Methodologies in measuring behaviour PDF

Methodologies in the behavioural sciences and neurosciences Approaches and Methods in the Study of Behaviour  Simple-system approach Simple behaviours as indices or markers Advantage: parsimony. Ease of replication and quantification. Reductionistic approach. Disadva...

Methodologies in the behavioural sciences and neurosciences Approaches and Methods in the Study of Behaviour  Simple-system approach Simple behaviours as indices or markers Advantage: parsimony. Ease of replication and quantification. Reductionistic approach. Disadvantages: missing subtle interactions. Definitions of terms (operational and especially, ostensive definitions) and meaningless statements. Examples: "we found that the rats behaved more". "it was determined that the animals in the experimental group were more active". Simple-system approach: Challenges Issues or questions: Easy to determine if the behaviour is present or not? Enough to count occurrences or frequencies? Expression of the behaviour? Intensity important? Other modulations? The 3 little P's (Fentress, 1988): what is the study of behaviour (neuroscience) about? Process Pattern Phenotype The straw house The stick house The brick house Organism Event A Genes X X X Environment Event B Experience/Environment The study of individual behavioural patterns Particularities of behavioural neuroscience (including clinical/medical neuroscience) over the other behavioural sciences and neurosciences: Direct observations of behaviour: often idiographic, longitudinal and descriptive studies. Interest in patterns of behaviour (more or less stereotyped movements or actions). Ethograms, sociograms/sociomatrices Ethogram: inventory, catalogue, written repertoire of all the behaviour patterns of a species. They tend to focus on the form of behaviour, in orders to study behaviour sequences. The brain is a sequencer of movements / motor actions Sociogram: Same, but for social behaviours. Ostensive de nitions: Definitions of the behaviours with detailed descriptions, examples, photos, drawings, etc. Make sure everybody on your team agrees with the categories: This can be a real challenge! fi Studying behaviour patterns: ethograms Choose the species Choose the individual(s), group(s), etc. Choose the behaviours, or signs Choose the measures Choose the sampling rules Choose the recording rules  Consistency » reliability Intra-observer consistency (with an intra-observer reliability measure): each observer records SIMILAR behaviours the SAME WAY. That includes you: Do you keep scoring consistently over time? Inter-observer consistency (with an inter-observer reliability measure such as an index of concordance, Kappa coefficient, Kendall coefficient): consistency of recording and scoring for between ALL OBSERVERS. This guarantees replicability for your team of investigators or any other team wanting to verify your results or expand on your ideas.  De ne the behaviour’s boundaries The issue: the segmentation/clustering of behaviour(s). The concept of “bout” (think of the French “un bout” for “portion”, “part”, “segment”). Where/when does it start, and where/when does it end? (temporal and spatial boundaries). Clusters of behaviours, acts, actions or events. fi  usually defined as a ratio of occurrences per unit of time Frequencies example: number of pecks per minute Measures of tendency usually defined as a ratio of occurrences per trials, bouts, sequences (acronym: "FOLD") Occurrences example: number of pecks per sequence This taxonomy of dependent variables was time between events, states, actions. first suggested by Russell, Mead & Hayes Latencies time between stimulus and reaction to the stimulus (e.g., reaction times). (1954) and points out the particular nature of the concept of "intensity" and distinguishes well between frequencies and occurrences of duration of a single occurrence. a behaviour. total duration. Durations mean duration. duration as proportion of time duration as percentage of total time Common definitions and uses: amplitude or physical quantity. Measures of intensity for psychologists, concept of “local rate”: number of component acts per Many potential operational or ostensive definitions. unit time spent on the activity. Sped-up or hurried nature of a behaviour. rating scales (Likert-like) extracting serial and temporal patterns in a sequence of events (actions Measures of spatio-temporal con guration: or movements) or states. sequences (the Lashley serial order problem) issues of predictability, stereotypy, rigidity (temporal, spatial), motor perseverations (repetitions of a behaviour), etc. fi Kinematics/dynamics } Velocity (v) Distance (d) d= vt Time (t) Acceleration (a) d = vi + ½ at 2 vf = vi + 2 at  Sequential analysis: methodology and issues Objective: Search for behaviour (spatio-temporal) patterns How stereotyped or predictable is the action sequence? Random: 0% Stochastic (probabilistic): > 0% and < 100% Deterministic: 100% Tools (quantitative): Markovian analysis, Information Theory analysis, Log-linear analysis, time series analysis, etc.  Corollaries How important is the serial order of behaviour? Lashley (1951)!* Addressing the temporal and spatial structure of the sequence? How preceding events influence or predict the current one or upcoming events? Analysis of transitional probabilities (with matrices and digrams) Frequency of trigrams, tetragrams, pentagrams, etc. * https://www.sciencedirect.com/science/article/abs/pii/S0167945707000280  Stochastic processes Main tools for our example Information theory Markovian chains For both approaches, there are a few basic steps that apply to the “number” (count or frequency) of behaviours (called events): First step, identify the “monograms” Example: if 2 behaviours, A and B, count how many times A occurs and how many times B occurs.  Step 1: Monograms Behaviour sequence: AABAABABAABAAABAB Behaviour A: 11 Behaviour B: 6  Step 2: Digrams and Transition matrices Sequence: AABAABABAABAAABAB A B A 5 6 B 5 0  Step 3: Trigrams Sequence: AABAABABAABAAABAB AAA: 1 AAB: 4 ABA: 5 BAA: 3 etc...  Step n... Tetragrams: e.g., AAAA, AAAB, AABA... Pentagrams: e.g., AAAAA, AAAAB... Many analyses will stop with digrams (based on the idea that preceding events in uence or predict the current one or upcoming events) Patterns tend to emerge. Then... fl  Interpretation 1 Predictive value: Detectable patterns? Structure? Patterns of behaviour as indicators of neural patterns (e.g., the concept of CPG or central pattern generator). Also: FAP’s, MAP’s, and other pre-programmed motor patterns, “Universal Grammar” of movements. Contextual analysis: Search for a meaning » chickadees and other animals are chatting / communicating. About motor cognition (frontal lobes)? Sequence learning?  Interpretation 2 Motor signatures: Cortical regions: Higher cognitive dimension (planning, optimisation of movements, problem solving, etc.) Cerebellum: Timing, 3D adjustments (3D environments), fine motor repetitive behaviours Basal ganglia: Sequencing (more so innate, or based on habit learning)  Sampling: Who or what is observed and when? Ad libitum ("whoever", "whatever", "whenever") Sampling methodologies: Behaviour or event sampling: focus on specific, target, behaviours (what?) Scan or time sampling: focus on specific time periods, intervals (when?) Focal or individual sampling: focus on one animal/ individual (who?)  Time sampling: How is the behaviour recorded and/or scored? Continuous recording: all-occurrences recording or scoring Time sampling: Instantaneous sampling: On beep: behaviour occurring? (yes or no). Good for relatively long duration events and common events. Sub-types: Point sampling or fixed interval point sampling One-zero sampling: behaviour occurring? (yes or no) in previous sample interval. Subtypes: One-zero convenience sampling; One-zero fixed time sampling; One-zero random time sampling The microstructure of behaviour  What is behaviour? Change in time in space It is about movement, a motion, an action. Reflects a change in the nervous or endocrine system (proximate immediate causes). Common terminologies carry this idea of “motion”: “Motivation”, “emotion”, “hormone” (from the ancient Greek participle ὁρμῶν, "setting in motion”), etc.  The linguistic and musical metaphors Both language and music are de ning changes in time (and space, e.g., dance, or simply “playing music” or singing — as behaviour) Some principles around the conceptualisation of language (as a behaviour, and more) and music apply to behaviour as a stochastic process. fi  The linguistic metaphor Linguistic metaphor (from semiotics): Historically important in ethology, neuroethology, etc. Syntax (grammar): Patterns, rules (= predictability) Semantics: Meaning (cues and signals) Pragmatics: Social (or physical) context Prosody: Amplitude and intensity of action Musical metaphor: More appropriate?  The expression of behaviour: The musical metaphor Melody Rhythm Harmony ··· Temporal organisation: Serial organisation: Parallel organisation: Rhythmic patterns, variations in intensity, combinatorial organisation, duration of events, time modulatory signature coordination, progressions signature New trends: Soni cation of data » Musi cation of data Detection patterns (e.g., rhythm) via sound or music. fi fi  Stochastic processes Probability in time, dynamic probabilities, transitional probabilities. Typical approaches: Markov chains, Information Theory, time series analysis. Use of transition matrices (for digrams): Digram table Search for frequency of: Monograms: A, B A B Digrams: AA, AB,... ➝ Trigrams: AAA, AAB,... A n(aa) n(ab) Tetragrams: AAAA, AAAB, … B n(ba) n(bb) Etc. The diversity of serial patterns Common mistake in behaviour analysis: Assume that the temporal organisation, the sequence, is static, non-dynamic, unmodulated » Here the musical metaphor AND the “prosody” dimension of the linguistic metaphor of behaviour make that point. Example: A is always followed by a random string, but: A (low intensity) is always followed by BAB. A... (long duration) is followed 78% of the time by BB. A (different tone) is followed 45% of the time by C.  Other factors Endogenous factors: Exogenous factors: Metabolism The stimulus (its Motivational status duration, intensity, etc.) Reproductive status The context/situation/ environment Immune system integrity Physical context Etc. Social context  Sequences 1 Simple sequence of mutually exclusive events. 1. Reciprocations (alternating events) in string 1. 2. Perseverations (repeating events) in string 2. Perseverations of “dark” events. String 1 String 2  Sequences 2 Behavioural events can be discrete but they often overlap and/or are temporally spaced: 1. Overlaps. 2. Gaps. Overlap Gap  Sequences 3 Temporal and spatial “invasions” in behavioural events: 1. Overshadowing. 2. Encapsulation. Overshadowing Encapsulation  Sequences 4 Sizes and shapes of circles represent: 1. Intensity, amplitude, magnitude: vertical. 2. Duration (time): horizontal. intensity of event Intensity duration of event Duration  Sequences 5 Ambiguous events or ambiguous ow of events: 1. Dimensional ambiguity (fuzziness; dashed boundaries) 2. Categorical ambiguity (uncertain identity: white or grey?) 3. Transitional ambiguity (under-de ned boundaries) 1 2 3 fi fl  Sequences 6 Very deterministic patterns can be found within sequences with a dominance of randomness and vice versa: High frequency tetragram  Sequences 7 Parallelism between two body part (head and tail) with synchrony (1) or asynchrony (2): 1 2 Head Tail  Sequences 8 The reality: A little bit of everything... Head Tail  Categories of behaviour Observational data on naturally occurring (and observable) behaviours (i.e., events, actions, movements): measures of latency, frequency, duration, intensity and sequencing. Learning and cognitive tests are also applied to test perceptual and cognitive abilities (perception, memory, decision making, problem solving, etc.) States, such as emotions, if well defined, can also be measures via electrophysiological measures (with GSR, ECG, etc.) » Autonomic nervous system activity  Clusters (categories) of behavioural assays 1. Simple-general 2. Simple-specific Slides with a yellow background 3. Acquired behaviours are only lists of 4. Developmental examples, nothing 5. Affective/conative to commit to 6. Socio-affective (interactive; dyads, triads, etc.) memory unless in bold and 7. Cognitive (beyond conditioning) underlined 8. Psychopathological / neuropathological  Cluster 1: Simple-general Simple re exes: Stretch, Knee jerk (humans), Sneezing, Eye blink, Pupillary contraction, Startle, etc. Postures: Standing, Rearing, Lying, Balancing, Sitting, Urination posture, etc. Includes static postures and postural changes. Locomotion: Walking, Creeping, Running, Crawling, Swimming, Stalking, Flying, Hopping, etc. Activity levels: Hyperactivity, hypoactivity: often using open- field tests (see next slide). fl  Locomotor assays Direct observation Photocell-based systems and other automatized systems Motor coordination and balance apparatus: Balance with rotarod Beam walking Footprint analysis  Cluster 2: Simple-specific Species-speci c (species-typical) action patterns (i.e., innate and instinctive behaviours). Simple action sequences (FAP’s, MAP’s) Complex action sequences (e.g., courtship in doves) This important category includes normal and abnormal stereotyped behaviours (e.g., perseverations). fi  Ingestion: tasting, chewing, biting, sipping, drinking, etc. Feeding: taste and food preference tests Drinking: taste preferences Courtship & sexual behaviour: sniffing, licking, chasing, retreating, mounting, etc. Grooming: washing, preening, licking, etc. Gestures: grimacing, tail erection, squinting, tooth baring, smiling, etc. Social & communicative behaviours: Emotional reactions: Defecations, urinations, freezing, etc. Escape & defence (from fear or anxiety): Hissing, spitting, submissiveness, etc. Aggressive behaviour: biting, chasing, dominant postures, etc. Affiliative behaviour: Play, huddling, parental behaviour, etc. Vocalisations: Audible & ultrasonic Scent-marking Note: aggressive & defensive behaviours are indexes of anxiety in the following tasks: defensive burying colony-intruder paradigm elevated plus maze  Cluster 3: Acquired behaviours Re ex / startle / orienting responses Learned responses / conditioned behaviours Simple, non-associative learning Habituation Sensitization Conditioning Classical / Pavlovian / respondent/ type I conditioning Instrumental / Skinnerian / operant/ type II conditioning fl  Pavlovian / classical / respondent conditioning CER (or conditioned emotional response) Avoidance conditioning: active (initiation of behaviour) or passive (suppression of behaviour) avoidance Signalled avoidance Unsignalled (or Sidman) avoidance Conditioned compensatory response Conditioned withdrawal: elicited by drug environment and drug-associated cues Conditioned drug tolerance Aversive conditioning: Taste/odour aversions; most common: Conditioned taste aversion Conditioned defensive burying Skinnerian / instrumental / operant conditioning, i.e., appetitive learning Maze learning: radial arm maze, water maze, etc. Stimulus control learning (learning about the stimulus properties of hormones): Discrimination learning Generalisation learning Brain self-stimulation/self-administration (based on the reinforcing value of a hormone) Specialized  Complex clusters Cluster 4: Developmental cluster Cluster 5: Affective (emotion) / conative (motivation) Cluster 6: Socio-affective (interactive), incl. parental (maternal, paternal). Cluster 7: Cognitive (attention, memory, problem solving, etc.) Cluster 8: Preclinical models of neurologic and psychiatric disorders* * neuropathologies and psychopathologies Developmental clusters Developmental milestones Confounding variables: A few examples handling time of day nutrition litter size post-natal and maternal effect  Developmental milestones in rodent Three main categories: Physical landmarks: Eye opening, incisor eruption, fur development, etc. Locomotor behaviours: elevation of head, elevation of the forelimb and shoulders, pivoting, crawling, walking, swimming, etc. Re exes: See next slide. fl  Milestones: Reflexes Surface righting Tactile startle Air righting Crossed extensor Negative geotaxis reflex Cliff avoidance Rooting reflex Visual placing Grasp reflex Limb placing Bar holding Vibrissa placing Level screen test Auditory startle Vertical screen test  Milestones: Assessment Assessments are species-specific based on known and well documented values. For each category and milestone, and for each species (rat vs. mouse) and strains (if applicable), we know: Average age for response (in days) Range for the response (in days)  Assays for motivation Eating/drinking Food intake measures Food preference tests Reproductive / sexual behaviour Mounts, intromissions, ejaculations, lordosis Hormone replacement Castration/ovariectomy experiments  Assays for parental and social behaviours 1 Observational / descriptive: Licking of pups Nest construction: see Actively hovering over pups gure Motionless positioning over Retrieval pups fi Assays for parental and social behaviours 2 Experimental: Hormonal induction of maternal/paternal behaviour Nest construction, etc. (previous slide) Maternal/paternal preferences (rewarding value of pups); Examples: Conditioned place preference paradigm T-maze and Y-maze tests “Pupomat”: Carousel preference test » Stress, anxiety, depression Measures: Hormones, Behaviours, etc. Experimental stressors: Restraint stressor Electric foot shock stressor Swim stressor Social isolation stressor Resident/intruder stressor Maternal separation stressor Sleep deprivation stressor  Stress, anxiety, depression Anguish What is the difference between these Arousal constructs? Fear Construct validity? Phobia Discriminant validity? Panic Other related Trauma / PTSD constructs: Grief  Rodent models of depression Use of stress to induce depression-like states Behavioural despair (immobility) and learned helplessness paradigms. Initially derived from a forced swim test Dogs: Seligman’s procedure, i.e., inescapable shocks in shuttle box Tail suspension test (mice) Uncontrollable shocks Forced sleep deprivation  Problems There are no animal models of depression that mimic all the symptoms of depression. All current models are models of reactive depression. They typically focus on one or a few of the following aspects of depression: Reduction in psychomotor activity Anhedonia Neuroendocrine responses Cognitive changes Eating dysfunctions Sleeping dysfunctions  Reactive depression Hypothalamus Triggered by stress (broadly defined: social or | CRF/CRH physical): Tends to involve by default the HPA axis. | Depression does naturally involve the HPA axis. Pituitary gland Depressed individuals: enlarged pituitary and | adrenal glands. ACTH | Many depressed individuals have higher levels of Adrenal cortex glucocorticoids (cortisol and corticosterone), | produce more glucocorticoids than non-depressed Cortisol or for the same increase in ACTH. corticosterone  Stress affects: Endocrine system: Adrenaline and noradrenaline (SAM axis) Norepinephrine (NE) Acetylcholine (ACh) GABA And they, in turn, regulate CRF from hypothalamic cells. Depressed individuals have… higher levels CRF levels in their CSF more CRF-producing cells in the hypothalamus... than non-depressed individuals. HPA axis (see right) activity » glucocorticoids (CORT) Both ECT and antidepressants reduce CRF levels.  Dexamethasone (Dex) Dexamethasone: synthetic glucocorticoid Dexamethasone challenge: should induce a strong down- regulation (negative feedback loop) of CRF and ACTH. This does not occur in depressed individuals. Non-responders to DEX treated successfully with anti- depressants are more likely to relapse than those that respond to DEX.  Cortisol and depression Dexamethasone (synthetic gluc.) challenge* * should induce a strong down-regulation (negative feedback loop) of CRF and ACTH  Animal tests of anxiety Tests of anxiogenic or anxiolytic drugs, hormones, situations, etc. Social interaction Light/dark exploration Elevated plus-maze (arm visits) Open field test (space use; activity levels) Defensive burying Thirsty rat conflict: Shocks when drinking  Cognitive testing in animals Major interest in this area in recent years: Dementias, other cognitive impairments* Movement disorders with dementia Other neuro-degenerative diseases (e.g., multiple sclerosis) Neurotoxins and endocrine disruptors Long tradition of tests in animal learning and cognition research. * e.g., ADHD  Cognitive processes Sensation and Perception; Psychophysics Attention / working memory / latent inhibition* / sign-tracking Learning and Memory: Managing information Acquisition; Retention (storage); Retrieval Consciousness levels: Implicit (unconscious / incidental) Explicit (conscious) Problem-solving, decision-making, etc. * latent inhibition: learning to ignore irrelevant stimuli; problematic with schizophrenia  Tests: A few examples T and Y mazes (typically for invertebrates and lower vertebrates) Radial-arm mazes (4, 6, 8, 12 arms) Morris water maze (see next slide) Detection learning: yes/no, go/no-go to presence/absence of S Discrimination learning: mAFC (e.g., 2AFC, 3AFC, … 6AFC) Identi cation* learning: matching-to-sample Rule learning / learning sets * naming task in adult humans fi  Preclinical models of neurologic and psychiatric disorders: a sample Parkinson’s Dural inflammation model Models: Mutant strains (migraine pain model) Lesions Painful diabetic neuropathy (STZ Toxins rat model) 6-OHDA Nociception: hot-plate, tail-flick, MPTP formalin tests Global/Focal Cerebral Ischemia Experimental autoimmune Photochemical cortical lesions encephalomyelitis (EAE) Fluid percussion model Amyotrophic lateral sclerosis (traumatic brain injury) (ALS) models Neuropathic pain Chemoconvulsant model of chronic spontaneous seizures