base knowledge Flashcards
Histological and molecular techniques
- Immunohistochemistry
- Western Blot
- qPCR
What does immunohistochemistry test for?
to visualise and quantify protein expression in tissue sections
what does Western Blot test for?
Measures protein levels in tissue homogenates or cultured cells
what does qPCR test for?
Gene expression
what is the Moris Water Maze test used for?
spatial memory & learning
what is the reversal Morris Water Maze used for?
behavioural flexibility
what is the T-maze alternation test for?
Working memory
what is the elevated plus maze used for?
anxiety levels
what is the open field test used for?
locomotion & anxiety
what is the rotarod test used for?
motor learning & co-odrination
what is the pole test used for?
parkinsons model motor assessment
what is the three-chamber sovial test used for?
social behaviour - in ASD models
what is the grooming behaviour analysis used for?
repetitive behaviour - ASD like phenotypes
How is immunohistochemistry visualised?
- flourescence microscopy (IF)
- Colorimetric staining (IHC)
what does a bands intensity indicate in Western blotting?
stronger band = more protein expression
how is protein loading controlled in Western blot?
housekeeping proteins like B-actin
why is WB better than IHC for quantifying protein levels?
- more precise level markers, but lacks spatial resolution
why would you use qPCR over Western Blot?
- if you were measuring mRNA levels, not protein levels
when would the Moris Water MAze be used?
evaluation of memory impairment in alzheimer’s models
What does perservation in a T maze alternation test show?
indicates a deficit in memory or decision making (cognitive deficits), as wild type mice have a natural tendency to alternate between arms when making choices.
what area of the brain does a positive T-Maze test indicate dysregulation is occuring?
prefrontal cortex, hippocampus, and basal ganglia
How the rotarod test works (3)
- mice are placed on a rotating rod
- time to fall (latency) is recorded over multiple trials
- Learning is shown by increased latency over trials
for which neuro-disorder would the rotarod test be a suitable test? (2)
Huntingtons’s and Parkinson’s
what does reduced social interaction with the novel mouse indicate in a three-chamber social interaction test?
Autism like phenotype
what neuro-divergent model of mice shows excessive grooming behvaiour?
autism models - linked to repetitive behaviours
what behaviours are mice models of austism linked to? (2)
- repetitive behaviours
- social deficits
what are the typical tests for mouse mdoels of ASD? (3)
- three chamber test - social behaviour
- Grooming - repetitive
- perseverance tests
what gene is important in ASD?
- UBE3A (angelman syndrome)
what is the important gene related to schizophrenia & psychosis (1)?
DISC1 gene
animal model tests for psychosis & schizophrenia
- Novel object recognition test - memory impairment
- T maze alternation test - working memory deficits
what are the animal model tests for mood disorders? (3)
- Forced swim test - Behvaioural despair in depression
- Sucrose preference test - Anhedonia, lack of pleasure response
- Elevated plus maze - anxiety related behaviour
what is the major gene related to mood disorders?
BDNF
what is the major gene related to epilepsy?
SCN1A
what are the neurotransmitters invovled in epilepsy:
- GABAergic dysfunction - reduce inhibition leads to excessive firing
- Glutamate overactivity - hyperexcitable neurons
animal model tests for epilepsy: (1)
- electroencephalography (EEG) in mice - detects abnormal neural firing
cause of parkinsons disease?
loss of dopaminergic neurons in the substantia nigra
what are the animal model tests used to test for Parkinsons?
- MPTP mouse model - induces PD-like neuro-degeneration
- Pole test- tests bradykinesia
- rotarod test - Motor coordination
what are the neurotransmitter involvement in PD?
- dopamine depletion in basal ganglia - leads to motor symptoms
- ACH imbalance - leads to cognitive dysfunction
what are the animal model tests used to test for alzheimers? (3)
- Morris Water Maze - spatial memory deficits
- T-maze alternation - working memory impairments
- Novel object recognition - recognition memory
what is the neurotransmitter involved in alzheimers?
ACH deficit - loss of cholinergic neurons
What is the cause of huntingtons disease?
expansion of CAG repeats in the HTT gene, leading to toxic protein aggregation
animal model tests for Huntington’s (2)
- Rotarod test - tests motor decline
- Gait analysis - coordination issues
