The use of atomic force microscopy and organotypic tissue culture to study the effect of a short arginine-containing peptide with analgesic potential on the sensory neuron membrane

Authors

  • Maksim M. Khalisov Pavlov Institute of Physiology, Russian Academy of Sciences
  • Valentina A. Penniyaynen Pavlov Institute of Physiology, Russian Academy of Sciences
  • Alexander V. Ankudinov Ioffe Institute, Russian Academy of Sciences
  • Svetlana A. Podzorova Pavlov Institute of Physiology, Russian Academy of Sciences
  • Boris V. Krylov Pavlov Institute of Physiology, Russian Academy of Sciences

DOI:

https://doi.org/10.33910/2687-1270-2022-3-1-58-68

Keywords:

sensory neurons, short peptides, Ac-RERR-NH2, atomic force microscopy, organotypic nerve culture method

Abstract

Opiate use to treat chronic pain is known to be associated with negative side effects. Therefore, the development of new safe and effective non-opioid analgesics is both important and urgent goal of modern science. Previously, we showed that tetrapeptide Ac-RERR-NH2 has potential as an analgesic drug substance, since it can reduce the excitability of nociceptive neurons responsible for encoding nociceptive signals. The effect of sub-nanomolar concentration of Ac-RERR-NH2 tetrapeptide on embryonic sensory neurons was investigated with the use of atomic force microscopy (AFM) and organotypic tissue culture method. The quasi-static PeakForce QNM AFM mode was used, which allows mapping of local mechanical cell properties. Following administration of the substance, sensory neurons tended to decrease their stiffness. This result is based on the analysis of the values of the apparent Young’s modulus of sensory neurons and their deformation, controlled for the slipping of the tip of the probe over the surface under study. The area index (AI) criterion showed that the studied tetrapeptide has pronounced neurite-stimulating properties. The data obtained suggest that Ac-RERR-NH2, acting at very low concentrations, is able to trigger an as yet unidentified intracellular cascade regulating the growth of neurites of sensory neurons.

References

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REFERENCES

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Haase, K, Pelling, A.E. (2015) Investigating cell mechanics with atomic force microscopy. Journal of The Royal Society Interface, vol. 12, no. 104, article 20140970. http://dx.doi.org/10.1098/rsif.2014.0970 (In English)

Hu, J., Chen, S., Huang, D. et al. (2020) Global mapping of live cell mechanical features using PeakForce QNM AFM. Biophysics Reports, vol. 6, no. 1, pp. 9–18. https://doi.org/10.1007/s41048-019-00103-9 (In English)

Hutter, J. L., Bechhoefer, J. (1993) Calibration of atomic-force microscope tips. Review of Scientific Instruments, vol. 64, no. 7, pp. 1868–1873. https://doi.org/10.1063/1.1143970 (In English)

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Sneddon, I. N. (1965) The relation between load and penetration in the axi-symmetric boussinesq problem for a punch of arbitrary profile. International Journal of Engineering Science, vol. 3, no. 1, pp. 47–57. https://doi.org/10.1016/0020-7225(65)90019-4 (In English)

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Published

2022-06-30

Issue

Vol. 3 No. 1 (2022)

Section

Experimental articles

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Copyright (c) 2022 Maksim M. Khalisov, Valentina A. Penniyaynen, Alexander V. Ankudinov, Svetlana A. Podzorova, Boris V. Krylov

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