Animal Models of AD

Question 1

2 novel models in AD research

Answer 1

virus based models

PLB transgenic knock in mice

Question 2

ideal gene delivery system

Answer 2

transfer genes into developing and mature animals
transduce cells with high efficiency
be cell type/tissue specific
mediate high level, long term expression
limited cytotoxicity
avoid over expression
sufficient DNA lengths

Question 3

2 viral gene delivery applications

Answer 3

disease modelling - mutated genes, introduce genes

treatment - gene therapy, functional genes

Question 4

receptors for adenovirus

Answer 4

coxsackie adenovirus receptor

CAR

Question 5

what determines cell types virus infects and targetting capacity?

Answer 5

serotype

Question 6

what are serotypes?

Answer 6

sub-groups of micro-organism which share similar antigens

Question 7

viral vector systems

Answer 7

retrovirus
adenovirus
AAV
HSV

Question 8

AD relevant AAV viruses

Answer 8

APP
BACe1
Tau

Question 9

immunoblot

Answer 9

show virus expressing the gene

transgene expression

Question 10

expression analyses

Answer 10

analysis under bright field, fluoresence and merged microscopy
vesicles taken up transgene

Question 11

amyloid transfected neurons

Answer 11

degeneration and death

Question 12

tau transfected neurons

Answer 12

form clusters

Question 13

quantification of cell death

Answer 13

count % healthy neurons

defined processes, no vacuoles

Question 14

what can increase % healthy neurones with tau/APP?

Answer 14

caffeine

Question 15

problems with transgenic AD mouse models

Answer 15

varied promoters, no suitable controls, difficult to compare

Question 16

PLB lines positives

Answer 16

compare triple/double/single lines
APP, tau, PS1, BACE1
instead of pronuclear injection use knock in
APP and tau - use cre/loxp to remove 1

Question 17

PLB4BACE1 knock in model rational

Answer 17

BACE rate limiting step in production of beta amyloid

drug discovery

Question 18

how to see if animal expressing hBACE1?

Answer 18

immunohistochemistry and western blot
increased compared to control
dentate gyrus and Ca1

Question 19

APP processing in PLB4Bace1

Answer 19

decreased full length APP

increased cleavage products

Question 20

PLB4 - brain inflammation

Answer 20

severe in hippocampus and cortex

GFAP astrocyte staining

Question 21

PLB4BACE1 mice behaviour

Answer 21

decreased activity, working memory, semantic memory, anxiety, spatial memory

Question 22

metabolic phenotypes of PLB4 mice

Answer 22

diet induced obesity and glucose intolerance

Question 23

PLB4 FDD-PET imaging

Answer 23

less uptake of glucose in PLB4 mice in brain

increased brown adipose tissue - subtype of diabetes

Question 24

type of PLB diabetes

Answer 24

lean type 2

Question 25

AD and diabetes - what is the link?

Answer 25

increased risk of type 2 diabetes in AD | BACE1

Question 26

CHOP

Answer 26

multifunctional and pro-apoptotic transcription factor

Question 27

PLB - food intake and energy regulation

Answer 27

both increased

Question 28

PTP1B

Answer 28

inhibitor of insulin receptor | increased in PLB4 mice

Question 29

ER stress leads to what?

Answer 29

upregulation of CHOP

Question 30

role of Ach

Answer 30

attention, arousal and memory

Question 31

delayed matching to place

Answer 31

rats learn to locate platform, delay or 30s or 1 hour to trial 2 platform changed each day

Question 32

what type of drug is scopolamine?

Answer 32

anticholinergic

Question 33

acquisition of DMTP under scopolamine

Answer 33

learn DMTP - impair long term memory

Question 34

amyloid cascade

Answer 34

APP, amyloid protein, amyloid deposits, neuronal toxicity

Question 35

beta amyloid + scopolamine

Answer 35

additional effects impairs memry in water maze

Question 36

where is tau found?

Answer 36

normally in neurons pathologically in glia trace amounts in heart, kidney, lung, muscle, pancreas

Question 37

normal function of tau

Answer 37

promote tubulin assembly into MT bundling of MT stabilises cytoskeleton

Question 38

tau in AD

Answer 38

Braak and Braak et al 1993 tau undergo conformational change early in AD abnormal tau accumulate in somatodendritic compartment of neuron

Question 39

3 types of tau pathology

Answer 39

Neurofibrillary tangles - pyramidal cells filled with paired helical and straight filaments of tau neuritic plaques - tau around amyloid core neuronal threads

Question 40

hyperphosphorylation of tau

Answer 40

abnormal in AD destabilises MT - aggregation okadoic acid blocks phosphatases

Question 41

Tau and PAPS

Answer 41

PAPS produces pores in cell allowing intracellular delivery of tau into hippocampal cells then use okadaic acid - via minipump for 7 days water maze and histology

Question 42

reference memory water maze

Answer 42

4 days, 4 trials per day platform same position reversal learning - in opposite quadrant

Question 43

3 types of animals in PAPS trial

Answer 43

only PAPS PAPS and tau PAPS, tau and okadaic acid

Question 44

triple animals in PAPS

Answer 44

slow learners reduced long term potentiation tau in CA1 of hippocampus phosphorylated tau in distal dendrites