Molecular Modulation by Lentivirus-Delivered Specific shRNAs in Endoplasmic Reticulum Stressed Neurons

874 Views

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10:50 min

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April 24th, 2021

DOI :

10.3791/61974-v

April 24th, 2021

•

874 Views

Carolina Morales¹, Mariano Bisbal¹, Mariana Bollo¹

¹Instituto de Investigación Médica M y M Ferreyra, INIMEC-CONICET, Universidad Nacional de Córdoba

Chapters

0:05

Introduction

0:50

Primary Rat Cortical Neuron Cultures

2:25

Lentivirus Production

4:41

Lentivirus Infection

6:23

Primary Neuron Cultures Stressed with Ganglioside GM2 Accumulation and Immunocytochemistry

7:43

Neurite Atrophy Analysis

8:38

Results: GM2 Accumulation-Induced Neuritic Atrophy Modulated by CN-Aα and CHOP Knock-Down in an Opposite Manner

10:07

Conclusion

Transcript

This approach may be used in cell system models of endoplasmic reticulum stress by manipulating the molecular expression of downstream pair components. The main advantage of this technique is to remove the impact of paired signaling by the visuali

Summary

In the present study, the expression is knocked down of two downstream signaling components of the PERK pathway, the cytoprotective calcineurin and the pro-apoptotic CHOP, by using specific shRNAs. In opposite ways, these modulate the susceptibility of primary cortical neurons to neurite atrophy after induction of endoplasmic reticulum stress.

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