Biyofizik 3

Öğr. Gör. Cemil ÖZGÜL

 İyon kanallarının araştırılmasında
kullanılan teknikler

1- Voltaj kenetleme Tekniği

2- Yama-menteşe (Patch-clamp) tekniği
Voltaj Kenetleme
 Voltage clamping: 3 prensibi
(1) Hücrelere pozitif akım verirsek depolarize,
negatif akım verirsek hiperpolarize olurlar.
Depolarizing response
Hyperpolarizing response
V V

I I
 Voltage clamping: 3 prensibi
(2) Hücrelere akım gönderildiği zaman depolarizasyon
ve hiperpolarizasyon için zaman geçer. Çünkü,
hücrelerin kapasitansının değiştirilmesi gereklidir.

+ + + + + + +
- - - - - - - -
+ + + +

I
 Voltage clamping: 3 prensibi
(3) Şayet iyonların net bir difüzyonu hücreye
yoksa, hücre zarı potansiyeli değişmeyecektir.

Nucleus
+
+ V

INa
 Voltage clamping: 3 prensibi
(3) Şayet iyonların net bir difüzyonu hücreye
yoksa, hücre zarı potansiyeli değişmeyecektir.

- -
Hyperpolarizing I
Nucleus
+
+ V

INa
 How the voltage clamp works:

-10 mV
V -65 mV

1 msec
 Voltage clamp: what is the behavior of
voltage dependent sodium current?

-10 mV
V -65 mV
In the presence of
tetraethylammonium
(TEA)

I
1 msec
 Voltage clamp: what is the behavior of
voltage dependent sodium current?

Na current

-10 mV
V -65 mV
 Voltage clamp: what is the behavior of
voltage dependent sodium current?

-10 mV
V -65 mV Na current

I
 Voltage clamp: what is the behavior of
voltage dependent potassium current?

In the presence of
-10 mV tetrodotoxin (TTX)
V -65 mV

I
 Voltage clamp properties of ionic currents
underlying the action potential

1. Na current activates and then, EVEN WHILE THE

COMMAND POTENTIAL IS MAINTAINED,
automatically inactivates
2. K current activates more slowly than Na current
3. K current does not inactivate as long as
depolarization is maintained (To close K channels,
it’s necessary to repolarize the cell).

4. Finally, it should be noted that the voltage clamp

method has been used to study many other types of
voltage-dependent channels, including calcium
channels, inactivating K channels, Ca-activated K
channels, etc.
 Changes in permeability accompanying action potential

At peak, high Na
permeability

0 mV
High K
permeability (even
higher than at
rest)

-65
mV GK

1 msec

GNa
Akım-Voltaj İlişkisi
Patch-Clamp
Patch-Clamp
Patch-Clamp
Patch-Clamp
Patch-Clamp
Patch-Clamp
Patch-Clamp
Patch-Clamp