Detektion der α-4, α-5 sowie δ Untereinheit von GABA-Rezeptoren in Thalamus und Kortex des Mäusehirns mittels Western Blot

Abstract

The aim of this research was to enable the detection of the α-4, α-5 and δ subunits of GABA-receptors in the cortex and thalamus of mice. The chosen method therefore was the proven analysis method Western Blot, for which the protocol should be created.
For this purpose, a series of experiments with slight variations concerning different parameters were performed.
One of these parameters was the blocking-buffer, meaning two different solutions were used in order to find the most suitable one. The utilised blocking-buffers were BSA and Roti-Block. It has been found that BSA is the preferable blocking buffer for the detection of the α-4 and α-5 subunit, as it exhibits less unspecific bindings, whereas no difference between the use of BSA and Roti- Block to detect the δ subunit could be perceived.
Another variation of parameters was the utilisation of α-4 knockout mice to confirm the previous detection as a specific binding on the basis of negative samples. Yet this turned out to be useless because the genetic modification of mice was only partly successful, in that there was still quite a high number of α-4 subunits
left in the knockout mice.
Nevertheless it was possible to detect the relevant subunits of the GABA-receptor, which implies that a functioning protocol for the detection of the α-4, α-5 and δ subunit using Western Blot has been created.