Week 3

Question 1

• different neurodegenerative diseases
• imparied proteostasis=

Answer 1

• different neurodegenerative diseases can have different protein aggregates
• impaired proteostasis leads to protein misfolding

Question 2

• α-synuclein behaves similarly
• what does the chaperone system do?
• function of ubiquitin-proteasomoes and autophagy-lysosome systems
• toxic oligers become what and lead to what
• misfolded proteins accumulate where and causes what

Answer 2

• a-synuclein behaves similarly to prions
• the chaperone system helps fold unfolded proteins into non-toxic oligomers
• ubiquitin-proteasomoes and autophagy-lysosome degrade proteins
• misfolded proteins = toxic oligomers become amyloid fibrils and lead to impaired proteostasis (formation of proteins) and saturated and overwhelmed pathways
• misfolded proteins accumulate in the ER and lead to ER stress and change in refulation of apoptosis and cell death.

Question 3

• Toxic oligomers & protein aggregate could be passively released
• oligomers form pores where that cause what
• toxic oligomers and prion similarity

Answer 3

• Toxic oligomers & protein aggregate could be passively released into the extracellular space
• oligomers form pores in nearby cells that cause an influx of Calcium ions that disrupt cell function
• toxic oligomers leave the affected cell through vesicles, exocytosis and enter a new cell to induce more misfolding. serves as a template

Question 4

model of alpha-synuclein oligomers:
- controls
- Results: Cells with two different
- GTP/alpha-synuclein constructs dimerized glowed how and what produced no light
- what was this model injected into
- mouse model injection in delivered via

Answer 4

• positive control: just GFP or a whole GFP attached to a-synuclein
• Results: cells with two different GFP/ a-synuclein constrcuts that were dimerized glowed green but not as much fluorensce as the control. When one dimer is paired with a-syn and another isnt, it does not glow= negative control
• this model was injected into C elegans
• mouse model is injected via virus vector

Question 4

Protein Fragment Complementation:
- marker
- what needs to be produced and how can it be produced

Answer 4

• 2 a-synuclein, one attached to N marker and another attached to C marker
• we need to produce GFP and in order to do that the two componenets need to dimerize, and come together

Question 5

• MPP+ and MPTP
• What does MPP+ do
• Rotenone

Answer 5

• MPTP is inactive, goes into mitochondria which makes it into MPP+ which is a toxic, active form.
• MPP+ => Inhibition of Complex 1 chain= ER stress, clinical features of PD, DP neurons in SNc, Lewy body like inclusions but not lewy body
• Rotenone: also complex 1 inhibitor = ER stress, PD symtoms, DP neurons in SNc, Lewy body like inclusions but not lewy body

Question 6

1- octen-3-ol
effects on flies

Answer 6

• volitile compound found in mould
• can be environmental effect leading to PD
• only minimal dosages (0.5 ppm) can lead to Drosophila lethality after 12 days and movement deficits after 7 days
• 1- octen-3-ol is a toxin that reduces dopamine levels

Question 7

Gal4, UAS, ElaV

Answer 7

Gal4- transcriptional activator
UAS: upstream activator sequence
ElaV activates Gal4, Gal4 activates UAS activates GFP=> fluoresence

Question 8

TH-Gal4; UAS-GFP transgenic model

Answer 8

TH activates Gal4, Gal4 activates UAS, UAS activates GFP and dopaminergic neurons fluoresce