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Carbachol blocks beta-amyloid fragment 31-35-induced apoptosis in cultured cortical neurons
Abstract:
We reported previously that many neurodegenerative changes characteristic of apoptosis could be induced by a short fragment of beta-amyloid protein, A(beta31-35), in cultured newborn mice cortical neurons, and that these changes were accompanied with alterations in expression of some genes. This study was designed to examine whether the apoptotic processes and related gene modulations in this model could be affected by coadministration of carbachol by electrophoretic analysis for DNA ladder formation and by RT-PCR assays for genomic modulation. The results showed that (1) simultaneous incubation with carbachol dose- and time-dependently blocked the specific DNA ladder formation induced by exposure to A(beta31-35) and (2) the A(beta31-35)-induced downregulation of bcl-2 and upregulations of bax, p53, and c-fos genes were reversed or ameliorated by the coadministration of carbachol. It is proposed that A(beta31-35)-induced apoptosis can be prevented by carbachol through mechanisms that modulate the expression of related genes.
Insights
Carbachol prevents beta-amyloid (Aβ31-35) induced neurodegeneration by blocking DNA damage and modulating gene expression. This finding offers potential therapeutic avenues for neurodegenerative diseases.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Beta-amyloid (Aβ) protein fragments are implicated in neurodegenerative diseases.
- Aβ31-35 peptide induces apoptosis and alters gene expression in neuronal cultures.
Purpose of the Study:
- To investigate the effect of carbachol on Aβ31-35-induced apoptosis and gene modulation in neuronal cells.
- To determine if carbachol can prevent neurodegeneration mediated by Aβ31-35.
Main Methods:
- Electrophoretic analysis for DNA ladder formation.
- Reverse transcription-polymerase chain reaction (RT-PCR) assays for gene expression analysis.
- Primary cortical neuron cultures from newborn mice.
Main Results:
- Carbachol dose- and time-dependently inhibited Aβ31-35-induced DNA ladder formation.
- Carbachol reversed or ameliorated Aβ31-35-induced changes in bcl-2, bax, p53, and c-fos gene expression.
- Aβ31-35 caused downregulation of bcl-2 and upregulation of bax, p53, and c-fos.
Conclusions:
- Carbachol can prevent Aβ31-35-induced apoptosis in cultured neurons.
- The protective effect of carbachol involves modulation of apoptosis-related gene expression.
- Carbachol represents a potential therapeutic agent for Aβ-related neurodegeneration.