The 11th Annual

WPI-IIIS Symposium~Deciphering the Mysteries of Instinctive Behaviors~

Date:Venue: 1F Hall, IIIS Building, University of Tsukuba

Wednesday, February 22, 2023, 8:30 – 18:00

Time Table
Opening |8:30 - 8:45

MC: Mayumi Kimura (WPI-IIIS, University of Tsukuba)
Welcome

Masashi Yanagisawa

Director, WPI-IIIS, University of Tsukuba

Opening Address 1

Akira Ukawa

WPI Academy Director, JSPS

Opening Address 2

Noriko Osumi

WPI Academy Officer / Vice President, Tohoku University

Session 1 | Chair: Yu Hayashi (WPI-IIIS, University of Tsukuba)
8:45 - 9:25

The evolution of sleep loss in Mexicancavefish

Alex Keene

Texas A&M University

Page 4

9:25 - 10:05

Metabolic control of sleep

Anissa Kempf

University of Basel

Page 5

10:05 - 10:20 BreakSession 2 | Chair: Arisa Hirano (WPI-IIIS, University of Tsukuba)
10:20 - 11:00

Sleep duration, sleep quality and energy
metabolim

Esra Tasali

University of Chicago

Page 6

11:00 - 11:40

Sleep and circadian rhythm disruption: fromgenes and environment to behaviour

Stuart Peirson

University of Oxford

Page 7

11:40 - 11:55

Lunch Preparation / BreakSession 3 | Chair: Kaspar Vogt (WPI-IIIS, University of Tsukuba)
11:55 - 12:35

Rapid eye movement sleep is initiated bybasolateral amygdala dopamine signaling inmice

Emi Hasegawa

Kyoto University

Page 8

12:35 - 13:15

Midbrain circuits linking social stress andsleep

Xiao Yu

Chinese Academy of Sciences, Shanghai

Page 913:15 - 13:55

Hypothalamic neural circuit mechanism
controlling female reproductive behaviors

Sayaka Inoue

Stanford University

Page 10

13:55 - 14:05 Photo / Break

Poster Session | Chair: Shoi Shi (WPI-IIIS, University of Tsukuba)
14:05- 14:45

Data BlitzPage 14-15

14:45 - 16:00 Poster Presentation

Page 14-15

Session 4 | Chair: Sakiko Honjoh (WPI-IIIS, University of Tsukuba)
16:00 - 16:40

The mechanisms and meaning of sleep
homeostasis

Vladyslav Vyazovskiy

University of Oxford

Page 11

16:40 - 17:20

Towards cortex-wide volumetric recording ofneuroactivity at cellular resolution

Alipasha Vaziri

The Rockefeller University

Page 12

17:20 - 18:00

A signaling pathway for transcriptional
regulation of daily sleep amount in mice

Qinghua Liu

National Institute of Biological Sciences, Beijing

Page 13

18:00

Closing3

Session 1

8:45 - 9:25

The evolution of sleep loss in Mexican cavefish
ABSTRACTAlex Keene
Texas A & M University, USA4Sleep is nearly ubiquitous throughout the animal kingdom,
yet its duration and timing varies dramatically betweenspecies. We have developed the Mexican tetra, Astyanax
mexicanus as a model to study evolved changes in sleep,and how naturally occurring genetic variation encodesfor sleep differences between individuals. These fish
exist as eyed-surface populations that inhabit the riversof northeast Mexico and multiple blind cave populations
that have converged on sleep loss. We have identified
neuromodulators that contribute to the evolution of sleep
loss in A. mexicanus cavefish including upregulation of
the wake-promoting neuropeptide Hypocretin/Orexin. In
addition, we have developed transgenic and gene-editing
methodology in this emergent model system allowing forsystematic investigation of the genes and neurons regulating
evolved differences in sleep. Systematic analysis has
identified convergence on wide-spread neuroanatomicaldifferences between surface fish and cavefish including
hypothalamic expansion that is accompanied by increased
sleep intensity. Current studies seek to identify how
naturally occurring genetic variation contributes do these
phenotypes. Investigating the mechanisms of sleep loss in
Mexican cavefish has potential to provide insights into thevariation in sleep need throughout the animal kingdom.
Further, the resources developed to study sleep can bebroadly applied to study other cavefish associated traits
including obesity, diabetes, and dysregulation of stressresponse.

Session 1

9:25 - 10:05

Metabolic control of sleep
ABSTRACT

Anissa Kempf
University of Basel, Switzerland

Sleep is essential, yet its function remains one of biology’s
biggest mysteries. A short or poor night of sleep is usuallyfollowed by longer and/or deeper periods of sleep. The
existence of such a compensatory mechanism suggeststhat our brain can monitor the amount of waking time
and trigger corrective action if needed. In the fly brain,
this behavioural change is reflected in electrical changesof sleep-promoting neurons that project to a region of thefly brain referred to as the dorsal fan-shaped body (dFB).
Previous work has shown that dFB neurons of sleepdeprived flies are more excitable than those of rested flies.
This suggests that the excitability state of dFB neurons tracks
the sleep need of the fly: as sleep need builds up, dFB
neurons upregulate their excitability, and as sleep debt getscleared, it reverts back to baseline. Understanding what
modulates the excitability of dFB neurons will undoubtedly
inform us about the molecular basis of sleep need. Here, I
will show that mitochondrial metabolism plays a key rolein this process: dFB neurons track their redox state to forma biochemical memory of sleep need and to adjust their
excitability state accordingly. Uncovering the molecularbasis of sleep homeostasis might shed light onto one of
biology’s biggest mysteries: the function of sleep.

5Session 2

10:20 - 11:00

Sleep duration, sleep quality and energy
metabolism
ABSTRACT

Esra Tasali
University of Chicago

6Sleep has been recognized as an important contributor tometabolic health. Insufficient sleep duration and reduced
sleep quality are associated with impairments in energy
metabolism. Insufficient sleep duration has been associatedwith an increased risk for obesity and diabetes. Sleep
apnea is a highly common sleep disorder. It has beenstrongly linked to adverse metabolic (e.g., impaired glucosemetabolism) and cardiovascular outcomes. This talk will
review the clinical and experimental human studies linking
reduced sleep duration/quality to impairments in energy
metabolism, discuss potential underlying mechanisms, andprovide highlights from interventional human studies aiming
to improve sleep patterns and metabolic outcomes.

Session 2

11:00 - 11:40

Sleep and circadian rhythm disruption: from genes
and environment to behaviour
ABSTRACTStuart Peirson
University of Oxford, UKThe 24h cycle of activity and sleep provides perhaps
the most familiar example of circadian rhythms. Inmammals, circadian activity rhythms are generated bya master biological clock located in the hypothalamicsuprachiasmatic nuclei (SCN). This clock is synchronized(entrained) to the external light environment via light input
from retinal photoreceptors. However, sleep is not a simple
circadian output and also is regulated by a homeostatic
process whereby extended wakefulness increases the needfor subsequent sleep. As such, the amount and distributionof sleep depends upon the interaction between bothcircadian and homeostatic processes. To understand thecomplex genetic regulation of sleep, we have applied
forward and reverse genetic approaches in mice. Firstly,using a forward genetic screen using behavioural sleep
screening, we have identified a novel sleep mutant. In
addition, using transcriptomic meta-analysis of sleep
deprivation data, we have been able to identify novel sleep
genes and subsequently verify sleep phenotypes. Finally,
the light environment also exerts effects on sleep andcircadian rhythms. The modern artificial light environment
leads results in exposure to dim light on an evening(DLE), which causes a misalignment of normal sleep andcircadian rhythms, with consequences for peripheral clocks,
metabolism and cognitive behaviour.

7Session 3

11:55 - 12:35

Rapid eye movement sleep is initiated bybasolateral amygdala dopamine signaling in mice
ABSTRACTEmi Hasegawa
Kyoto University, Japan

8

Abstract: The sleep cycle is characterized by alternating
non-rapid eye movement (NREM) and rapid eye movement(REM) sleeps. The mechanisms by which this cycle isgenerated are incompletely understood. We found thata transient increase of dopamine (DA) in the basolateral
amygdala (BLA) during NREM sleep terminates NREM sleep
and initiates REM sleep. DA acts on dopamine receptor
D2 (Drd2)-expressing neurons in the BLA to induce theNREM-to-REM transition. This mechanism also plays a rolein cataplectic attacks, which are a pathological intrusionof REM sleep into wakefulness, in narcoleptics. Theseresults show a critical role of DA signaling in the BLA in
initiating REM sleep and provide a neuronal basis for sleep
cycle generation. Furthermore, this mechanism involved
serotonergic neurons in the dorsal raphe and serotonin
modulates DA signaling in the BLA to induce the NREM-toREM transition.

Session 3

12:35 - 13:15

Midbrain circuits linking social stress and sleep
ABSTRACT

Xiao Yu
Chinese Academy of Sciences,
Shanghai, China

Stress correlates with an increased risk of clinical anxiety.
Sleep has been suggested to be one of the mechanismsfor alleviating the malign effects of stress. Nevertheless,the function and benefits of sleep are poorly understood.
In mice, social defeat stress, an ethological model for
psychosocial stress, induces sleep. Such sleep could enable
resilience, but how stress promotes sleep is unclear. In
this lecture, I will talk about how we discovered some key
cells in the midbrain that regulate sleep in mice, and how
these cells play an important role in mood control. I will
substantially demonstrate the mechanisms underlying how
stress-induced sleep was produced and how this sleep helps
the mental processing of stressful events by discoveringa specific circuit in the mouse midbrain dedicated todetecting stress and inducing restorative sleep. We foundthese specialized cells in the midbrain receive stress inputs
from stress-detecting centers in the brain and induce sleep
through the lateral hypothalamus. Stress-induced sleep
generated by this pathway mitigated stress-induced anxiety,and alleviated stress levels, restoring mental and bodyfunctions. This discovery potentially provides a refinedroute for treating anxiety disorders, helping to understate
the essential mechanisms of how sleep benefits in reducing
anxiety in the brain, and strengthening the fundamental
function of sleep.

9Session 3

13:15 - 13:55

Hypothalamic neural circuit mechanism controlling
female reproductive behaviors
ABSTRACT

Sayaka Inoue
Stanford University, USA

10Behaviors are associated to the internal physiological state.
In many species, including mice, females dramaticallychange their sexual behaviors along the state of ovulation.
Females of many species are sexually receptive exclusively
during the estrus, ovulatory phase of the estrous cycle, while
they are not receptive during other phases. Sex hormonessuch as estrogen and progesterone are required for bothovulation and female sexual behavior. Although central andperipheral mechanisms of ovulation is well characterized,
neural circuit mechanisms underlying the estrus-associatedchange of the behavior is poorly understood. We havepreviously shown that progesterone receptor (PR) expressing
neurons in the ventromedial hypothalamus (VMH) areessential for female sexual behavior. Targeted ablationof PR+ VMH neurons abolished female sexual behavior
while it kept the estrous cycle intact. However, whether
PR+ VMH neurons play a role in the estrus-associatedchange of female sexual behavior or they control thebehavior independently from the estrus is unclear. We
examined whether PR+ VMH neurons play a role in theestrus-associated change of female sexual behavior. We
find that PR+ VMH neurons significantly strengthen theirfunctional connections during estrus with the anteroventral
periventricular nucleus. This cyclic increase in connectivity
depends on estrogen signaling in PR+ VMH neurons. We
further find that these projections are essential for female
sexual receptivity during estrus because optogenetic
inhibition of these connections abolishes female sexualbehavior. These findings demonstrate that periodic
remodeling in this behaviorally salient connections playa critical role in associating female sexual behavior
with internal physiological state. We further identified
a transcriptomically-defined homogeneous population
expressing Cckar within the PR+ VMH population. The
Cckar+ VMH neurons are the only neurons within the PR+
VMH neurons that regulate female sexual behavior. I will
present these and recent findings to discuss hypothalamic
circuit mechanisms that control the timing of female sexualbehavior.

Session 4

16:00 - 16:40

The mechanisms and meaning of sleep
homeostasis
ABSTRACT

Vladyslav Vyazovskiy
University of Oxford, UK

According to traditional view, the need for sleep
accumulates during wakefulness and dissipates during sleep.
Despite decades of research, it is still uncertain precisely
which biological variables form the substrate of sleep
need, what characteristics of wake challenge their stability,
how information about sleep-wake history is integrated
over time, and how sleep mediates the restoration ofhomeostasis. An even more fundamental question therefore
remains whether homeostatic sleep regulation reflects anactive process, dynamically shaping daily sleep architecture
in response to a physiological need for the homeostatic
regulation of specific variables, or whether it corresponds
instead to an unknown innate time-keeping process which
ensures only that a certain daily quota of sleep is obtained.
While the earliest theories of sleep homeostasis supposed
the existence of a single variable, termed Process S, which
describes sleep drive at the global level, current views
emphasise the importance of a local and activity-dependent
component. Many candidate mechanisms have been
proposed for the substrate of sleep homeostasis, including
processes occurring at a cellular or local network level.
These include the maintenance of cellular homeostasis,the replenishment of energy stores, the influence of sleeprelated signalling molecules such as adenosine or cytokines,
and the regulation of imbalanced synaptic strengths. The
preference for global sleep, despite its fundamentally local
mechanisms, may be evidence that sleep homeostasis
ultimately does not serve a single specific local function.
An alternative view is that the recent history of neuronal
activity levels may regulate the local tendency to generate asignal (reflected in Process S) which is integrated over larger
neuronal populations through intrinsic network mechanisms
in order to produce a global sleep propensity signal that
estimates the total time spent awake with great accuracy.
This mechanism would enable the brain to enforce a daily
quota of sleep, which could have many benefits at thephysiological and ecological level, rather than to initiate
sleep in response to the homeostatic need of one specificregulated variable.

11Session 4

16:40 - 17:20

Towards cortex-wide volumetric recording ofneuroactivity at cellular resolution
ABSTRACT
Understanding how sensory information is represented,
processed and leads to generation of complex behavior
is the major goal of systems neuroscience. However, theability to detect and manipulate such large-scale functional
circuits has been hampered by the lack of appropriate
tools and methods that allow parallel and spatiotemporally
specific manipulation of neuronal population activity while
capturing the dynamic activity of the entire network at highspatial and temporal resolutions.

Alipasha Vaziri
The Rockefeller University, USA

A central focus of our lab is the development andapplication of new optics-based neurotechnologies for largescale, high-speed, and single-cell resolution interrogationof neuroactivity across model systems. Through these, wehave consistently pushed the limits on speed, volume size,and depth at which neuronal population activity can beoptically recorded at cellular resolution. Amongst others
have demonstrated whole-brain recording of neuroactivityat cellular resolution in small model systems as well as more
recently near-simultaneous recording from over 1 million
neurons distributed across both hemispheres and different
layers of the mouse cortex at cellular resolution.
I will present on our efforts on neurotechnology
development and how the application of some of these
optical neurotechnologies could enable solving aqualitatively new range of neurobiological questions thatare beyond reach of current methods. Ultimately, our aim isto uncover some of the computational principles underlying
representation of sensory information at different levels,
its processing across the mammalian brain, and how its
interaction with internal states generates behavior.

12Session 4

17:20 - 18:00

A signaling pathway for transcriptional regulationof daily sleep amount in mice
ABSTRACT

Qinghua Liu
National Institute of BiologicalScience, Beijing, China

Different mammals vary widely in daily sleep quotas,
ranging from 2 to 5-h in giraffes to 18 to 22-h in koalas1,2,
reflective of a specific homeostasis between sleep andwakefulness in each species. In mice and humans, sleep
quantity of individuals is governed by genetic factors andexhibit age-dependent variations3,4. However, the coremolecular pathways and effector mechanisms that regulate
sleep quantity in mammals remain unclear. Here, wecharacterize a major signaling pathway for transcriptional
regulation of sleep amount in mice by adeno-associatedvirus-mediated somatic genetics analysis5. Adult brain
chimeric-knockout of LKB1 kinase, an activator of AMPKrelated protein kinase SIK3/SLEEPY6-8, markedly reduces
non-rapid eye movement sleep (NREMS) amount anddelta power–a measure of sleep depth. Downstreamof LKB1-SIK3 pathway, gain/loss-of-function of histone
deacetylases HDAC4/5 in adult brain neurons causes
bidirectional changes of NREMS amount and deltapower. Phosphorylation of HDAC4/5 is regulated inrelation to sleep need, and HDAC4/5 specifically regulate
sleep amount in posterior hypothalamus. Genetic andtranscriptomic studies reveal that HDAC4 cooperates with
CREB in both transcriptional and sleep regulation. Thesefindings introduce the concept of signaling pathways
targeting transcription modulators to regulate daily sleep
need and demonstrate the power of somatic genetics inmouse sleep research.

13

Poster Session
The 11th Annual WPI-IIIS Symposium~Deciphering the Mysteries of Instinctive Behaviors~

Data Blitz by Poster Presenters

14:05 – 14:45

Poster Presentation

14:45 – 16:00

Poster List
#PresenterTitle

1

Yusuke Iino

Synaptic plasticity in prefrontal cortex regulates sleep

2

Ayako Imamura

Both ablation and activation of POA cause sleep fragmentation

3

Kseniia Prokofeva

Regulation of Sleep and Wakefulness by Neuronal Circuits linking Ventrolateral Preoptic Nucleus andLateral Hypothalamic Area

4

Zhongwen Zhang

Mechanism by which the BNST→DpMe pathway induces arousal

5Ruth Li

Role of Gastrin-releasing hormones-producing neurons in regulation of the circadian rhythms

6

Tohru Takahashi

24 hours QIH induction by OPN4-based optogenetics

7

Ai Miyasaka

Male sexual behaviors are regulated by dopaminergic signaling in NAc in mice

8

Susumu Umezawa

Ventral tegmental area dopaminergic neurons mediate sleep reduction induced by ablation of thesurrounding GABAergic neurons

9

Koustav Roy

Opto-chemical control of slow-wave sleep in the nucleus accumbens by a photoactivatable allostericmodulator of adenosine A2A receptors

10

Rintaro Otani

Development of photocaged drugs targeting adenosine 2A receptor

11

Keita Kajino

Design and synthesis of novel κ opioid receptor agonists with bicyclo [2.2.2] octane skeleton

12

Kao Hashimoto

Analysis of orexinergic function in chronic pain-like states

13

Insung Park

Orexin receptor antagonist modulates sleeping energy metabolism without overt effects on sleep
architecture

14

Yue Cao

Does different-intensity exercise affect the rest-activity rhythms among older adults?

15

Saki Tsumoto

A Study on Automatic Sleep Analysis for a Variety of Sleep Patterns

16Javier DiazAssessing Wakefulness Intensity as a Counterpart to Sleep Depth During the Sleep-Wake Cycle

17

Zhiwei Fan

Binaural beats at slow frequencies less than 1 Hz shorten the latency of slow-wave sleep

18

Kazuki Sato

Targeted memory reactivation during REM sleep selectively enhances brain responses to unpleasantimages

19

Hibiki Okamura

Exploration of a novel mechanism of sleep regulation that depends on feeding condition

20

Ami Kaneko

Analysis of the mechanism of REM sleep with focus on Parkinson’s disease

21

Srinivasan Sakthivel

Transient recruitment of adult-born neurons for fear memory consolidation in REM sleep

22

Vergala Pablo

CaliAli: neuronal tracing utilizing vasculature in calcium imaging

23

Noriko Hotta-HirashimaSex difference in sleep/wakefulness from infancy to adulthood in a hypersomnia model, Sleepy
mutant mouse24

Liqin Cao

Sleep disturbances in mice with a CDKL5 kinase-dead missense mutation, a novel mouse model ofneurodevelopmental disorder

25

Tomoyuki Fujiyama

NALCN in the forebrain and pons-medulla regions have distinct roles in REM sleep regulation

26

Minjeong Park

Sleep/wake behavior of mice lacking PKA-phosphorylation sites of SIK327

Tomohiro Kitazono

Novel upstream and downstream pathways of SIK3 in the regulation of sleep and wakefulness

28

Shinya Nakata

Molecular mechanisms for sleep/wake regulation by SIK3 kinase signaling

29

Fuyuki Asano

SIK3-HDAC4 in the suprachiasmatic nucleus regulates the timing of arousal at the dark onset andcircadian period in mice

30

Kanako Iwasaki

SIK3 regulates sleep amount through hypothalamic nuclei

15

MEMO