Flashcards: Animal Research Regulation and Techniques — 24 cartões

UK animal ethics law

Animals (Scientific Procedures) Act 1986 regulates animal research.

Directive 2010/63/EU

EU law updated animal research regulation, enacted in UK 2013.

Regulated animals

Vertebrates and cephalopods are covered; invertebrates like C. elegans are excluded.

Procedure definition

Any act likely causing pain, suffering, distress, or lasting harm above a needle threshold.

Three licences required

Establishment (PEL), project (PPL), individual (PIL).

Cosmetic testing ban

Complete ban under UK animal research framework.

3Rs principle

Replacement, Reduction, Refinement to minimize animal use and suffering.

Replacement example

Use in vitro or less sentient organisms instead of higher animals.

Reduction example

Design studies to use fewer animals with proper controls.

Refinement example

Improve handling, anesthesia, and monitoring to reduce pain.

Humane endpoints

Predefined criteria to end experiments early, minimizing suffering.

Stereotaxic landmarks

Bregma and Lambda are key skull landmarks for targeting brain regions.

Stereotaxic workflow

Identify Bregma, level skull with Lambda, drill burr hole, lower micro-syringe.

Rodent behavioural tests

Rotarod (motor), Cylinder (sensorimotor), NOR (memory), Sucrose (anhedonia).

C. elegans habituation

Decreased response after repeated stimuli, reflecting learning.

Knock-Out vs Knock-In

KO deletes gene; KI inserts or modifies gene.

CRISPR sgRNA

A 20-base-pair sequence guiding Cas9 to target DNA.

PCR thermocycle steps

Denaturation, annealing, extension.

DNA template in PCR

Starting DNA strand(s) to be amplified.

RT-PCR purpose

Convert RNA to cDNA then amplify.

qPCR Ct value

Cycle at which fluorescence crosses threshold; lower means more target.

Relative vs absolute qPCR

Relative compares Ct between samples; absolute uses standards for copy number.

Agarose gel migration

DNA moves toward positive; smaller fragments migrate farther.

Gel visualization limit

Band intensity reflects relative DNA amount, size estimated with ladder.