Boys Town National Research Hospital (BTNRH)
Genetic & Molecular Biology Labs
Auditory Neurobiology
Vestibular Neurogenetics
Neurochemistry
Retinal NeurobiologyGene Expression
Lymphatic Research
Gene Marker & Identification
Cell Signaling & Angiogenesis
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 LabEdward Walsh, JoAnn McGee
Hz
dB SPL
Techniques Single cell electrophysiology Evoked potential Sensory cell transduction Inner ear and CNS histology and
ultrastructure Immunocytochemistry and histochemistry Otoacoustic emissions Microiontophoresis
Auditory Neurobiology LabEdward Walsh, JoAnn McGee
Goal is to identify the molecular and biochemical mechanisms by which endogenous angiogenesis inhibitors prevent tumor development and growth
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 hypoxia-associated 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 LabSudhakar 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
ImmunohistochemistrySudhakar et al. (2005)
Cell Signaling & Angiogenesis LabSudhakar Akulapalli
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 LabDominic Cosgrove, Velidi Rao
Gene Expression LabDominic 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
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)
Gene Marker & Identification LabBill Kimberling, Dana Orten, Phil Kelley
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 LabBill 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
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)
Lymphatic Research Laboratory Richard Tempero
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 LabBarbara Morley
Techniques Pharmacological and immuno-
cytochemical 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
Neurochemistry LabBarbara Morley
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 LabYou-Wei Peng
Retinal Neurobiology LabYou-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)
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.
Vestibular Neurogenetics LabYesha Lundberg
Deletion of Oc90 gene leads to abnormal
otoconia formation and balance deficits
Vestibular Neurogenetics LabYesha Lundberg
Techniques Gene targeting Protein biochemistry Histological and ultrastructural analyses Cell culture Physiological studies (collaborative)
Lymphatic Research R Tempero
Neurochemistry B Morley
Retinal Neurobiology Y Peng
Vestibular Neuro-genetics
Y Lundberg
BTNRH: Genetic and Molecular Biology Laboratories
Auditory Neurobiology E Walsh, J McGee
Cell Signaling andAngiogenesis
S Akulapalli
Gene Expression D Cosgrove, V Rao
Gene Mapping and Identification
B Kimberling, D Orten, P Kelley