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Characterization of polyclonal antibodies against GABA-A receptor subunits
Austria (AMSA) - University of Vienna, Vienna
Department of Pathobiology of the Nervous System; Center for Brain Research
Univ. Prof. Dr. Johannes Berger
Assoc. Prof. Dr. Petra Scholze
Type of Research Project
- Basic science
What is the background of the project?
GABAA receptors are ligand-gated chloride channels composed of five subunits, and play an important role in neurotransmission. The main endogenous ligand is gamma-aminobutyric acid, the major inhibitory neurotransmitter in the central nervous system. Further ligands are also benzodiazepines, a class of tranquilizer drugs. However, the existence of six , three β, three γ, one δ, ε, θ, π, and three ρ subunits gives rise to an enormous diversity of receptor subtypes with distinct subunit composition and therefore pharmacological properties. For example, the majority of GABAA receptors (those containing α1-, α2-, α3-, or α5-subunits) are benzodiazepine sensitive, there exists a minority of GABAA receptors (α4- or α6-subunit containing) which are insensitive to classical 1,4-benzodiazepines, but instead are sensitive to other classes of GABAergic drugs such as neurosteroids and alcohol. To study these receptors, subunit specific antibodies are needed. Our lab has generated a diversity of subunits, which now need to be tested for their specificity, by using these specific antibodies.
What is the aim of the project?
In the current project we aim to identify binding sites unique for 6 containing GABAA receptors to pave the way for future drug developmental research.
What techniques and methods are used?
First, the student will learn the following lab techniques, and when confident enough, will perform them by himself/herself: Immunoprecipitation: to precipitate a protein antigen out of solution using an antibody that specifically binds to it. Western – Blot: to detect specific proteins in a sample od tissue homogenate or extract. As well, statistical analysis will be performed: all results will be expressed as mean ± SEM (standard error of the mean), data will be analyzed with GrapdPad Prism 6.0 program and evaluated with one-way ANOVA with Dunnett’s multiple comparison test.
What is the role of the student?
What are the tasks expected to be accomplished by the student?
Perform western blot experiments with different samples and judge the quality of the antibody. After a short training period the student will be expected to perform those experiments independently. Techniques of immunoprecipitation will be explained following established protocols, which will also be expected from the student to work independently on these. Data analysis will be done based on the student’s performance and contribution.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
The student will be actively involved in our weekly lab meetings, where he/she will in the end be expected to give a final presentation of his/her results (see below).
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- The student will prepare a poster - The student will prepare a presentation
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Excellent english language skills; very dedicated students only.
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months) - Students in biomedical fields
- Werner Sieghart “Allosteric Modulation of GABAA Receptors via Multiple Drug Binding Sites” Advances in Phramacology 72:53-96
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