This document summarizes a seminar presented by Mariana Hoyos and Daniela Lara on the role of IDH1 in granulosa cell senescence through reactive oxygen species (ROS) activated MAPK signaling pathways. The seminar introduction discusses MAPK signaling pathways and IDH1 enzyme's importance. The objective was to study IDH1's role in ROS-induced granulosa cell dysfunction under chronic stress. Materials and methods included ELISA, immunohistochemistry, real-time PCR, and Western blot. Results showed chronic stress decreases IDH1 expression and increases oxidative stress, and IDH1 knockdown decreased proteins related to autophagy and cell cycle. The discussion supported the results. In conclusion, chronic
2. Table of Contents
Title Introduction
Materials and
Methods
Objective
01 02 03
04
Results
05 06
Discussion
07
Conclusion
3. TITLE
Daniela lara Patiño and Mariana Hoyos
3rd semester students of the Pontifical Bolivarian University
4. INTRODUCTION
#1 MAPK SIGNALING PATHWAYS
Mitogen-activated protein kinase
(MAPK) cascades have been shown to
play a key role in transduction
extracellular signals to cellular
responses
5. INTRODUCTION
#2 What is IDH1 and why it is important
● Important enzyme in the
tricarboxylic acid cycle
● Decreased expression of IDH1
disrupted NADPH homeostasis
which induced oxidative stress
damage or increased cell
sensitivity to oxidative stress
● However, whether IDH1 plays a
role in oxidative stress induced
dysfunction of GCs under CUS
was unknown.
6. CREDITS: This presentation template was
created by Slidesgo, including icons by
Flaticon, infographics & images by Freepik
Study the importance of IDH1 in
the acceleration of granulosa
cells senescence through ROS
activated MAPK signaling
pathways
OBJECTIVE
7. MATERIALS AND METHODS
#1 Enzyme-linked immunosorbent assay (ELISA)
The elisa is a test that is within the
immunoassays, which means that
it evaluates a reaction between an
antigen and an antibody.
The reaction between an antigen
and an antibody cannot be seen by
itself, so, in the case of ELISA an
enzyme is used.
9. #3 Real time PCR
Amplification and detection
processes occur
simultaneously in the same
closed vial.
Through fluorescence
detection , the amount of
synthesized DNA can be
measured every moment ,
since the fluorescent emission
in the reaction is proportional
to the amount of DNA formed.
10. #4 Western Blot
● SDS-PAGE
polyacrylamide
● (SDS) Sodium
dodecyl
detergent
● Electrophoresis
11. RESULTS
PANEL A and B
CUS induces oxidative stress of GC and
decreases the expression of IDH1 in antral
follicle granulosa cells in mice
13. PANEL A and G
Western blotting tested the
silencing efficiency of IDH1
knockdown by two shRNAs
IDH1 knockdown decreased the
protein expression level of
CCNB1 and
CDK1 in KGN cells by Western
blotting analysis.
14. PANEL B and E
Western blotting
analysis showed the
levels of proteins
related to autophagy,
including Beclin1,
LC3II.
The levels
of
phosphory
lated and
non-phosp
horylated
pathways
using
specific
antibodies
15. DISCUSSION
Authors Arguments Supports or is
against the
articles results
Y.K Hsu, et al
When oxidative damage was cumulated too
huge, these ROS will accelerate cellular
senescence
Z.Zhang, et al
Previous study reported that α-LG existed in
follicle fluid and decreased with age, could
delay age-related fertility decline
H, Xing, et al
Autophagy participates in regulation of cell
homeostasis and protecting cell against
oxidative stress
16. CONCLUSIONS
01
02
03
It proves that chronic
stress can cause ovarian
dysfunction
ROS accumulation triggered
MAPK signaling pathways
activation
The improvement
of IDH1 activity
might be a
beneficial
therapeutic
strategy for ovarian
insufficiency
