What if APP is not up at all?
- 1) we know APP mRNA is up in the transcriptome in response to motor learning.
- 2) if we don't see APP protein up in our experiment, we likely need to add 1 more timepoint (17days) because the time from transcription to translation can take up to 3 days in vivo.
What if APP is not alternatively spliced?
It is not alternatively spliced in response to learning.
What is GAPDH?
It is a house keeping gene. House keeping genes are used as a control. For example, if you show APP protein goes up in your western, how do we know you didn't cheat? you use a house keeping gene to show that you loaded equal amount of proteins in each well during a western blot, because house keeping genes usually don't go up or down.
Why do you want to do the BS/MS?
I want to partake in the BS/MS program because I recently took the MCAT, and I will be applying to medical school this cycle. When I go into medicine one day, I plan to also do some form of research. Hence, by having a master’s degree, it will give me leverage among my class.
Stuff we talked about in your office:
- Upon training mice, we see an increase in boutons and dendritic complexity.
- When looking at the transcriptome throughout training, from the interval 1-7 days, we see everything as being fairly neutral. However, at around day 14, we see an upregulation. For this reason, we are interested to know if APP plays a role in motor learning in healthy subjects. APP does play a role in Alzheimer’s, a disease where memory loss is an associated pathology.
- 1) Forelimb reach to grasp task – focusing on days 1, 4, 7 & 14
- 2) Extract the APP protein and subject it to western blot analysis
- 3) Histology
- ) Perfusion – fixative to replace blood and fix tissue
- ) Tissue processing by cutting the brain
- ) Immunolabeling - antibodies against APP, then we will use Neun = Neuron Marker and a retrograde tracer