Meeting the Lab

Finally went to work today :) It was a half hour uphill trek to get there but I made it :P My SIM card is still not working so Sven called me at around 8:30 to make sure I was still alive and coming to work :P Such a worry wart. When we finally met up at the bus stop, I told him about my weekend (the fact that I got lost and that I watched a film at the cinema) and he totally laughed at me. I guess it is pretty weird that I watched a German film without knowing any German :P.

First thing we did when we got to the office at 9am, was coffee break! I get my own mug and desk at the office :) The PI (Tina) also brought some chocolate croissant like thing for all the people in the lab and she got me a travel book for Mainz. She joked that it was perfect for me after my trek to school since it was how to tour Mainz by foot. The lab was pretty small, including only 2 PhD students (Sven and Florian), the PI, and myself.

Sven gave me a tour of the areas that I will be working in (all the equipment spread out in a bajillion rooms -___-). Did I mention I work at the Clinic of Psychiatry and Psychotherapy in the University of Medicine? Yea, slightly weird. So this means that all doors are automatically locked and can only be opened via keycard. If you turn the knob without a keycard, it just keeps turning but does not lock on to the locking mechanism to actually open the door. For some reason, my keycard didn't work the first time (everything Sven gives me doesn't seem to work...first the SIM card and now the keycard. lol. I kid.) but luckily I got it to work after another visit to the office.

As for techniques I learned today, Sven taught me how to subculture a overcrowded cell plate in a sterile manner (he demonstrated and then I did one myself) and I learned to prepare a cell medium for my cells :). I also observed as he measured ADAM10 expression from cells treated with miRNAs. Expression was measured through the reporter gene luciferase with a machine that measured luminescence (or absorbance if it has to). Everything was pretty good today and I didn't screw up yet. So let's cross my fingers and hope I don't majorly screw up any time soon :)

If you guys are wondering what the hell my project is on, read on. So it is widely accepted that Alzheimer's disease is caused by plaques and tangles within the brain. My lab focuses on the plaque component of the disease. Amyloid precursor protein (APP) is the initial protein within the brain. A beta secretase (BACE1) followed by a gamma secretase cleaves and releases three separate proteins from the cell membrane. The protein of interest is AB-peptide, which clumps together to form the plaques.

The previous lab my PI worked in discovered another protease, ADAM10, that cleaves APP in such a way that that AB-peptide would not be formed after gamma secretase cleavge. My project specificially studies how two particular transcription factors affect the expression of ADAM10 and BACE1. The promoter activity for each protein is measured with a reporter gene, luciferase and the ratio for promoter activities will be calculated (ADAM10 promoter activity/BACE1 promoter activity) where a high ratio means that more ADAM10 is produced while BACE1 production would be decreased. This section measures specifically the transcription of ADAM10 and BACE1.

The second section of my project studies the effects of the transcription factors on ADAM10 and BACE1 translation. Protein expression would be measured directly via western blot or ELISA. And that is the general basics of my lab.