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