Gedeon, Jeremy
(2024)
In-vivo Calcium Imaging of Sensory Neurons in the Rat Trigeminal Ganglion.
Master's Thesis, University of Pittsburgh.
(Unpublished)
Abstract
Genetically encoded calcium indicators (GECIs) enable the use of imaging techniques to monitor changes in calcium in targeted cell populations. Their large signal-to-noise ratio makes GECIs a powerful tool to detect stimulus-evoked activity in sensory neurons. GECIs facilitate population level analysis of stimulus encoding with the number of neurons that can be studied simultaneously. This population encoding is most appropriately done in vivo. Dorsal root ganglia (DRG), that house the soma of sensory neurons innervating somatic and visceral structures below the neck, are used most extensively for in vivo imaging because these structures are accessed relatively easily. More recently, this technique was used in mice to study sensory neurons in the trigeminal ganglion (TG), that innervate oral and craniofacial structures. There are many reasons to study TG rather than DRG, including the long list of pain syndromes specific to oral and craniofacial structures that appear to reflect changes in sensory neuron activity, such as trigeminal neuralgia. Mice are used most extensively in the study of DRG and TG neurons because of the availability of genetic tools. However, differences in size, ease of handling, and potentially important species differences, there are reasons to study rat rather than mouse TG neurons. Thus, I developed an approach for imaging rat TG neurons in vivo. I injected neonatal pups (p2) intraperitoneally with an AAV encoding GCaMP6s resulting in >90% infection of both TG and DRG neurons. TG were visualized in the adult following craniotomy and decortication, and changes in GCaMP6s fluorescence monitored in TG neurons following stimulation of mandibular and maxillary regions of the face. I confirmed that increases in fluorescence were stimulus evoked with peripheral nerve block. While there are many potential uses for this approach, I are using it to characterize the subpopulation(s) of TG neurons changed following peripheral nerve injury.
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| Item Type: |
University of Pittsburgh ETD
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| Status: |
Unpublished |
| Creators/Authors: |
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| ETD Committee: |
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| Date: |
14 October 2024 |
| Date Type: |
Publication |
| Defense Date: |
31 July 2024 |
| Approval Date: |
14 October 2024 |
| Submission Date: |
30 July 2024 |
| Access Restriction: |
No restriction; Release the ETD for access worldwide immediately. |
| Number of Pages: |
59 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
School of Medicine > Neurobiology |
| Degree: |
MS - Master of Science |
| Thesis Type: |
Master's Thesis |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
surgery, pain, nervous, system, technique, local, translation, cytokine, inflammatory mediator, injury-induced plasticity, neurotrophin, nociceptor sensitization |
| Date Deposited: |
14 Oct 2024 15:47 |
| Last Modified: |
14 Oct 2024 15:47 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/46782 |
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