Astrocyte senescence promotes glutamate toxicity in cortical neurons by GetAssignment.com.au

1. ASTROCYTE SENESCENCE
PROMOTES GLUTAMATE TOXICITY IN
CORTICAL NEURONS
By GetAssignment.com.au

2. The article by Limbad et al.
investigates the effects of
astrocyte senescence on
glutamate toxicity in cortical
neurons. The study uses a
variety of lab techniques to
explore this research question.
INTRODUCTION

3. CELL CULTURE
The study uses primary cortical
neurons and astrocytes cultured
from postnatal day 1-2 mice (Prah, et
al., 2019). The cells are cultured in
media containing fetal bovine serum
and other supplements to support
their growth

4. SENESCENCE ASSAY
To induce senescence in astrocytes,
the study uses a technique called
senescence-associated beta-
galactosidase (SA-β-gal) staining.
This assay detects the accumulation
of the SA-β-gal enzyme, which is a
marker of senescence.

5. To assess the effects of senescent astrocytes on cortical neurons, the study
uses a glutamate toxicity assay. This involves exposing the neurons to various
concentrations of glutamate and measuring their viability using a colourimetric
assay.
GLUTAMATE TOXICITY ASSAY
JAN

6. IMMUNOCYTOCHEMISTRY
To visualize the expression of
various proteins in the cells, the
study uses immunocytochemistry
(Srebotnik Kirbiš, et al., 2020).
This technique involves labelling
proteins with specific antibodies
and visualizing them using
fluorescent microscopy.

7. WESTERN BLOTTING
To quantify the expression levels of
specific proteins, the study uses
western blotting. This involves
separating proteins using gel
electrophoresis and then
transferring them onto a membrane.
The membrane is probed with
specific antibodies and the protein
bands are visualized using
chemiluminescence.

8. To measure gene expression levels, the study uses RNA isolation and
quantitative polymerase chain reaction (qPCR). RNA is extracted from the
cells and then converted to cDNA, which is used as a template for qPCR
(Vogels, et al., 2021). The expression levels of specific genes are measured
using fluorescent probes.
RNA ISOLATION AND QPCR

9. CELL TRANSFECTION
To manipulate gene expression
levels in the cells, the study uses
cell transfection. This involves
introducing foreign DNA or RNA into
the cells using various transfection
reagents. The study uses this
technique to overexpress or
knockdown specific genes of
interest.

10. STATISTICAL
ANALYSIS
To determine the significance of the results, the study
uses various statistical tests, including t-tests,
ANOVA, and linear regression. The data are
analyzed using software such as GraphPad Prism.

11. CONCLUSION
The study by Limbad et al. uses a variety of lab techniques to
investigate the effects of astrocyte senescence on glutamate
toxicity in cortical neurons. The results suggest that senescent
astrocytes promote glutamate toxicity in neurons, potentially
contributing to neurodegeneration in aging and age-related
diseases.

12. THANK YOU
www.GetAssignment.com.au