Neurobiol

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Nitric oxide (NO) regulates proliferation, differentiation and survival of neurons. Although NO is reported to involve in NGF-induced differentiation of PC12 cells, the role of NO has not been characterized in primary neuron cells. Therefore, we investigated the role of NO in neuronal differentiation of primary cortical neuron cells. Primary cortical neuron cells were prepared from rat embryos of embryonic day 18 and treated with NMMA (NOS inhibitor) or PTIO (NO scavenger). Neurite outgrowth of neuron cells was counted and the mRNA levels of p21, p27, c-jun and c-myc were measured by RT-PCR. Neurite outgrowth of primary cortical neuron cells was inhibited a little by NOS inhibitor and completely by NO scavenger. The mRNA levels of p21 and p27, differentiation-induced growth arrest genes were increased during differentiation, but they were decreased by NOS inhibitor or NO scavenger. On the other hand, the level of c-jun mRNA was not changed and the level of c-myc mRNA was increased during differentiation differently from previously reported. The levels of these mRNA were reversed in NOS inhibitor- or NO scavenger-treated cells. The level of nNOS protein was not changed but NOS activity was inhibited largely by NOS inhibitor or NO scavenger. These results suggest that NO is an essential mediator for neuronal differentiation of primary cortical neuron cells.

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Neuronal loss in primary long-term cortical culture involves neurodegeneration-like cell death via calpain and p35 processing, but not developmental apoptosis or aging

Cited by 31 publications

References 49 publications

Inhibition of Calpain Prevents N-Methyl-d-aspartate-Induced Degeneration of the Nucleus Basalis and Associated Behavioral Dysfunction

Inhibition of Calpain Prevents N-Methyl-d-aspartate-Induced Degeneration of the Nucleus Basalis and Associated Behavioral Dysfunction

Neurodegeneration and Neuroinflammation in cdk5/p25-Inducible Mice

Nitric Oxide Is an Essential Mediator for Neuronal Differentiation of Rat Primary Cortical Neuron Cells

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