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Genotyping of androgen and estrogen receptor knockout mouse models
Belgium (BeMSA) - KU Leuven, Leuven
Department of Chronic Diseases, Metabolism and Ageing
Prof. Dirk Vanderschueren
English, Dutch, French, and Spanish
Type of Research Project
- Basic science
What is the background of the project?
Androgens and estrogens act through the androgen receptor (AR) and the estrogen receptor (ER) to play crucial physiological roles in the development and maintenance of the reproductive organs. In addition, they have pleiotropic effects on several extragenital structures including muscle, bone, kidney and brain. Pharmacological targeting of AR and ER is used in the clinic e.g. in prostate and breast cancer but also in osteoporosis and frailty. However, these treatments are hampered by important side effects. Therefore, an in-depth understanding of the biology of these transcription factors is of utmost importance to develop better, tissue- or disease-specific AR and ER targeting. In this context, our laboratory has generated various mouse models to unravel androgen and estrogen action on different tissues by studying the phenotype of mice in which the AR or the ER has been selectively ablated in a specific tissue (= AR or ER knockout mouse models).
What is the aim of the project?
The aim of the present project will be to determine the genotype of mice issued from the breeding of kidney-specific AR knockout mice and neuron-specific ER knockout mice.
What techniques and methods are used?
The phenotyping of AR and ER knockout mouse models is preceded by a very important step, which is the genotyping of the mice. This procedure enables us to assign them to the correct experimental group i.e. knockout or control by performing a polymerase chain reaction (PCR) on their DNA Genotyping is performed in several steps: DNA extraction from mouse tissue, Polymerase chain reaction (PCR), Agarose gel electrophoresis
What is the role of the student?
- If the project includes “lab work”
- the student will take active part in the practical aspect of the project
- The tasks of the student will be performed on his/her own
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
The student will be actively involved in projects assessing androgen action on kidney and estrogen indirect action on bone and body composition via the brain and the different steps of genotyping listed above. Concretely, they will first observe the different techniques. Next, when they are ready for it, they will execute the different techniques by himself, of course under close supervision. At all times, they will be involved in the analysis of the data. Indeed, in the last step i.e. agarose gel electrophoresis, the PCR product has to be loaded on a gel and visualized, after which interpretation of the obtained findings is needed to correctly determine the genotype of each animal.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
During the first week of the exchange, the student will be taught some theoretical background around the knockout mouse technology and the different technical steps of the genotyping procedure. This will be done by the tutor.
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 presentation - The student will prepare a scientific report
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Eagerness to learn, motivation
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) - Pre-Medical students from the American-British system - Students in biomedical fields - Dental medicine students (IADS members)
- Dubois V; Laurent MR; Sinnesael M; Cielen N; Helsen C; Clinckemalie L; Spans L; Gayan-Ramirez G; Deldicque L; Hespel P; Carmeliet G; Vanderschueren D; Claessens F. A satellite cell-specific knockout of the androgen receptor reveals myostatin as a direct androgen target in skeletal muscle. FASEB J. 2014 Jul;28(7):2979-94.
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