Boys Town National Research Hospital (BTNRH) Genetic & Molecular Biology Labs Auditory Neurobiology Vestibular Neurogenetics Neurochemistry Retinal Neurobiology

Boys Town National Research Hospital (BTNRH)

Genetic & Molecular Biology Labs

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


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dB

SP

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Techniques Single cell electrophysiology Evoked potential Sensory cell transduction Inner ear and CNS histology and

ultrastructure Immunocytochemistry and histochemistry Otoacoustic emissions Microiontophoresis



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)



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




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%)



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)



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



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




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



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

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

Documents

Boys Town National Research Hospital (BTNRH) Genetic & Molecular Biology Labs Auditory Neurobiology Vestibular Neurogenetics Neurochemistry Retinal Neurobiology