Infrared neural modulation is an optical technique to stimulate or inhibit neural activity with pulsed infrared light (IR). This study focuses on effects of IR and heat on microglial calcium signaling, a main indicator of physiological changes within the immune-like cell. Widefield fluorescence microscopy quantified the intracellular calcium fluxes, cell death, and early apoptosis fluorescent indicators in BV2 immortalized microglia cell culture after irradiation. We have found that unique cellular responses are highly associated with different temperature increases, and the irradiance thresholds for these responses do not drive cells to Caspase-3 apoptosis nor necrotic cellular death.
Significance: Infrared (IR) inhibition can selectively block peripheral sensory nerve fibers, a potential treatment for autonomic-dysfunction-related diseases (e.g., neuropathic pain and interstitial cystitis). Lowering the IR inhibition threshold can increase its translational potentials.
Aim: Infrared induces inhibition by enhancing potassium channel activation. We hypothesized that the IR dose threshold could be reduced by combining it with isotonic ion replacement.
Approach: We tested the IR inhibition threshold on the pleural-abdominal connective of Aplysia californica. Using a customized chamber system, the IR inhibition was applied either in normal saline or in isotonic ion-replaced saline, which could be high glucose saline, high choline saline, or high glucose/high choline saline. Each modified saline was at a subthreshold concentration for inhibiting neural conduction.
Results: We showed that isotonically replacing ions in saline with glucose and/or choline can reduce the IR threshold and temperature threshold of neural inhibition. Furthermore, the size selectivity of IR inhibition was preserved when combined with high glucose/high choline saline.
Conclusions: The present work of IR inhibition combined with isotonic ion replacement will guide further development of a more effective size-selective IR inhibition modality for future research and translational applications.
Stimulated Raman scattering (SRS) offers a drastic speed advantage over conventional vibrational spectroscopic imaging techniques – making it ideal for studying fast biochemical dynamics. We developed an experimental paradigm that applies spectral stimulated Raman scattering (SRS) imaging to study the mechanisms of infrared (IR) photostimulation of neuronal cells. Infrared neural stimulation (INS) is a label-free optical neuromodulation technique with high spatial and temporal precision. Using SRS, changes in lipid and water vibrational signatures in live cells during INS were observed, suggesting that lipid membrane deformation accompanies IR exposure. The speeds afforded by SRS enables unprecedented observation of fast cellular biophysical dynamics.
Infrared neural inhibition (INI) is a relatively new modality of neural control which has potential as a novel pain therapy due to its high spatial specificity and selective inhibition of small diameter neurons at lower temperatures. Computational modeling using a modified Hodgkin-Huxley model in the squid giant axon has shown that temperature sensitive potassium currents mediate INI’s thermal block. This model was modified to reflect Aplysia parameters which have smaller unmyelinated axons on the order of mammalian C fibers, and simulated results were validated in vitro. Results support he hypothesis that potassium currents are needed to create a thermal block.
We have shown previously that astrocytes are independently sensitive to IR exposure in the context of infrared neural stimulation (INS). More recently, our advances have identified some physiological basis of astrocytic sensitivity to INS. Single pulse INS evokes intracellular calcium signaling primarily mediated through inositol-triphosphate receptor (IP¬3R) activation, rather than thermally sensitive transient receptor protein (TRP) pathways. Interestingly, different dosing strategies for IR appear to evoke different physiology. Our findings suggest that tailoring irradiation parameters with INS may be used for preferentially drive separate physiological responses in astrocytes, and possibly other cell types.
Infrared neural inhibition (INI) is a method of blocking the generation or propagation of neural action potentials through heating with infrared lasers that target peaks in the water absorption spectrum (1460 nm, 1875 nm). Recent work showed that the length of axon heated modulates the temperature needed for inhibition. Different irradiation lengths for INI were tested in Aplysia nerves (n=7). Once 1 mm of irradiation was exceeded, longer irradiation lengths resulted in higher temperatures at inhibition threshold. Optimizing irradiation lengths of nerve to minimize temperatures will help inform the design of devices for longitudinal animal studies and human implementation.
Thermal block of unmyelinated axons may serve as a modality for control, suggesting a means for providing therapies for pain. Computational modeling predicted that potassium channels are necessary for mediating thermal block of propagating compound action potentials (CAPs) with infrared (IR) light. Our study tests that hypothesis. Results suggest that potassium channel blockers disrupt the ability of IR to block propagating CAPs in Aplysia californica nerves, whereas sodium channel blockers appear to have no significant effect. These observations validate the modeling results and suggest potential applications of thermal block to many other unmyelinated axons.
Pulsed infrared (IR) light has been used in multiple animal models to inhibit neural activity. Duke et al.
reported inhibition associated with a temperature increase of ~8°C in Aplysia californica buccal nerve 2
(BN2). There is no evidence that the current irradiation schemes alters nerve functionality, however lower
temperatures provide a safer environment for sustained inhibition. Inhibition paradigms use a single optical
fiber to deliver IR light, resulting in a single hotspot within the nerve. One proposed method for decreasing
peak temperatures is to use a lower radiant exposure over a greater area, effectively heating the nerve more
evenly. Preliminary computational modeling suggests that using two axially adjacent optical fibers reduces
peak temperatures required for infrared neural inhibition (INI). This hypothesis is being validated in vitro in
Aplysia. Pleural abdominal nerves were dissected out, and suction electrodes were applied to electrically
stimulate and record neural activity. A custom probe (core diameters= 400 μm) was used to simultaneously
apply IR light from two diode lasers (Lockheed-Martin, λ=1875nm) to the nerve and monitor the radiant
exposure out of each. Radiant exposures required for inhibition using a single fiber were reduced by ~37.4%
by using two axially adjacent optical fibers. While mechanisms behind infrared inhibition are not fully
understood, data suggests that a threshold temperature is required. By reducing peak temperatures, neural
block using IR light will subject nerves to lower peak temperatures and provide a more research and clinically
relevant technology.
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