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Advanced molecules, membranes and cells > Animal Models of AD > Flashcards

Animal Models of AD Flashcards

1
Q

2 novel models in AD research

A

virus based models

PLB transgenic knock in mice

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2
Q

ideal gene delivery system

A

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

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3
Q

2 viral gene delivery applications

A

disease modelling - mutated genes, introduce genes

treatment - gene therapy, functional genes

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4
Q

receptors for adenovirus

A

coxsackie adenovirus receptor

CAR

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5
Q

what determines cell types virus infects and targetting capacity?

A

serotype

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6
Q

what are serotypes?

A

sub-groups of micro-organism which share similar antigens

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7
Q

viral vector systems

A

retrovirus
adenovirus
AAV
HSV

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8
Q

AD relevant AAV viruses

A

APP
BACe1
Tau

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9
Q

immunoblot

A

show virus expressing the gene

transgene expression

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10
Q

expression analyses

A

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

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11
Q

amyloid transfected neurons

A

degeneration and death

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12
Q

tau transfected neurons

A

form clusters

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13
Q

quantification of cell death

A

count % healthy neurons

defined processes, no vacuoles

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14
Q

what can increase % healthy neurones with tau/APP?

A

caffeine

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15
Q

problems with transgenic AD mouse models

A

varied promoters, no suitable controls, difficult to compare

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16
Q

PLB lines positives

A

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

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17
Q

PLB4BACE1 knock in model rational

A

BACE rate limiting step in production of beta amyloid

drug discovery

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18
Q

how to see if animal expressing hBACE1?

A

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

19
Q

APP processing in PLB4Bace1

A

decreased full length APP

increased cleavage products

20
Q

PLB4 - brain inflammation

A

severe in hippocampus and cortex

GFAP astrocyte staining

21
Q

PLB4BACE1 mice behaviour

A

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

22
Q

metabolic phenotypes of PLB4 mice

A

diet induced obesity and glucose intolerance

23
Q

PLB4 FDD-PET imaging

A

less uptake of glucose in PLB4 mice in brain

increased brown adipose tissue - subtype of diabetes

24
Q

type of PLB diabetes

A

lean type 2

25
Q

AD and diabetes - what is the link?

A

increased risk of type 2 diabetes in AD

BACE1

26
Q

CHOP

A

multifunctional and pro-apoptotic transcription factor

27
Q

PLB - food intake and energy regulation

A

both increased

28
Q

PTP1B

A

inhibitor of insulin receptor

increased in PLB4 mice

29
Q

ER stress leads to what?

A

upregulation of CHOP

30
Q

role of Ach

A

attention, arousal and memory

31
Q

delayed matching to place

A

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

32
Q

what type of drug is scopolamine?

A

anticholinergic

33
Q

acquisition of DMTP under scopolamine

A

learn DMTP - impair long term memory

34
Q

amyloid cascade

A

APP, amyloid protein, amyloid deposits, neuronal toxicity

35
Q

beta amyloid + scopolamine

A

additional effects impairs memry in water maze

36
Q

where is tau found?

A

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

37
Q

normal function of tau

A

promote tubulin assembly into MT
bundling of MT
stabilises cytoskeleton

38
Q

tau in AD

A

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

39
Q

3 types of tau pathology

A

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

40
Q

hyperphosphorylation of tau

A

abnormal in AD
destabilises MT - aggregation
okadoic acid blocks phosphatases

41
Q

Tau and PAPS

A

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

42
Q

reference memory water maze

A

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

43
Q

3 types of animals in PAPS trial

A

only PAPS
PAPS and tau
PAPS, tau and okadaic acid

44
Q

triple animals in PAPS

A

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

Advanced molecules, membranes and cells (20 decks)

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