Focused ultrasound (FUS) mediated neuromodulation has become a trending topic due to its promising attributes that enable precise and transcranial neuromodulation. Despite multiple reports of FUS effects on neurons, nervous systems, and the human brain, the mechanisms underlying such excitation or inhibition remain controversial. In our previous study, we showed that 2.1 MHz FUS induced membrane depolarizations on single crayfish motor axons in the presence of voltage-gated channel blockers, which led to a nanopore hypothesis: FUS triggered lipid molecule reconfiguration and form ion-permeable nanopores on the axonal membrane. Based on this hypothesis, stretching of the axonal membrane due to swelling in low osmolarity should increase the probability of nanopore formation under FUS. As predicted, exposure to 75% hypotonic saline induced significant increases in amplitude and frequency of occurrence of those FUS-induced depolarizations (FUSD) while the onset latency of the FUSD showed a significant decrease. Those results support the hypothesis that FUSD can be modulated by mechanically altering membrane properties.
Since FUS inevitably perturbs cell membranes, we examined the role of mechanosensitive K2P channels at the crayfish opener neuromuscular junction. At ultrasound intensity lower than those used to evoke FUSD, FUS consistently induced membrane hyperpolarization (FUSH) in motor axons but not muscle fibers, which may lack K2P. Since K2P channels are also thermosensitive, we varied the temperature from 12 to 32 °C. However, there was no significant correlation between FUSH amplitudes and temperature. Furthermore, FUSH was not inhibited by the K2P channel blockers, although the presence of the channels was confirmed by K2P blockers which increased input resistance and depolarized axonal resting membrane potential. Thus, it is unlikely that K2P channels underlie FUSH.
We also studied the impact of FUS on propagating action potentials (APs) in the crayfish motor axons. APs recorded during FUS took off from a hyperpolarized membrane potential and exhibited larger amplitudes and shorter duration. Three hypotheses were examined and eliminated. The US modulated AP shape changes cannot be due to: (1) alterations in microelectrode characteristics, (2) the increase in the fraction of sodium channels in the closed and not-inactivated state due to the hyperpolarization and (3) US activation of K2P channels which in turn altered AP shapes. One potential mechanism that requires further investigation is that FUS may accelerate the activation of sodium channel opening. Other factors that may indirectly modulate AP shapes are discussed.
In summary, results presented in this thesis suggest that FUS-mediated membrane responses in a single cell could vary depending on the FUS intensity and the type of ion channel a given cell expresses. Furthermore, ultrasound not only evokes changes membrane potential but also modulates action potentials. Collectively, these results represent significant contribution to the understanding of mechanisms underlying ultrasound neuromodulation at the cellular level.
Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/48049 |
Date | 08 February 2024 |
Creators | Yu, Feiyuan |
Contributors | Lin, Jen-Wei |
Source Sets | Boston University |
Language | en_US |
Detected Language | English |
Type | Thesis/Dissertation |
Rights | Attribution 4.0 International, http://creativecommons.org/licenses/by/4.0/ |
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