Keynote Lecture
Novel Neuronal-Glial Signaling in Magnocellular and Parvocellular Neurons of the Hypothalamic
Paraventricular Nucleus
Jeffrey G. Tasker, Juhee Haam, Zhiying Jiang, and Chun Chen
Department of Cell and Molecular Biology, Neuroscience Program, Tulane University, New Orleans,Louisiana, USA
It is increasingly clear that glial cells can no longer be relegated to the role of supporting castmembers of the central nervous system, and that they perform critical signaling functions that
contribute to signal transmission in neural circuits. The notion of the tripartite synapse, according towhich signal transmission occurs among pre- and postsynaptic neural elements and astrocytic
partners, is now well accepted. However, this neuronal-glial signaling is highly localized to thesynapse, where intercommunication between astrocytes and neurons modulates presynaptic releaseprobability and postsynaptic responses. We recently discovered a novel form of neuronal-glial
signaling in the hypothalamic paraventricular nucleus (PVN) that exploits the full spatial domain ofastrocytes to transmit retrograde signals to distal upstream neuronal targets. Thus, vasopressin andcorticotropin releasing hormone (CRH) neurons in the PVN transmit retrograde signals to distal
glutamate and/or GABA neurons via dendritic neurotransmitter/neuropeptide release and activationof an astrocyte signaling intermediate. The upstream glutamate and GABA neurons project back to
the vasopressin and CRH neurons to form a closed neuronal-glial-neuronal signaling loop. This novel
form of retrograde trans-neuronal-glial transmission allows PVN neurons to regulate their synaptic
inputs by controlling presynaptic neuron firing, thus providing a powerful means of influencing theirhormonal output. This work was supported by NIH grants NS042081 and MH066958, and by thePierson Chair in Neuroscience.

Jeffrey G. Tasker, Ph.D.
Catherine and Hunter Pierson Chair in Neuroscience
Professor of Cell and Molecular Biology
Tulane University
Jeffrey Tasker received his bachelor’s degree in biology from the Universityof Colorado in 1981 and his doctorate in neuroscience from the University of
Bordeaux, France in 1986. He received postdoctoral training in the
Physiology Dept at the Tulane University Health Science Center and in the
UCLA Neuropsychiatric Institute. He joined the faculty of Tulane
University as an assistant professor in the Department of Cell andMolecular Biology in 1991, and currently holds the rank of professor. He
has had continuous research funding from the National Institutes of Healthfor over 20 years and has published over 75 research papers, reviews andbook chapters. He became the Catherine and Hunter Pierson Chair in
Neuroscience in 2005, served as director of the Tulane University
Neuroscience Program from 2006 to 2014, and is currently director of theDivision of Neurobiology of the Cell and Molecular Biology Department.
He has served on several NIH and NSF grant review panels and on theeditorial board of the journals Endocrinology, Stress, and Steroids. The
research in his laboratory uses electrophysiology and molecular biology
approaches to study the electrical activity and molecular signaling of
neuroendocrine cells of the hypothalamus and neurons of the amygdala,with implications for the understanding and future treatment of such
central nervous system conditions as stress, depression, obesity, andposttraumatic stress disorder.

Formation of Endoplasmic Reticulum-Associated Compartment (ERAC) in Vasopressin Neurons: A
Mechanism by Which ER Stress Is Reduced
Hiroshi Arima, Yoshinori Azuma, Yoshiaki Morishita, Masayuki Hayashi, and Daisuke Hagiwara
Department of Endocrinology and Diabetes, Nagoya University Graduate School of Medicine,
Nagoya, Japan
Analyses of a mouse model for familial neurohypophysial diabetes insipidus (FNDI), characterized
by progressive polyuria due to progressive decreases in arginine vasopressin (AVP) release, revealed
that mutant proteins are accumulated in a sub-compartment of endoplasmic reticulum of AVP
neurons. By forming such a structure called ER-associated compartment (ERAC), AVP neurons arelikely to reduce ER stress. However, the formation of ERAC is hampered in relatively old FNDI miceor in the mice subjected to chronic dehydration. Failure of ERAC formation induces autophagy in the
AVP neurons, which finally die through the mechanism called autophagy-associated cell death.
ATF6α, one of three ER stress sensors, seems to contribute to the formation of ERAC through theupregulation of ER chaperone such as BiP. It is also of note that, while enlargement of a
sub-compartment of ER, a similar structure to ERAC, was observed in the AVP neurons in wild-typemice subjected to dehydration, dilation of ER was diffused in the AVP neurons in dehydrated ATF6α
knockout mice. Thus, our data suggest that misfolded proteins are confined to the ERAC in the AVP
neurons of not only FNDI but also wild-type mice, in order to reduce ER stress.

Hiroshi Arima, M.D., Ph.D.
Professor, Department of Endocrinology and Diabetes, Field ofInternal Medicine, Nagoya University Graduate School of Medicine
Hiroshi Arima is a graduate of Nagoya University in 1988. After the
M.D. degree, he obtained his Ph.D. degree from Nagoya University in
1997. Following an appointment as a clinical and research fellow at
Nagoya University, he worked at Section on Endocrine Physiology,
Developmental Endocrinology Branch, National Institute of Child
Health and Human Development, National Institutes of Health, as avisiting fellow from 1998 through 2001. After coming home, he wasappointed a clinical and research associate, assistant professor andthen associate professor at Nagoya University. He became Professor of
Endocrinology and Diabetes at Nagoya University Graduate School of
Medicine in 2015. He is a clinical endocrinologist and specialized inwater and electrolytes balance as well as energy balance of anorganism.

Study of the Molecular and Physiological Mechanism Regulating Social Memory by Neurons Expressing
Oxytocin Receptor
Katsuhiko Nishimori
Laboratory of Molecular Biology, Graduate School of Agricultural Science, Tohoku University, Sendai,
Japan

Interest to neurohypopysial hormon oxytocin (OXT), basically synthesized in PVN magnocellular
neurons and parvocellular neurons and SON, is now quickly growing with its curing effect, whenadministrated to ASD patients, as a new pharmaceutical to improve social behaviors of them. We
generated oxytocin gene deficient and oxytocin receptor gene deficient mice lines, observed impaired
social behaviors with them, and considered these line as a mice-level ASD model animal. We are now
trying to clarify the mechanism, how deletion of oxytocin receptor gene leads to the onset ofabnormal social behaviors, and the localization of neurons expressing oxytocin receptor, of which
activation gives rise to normalization of social behaviors in mice. With this strategy, we design
experiments to elucidate the pharmaceutical mechanism by which the administration of OXT has atherapeutic effect on ASD patients. In the analysis of the brain prepared from oxtr-venus knock-in
mice, we found that various nuclei related to social behaviors expressed OXTR in different typeneurons, such as serotonergic, GABAergic, Glutaminergic and dopaminergic in nuclei-specificmanners. OXTR-expressing neurons in medial amygdaloid nucleus (MeA) and lateral septal nucleus(LS) might have a higher contribution on the regulation of social memory. On the other hand, higher
population of OXTR-expressing neurons in MeA (30%) and in LS (~100%) were characterized to be
GABAergic and calretinin(+) interneurons. With the hypothesis that the major population of neurons
expressing OXTR were GABAergic, we generate GABAergic neuron-specific OXTR gene KO mice, bycrossing Vgat-IRES-Cre line with Oxtr(fx/fx) line. Resultant mice showed impaired social novelty.
We continuously analyzed the responsible neurons expressing OXTR on social behaviors andespecially social novelty, in LS and MeA.

Katsuhiko Nishimori, Ph.D.
Professor, Laboratory of Molecular Biology, Graduate School of
Agricultural Science, Tohoku University
Katsuhiko Nishimori is a graduate of University of Tokyo in 1977, andhe also obtained his Ph.D. degree from University of Tokyo in 1982.
After working as a post-doc at Dr. Teruhiro Beppu’s lab at Universityof Tokyo, he became an assistant professor at Laboratory ofEnzymology, Institute of Applied Microbiology, University of Tokyo.
Then, he was appointed as associate professor at Graduate Program in
Agricultural Science, Tohoku University. He worked as a visiting
associate professor at Department of Pathology, Baylor College ofMedicine at Huston from 1994 through 1996. He participated ingenerating the oxytocin knockout mouse during his stay at Baylor.
After coming back to Sendai, he was appointed Professor of Molecular
Biology in 2001 at Graduate Program in Agricultural Science, Tohoku
University. He developed many novel mouse lines, and among them, isthe oxytocin receptor knockout mouse which is of great value inelucidating the mechanism of oxytocin.

Neural, Hormonal and Experiential Control of Social and Emotional Behavior
Sonoko Ogawa
Laboratory of Behavioral Neuroendocrinology, University of Tsukuba, Tsukuba, Japan
We have been studying brain mechanisms of social behavior, particularly regulation of sex-specificsexual and aggressive behavior by gonad steroids. In our recent studies, we also have focused onsocial interactive behaviors including social preference, social recognition and social memory as wellas emotional and anxiety-related behavior in social context. Behavioral analysis in knockout mice of
two types of estrogen receptors, ER or ER, revealed that ER might be involved in ‘modulatory’regulation of various social and emotional behaviors. Considering differential localization between
two types of ERs, we are currently testing several hypotheses that ER in the paraventricular
nucleus of hypothalamus, medial amygdala, dorsal raphe nuclei, and locus coeruleus may mediate
genomic action of estradiol. In this talk, we will overview our recent findings from behavioral
analysis in mice with brain site-specific manipulation of ER gene expression and discuss possible
neural, hormonal and experiential control of social and emotional behavior.

Sonoko Ogawa, Ph.D.
Professor, Laboratory of Behavioral Neuroendocrinology, University ofTsukuba
Sonoko Ogawa is a graduate of the University of Tsukuba in1979, andshe also obtained her M.A. degree at the same university. She obtained
the Ph.D. degree at the University of Connecticut in 1988. She moved
to the Rockefeller University, New York, and was appointed a
postdoctoral associate, research associate, assistant professor, andthen associate professor in the Laboratory of Neurobiology and
Behavior. In 2004, she became a full professor in the Laboratory of
Behavioral Neuroendocrinology at the University of Tsukuba. She isalso a visiting professor in the School of Medicine and Health Sciencesat Monash University Sunway Campus, Malaysia, as well as an
adjunct faculty member in the Laboratory of Neuroendocrinology at
the Rockefeller University. She is specialized in behavioral
neuroendocrinology and has been studying the brain mechanisms ofsocial and emotional behaviors. She is an awardee of Young
Psychologist Award at the 24th International Congress of Psychology
in 1988, UConn Alumnae Who Have Made a Difference by the
University of Connecticut Alumni Association in 1988, the Rockefeller
University/Karolinska Institute Nicholson Award, 2003, andUniversity of Tsukuba, Academic Year 2012 Best Faculty MemberAward in 2013.

Noxious or Non-Noxious Inputs to Oxytocin Neurons: Possible Roles in the Control of Behaviors
Tatsushi Onaka, Yuki Takayanagi, Masahide Yoshida, and Shota Okabe,
Division of Brain and Neurophysiology, Department of Physiology, Jichi Medical University,
Shimotsuke, Japan
Oxytocin-plays an essential role in milk ejection and parturition in mammals. Oxytocin has also
been shown to be involved in the control of various behaviors, including anxiety-related behaviors,
eating behaviors and affiliative behaviors. We have previously shown that stressful stimuli such as
noxious stimuli and conditioned fear stimuli, activate hypothalamic oxytocin neurons via activationof brainstem catecholaminergic/prolactin-releasing peptide (PrRP)-postive neurons. Oxytocin
neurons are activated not only by aversive stimuli but also possibly by pleasant stimuli. Food intake
has been shown to induce activation of oxytocin neurons. We found that PrRP neurons are also
involved in activation of oxytocin neurons following food intake. Social contact may also affectactivity of oxytocin neurons. Non-noxious pleasant tactile stimuli induced 50-kHz ultrasonicvocalization, an index of positive states in rats, and activated hypothalamic oxytocin neurons.
Physiological roles of oxytocin released during stressful or pleasant stimuli remain to be clarified.
However, application of oxytocin has been shown to have anxiolytic, anorexic and pro-social actions.
In fact, we have shown that endogenous oxytocin reduces anxiety-related behaviors, decreases
amounts of food intake per meal and facilitates social recognition via various neural pathways.

Tatsushi Onaka, M.D., Ph.D.
Professor of Physiology, Jichi Medical University
Tatsushi Onaka obtained his M.D. degree from the University of Tokyoin 1985, and his Ph.D. degree from Jichi Medical University in 1990.
He became an assisitant professor at Jichi Medical University, andthen worked at Babraham Institute, Cambridge, UK, as a British
Council Fellow from 1992 through 1994. After coming home, he wasappointed an associate professor, and in 2006, he became Professor of
Physiology at Jichi Medical University. He is specialized in thephysiological and endocrinological regulatory mechanisms ofneuroendocrine peptides as well as their implications in animal
behavior. Among other neuroendocrine peptides, he is especiallyinterested in oxytocin and prolactin-releasing peptide. He is anawardee of Kawakami Prize of Japan Neuroendocrine Society in 2003.
He gave a Mortyn Jones Memorial Lecture of British Neuroendocrine
Society in 2011.

Optogenetic Approach to Regulate Neuronal Activity in Rat Vasopressin Neuron
Yoichi Ueta
Department of Physiology, School of Medicine, University of Occupational and Environmental
Health, Kitakyushu, Japan
Optogenetic approach is a powerful tool to regulate neuronal activity by light-sensitive ion channels
and investigate the interaction between neuronal network and behavioral change. Here, we havegenerated a transgenic rat that expresses the arginine vasopressin (AVP)-channelrhodopsin 2(ChR2)-eGFP fusion gene. The eGFP that indicates the expression of the ChR2 gene was observed in
the supraoptic nucleus (SON), the magnocellular divisions of the paraventricular nucleus (PVN) andthe suprachiasmatic nucleus (SCN) that are known to localize AVP-containing neurons. Confocal
laser scanning microscopic observation revealed that ChR2-eGFP was mainly localized in themembrane of magnocellular neurosecretory cells (MNCs) in the SON and the PVN of transgenic rats.
The eGFP was also observed in the fibers originated from AVP neurons and posterior pituitary gland.
The intensities of eGFP in those nuclei were marked increased after chronic salt loading (2% NaCl todrink for 5 days). Using whole cell patch-clamp recordings in in vitro preparations such as a single
cell isolated from the SON and brain slice including the SON of AVP-ChR2-eGFP transgenic rats,
repeated blue light evoked action potentials repetitively in a current clamp mode, and caused inward
currents in a voltage clamp mode. Optogenetic manipulations to regulate neuronal activity of AVP
MNCs have come true, using transgenic techniques.

Yoichi Ueta, M.D., Ph.D.
Professor of Physiology, School of Medicine, University of Occupational
and Environmental Health
Yoichi Ueta is a graduate of University of Occupational andEnvironmental Health, from which he obtained his M.D. degree in
1987, and his Ph.D. degree in 1991. Then he worked at Babraham
Institute (Dr. Gareth Leng), Cambridge, UK, as a research fellow.
After coming home, he was appointed an instructor at University ofOccupational and Environmental Health. He worked at Department ofMedicine (Prof. Stafford Lightman), University of Bristol, as a visiting
researcher from 1993 through 1995. After serving as a lecturer, hebecame Professor of Physiology, University of Occupational andEnvironmental Health in 2000. He is specialized in the physiologicalmechanisms of neurohypophysial hormones and neuropeptides related
to stress response. Recently, he developed rat lines whose
vasopressinergic or oxytocinergic neurons are labeled by fluorescentproteins. He is an awardee of Kawakami Prize of Japan
Neuroendocrine Society in 1998.

Characterization and Future Perspectives of the Corticotropin-Releasing Factor-Modified Yellow
Fluorescent Protein-Knock-In Mouse
Keiichi Itoi
Graduate School of Information Sciences and Graduate School of Medicine, Tohoku University,
Sendai, Japan
Corticotropin-releasing factor (CRF) produced in the paraventricular nucleus of the hypothalamus(PVH) plays a pivotal role in the regulation of the hypothalamic-pituitary-adrenal (HPA)-axis. All
stress inputs are conveyed to hypothalamic CRF neurons via neural pathways and humoral factors.
It has not been fully understood how CRF neurons are regulated by these inputs because
experimental animals in which CRF neurons can be visualized with fluorescent proteins have notbeen available until recently. We developed a mouse line in which Venus (modified yellow
fluorescent protein) gene was inserted into the CRF gene in frame by homologous recombination(CRF-Venus). Although Venus was colocalized with CRF in the PVH in the CRF-Venus mouse, aconsiderable discrepancy was observed. By bilateral adrenalectomy, most Venus-expressing neurons
became expressing CRF, so the discrepancy may partly be explained by the suppressive
glucocorticoid effect on the CRF promoter. Another mouse line was generated by deletions of the
pgk-Neo cassette from the CRF-Venus genome (CRF-VenusNeo); Venus fluorescence became more
prominent, and most CRF neurons expressed Venus in physiological glucocorticoid states in the
CRF-VenusNeo. Most Venus-expressing neurons also expressed CRF mRNA in brain regions
within and outside the PVH. Direct neural inputs to the Venus neurons, containing glutamate or-aminobutyric acid (GABA), were demonstrated by patch-clamp recordings. Effects of varioustransmitters on the glutamatergic or GABAergic inputs to the CRF neurons are now underinvestigation. With the same strategy for generating the CRF-Venus, CRF-iCre knock-in mouse was
generated; it contains iCre sequence in frame instead of Venus. The iCre recombination took place
selectively in brain regions that express CRF. CRF-Venus, CRF-VenusNeo, and CRF-iCre will be
useful tools for studying the regulatory mechanisms of CRF neurons by cellular and physiological
viewpoints.

Keiichi Itoi, M.D., Ph.D.
Professor of Information Biology and Neuroendocrinology
Graduate Schools of Information Sciences and Medicine
Tohoku University
Keiichi Itoi is a graduate of Tohoku University in 1980. After obtaining
his Ph.D. degree from Tohoku University in 1987, he became apost-doc at Department of Pharmacology (Prof. Thomas Unger),
University of Heidelberg, Germany, and next year, he was appointed
an assistant professor at the same institute. After coming back toJapan in 1990, he became an assistant professor at Tohoku University
Hospital, and later, an associate professor. He worked at Mental
Health Research Institute, University of Michigan (Co-directed by
Stan Watson and Huda Akil), from 1996 through 1998 as a visiting
scholar. In 2001, he was appointed Professor of Information Biology at
Graduate School of Information Sciences, Tohoku University, and also
Professor of Neuroendocrinology at Graduate School of Medicine. He isan awardee of Kawakami Prize of Japan Neuroendocrine Society in
1995.

‘Volume Transmission’ Role of Oxytocin in the Regulation of the Male Sexual Function in the Spinal Cord
Hirotaka Sakamoto
Ushimado Marine Institute, Graduate School of Natural Science and Technology, Okayama
University, Okayama, Japan
A novel concept is proposed that certain neurons in the central nervous system may use bothsynaptic (or wiring) transmission and non-synaptic (or volume) transmission communications. We
previously demonstrated that the gastrin-releasing peptide (GRP) system mediates spinal centerspromoting penile reflexes in rats. A group of oxytocin neurons situated in the parvocellular part ofthe paraventricular nucleus (pPVN) and projecting to the spinal cord controls penile reflexes.
Therefore, the hypothesis that oxytocin, which is transported by long descending
paraventriculospinal pathways, activates proerectile spinal centers has been proposed.
Subsequently, we have found that the axonal distribution of oxytocin in the lumbar spinal cord
exhibits a male-dominant sexual dimorphism in rats. Furthermore, oxytocin binding andexpression of the specific oxytocin receptor were both observed in the spinal GRP neurons.
Consequently, oxytocin efferents might secrete oxytocin from the spinal axonal terminals, andregulate male sexual function through an oxytocin receptor-mediated mechanism in spinal GRP
neurons. However, it has been reported that there are few functional synaptic contacts between
oxytocin neurons in the pPVN and spinal GRP neurons, because most pPVN neurons remained
unlabeled by retrograde trans-synaptic tracing with pseudorabies virus from the bulbocavernosus
muscle in experiments performed to identify neural circuits controlling male sexual functions.
Taken together, these results suggest that hypothalamic oxytocin projections in the lumbar spinal
cord may function, at least partly, via a non-synaptic mechanism: a volume transmission.

Hirotaka Sakamoto, Ph.D.
Associate Professor, Ushimado Marine Institute, Graduate School of
Natural Science and Technology, Okayama University
Hirotaka Sakamoto is a graduate of Hiroshima University in 1997.
After obtaining the M.A. degree (neuroscience) in 1999, he also
obtained his Ph.D. degree (neuroscience) in 2002 from Hiroshima
University. He also obtained another Ph.D. degree (medical science)from Kyoto Prefectural University of Medicine in 2009. After a JSPS
research fellow and post-doctoral fellow, he was appointed a research
associate at Kyoto Prefectural University of Medicine. In 2009, hebecame Associate Professor of Natural Science at Ushimado Marine
Institute, Graduate School of Natural Science and Technology,
Okayama University. His research interests are sexual differentiationof the brain and spinal cord, genomic and non-genomic effects ofhormones on neurons, and the effects of severe psychological stresseson neurons. He is a recipient of many awards for young investigators
including those from Japan Neuroendocrine Society (2008), Japanese
Association of Anatomists (2009), and Zoological Society of Japan(2012). He is an awardee of 2010 Young Scientists’ Prize, theCommendation for Science and Technology from the Minister of
Education, Culture, Sports, Science and Technology of Japan in 2010.

A New Hypothalamic Area Enriched with Perineuronal Nets Having Bidirectional Connections between
the Lateral Septum
Mayumi Nishi, Noriko Horii-Hayashi, and Takayo Sasagawa
Department of Anatomy and Cell Biology, Nara Medical University, Kashihara, Japan
The paraventricular hypothalamic nucleus (PVN) is a well-studied region as the center of regulating
the hypothalamic-pituitary adrenal axis. The present study found a new in the anterior
hypothalamus (AH) of mice, a triangular-shaped area between the PVN and the fornix. This area
was clearly stained with the WFA lectin that is broadly used for labeling the specialized extracellularmatrix structure called perineuronal nets (PNNs). WFA stained this region with a triangularmorphology in coronal brain sections, we named this region perifornical area of the anterior
hypothalamus (PeFAH). The aim of this study was to characterize the PeFAH in terms of its
expressing neuropeptides, neural connections, and functions. Histological and DNA microarray
analyses demonstrated that many of PeFAH neurons were non-GABAergic and highly expressed
enkephalin (Enk) neuropeptides. Moreover, PeFAH neurons expressed calretinin, but notparvalbumin and calbindin. Neuronal tracing revealed that both enkephalin- andcalretinin-expressing neurons in the PeFAH massively projected to the lateral septum. The PeFAH
was received inputs from the lateral septum (LS) neurons expressing calbindin, indicatingbidirectional neural connection between the PeFAH and LS. The c-Fos expression analysis indicatedthat activity of PeFAH neurons was increased by psychological stressors such as restraint, novelty,
and aggression, but not induced by homeostatic stressors of dehydration and fasting. Considering
neuronal subtype and projection, the region of a compact cluster of Enk-positive neurons within the
PeFAH is comparable with the perifornical nucleus previously identified in rats.

Mayumi Nishi, M.D., Ph.D.
Professor, Department of Anatomy and Cell Biology, Nara Medical
University
Mayumi Nishi is a graduate of Kyoto University, and obtained herbachelor’s degree in pharmaceutical Science in 1980. She obtained her
degree of M.D. from Kyoto Prefectural University of Medicine in 1991.
She also obtained her Ph.D. degree from the same university in 1997.
She worked at Department of Biology and Center for Neural Science,
New York University at New York (Prof. Efrain C. Azmitia) as avisiting scientist from 1994 through 1996. After coming back to Japan,
she was appointed an assistant professor, lecturer, and associate
professor at Department of Anatomy and Neurobiology, Kyoto
Prefectural University of Medicine.
In 2009, she became Professor of Anatomy and Cell Biology at Nara
Medical University. Her major interest is the neuroendocrinology ofstress and emotion and her research is aimed at understanding themechanisms by which environmental factors interact with the genome
to influence brain development and to produce diverse forms of neuralplasticity over a lifetime. She is a 1999 awardee of Best Imaging
Award of Japan Bioimaging Society.

Paraventricular Nucleus Mechanisms of Stress Integration
James P. Herman, Matia B. Solomon, Sriparna Ghosal, and Jeffrey G. Tasker
Department of Psychiatry and Behavioral Neuroscience, University of Cincinnati, Cincinnati, Ohio,
USA and Department of Cellular and Molecular Biology, Tulane University, New Orleans, Louisiana,
USA
Parvocellular paraventricular nucleus (PVN) neurons control activation of the
hypothalamo-pituitary-adrenocortical (HPA) axis. These neurons synthesize and secrete
corticortropin-releasing hormone (CRH), which is responsible for ACTH secretion and subsequently,
glucocorticoid release. Our laboratory has used expression of Cre recombinase driven by asimpleminded-1 (Sim1) promoter to explore the impact of selective PVN (and SON)-targeted genedeletion on HPA axis activity. Use of Sim1 to delete the glucocorticoid receptor (GR) in PVN (andSON) neurons results in exaggerated ACTH and corticosterone responses to acute stress in malemice. In combination with electrophysiological data from the Tasker group, these results indicatethat the canonical GR is required for fast feedback inhibition of the HPA axis. There was no effect of
PVN GR deletion on responses to chronic variable stress exposure, indicating that the GR is not
involved in limiting the impact of repeated stress exposure. To test mechanisms of PVN activation,
we bred Sim1-Cre with glucagon-like peptide 1 receptor (GLP1R) flox mice to selectively block theactions of the putative stress-excitatory neuropeptide GLP1 on HPA activity. Deletion of GLP1R in
PVN decreases both ACTH and corticosterone responses to stress and attenuates parvocellular PVN
Fos activation, consistent with a role for PVN GLP1 in stress activation. Notably, Sim1-mediatedPVN gene deletion also attenuated heart rate and blood pressure responses to acute stress, perhaps
via reducing activation of PVN preautonomic neurons. Collectively, these data provide evidence for
PVN GR and GLP1 in stress control, the former essential for stress inhibition, the latter for stressexcitation.

James P. Herman, Ph.D.
Professor of Psychiatry and Behavioral Neuroscience and Director of theLaboratory of Stress Neurobiology, University of Cincinnati
James P. Herman earned his B.S. in Chemistry/Psychology at Hobart
College in 1979 and his Ph.D. from the University of Rochester in 1987,
from the Department of Neurobiology and Anatomy. He worked as a
post-doc at the Mental Health Research Institute, University of Michigan.
He started his academic career in the Department of Anatomy and
Neurobiology at the University of Kentucky, where he was an Associate
Professor and the James and Barbara Holsinger Chair of Anatomy and
Neurobiology. He joined the University of Cincinnati faculty in 2000 and iscurrently the Donald C. Harrison Professor and Vice-Chair for Basic
Research in the Department of Psychiatry and Behavioral Neuroscience.
He is also Director of the University of Cincinnati Neurobiology ResearchCenter. He currently serves as Editor-in-Chief of Stress, as a section editor
for the European Journal of Neuroscience, and on several editorial boards.
Dr. Herman’s major research interests include structural, functional andmolecular biological principles underlying brain stress integration, with
an emphasis on delineating mechanism linking stress with mental illness,
neurodegeneration and metabolic disease. He has made major
contributions to our understanding of the role of limbic neurocircuitry instress adaptation and stress-related pathologies, and has applied tostate-of-the-art approaches to delineate molecular mechanisms underlying
stress hormone signaling in the brain.

Induction of Hypothalamic Neurons from Pluripotent Stem Cells
Hiroshi Nagasaki
Department of Physiology I, School of Medicine, Fujita Health University, Toyoake, Japan
Recently, various hypothalamic neurons have been successfully engineered from pluripotent stem
cells including mouse and human embryonic stem cell lines. Because pluripotent cells have to follow
the stepwise changes in the organogenesis, developmental analyses on hypothalamus have been
inevitable that the numerous transcription factors determine specification, survival, and migration
during the formation of specific neurons. Hypothalamic progenitor cells arise from ventricular zone
at E10.5, which express Rax (retina and anterior neural fold homeobox) gene. Then Otp (orthopedia)
and SF-1 (steroidogenic factor 1) respectively appear in the dorsal and ventral regions at E13.5,
which subsequently produce various transcription factors required for final maturation of the
hypothalamic neurons. In the pluripotent stem cells, rostral-dorsal hypothalamic–like progenitors(Rax+) are generated from floating aggregates in serum-free and chemically defined medium
conditions with minimized exogenous patterning signaling. A certain population of the Rax+
progenitors eventually generate Otp+ neuronal precursor, and subsequently develop to various dorsal
and lateral hypothalamic neurons including vasopressin, oxytocin, melanin-concentrating hormone,and orexin. The magnocellular vasopressinergic neurons release the hormone upon stimulation. On
the other hand, treatment with sonic hedgehog on the floating culture induces differentiation
markers including SF-1 that is specific for rostral–ventral hypothalamic-like precursors. These cells
eventually produce NPY and POMC. Now it is possible to induce most types of therostral-hypothalamic neuron from pluripotent stem cell, that the application of the cells would haveadvantages on the studies on specification, migration, drug development, and regenerative medicine.

Hiroshi Nagasaki, M.D., Ph.D.
Professor of Physiology, School of Medicine, Fujita Health University
Hiroshi Nagasaki is a graduate of Nagoya University Medical School
in 1989. He obtained his Ph.D. degree at Nagoya University in 1997.
After working as a postdoctoral research fellow, he was appointed a
research associate at Department of Pharmacology, University ofCalifornia, Irvine (Dr. Civelli lab), in 2000. After coming back toJapan, he was appointed an assistant professor, and then associate
professor at Department of Metabolic Medicine, Nagoya University,
Graduate School of Medicine. He moved to Department of Physiology,
Fujita Health University in 2011 and served as an associate professor.
In 2015, he became Professor of Physiology, at Fujita Health
University School of Medicine. He is specialized in the development of
the hypothalamus, and he recently succeeded in differentiating
pluripotent stem cells into specific classes of neuroendocrine neuronsof the hypothalamus.

Transcriptional Regulation of Vasopressin Gene: Update in 2015
Yasumasa Iwasaki and Keiichi Itoi
Health Care Center and Department of Endocrinology, Metabolism, Nephrology, Kochi University,
Kochi, Japan and Laboratory of Information Biology, Graduate School of Information Sciences,
Tohoku University, Sendai, Japan
Expression of arginine vasopressin (AVP) in magnocellular neurons of the supraoptic/paraventricular nuclei of the hypothalamus is known to be regulated by osmotic stress. A part of theosmoregulation of AVP synthesis/release is under the control of osmoreceptors residing in thecircumventricular organs (subfornical organ and organum vasculosum of the lamina terminalis), andthe molecular mechanism of osmosensation has been unraveled in recent years. On the other hand,
long-term regulation of AVP neurons including that of the AVP gene expression appears to be
mediated by the osmosensitive mechanisms within the vasopressinergic neurons themselves as wellas the neural inputs from the aforementioned osmoreceptors. Our research group identified
previously the transcription factors, CREB and AP1 (Fos/Jun), activated through the intracellularcAMP-dependent protein kinase A and calcium-dependent signaling, respectively, as positive
regulators of the AVP gene expression. Unexpectedly, other CREB-family transcription factors
related to the endoplasmic reticulum (ER) stress were recently shown to be involved in theregulation of the AVP gene, based on an emerging concept that osmotic stress triggers the ER stress.
Regarding the AVP expression in the parvocellular neurons of the paraventricular nucleus,
regulating the hypothalamo-pituitary-adrenal axis, another transcription factor, FosB, which has
much longer intracellular half-life than c-Fos, may play an important role, especially during low
glucocorticoid states.

Yasumasa Iwasaki, M.D., Ph.D.
Professor, Director of Health Care Center, Kochi University
Yasumasa Iwasaki is a graduate of Tohoku University, Japan, in 1981.
He worked as a post-doc fellow at University of Chicago (Prof. Gary
Robertson) from 1988 through 1990, and then as a research associate
at Harvard University (Prof. Joseph Majzoub) from 1990 through1992. After coming back to Japan, he obtained his Ph.D. degree from
Nagoya University in 1997, and was appointed an assistant professor
and then lecturer at Nagoya University School of Medicine. In 2004,
he moved to Kochi Medial School, Kochi University, and served as alecturer. In 2009, he got promoted to Professor, Director of Health Care
Center, Kochi University. He is specialized in the molecular biology ofvasopressinergic neurons. He is an awardee of Kawakami Prize ofJapan Neuroendocrine Society in 1999.

Molecular Regulation of CRH Gene Expression in Parvocellular Neurons
Greti Aguilera
National Institute of Child Health and Human Development, NIH, Bethesda, Maryland, USA
Corticotropin releasing hormone (CRH) is essential for stress adaptation, mediating
hypothalamic-pituitary-adrenal axis, behavioral and autonomic responses to stress. Activation of
hypothalamic parvocellular CRH neurons depends on neural afferents from the brain stem andlimbic system, leading to CRH release and synthesis. CRH gene transcription is required forrestoration of mRNA levels following translation to protein. However, termination of this response isalso essential for preventing pathology associated with chronic elevations of CRH and HPA axis
activity. Transcriptional activation of the CRH gene depends on cyclic AMP/protein kinase A
signaling and binding of phospho-CREB to a CRE at –270 in the CRH promoter. This CRE isessential for activation of the CRH promoter, and DNA methylation at its internal CpG reduces
CREB binding to the promoter affecting CRH expression. Phospho-CREB alone cannot drive CRH
transcription but it requires nuclear translocation and activation of the CREB co-activator,
Transducer Of Regulated CREB activity (TORC) and its recruitment by the CRH promoter. Cyclic
AMP activates TORC by inhibiting salt induced kinase (SIK) 2 allowing TORC dephosphorylationand nuclear translocation. The magnitude and duration of CRH neuron activation andtranscriptional response is limited by glucocorticoids feedback inhibition, mainly throughmodulation of afferent pathways to the hypothalamic CRH neuron, as well as intracellular
production of the repressor, Inducible Cyclic AMP Early Repressor (ICER). In addition, parallel toactivating TORC, cyclic AMP induces SIK 1 expression, which may mediate nuclear TORC
phosphorylation and inactivation, providing an additional mechanism for terminating CRH
transcription.

Greti Aguilera, M.D.
Scientist Emerita, National Institute of Child health and Human Development,
National Institutes of Health
Greti Aguilera received her MD degree from the University of Chile in 1968. She
was trained in Clinical Endocrinology, Department of Medicine, University ofChile, and in basic research on steroid biochemistry at the Department of
Physiology and Biophysics. In 1975 she joined Dr. Kevin Catt’s laboratory at
NICHD as an International Fogarty Fellow, and in 1977 was appointed as avisiting scientist, and then as a senior researcher in 1984. From 1989 to 2014, she
was chief of the Section of Endocrine Physiology, at the Developmental
Endocrinology Branch in NICHD, leading a research program on theneuroendocrine mechanisms controlling homeostasis. Since 2014 she is scientist
emerita at the same institution. Research contributions include identification offactors responsible for regulation of adrenal glomerulosa responsiveness toangiotensin II; characterization of the relative role of corticotropin-releasing
hormone and vasopressin on the HPA axis adaptation during stress;
characterization of the properties, signaling-transduction and regulation of thereceptors for these peptide hormones, and the identification of novel mechanisms
for the transcriptional regulation of CRH. Current work in Dr. Aguilera’slaboratory focuses on the molecular mechanisms of positive and negative
regulation of CRH transcription responsible for the rapid but transient responses
to stress, the consequences of early life experiences and circadian and ultradianrhythms of glucocorticoid secretion on gene transcription, and the mechanisms
determining pulsatile glucocorticoid secretion by the adrenal. She has about 250
publications in leading journals and books in endocrinology andneuroendocrinology.