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Boys Town National Research Hospital (BTNRH) Genetic & Molecular Biology Labs. Auditory Neurobiology. Lymphatic Research. Neurochemistry. Cell Signaling & Angiogenesis. Retinal Neurobiology. Gene Expression. Vestibular Neurogenetics. Gene Marker & Identification.

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Boys Town National Research Hospital (BTNRH) Genetic & Molecular Biology Labs

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Boys town national research hospital btnrh genetic molecular biology labs

Boys Town National Research Hospital (BTNRH)Genetic & Molecular Biology Labs

Auditory Neurobiology

Lymphatic Research

Neurochemistry

Cell Signaling & Angiogenesis

Retinal Neurobiology

Gene Expression

Vestibular Neurogenetics

Gene Marker & Identification


Boys town national research hospital btnrh genetic molecular biology labs

Auditory Neurobiology LabEdward 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

dB SPL

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Boys town national research hospital btnrh genetic molecular biology labs

Auditory Neurobiology LabEdward Walsh, JoAnn McGee

Techniques

  • Single cell electrophysiology

  • Evoked potential

  • Sensory cell transduction

  • Inner ear and CNS histology and ultrastructure

  • Immunocytochemistry and histochemistry

  • Otoacoustic emissions

  • Microiontophoresis


Boys town national research hospital btnrh genetic molecular biology labs

Cell Signaling & Angiogenesis LabSudhakar Akulapalli

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


Boys town national research hospital btnrh genetic molecular biology labs

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

  • Immunohistochemistry

Sudhakar et al. (2005)


Boys town national research hospital btnrh genetic molecular biology labs

Gene Expression LabDominic 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


Boys town national research hospital btnrh genetic molecular biology labs

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


Boys town national research hospital btnrh genetic molecular biology labs

Gene Marker & Identification LabBill 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%)


Boys town national research hospital btnrh genetic molecular biology labs

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


Boys town national research hospital btnrh genetic molecular biology labs

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


Boys town national research hospital btnrh genetic molecular biology labs

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)


Boys town national research hospital btnrh genetic molecular biology labs

Neurochemistry LabBarbara 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.


Boys town national research hospital btnrh genetic molecular biology labs

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


Boys town national research hospital btnrh genetic molecular biology labs

Retinal Neurobiology LabYou-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


Boys town national research hospital btnrh genetic molecular biology labs

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)


Boys town national research hospital btnrh genetic molecular biology labs

Vestibular Neurogenetics LabYesha 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


Boys town national research hospital btnrh genetic molecular biology labs

Vestibular Neurogenetics LabYesha Lundberg

Techniques

  • Gene targeting

  • Protein biochemistry

  • Histological and ultrastructural analyses

  • Cell culture

  • Physiological studies (collaborative)


Boys town national research hospital btnrh genetic molecular biology labs

BTNRH: Genetic and Molecular Biology Laboratories

  • Lymphatic Research

    • R Tempero

  • Neurochemistry

    • B Morley

  • Retinal Neurobiology

    • Y Peng

  • Vestibular Neuro-genetics

    • Y Lundberg

  • 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


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