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Core Course MM # 707 Electrophysiology: Patch Clamp. Dr. Sonia Siddiqui Dr. Panjwani Centre For Molecular Medicine and Drug Research (PCMD). 3T3 cells. 3T3 cells. Butanol extracts (10 ug/ml). Butanol extracts (20 ug/ml). Control. - PowerPoint PPT Presentation
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Dr. Sonia Siddiqui
Dr. Panjwani Centre For Molecular Medicine and Drug Research (PCMD)
Core Course MM # 707
Electrophysiology: Patch Clamp
3T3 cells
3T3 cells
Results: Effect of cardamom extracts (Butanol) on 3T3 cell line
Control Butanol extracts (10 ug/ml)
Butanol extracts (20 ug/ml)
Original recording graphs showing the same duration (100 msec) of the opening of the channels in B1 and B2 when compared to controls.
However B2 increases outward rectifying potassium currents
Results: Effect of cardamom extracts (D1-D3 hydroalcoholic phase) on 3T3 cell line
C D1 (10 ug/ml) D2 (15 ug/ml) D3 (20 ug/ml)
Results: Effect of cardamom extracts (EthylAcetate) on 3T3 cell line
C EA1 EA2
Results: Effect of cardamom extracts (n-Hexane) on 3T3 cell line
C n-Hexane-1 n-Hexane-2
Postnatal Hippocampal neurons
Identification of hippocampal neurons
Pattern of hippocampal firing
Pattern of cortical neurons firing
Effect of size of the cell body of a neuron
Effects from Differentiated neurons on the currents
Identification of 3T3 cell line
Pulse Protocol for Na and K+ currents
-110mV -60mV
60mV
-55mV /holding potential
Results: Hippocampal neurons
Results: Effect of peptide (P1 ) on Cortical neurons
C After 5 mins After 10 mins
Results: Effect of peptide (P2 ) on Cortical neurons
C After 5 mins After 10 mins
Results Effect of peptides isolated from bacteria on cortical neurons
Control After 5min After 10min
Results: Effect of peptide (P3) on Hippocampal neurons
K+ channels
C After 5 mins After 10 mins
K+ channels
C After 5 mins After 10 mins
Results: Effect of peptide (P4 ) on hippocampal neurons
Results: Methonolic Nicotinum Tobacom Extract (0.5mg/ml)
Methonolic Extract was tested on hippocampal cultured neurons. We found that it blocks delayed rectified Potassium currents
NAControl
Cur
rent
s w
ere
norm
aliz
ed b
y ce
ll ca
paci
tanc
e (p
A/p
F)
Results Centella Asiatica Extract (0.5mg/ml) On K+ and Na+ Currents
Asiaticoside, an active constituent in methanol and ethylacetate extracts of C. asiatica, has anxiolytic activity.The extracts of C. asiatica, except Aqueous extract, possess anticonvulsant and neuroprotective activity and thus can be used for effective management in treatment of epileptic seizures.
Methonolic Extract was tested on hippocampal cultured neurons. We found that it does not effect delayed rectified Potassium currents
Control
Control
Control
CA
CA
CAControlNa currents
Na currents
2
Results Centella Asiatica Extract (0.5mg/ml) On Sodium Currents
Methonolic Extract was tested on hippocampal cultured neurons. We found that it reduces Sodium currents. This is in consistent with the previous reported studies by Visweswari et al., 2011. This medicinal herb used as an anticonvulsant and for enhanced memory. The most important ingredients in methonolic extract is Asiaticoside which has an anxiolytic activity.
Control CA Wash
Centella Acasia Extract on Sodium currents
-300
-250
-200
-150
-100
-50
0
-80 -60 -40 -20 0 20 40 60 80
voltage (mV)
Cu
rre
nts
(p
A)
Control
C.acasia
Results Centella Asiatica Extract (0.5mg/ml) On Sodium Currents
Results: Serpindus Trifalitus extract (0.5mg/ml) On hippocampal K Currents
Serpindus Trifalitus Extract on Transient Potassium currents
-150
-100
-50
0
50
100
150
200
250
300
-80 -60 -40 -20 0 20 40 60 80
voltage (mV)
Cu
rre
nts
(p
A)
Control
S. trifalitus
Control ST
Transient K Currents
Serpindus Trifalitus Extract on Sustained Potassium currents
-50
0
50
100
150
200
250
-80 -60 -40 -20 0 20 40 60 80
voltage (mV)
Cu
rre
nts
(p
A)
Control
S. trifalitus
Sustained K Currents
Serpindus Trifalitus Extract on Sodium currents
-500
-400
-300
-200
-100
0
100
200
300
-80 -60 -40 -20 0 20 40 60 80
voltage (mV)
Cu
rre
nts
(p
A)
Control
S. trifalitus
Results: Serpindus Trifalitus extract (0.5mg/ml) On hippocampal Na Currents
Control STWe observed that after 2-3 min application of Serpindus Trifalitus extract all cells were died in culture plate
Results: Co-cultured Hippocampal neurons