Measuring Animal Behavior

and Evidence for Behavior

Lecture 4
 Factorial Design Example
Fly covered in mold
• Some fungi are delicious and nutritious, others
are lethal
• Flies may avoid locations where they might be
infected with deadly mold
• Females feed more, so male and female
avoidance may differ

Have:
• Mold possible treatments:
• mold
• no mold
• Sex possible treatments:
• female
Fungus enters through
• male weak points in cuticle (e.g.,
segmentation, hair sockets)
 Factorial Design Example
• For a factorial design, need all possible
combinations of two or more variables

Sex
males females

yes N flies N flies

Mold N = sample size

no
N flies N flies
Flies and Fungal Spore Exposure
• Do male or female flies
avoid fungal spores (aka
conidiospores)?

Dr. C. Nick Keiser

More info: https://www.keiserlab.com/
Keiser et al 2019. Sex differences in disease avoidance
behavior vary across modes of pathogen exposure. Ethology
126:5
Flies and Fungal Corpses
• Do male or female flies
Fly corpse killed by avoid dead fly corpses
the covered in fungus?
entomopathogenic
Fly fungus
(Metarhizium
robertsii)

Dr. C. Nick Keiser

More info: https://www.keiserlab.com/
Keiser et al 2019. Sex differences in disease avoidance
behavior vary across modes of pathogen exposure. Ethology
126:5
Determining What Question to Ask

• The question is important and can

lead to different understandings of
organism behavior
 Harnessing Animal Behavior for
Potential Applications
• Aphids avoid leaf surfaces with bacteria Pseudomonas syringia
• The efficacy of infection may differ with bacterial strain
• This behavior can be harnessed for future potential applications of
Psuedomonas to reduce aphid pest pressures in agricultural crops

Hendry et al. 2018. Current Biology 28.; Smee et al. 2021. Appl Environ Microbiol 87(11).
 Summary

• Measuring behavior requires considerations

inherent to conducting experiments
• Plus an understanding of observer effects and
appropriate sampling techniques
• The “correct” way to do an experiment depends on
the question being asked
• Experiments do not “prove” anything, they provide
support for one or more hypotheses
 Mechanisms:
Hormones & Behavior
Lecture 4
 Overview

• Big picture thoughts on investigating mechanisms

of behavior
• Hormones and their effects
• Vole example from the book
• Feedbacks between hormones and behavior
• Assigned paper
• Effects of one individual’s hormones on other
individuals
• Intrauterine mice
 MECHANISMS OF BEHAVIOR
Neurobiological mechanism: activation of dopamine neurons

VTA: ventral tegmental area

OFC: orbitofrontal cortex

Reduced behavior with foot shock Similar behavior with foot shock
 Approaching Mechanisms

When Looking for Mechanisms

• What mechanisms are responsible for behavioral differences:
• Among species?
• Among populations?
• Among sexes?
• Among individuals within the same population?
• Within a particular individual when they age, change ‘life stage’ (e.g.,
puberty, transition to parenthood), or even state (e.g., stressed vs
not)
When Starting with a Mechanism
• What does it “do”:
• When the mechanism is removed?
• When the mechanism is blocked at a particular time/tissue?
• When the action of the mechanism is augmented?
• When the mechanism evolves?
 Same Hormone, Same Behavior?

Hormones are integrated with other mechanisms of

behavior
• Hormones act on receptors
• Receptors activate functions in cells/tissues
• Changing one part of this “chain” of events (e.g., the
concentration of circulating hormone) will not always
have the same effect
• For voles, hormone concentration does not matter if there are
no receptors that respond to the hormone
 Same Hormone, Different
Behavior?

There can be more than one way to produce (or

inhibit) a behavior
• Functional redundancy—manipulating some mechanism
(e.g., injecting hormone) may have no effect even if the
mechanism is in fact important to the behavior
• Variation—different populations/species/sexes have
similar behaviors, but generated by different
mechanisms
Approaching Hormone Actions

• What question is being asked in a particular study

• Avoid assumptions. Verify that “the whatever
hormone” occurs in multiple studies at multiple
different biological scales and all point to the same
mechanism
• If different studies point to different hormones, that is
also valid
• The mechanisms of hormones can differ and/or are
complicated
 Key Points about Hormones
• We all have them (hormones)
• Hormones are chemically-diverse signaling
molecules
• Hormones are secreted by glands and can move
throughout the brain and body
 Glands and Hormone Secretion
• Hormones are secreted often by glands and can move all throughout the brain and
body
• Hormones are good candidates for coordinating multiple behavioral and
physiological changes or differences
• E.g., sex differences, state transitions (puberty, pupation, parenthood), “fight
or flight”, etc.
• Hormones can have very specific effects at target cells

Endocrine Cells found in Endocrine

glands secrete hormones
 Hormone Circulation
• Hormones are secreted often by glands and can move all throughout the brain and
body, but affect specific target cells that contain receptors
 Neurotransmitters
• Neurotransmitters usually stay in a very local
area, e.g., the postsynaptic cleft, but some can
serve as hormones
• If secreted into the blood stream then they act
as neurohormones
Neurohormones a type of neurotransmitter
 Day Length,
Hormones, and
Behavior
• Day length triggers changes in
testosterone secretion
• Greater lengths of light result in
increased T(estosterone)
secretion
• Greater T results in greater
likelihood of aggression towards
other males
• This may allow for males to gain
better access to mates, nest
defense, and brood defense
 Hormones and Timescales

Hormones can act on a wide variety of timescales

• Immediate e.g., epinephrine/adrenaline
• Epinephrine was extracted by John Abel in the US and he
named it
• Adrenaline was marketed by Parke, Davis & Co and given this
propriety name
• So now both names are used interchangeably
• Medium-term (‘tonic’) e.g., cortisol and stress
• Long-term e.g., menopause
• “Latent”—hormones may act at one time to “prepare” animals to
perform a behavior in response to a cue that may come later
• E.g., migration, mating, building neural circuits, etc
 Hormones and Timescales

The time of day at which

circulating levels of key
endocrine factors peak in
humans.

Abbreviations: F, female
individuals only; M, male
individuals only

Gamble et al. 2014. Nature Reviews in Endocrinology 10(8).

 Testing Hypotheses About
Hormones and Behavior With Voles
Vole Species Differ in Parental Care
Prairie voles
• Monogomous
• usually one mate per
breeding season
• Males provide parental
care

Meadow voles
• Polygamous
• usually multiple mates
per season
• No male care

• These species are closely

related
• Most recent common
VP = name of a brain region
ancestor (MRCA) ~6 MYA
Dark areas = stained receptors
p. 72 of PAB 4th ed.
Vasopressin receptors distinguish
meadow and prairie voles

Prairie voles spend _____ time with partners

than meadow voles.

VP = name of a brain region

Dark areas = stained receptors
p. 72 of PAB 4th ed.
Activity: Think Pair Share
• Prairie voles spend _____ time with
partners than meadow voles.

• The number of vasopressin receptors is

______ for prairie voles.

• What experimental approach was used

(observational, experimental,
phylogenetic, theory)?

• Is this an interaction? If so, between

which factors?
 Meadow Voles have
Vasopressin Receptors
Genetically Added

What does these findings

VP = name of a brain region tell us?
Dark areas = stained receptors
p. 72 of PAB 4th ed.
Feedbacks
Between
Hormones
and
Sometimes
Behaviors
Glucocorticoids are steroid hormones that are anti-inflammatory
Dr. Safran – UC Boulder

Dr. Hau – Max Plank

ornithology
 Journal Article Formats and
Finding Information
• Some journals (and some article formats) are
shorter and don’t have the same structure as more
typical papers
• Do careful reading to figure out which parts are
which
• Also examine the Supplemental Material
Yellow highlighted
text is methods
Purple highlighted
text is results
 Establishing a Baseline
and Color Alteration
• Before experimental manipulation, darker males
had higher androgen concentrations and heavier
males had lower concentrations of androgens
• Those initially selected for treatment groups did
not differ significantly in plumage color, androgen
concentrations or body mass
• Color was modified using a non-toxic permanent
marker to match dark feather color
 Effects of Color Enhancement
After one week:
• Coloration
• Males from the color-enhanced group showed
increased concentrations of circulating androgens
• Males in the control group showed decreased
androgen concentrations
• Body Mass
• Males in the color-enhanced treatment group lost
body mass
• Males in the control groups gained weight between
the two sampling periods
Effect of Hormones on Male Barn
Swallows

body mass

androgens

Plumage color
 How might plumage
body mass color influence
androgens?

androgens

Behavior
of other
Plumage
barn
color swallows
Safran et al 2008

Lighter plumage color Darker plumage color

Testosterone Levels in Animals

body mass

androgens

plumage

PAB 4th ed p. 66
Safran et al 2008
The Effects of One Individual’s
Hormones on Other Individuals
Effect of one individual’s hormones on
another individual’s
 Neighbors and Hormones

• Intrauterine “neighbors” influence later behavior in mice

• Male mice gestating next to female siblings have lower exposure to testosterone
• Result: less aggressive and less sexually active than males gestating by male siblings
 Neighbors and Hormones
Similar results with gerbils

• Intrauterine “neighbors” influence later behavior in mice

• Male mice gestating next to female siblings have lower exposure to testosterone
• Result: less aggressive and less sexually active than males gestating by male siblings
 Activity: Think Pair Share

• Whose behavior was compared?

• What were the hypotheses?

• What predictions were tested?

• How were they able to focus on intrauterine position?

• Could the mice differ in some other way that

influences behavior?
 Summary
• Hormones travel through the body and across tissues
• Hormones have the potential to coordinate the expression
of multiple behaviors and other traits
• Differences in hormones and/or their receptors can
contribute to behavioral differences among individuals or
species
• Hormones can influence behavior AND behavior can
influence hormones
• Social interactions can also influence hormones
 Next Class
• Neurobiology

Assignments
• Reading: PAB ch3 neuro
section
• Variable channel expression
in identified single and
electrically coupled neurons
in different animals