[Authors]. [Abstract Title].

1685 Pages·2014·6.76 MB·English

by  Michele Appley

When citing an abstract from the 2014 annual meeting please use the format below.

[Authors]. [Abstract Title]. Program No. XXX.XX. 2014 Neuroscience Meeting Planner.
Washington, DC: Society for Neuroscience, 2014. Online.

2014 Copyright by the Society for Neuroscience all rights reserved. Permission to republish any
abstract or part of any abstract in any form must be obtained in writing by SfN office prior to
publication.
Poster
029. Neural Lineage Specification and Plasticity
Location: Halls A-C
Time: Saturday, November 15, 2014, 1:00 PM - 5:00 PM
Program#/Poster#: 29.01/A1
Topic: A.02. Neurogenesis and Gliogenesis
Support: CIRCNS
SIUC Research Rookies
Title: Rhox8 expression in rodent brains
Authors: H. G. HUFFMAN1,2, E. D. GRISLEY1,2, J. A. MACLEAN, II2, *J. L.
CHEATWOOD1;
1Anat., SIU Sch. Med., CARBONDALE, IL; 2Physiol., SIU Sch. Med., Carbondale, IL
Abstract: Homeobox genes encode transcription factors that govern many processes during
development. This 60-amino acid DNA-binding motif associates with promoters by either
activating or suppressing the transcription of downstream target genes. Recently, the Rhox, -(X-
linked reproductive Homeobox)-, genes were discovered. The Rhox gene set is expressed during
embryonic development in the testis, but a select few of these genes continue to show high
expression after birth which makes them candidates for controlling postnatal and adult
developmental events. At their peak expression in the testis (postnatal day 12), all Sertoli cells
express Rhox5, Rhox8, and Sox9, but Rhox5 expression become more restrictive after postnatal
day 30. RHOX8 protein is abundant in the testis, epididymis, ovary, and it is weakly detected in
the placenta. Of all 33 mouse Rhox genes, Rhox8 is the only one to show expression in somatic
cells in the embryonic testis. All others are expressed solely in germ cells. It is a common
phenomenon that testis expressed genes exhibit brain-specific splicing or transcripts from
alternative promoters. Given Rhox8’s potentially unique transcriptional control in the testes, we
wanted to determine if Rhox8 was similarly uniquely expressed in the brain. Thus, we examined
whether the mRNA of the somatic transcription factors Rhox8, Sox9, and Rhox5 could be
detected in adult rat and mouse brain tissue. Rats were euthanized, and brains were removed and
flash-frozen. To study the basic expression of the Rhox8, Sox9, and Rhox5 gene in brain tissue,
we extracted RNA rat and mouse brains, and then converted the RNA to cDNA in order to
perform qPCR (quantitative real-time polymerase chain reaction). Rhox8, Rhox5, and Sox9 were
all found to be highly expressed in positive control adult mouse testis tissue. However, only
Rhox8 had moderate expression in mouse cortex and hippocampus. As expected we found
Rhox8 to have moderate expression in adult rat cerebellum, Sox9 had minimal expression, but
Rhox5 had no detectable expression. These low levels may be due to the use of whole
cerebellum in the rat, which contains many cell types which do not appear to express Rhox8 in
addition to neurons, some of which were immunopositive for RHOX8 in the mouse brain.
Further characterization of RHOX8 expression in adult rats and adult and developing mice was
ongoing at the time of abstract submission.
Disclosures: H.G. Huffman: None. E.D. Grisley: None. J.A. MacLean: None. J.L.
Cheatwood: None.

Poster
029. Neural Lineage Specification and Plasticity
Location: Halls A-C
Time: Saturday, November 15, 2014, 1:00 PM - 5:00 PM
Program#/Poster#: 29.02/A2
Topic: A.02. Neurogenesis and Gliogenesis
Support: NIH Grant MH099628
Title: Spatiotemporal dimorphic expression of Xlr3 genes in the developing male and female
cortex
Authors: *A. MCDONOUGH1, J. BERLETCH2, C. DISTECHE2, R. HEVNER3,1;
1Ctr. for Integrative Brain Res., Seattle Childrens Res. Inst., Seattle, WA; 2Dept. of Pathology,
3Neurolog. Surgery and Pathology, Univ. of Washington, Seattle, WA
Abstract: The high occurrence of neurodevelopmental disorders in males may in part be
explained by the inheritance of their X-chromosome from their mothers, particularly if certain
genes are maternally imprinted and those genes influence cognitive function. This hypothesis is
supported by comparative studies on women with Turner syndrome who have inherited solely a
maternal or paternal X-chromosome, who exhibit parent-of-origin effects correlating with
differing types of cognitive impairment. Further studies on mouse models of Turner syndrome
suggest some X-linked genes may be imprinted and expressed solely from one X-chromosome.
The observed sex biases in susceptibility for many neurodevelopmental and neuropsychiatric
disorders, and studies on transgenic mice that separate the sex chromosome complement from
gonadal sex, suggest differences in brain structure, and thus development and plasticity, may be
due to the sex chromosomes rather than gonadal hormones. In preliminary studies we saw high
expression of Xlr3 genes in the developing mouse cortex. Based on these findings and previously
published research, we hypothesize that these genes function as transcriptional regulators and
they are expressed from the maternal X-chromosome in both males and females, and repressed
on the paternal X-chromosome in females. Using in situ hybridization and q-PCR we have
characterized the expression patterns and levels in the developing male and female cortex during
multiple stages of neurogenesis. This knowledge may help explain observed clinical differences
in susceptibility to autism and other neurodevelopmental and neuropsychiatric disorders.
Disclosures: A. McDonough: None. J. Berletch: None. C. Disteche: None. R. Hevner: None.

Poster
029. Neural Lineage Specification and Plasticity
Location: Halls A-C
Time: Saturday, November 15, 2014, 1:00 PM - 5:00 PM
Program#/Poster#: 29.03/A3
Topic: A.02. Neurogenesis and Gliogenesis
Support: NSF W05601
Joe and Marie Field Foundation
Title: Stroke injury enhances neurogenesis in multiple new stem cell niches along the ventricular
system at sites of high blood-brain-barrier permeability
Authors: *R. LIN1, J. CAI1, C. NATHAN1, X. WEI1, S. SCHLEIDT1, R. ROSENWASSER2, L.
IACOVITTI1;
1Farber Inst. of Neuroscience, Dept. of Neurosci., 2Dept. of Neurolog. Surgery, Thomas
Jefferson Univ., Philadelphia, PA
Abstract: Previous studies have established the subventricular (SVZ) and subgranular (SGZ)
zones as sites of neurogenesis in the adult forebrain (Doetsch et al., 1999a). Recently, our group
also suggested that circumventricular organs (CVOs) serve as adult neural stem cell niches
(Bennett et al., 2009, 2010). Since NSC proliferation and differentiation is relatively low in all of
these sites in the quiescent brain, in this study, we sought to determine whether these processes
might be amplified following perturbation. To do so, rats were subjected either to focal ischemic
injury (MCAO) or sustained intraventricular infusion of the cell mitogen bFGF (via Alzet pump).
Our data show that both stroke and bFGF induce a long-lasting (14 days) rise in the proliferation
(BrdU+) of nestin+, Sox2+, GFAP+ NSCs and increase their differentiation into Olig2+ glial
progenitors and Dcx+ neuroblasts in the SVZ and CVOs but also in several other novel sites
along the third (3V) and fourth (4V) ventricles. Importantly, all these sites were supplied by a
rich vasculature of fenestrated capillaries and a blood-brain-barrier (BBB) that was highly
permeable to systemically injected sodium fluorescein. These data indicate that stem cell niches
exist not only near the lateral ventricle but at multiple sites along the entire ventricular system,
consistent with the potential for widespread neurogenesis and gliogenesis in the adult brain. We
further suggest that because of their leaky BBB, all of these niches are well positioned to respond
to systemic cues following injury which could be important for brain repair.
Disclosures: R. Lin: None. J. Cai: None. C. nathan: None. X. Wei: None. S. Schleidt:
None. R. Rosenwasser: None. L. Iacovitti: None.

Poster
029. Neural Lineage Specification and Plasticity
Location: Halls A-C
Time: Saturday, November 15, 2014, 1:00 PM - 5:00 PM
Program#/Poster#: 29.04/A4
Topic: A.02. Neurogenesis and Gliogenesis
Title: Differences in proliferation behaviour and expression of phenotypic markers in embryonic
mouse adrenal chromaffin cells and sympathetic neuroblasts
Authors: *W. CHAN1, D. G. GONSALVEZ1, H. M. YOUNG1, E. M. SOUTHARD-SMITH2,
K. N. CANE1, C. R. ANDERSON1;
1Dept. of Anat. and Neurosci., Univ. of Melbourne, Melbourne, Australia; 2Div. of Genet.
Medicine, Dept. of Med., Vanderbilt Univ. Sch. of Med., Nashville, TN
Abstract: Adrenal medullary chromaffin cells and peripheral sympathetic neurons originate
from a common sympathoadrenal (SA) progenitor cell. The time course and signalling
mechanisms underlying this lineage separation during embryonic development are not fully
understood. The present study investigates the expression patterns of phenotypic markers as well
as cell cycle dynamics in the embryonic murine adrenal medulla and the neighbouring suprarenal
ganglion by immunohistochemical analysis. Transverse sections of the adrenal region in E10.5-
16.5 C57BL/6 mice were processed with antibodies against Sox10, tyrosine hydroxylase (TH),
phenylethanolamine N-methyltransferase (PNMT) and cocaine and amphetamine regulated
transcript (CART). Ki67 immunohistochemistry, coupled with dual-labelling with S-phase
markers, bromodeoxyuridine (BrdU) and 5-ethynyl-2’-deoxyuridine (EdU), allowed cell cycle
analyses. The noradrenergic marker, TH was detected in cells in both the adrenal and
sympathetic region, but with much stronger immunoreactivity (IR) for TH in the adrenal. There
was strong CART immunostaining in most neuroblasts, whereas it was detected in only a
handful of adrenal chromaffin cells. Comparison of CART-IR versus TH-IR showed that
neuroblasts have low intensity of TH-IR but high intensity of CART-IR, while chromaffin cells
in the adrenal medulla had high intensity TH-IR without detectable CART-IR. This phenotypic
segregation appeared as early as E12.5 when anatomical segregation of the two cell types is
difficult. The adrenergic marker, PNMT, was first detected in a subpopulation of chromaffin
cells from E14.5. Chromaffin cell and neuroblasts lineages showed differences in proliferative
behaviour from their earliest appearance at E12.5. Initially, 70% of Sox10+ neural crest
progenitors were proliferating at E10.5 but dramatically withdrew from the cell cycle at E11.5 as
they started to express TH and differentiate into neuroblasts and chromaffin cells. Whereas 88%
of TH+ neuroblasts were back in the cell cycle at E12.5, only around 20% of TH+ chromaffin
cells were in the cell cycle at E12.5 and subsequent days. Cell cycle length increased modestly
for TH+ cells in the ganglion and the adrenal medulla from E12.5. This study identifies the
intensity of TH-IR and the expression of CART as phenotypic markers for early discrimination
of the chromaffin cells and sympathetic neuroblasts. Developing chromaffin cells also show
strikingly less proliferative activity relative to sympathetic neuroblasts.
Disclosures: W. Chan: None. D.G. Gonsalvez: None. H.M. Young: None. E.M. Southard-
Smith: None. K.N. Cane: None. C.R. Anderson: None.

Poster
029. Neural Lineage Specification and Plasticity
Location: Halls A-C
Time: Saturday, November 15, 2014, 1:00 PM - 5:00 PM
Program#/Poster#: 29.05/A5
Topic: A.02. Neurogenesis and Gliogenesis
Support: 1F30MH102002-01
Title: The developmental genetic basis of cortical interneuron identity - the case of chandelier
cells
Authors: *S. M. KELLY1,2, M. HE1, Z. HUANG1;
1Cold Spring Harbor Lab., Cold Spring Harbor, NY; 2Stony Brook Univ., Stony Brook, NY
Abstract: Neuronal firing activity within the cerebral cortex is regulated by a diverse assortment
of inhibitory GABAergic interneurons, which are able to finely tune circuit activity through a
vast array of morphological, molecular, synaptic, and electrophysiological properties. Most
cortical interneurons in rodents arise predominantly from three neurogenic zones: the medial and
caudal ganglionic eminences (MGE, CGE), and the pre-optic area (POA). There is evidence that
interneuron class identity is determined in part in part by spatial and temporal origin within the
subpallium. Furthermore, intrinsic genetic programs and extrinsic cortical signals are both likely
to play a role in defining different neuronal subtypes within each class and distributing these
cells to cortical layers and regions in the proper density to allow the formation of meaningful
circuit motifs. To approach a complete understanding of interneuron development, our studies
have focused on characterizing the entire life history of the chandelier cell, arguably the most
distinct and uniform GABAergic interneuron yet to be described. Chandelier cells have an
unmistakable axon arbor of hundreds of vertically arranged cartridge synapses, each of which
specifically innervates the axon initial segment of a pyramidal cell, the site where action
potentials are generated. Neocortical chandelier cells (ChCs) are generated between mid-
gestation and birth from progenitors in the MGE expressing the homeodomain transcription
factor Nkx2.1. Here, using intersectional genetic fate mapping and novel birth dating techniques,
we provide an experimental system to examine for the first time the cellular and molecular
nature of progenitors that produce several well-defined MGE cell subtypes in cerebral cortex.
Further, we have compared the birth timing and progenitor origin of ChCs found in various
regions of the cerebrum, such as the hippocampus, piriform cortex, and neocortex. Through these
studies, we hope to discover and characterize the mechanism by which Nkx2.1+ progenitors
produce and distribute a diverse family of interneurons to be integrated into specific types of
cognitive circuits throughout the cerebral cortex.
Disclosures: S.M. Kelly: None. M. He: None. Z. Huang: None.

Poster
029. Neural Lineage Specification and Plasticity
Location: Halls A-C
Time: Saturday, November 15, 2014, 1:00 PM - 5:00 PM
Program#/Poster#: 29.06/A6
Topic: A.02. Neurogenesis and Gliogenesis
Title: Molecular characterization of human cortical interneurons
Authors: *J. L. CLOSE1, B. LEVI1, V. MENON1, S. ANDERSON2, S. RAMANATHAN3;
1Allen Inst. For Brain Sci., Seattle, WA; 2Univ. of Pennsylvania, Philadelphia, PA; 3Harvard
Univ., Cambridge, MA
Abstract: We have established a robust protocol for generating cortical interneuron progenitors
from human embryonic stem cells. The gene expression profile of these cells has been assayed at
the population, subpopulation and single-cell level to determine progenitor diversity and lineage
relationships within heterogeneous pools of interneuron precursors and differentiating neurons.
Using reporter lines developed to visualize specific populations and developmental stages, we
have determined the gene regulatory networks that underlie the production and maturation of
human interneuron subclasses that are crucial to cortical processing.
Disclosures: J.L. Close: None. B. Levi: None. V. Menon: None. S. Anderson: None. S.
Ramanathan: None.

Poster
029. Neural Lineage Specification and Plasticity
Location: Halls A-C
Time: Saturday, November 15, 2014, 1:00 PM - 5:00 PM
Program#/Poster#: 29.07/A7
Topic: A.02. Neurogenesis and Gliogenesis
Support: NIH NRSA Fellowship 1F31DK097939
NIH R01 Grant 2R01DK078158-04A1
NIH U01 Grant 1U01DK101038-01
Title: Serotonin receptor 5-HT3A in the development of autonomic and sensory innervation of
the lower urinary tract
Authors: *E. RITTER1,3, D. P. BUEHLER2, H.-H. WU4, E. SOUTHARD-SMITH2;
2Genet. Med., 1Vanderbilt Univ., Nashville, TN; 3Neurosci. Grad. Program, Vanderbilt Brain
Inst., Nashville, TN; 4Dept. of Pediatrics, Keck Sch. of Med. of Univ. of Southern California,
Los Angeles, CA
Abstract: Decades of research demonstrate the importance of serotonin (5-HT) signaling in the
fetal development and postnatal maturation of various neuronal populations. While most of these
studies focused on the brain and enteric nervous system of the gut, very few have inspected the
role of 5-HT in development of other peripheral nervous system components. In a gene
expression analysis conducted in our lab, we noted increased expression of the type 3A serotonin
receptor (5-HT3A) in fetal pelvic ganglia that provide autonomic innervation to the bladder and
urethra. This finding, along with reports of 5-HT3A in adult urinary function and visceral pain,
led us to pursue the role of this receptor in the development of bladder autonomic and sensory
innervation. We aim to define the temporal, spatial, and cell-type specific expression patterns of
the 5-HT3A receptor throughout lower urinary tract development as a basis for interrogating
gene function in this system. By employing the Htr3a-EGFP transgenic reporter mouse line and
immunohistochemistry, we have characterized expression of the 5-HT3A receptor in a variety of
both autonomic and sensory neuronal subtypes within the pelvic and dorsal root ganglia.
Expression of 5-HT3A begins early in the development of bladder innervation at 12 days post
coitus, which is the earliest time point 5-HT3A has been identified in mouse embryos. 5-HT3A
co-localizes with both sympathetic and parasympathetic markers in the pelvic ganglia throughout
fetal development, as well as several markers of nociceptive neurons in the dorsal root ganglia
and bladder urothelium. Ongoing experiments are focused on determining roles for 5-HT3A
signaling in urinary tract function and acute neurogenic inflammation in knockout mice. Given
that very few treatment options are available for urinary incontinence and pelvic pain syndromes,
elucidating the mechanisms of neurogenesis and maturation in the lower urinary tract lends itself
to the identification of novel and specific therapeutic targets for treating these prevalent
disorders.
Disclosures: E. Ritter: None. D.P. Buehler: None. H. Wu: None. E. Southard-Smith: None.

Poster
029. Neural Lineage Specification and Plasticity
Location: Halls A-C
Time: Saturday, November 15, 2014, 1:00 PM - 5:00 PM
Program#/Poster#: 29.08/A8
Topic: A.02. Neurogenesis and Gliogenesis
Support: NIH Grant RO1HD055655
NIH Grant R01MH056524
Title: Modeling neurodevelopmental disrutpion and glioblastoma with crispr/cas9 system
Authors: *F. CHEN1, A. CHE2, A. BECKER4, J. LOTURCO3;
1Dept. of Physiol. and Neurobio., Univ. of Connecticut, STORRS MANFLD, CT; 2PNB,
3Physiol. and Neurobio., Univ. of Connecticut, Storrs, CT; 4Dept. of neuropathology, Univ. of
Bonn Med. Sch., Bonn, Germany
Abstract: The newly developed CRISPR/Cas9 system offers fast, convenient, specific yet
efficient way for genome editing. Here we report successful modeling of neurodevelopmental
disorder as well as gliaoblastoma multiforme in rat using CRISPR/Cas9 system. In vivo
transfection of radial glia with sgRANs targeting PTEN successfully abolished PTEN expression
in cells in radial glia lineage i.e. neurons, astrocytes and oligodendrocytes. PTEN-null neurons
showed increased soma size, hypertrophic dendrites and axons. PTEN null neurons also showed
altered electrophysiological properties such as decreased input resistance, increased sEPSC and
mEPSC. These results indicate CRISPR/Cas9 can be used in vivo to manipulate gene expression.
Moreover, electroporation of three sgRNAs targeting PTEN, NF1 and P53, respectively, into rat
radial glia and successfully induced glioblastoma multiforme formation. To our knowledge, this
is the first successful demonstration of modeling neurodevelopmental disorder and glioblastoma
using CRISPR/Cas9 system.
Disclosures: F. Chen: None. A. Che: None. A. Becker: None. J. LoTurco: None.

Poster
029. Neural Lineage Specification and Plasticity
Location: Halls A-C
Time: Saturday, November 15, 2014, 1:00 PM - 5:00 PM
Program#/Poster#: 29.09/A9
Topic: A.02. Neurogenesis and Gliogenesis
Support: CIHR Grant
Title: Primitive neural stem cells at the top of the neural stem cell hierarchy
Authors: *S. YAMMINE1, R. LEEDER2, D. VAN DER KOOY1;
1Mol. Genet., 2Inst. of Med. Sci., Univ. of Toronto, Toronto, ON, Canada
Abstract: Primitive neural stem cells (pNSCs) are Oct4+GFAP- cells that can be isolated from
the mouse embryo as early as E5.5, and persist throughout development into adulthood where
they are a rare population residing in the subependymal zone of the lateral ventricles. pNSCs are
LIF-dependent and can be clonally passaged in vitro to self-renew or give rise to definitive
(d)NSCs, which are GFAP+ and dependent on EGF and FGF. Given that pNSCs arise earlier in
development than dNSCs and are upstream dNSCs in vitro, we asked whether this hierarchy is
true in vivo, and what the role of an upstream pNSC population is in the embryo and adult mouse
brain. We have previously shown that expression of the pluripotency gene Oct4 is an exclusive
marker for pNSCs, so we used Oct4fl/fl;Sox1Cre mice to assess the requirement of Oct4 in
pNSCs. The forebrain of adult mice homozygous for the Oct4 conditional knockout did not give
rise to any pNSC-derived neurospheres in vitro, while dNSC-derived neurospheres were not
affected by Oct4 loss. This indicates a requirement for Oct4 expression in either the survival or