Techniques and Methods
Surgical Modeling and Gene Therapy
We work with mice and rats to induce mid-thoracic spinal cord injuries which closely replicates much of the pathology observed in humans. Treating the injured spinal cord to facilitate regeneration of damaged neurons has been exceedingly complicated. Current drug-delivery approaches tend to focus on systemic delivery of pharmaceuticals or use of gene-therapy approaches targeting one spinal tract at a time. Both delivery approaches have strengths and limitations, but ultimately there is a need for an approach that confers the specificity of gene-therapy, with the expansive and temporal advantages of pharmaceuticals. We are developing an approach using available genetic tools that allows for a single spinal injection to target almost all neurons implicated in the SCI pathology, with little-to-no off target effects. Using our developed approach, we can manipulate whatever molecular target or pathway of interest to drive improvements in neuronal function and axon regeneration.
Genetic Engineering
The ability to target only the neurons implicated in the SCI pathology provides a prospectively clinically viable approach to treating the damaged spinal cord. However, which molecular targets can best facilitate repair and functional improvements remain an ongoing investigation. Our lab targets specific cell-signaling pathways by expressing or knocking down key proteins that may elicit a beneficial response in chronically injured neurons. Further, we create constitutively active mutants that can not only induce the expression of key reparative proteins, but can force the activation of molecular pathways vital for repair. Finally, the non-protein coding elements of gene-therapy also play critical roles in the subcellular localization of expressed proteins. Our lab continuously improves upon the constitution of AAV vectors to enable greater cellular and sub-cellular specificity to target protein localization to desired areas like the axon.
Cell Culture
Cell culture remains an ongoing endeavor in our lab. We utilize various immortalized, stem cell, and primary cell lines to interrogate neurobiology. We utilize various cell lines to explore the biological effects of our expressed transgenes across molecular neuroscience. In addition, we utilize genetic engineering of stem cells to enhance their regenerative properties.
Our lab has recently created a novel line of HEK293’s, we call Neuro293’s, that enable the ability for high-throughput testing of neurobiology of near-all mature neuronal proteins. Neuro293’s are a stable cell line of HEK293’s that express most mature neuronal proteins including proteins related to the axon cytoskeleton, neurotransmitter synthesis and release, pre-and post-synaptic functions, and a vast array of different ion channels related to excitable membranes.
Surgical modeling and Gene Therapy

Genetic Engineering

Molecular Biology
Standard molecular biological assessments are frequently used in our lab including western blots, qPCR/PCR, and immunoprecipitation. Further, we leverage high-throughput molecular techniques when needed such as RNA-sequencing and proteomics/phosphoproteomics to provide a more comprehensive assessment of chronic neuropathology.
Locomotor Analysis
As expected, assessing hind-limb locomotor abilities is core to our experimental data. We leverage established methods to assess hind-limb recovery in rodents with SCI, as well as develop novel approaches to quantify recovery of severely injured mice using existing technology such as the Catwalk XT. Our lab is currently adopting AI analysis such as the Automated Limb Motion Analysis (ALMA) for murine models of neurological disorders to provide a more continuous, objective, and sensitive assessment of hind limb functional abilities in rodents with severe SCI (we did not develop this but it sure seems like a good tool for the job!).
Immunohistochemistry and Tissue Analysis
Ultimately seeing is believing. Our lab specializes in the histological analysis of SCI pathology and axon regeneration. We have developed a high-throughput pipeline to quickly, efficiently, and accurately quantify tissue pathology and axon regeneration in our tissue samples. We apply machine learning algorithms, a refined immunolabeling process, and automated microscopy to expedite the time-consuming process that was immunohistochemistry.
Immunohistochemistry and Tissue Analysis
