Transcript Boys Town National Research Hospital (BTNRH)

Boys Town National Research
Hospital (BTNRH)
Genetic & Molecular Biology Labs
Auditory
Neurobiology
Cell Signaling &
Angiogenesis
Gene Expression
Gene Marker &
Identification
Lymphatic Research
Neurochemistry
Retinal
Neurobiology
Vestibular
Neurogenetics
Auditory Neurobiology Lab
Edward Walsh, JoAnn McGee
Goal is to understand the basic
mechanisms underlying auditory
system development in normal
and pathological systems.
 Role of thyroid hormone in
cochlear development
 Role of Vlgr1 in stereociliary
bundle development
 Development of amino acid
neurotransmission among
neurons of the caudal
brainstem
Auditory Neurobiology Lab
Edward Walsh, JoAnn McGee
Techniques
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Single cell electrophysiology
Evoked potential
Sensory cell transduction
Inner ear and CNS histology and
ultrastructure
 Immunocytochemistry and histochemistry
 Otoacoustic emissions
 Microiontophoresis
Cell Signaling & Angiogenesis Lab
Sudhakar Akulapalli
Goal is to identify the molecular and biochemical
mechanisms by which endogenous angiogenesis
inhibitors prevent tumor development and growth
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Identify the mechanism through which the carboxy terminal of
type IV collagen, specifically, the noncollagenous domain 1 of the
1 chain of type IV collagen (1(IV)NC1), inhibits angiogenesis
and tumor growth (type IV collagen is the major component of
vascular basement membranes)
Determine mechanism whereby 1β1 integrin regulates hypoxiaassociated factors associated with the antiangiogenic activity of
1(IV)NC1
Identify the cell signaling pathway involved in the non-integrin
mediated antiangiogenic activity of a1(IV)NC1
Identify natural proteins that can be used as therapeutic agents
for the treatment of cancer
Cell Signaling & Angiogenesis Lab
Sudhakar Akulapalli
Techniques
 In vivo tumor burden studies
 Cell culture systems (mouse lung
endothelial cells, mouse lung
fibroblast preparations)
 Migration and proliferation assays
 Immunoprecipitation and
immunoblotting to study protein
interactions
 Immunohistochemistry
Sudhakar et al. (2005)
Gene Expression Lab
Dominic Cosgrove, Velidi Rao
Study of molecular mechanisms underlying
progressive pathologies resulting from
mutations in basement membrane proteins;
the role(s) of cell adhesion and integrin
signaling proteins, cytokines and chemokines
 Alport Syndrome (Type IV collagen)
 Renal pathology
 Inner ear pathology
 Usher Syndrome Type IIa (Usherin)
 Retinal pathology
 Inner ear pathology
Gene Expression Lab
Dominic Cosgrove, Velidi Rao
Techniques
 Gene knockout mouse models for Alport
syndrome and Usher syndrome type IIa
 Immortalized cell lines derived from the
retina, the stria vascularis, and the renal
glomerulus of both normal and mutant
mice
 Biochemical and molecular techniques
 Treatment strategies
Gene Marker & Identification Lab
Bill Kimberling, Dana Orten, Phil Kelley
Identification of genes associated
with hearing loss and localization
to the human gene map
 Syndromic:
 Usher Syndrome (Type I, Type II and
Type III)
 Branchio-Oto-Renal (BOR) Syndrome
 Cleft Palate
 Nonsyndromic:
 Dominant Progressive Hearing Loss
 Autosomal Recessive Hearing Loss
(Auditory Neuropathy – otoferlin)
Usher Syndrome
• USH1b – myosin VIIa
• USH1c – harmonin
• USH1d – cadherin 23
• USH1f – protocadherin 15
• USG1g - sans
• USH2a – usherin
• USH2c – vlgr1
• USH3 – clarin1
Hereditary Hearing Loss
• Syndromic (20%)
• Usher Syndrome
• Nonsyndromic (80%)
• Dominant (15%)
• Recessive (80%)
Gene Marker & Identification Lab
Bill Kimberling, Dana Orten, Phil Kelley
Techniques
 Construct physical maps of the critical region of the
human genome associated with hearing loss,
identify candidate genes in that region, and
search for mutations in candidate genes
 Linkage mapping and haplotype mapping to identify
regions of human genome linked to hearing loss
genes
 Positional candidate cloning strategy to identify
genes causing hearing loss
 Treatment strategies
Lymphatic Research Laboratory
Richard Tempero
Study of the lymphatic system as it relates to
human inflammatory and malignant disease.
Study of the molecules and mechanisms that
contribute to lymphatic growth assessed by
lymphangiogenesis
Lymphatic vessels drain extracellular fluid and
improve the capacity for pathogen surveillance
by directing fluid towards regional lymph nodes
Lymphatic Research Laboratory
Richard Tempero
Techniques
 Murine model of
respiratory inflammation
(e.g., Mycoplasma
pulmonis infection)
 Study molecules and
mechanisms important for
lymphangiogenesis
Confocal image of normal
mouse trachea
• lymphatics (LYVE-1 red)
• blood vessels (CD31 green)
Neurochemistry Lab
Barbara Morley
The role of neurotransmitters and receptors in the
establishment of neural pathways and the
formation of maintenance of synaptic
connections.
 The role of acetylcholine and acetylcholinesterase in
the development and maturation of the primary
neurotransmitters in the auditory brainstem.
 Understanding how acetylcholine drives the auditory
system before the onset of hearing.
 Understanding how nicotinic acetylcholine receptors
function in outer hair cells and spiral ganglion cells.
Neurochemistry Lab
Barbara Morley
Techniques
 Pharmacological and immunocytochemical characterization of
neurotransmitter receptor subunits
and assembled receptors
 Expression of receptor subunits
using in situ hybridization and real
time RT-PCR
 Construction of mice and cell lines
that are have null mutations for
cochlea-specific genes
Retinal Neurobiology Lab
You-Wei Peng
Molecular mechanisms of hereditary retinal
degeneration, particularly retinitis
pigmentosa
 Mechanism of retinal degeneration in Usher
syndrome type IIa; the role of usherin in cell
signaling
 Mechanism by which rod-specific genetic
disorders lead to the degeneration of
genetically-normal cone photoreceptors
 Role of plasticity in bipolar cell connections
during retinal degeneration
Retinal Neurobiology Lab
You-Wei Peng
Techniques
 Immunofluorescence
 In situ hybridization
 Real time RT-PCR
 Retinal histology and
ultrastructure
Rod marker, rhodopsin (red), Cone
marker, cone transducin γ (green)
Vestibular Neurogenetics Lab
Yesha Lundberg
Study of the molecular and
cellular mechanisms of
otoconia development in the
vestibular system
Otoconia are composite
crystals that overlie and
provide optimal stimulus
input to the sensory
epithelium of the balance
system. The crystals are
critical for spatial orientation
and balance.
Deletion of Oc90 gene
leads to abnormal
otoconia formation and
balance deficits
Vestibular Neurogenetics Lab
Yesha Lundberg
Techniques
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Gene targeting
Protein biochemistry
Histological and ultrastructural analyses
Cell culture
Physiological studies (collaborative)
BTNRH: Genetic and Molecular
Biology Laboratories
 Auditory Neurobiology
 E Walsh, J McGee
 Lymphatic Research
 R Tempero
 Cell Signaling and
Angiogenesis
 S Akulapalli
 Neurochemistry
 Gene Expression
 D Cosgrove, V Rao
 Gene Mapping and
Identification
 B Kimberling, D Orten,
P Kelley
 B Morley
 Retinal Neurobiology
 Y Peng
 Vestibular Neuro-
genetics
 Y Lundberg