laura gallardo_nurr1 project

8/16/2019 Laura Gallardo_Nurr1 Project

1/13

Role of Nurr1 on excitatosynaptic structures in prim

hippocampal neuronsMaster's Degree in Biochemistry, Molecular Biology and Biomedicine

Lau


2/13

Nurr1 is involved in excitatory transmission-desurvival of glutamatergic neurons by actdownstream CREB and upstream of BDN

NMDAR

Ca2+Protein

kinases

CREB Nurr1

Nurr1

Nucleus

CREB

P

Bdnf !ro"


3/13

Synaptic plasticity processes play an important role in brain developmencognitive functions like learning and memory.

In pathological conditions (l!heimer disease" those plasticity processes be affected causing a reduction on synapse si!e and density.

#he number of $urr% e&pressing cells significantly declines in an agedemanner, accompanied by increased plaue deposition. #hus, it suggests altered e&pression of $urr% is associated )ith l!heimer*s disease progr

$MD+ stimulation is necessary for neuronal survival and developmendifferent cerebral areas.

$urr% could be implicated in synapse regulation and could be a candidarevert negative effects of amyloid on synapses during early stages of

l!heimer*s disease.


4/13

-vere&pressionof $urr%

ctivation ofdo)nstream

effectors

hange&citasynapincreasi!e a

densit

spineshippoca

neuro


5/13

%. Designing and producing $urr% silencing (sh+$" anovere&pression constructs in a lentiviral vector.

/. #o produce lentiviral viruses and titrate in 012/34#.

4. #o prepare hippocampal primary neurons to checklentiviral infection by +#5+ and )esternblot.

6. #o infect primary neurons to do immunocytochesmistryagainst 5SD3789:;I soft)are.


6/13

$ di%erent constructions

s%Nurr1 s%Nurr1 +cDNA N


7/13

$ di%erent constructions

cDNA Nurr1


8/13

Infecting hippocampal primary neurons (mouse" )ith sh+$ $urr


9/13

&here am ' now(

eckin' lentiviral infection (s%RNA a'ainst Nurr1) * ,estern-*loectin' %i!!oca"!al !ri"ar neurons ,it% s%RNA of Nurr1&tainin' cDNA of Nurr1 to !roduce t%e overe0!ression construct


10/13

1) Chec!ing lentiviral infection *shRN+ against Nurr1, by wesin cortical neurons&

s%Nurr1 (Rat) Control s%.Nurr1 s%1Nurr1

s%2Nurr1 s%.Nurr1


11/13

1) Chec!ing lentiviral infection *shRN+ against Nurr1, by wesin cortical neurons&


12/13

) 'nfecting hippocampal primary neurons with shRN+ of Nurr


13/13

.han! you

laura gallardo_nurr1 project

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