Microsystem Technologies

, Volume 23, Issue 8, pp 3671–3683 | Cite as

Sustained elevation of activity of developing neurons grown on polyimide microelectrode arrays (MEA) in response to ultrasound exposure

  • Massoud L. Khraiche
  • William B. Phillips
  • Nathan Jackson
  • Jit Muthuswamy
Technical Paper

Abstract

High frequency ultrasound (HFUS) is an attractive modality for noninvasive clinical applications such as imaging, diagnostics and more recently for stimulation of the central nervous system. The aim of this study was to investigate the modulation in the electrical activity of developing neurons due to the application of HFUS using polyimide based microelectrode array (MEA) that is acoustically transparent in order to allow ultrasound waves to transmit through the substrate and reach the growing neural layer. High frequency tone bursts of ultrasound were applied to a monolayer of developing primary neurons grown on an acoustical transparent polyimide MEA. HFUS was applied to primary neuronal culture at two frequencies (4.4 and 96 MHz) with spatial peak-temporal average intensities of 100 and 10 mW/cm2. Exposures were found to increase the spike rate of neurons in culture up to 20-fold in some cases and induce silent or still developing neurons to fire at a maximum rate of up to three new units per recording microelectrode. Another new observation reported in this study is that the increase in spike rate was sustained for over 6 min post stimulation. Our results also suggest that mechanical and not thermal effects of ultrasound largely mediate the increase in electrical excitability without any discernible spatial pattern or preference across the monolayer for the US parameters used in this study. The accessibility of the disassociated neuronal cultures to stimulation, imaging and recording provides a useful model for investigating the exact mechanisms behind the effect of ultrasound on neuronal excitability.

References

  1. Ang ES Jr, Gluncic V, Duque A, Schafer ME, Rakic P (2006) Prenatal exposure to ultrasound waves impacts neuronal migration in mice. Proc Natl Acad Sci USA 103:12903–12910CrossRefGoogle Scholar
  2. Bachtold MR, Rinaldi PC, Jones JP, Reines F, Price LR (1998) Focused ultrasound modifications of neural circuit activity in a mammalian brain. Ultrasound Med Biol 24:557–565CrossRefGoogle Scholar
  3. Bello SO (2006) How we may be missing some harmful effects of ultrasound—a hypothesis. Med Hypotheses 67:765–767CrossRefGoogle Scholar
  4. Boppart SA, Wheeler BC, Wallace CS (1992) A flexible perforated microelectrode array for extended neural recordings. IEEE Trans Biomed Eng 39:37–42CrossRefGoogle Scholar
  5. Chang JC, Brewer GJ, Wheeler BC (2001) Modulation of neural network activity by patterning. Biosens Bioelectron 16:527–533CrossRefGoogle Scholar
  6. Chiappalone M, Bove M, Vato A, Tedesco M, Martinoia S (2006) Dissociated cortical networks show spontaneously correlated activity patterns during in vitro development. Brain Res 1093:41–53. doi:10.1016/J.Brainres.2006.03.049 CrossRefGoogle Scholar
  7. Cohen AS, Pfister BJ, Schwarzbach E, Grady MS, Goforth PB, Satin LS (2007) Injury-induced alterations in CNS electrophysiology. Prog Brain Res 161:143–169. doi:10.1016/S0079-6123(06)61010-8 CrossRefGoogle Scholar
  8. Duck FA (2008) Hazards, risks and safety of diagnostic ultrasound. Med Eng Phys 30:1338–1348CrossRefGoogle Scholar
  9. Fleischer AC, Fleischer A, Toy EC, Toy E, Lee W, Manning FA (2011) Sonography in obstetrics and gynecology: principles and practice by 6 edition. McGraw-Hill Professional, New York CityGoogle Scholar
  10. Geddes-Klein DM, Schiffman KB, Meaney DF (2006) Mechanisms and consequences of neuronal stretch injury in vitro differ with the model of trauma. J Neurotrauma 23:193–204. doi:10.1089/neu.2006.23.193 CrossRefGoogle Scholar
  11. Hachiya H (2006) Safety of ultrasound diagnosis. J Med Ultrason 33(4):195CrossRefGoogle Scholar
  12. Hu JH, Ulrich WD (1976) Effects of low-intensity ultrasound on the central nervous system of primates. Aviat Space Environ Med 47:640–643Google Scholar
  13. Hu Y, Zhong W, Wan JM, Alfred C (2013) Ultrasound can modulate neuronal development: impact on neurite growth and cell body morphology. Ultrasound Med Biol 39:915–925CrossRefGoogle Scholar
  14. Ibsen S, Tong A, Schutt C, Esener S, Chalasani SH (2015) Sonogenetics is a non-invasive approach to activating neurons in Caenorhabditis elegans. Nat Commun 6:8264. doi:10.1038/ncomms9264 CrossRefGoogle Scholar
  15. Jackson N, Muthuswamy J (2009) Flexible chip scale package and interconnect for implantable MEMS movable microelectrodes for the brain. J Microelectromech Syst 18:396–404. doi:10.1109/JMEMS.2009.2013391 CrossRefGoogle Scholar
  16. Khraiche ML, Phillips WB, Jackson N, Muthuswamy J (2008) Ultrasound induced increase in excitability of single neurons. Conf Proc IEEE Eng Med Biol Soc 2008:4246–4249. doi:10.1109/IEMBS.2008.4650147 Google Scholar
  17. King RL, Brown JR, Newsome WT, Pauly KB (2013) Effective parameters for ultrasound-induced in vivo neurostimulation. Ultrasound Med Biol 39:312–331. doi:10.1016/J.Ultrasmedbio.2012.09.009 CrossRefGoogle Scholar
  18. Korb AS, Shellock FG, Cohen MS, Bystritsky A (2014) Low-intensity focused ultrasound pulsation device used during magnetic resonance imaging: evaluation of magnetic resonance imaging-related heating at 3 Tesla/128 MHz. Neuromodul Technol Neural Interface 17:236–241. doi:10.1111/ner.12075 CrossRefGoogle Scholar
  19. Legon W, Sato TF, Opitz A, Mueller J, Barbour A, Williams A, Tyler WJ (2014) Transcranial focused ultrasound modulates the activity of primary somatosensory cortex in humans. Nat Neurosci 17:322–329. doi:10.1038/nn.3620 CrossRefGoogle Scholar
  20. Maeda K, Kurjak A (2012) The safe use of diagnositic ultrasound in obstetrics and gynecology, Donald School. J Ultrasound Obstet Gynecol 3:313–317CrossRefGoogle Scholar
  21. Marinac-Dabic D, Krulewitch CJ, Moore RM Jr (2002) The safety of prenatal ultrasound exposure in human studies. Epidemiology 13(Suppl 3):S19–S22  CrossRefGoogle Scholar
  22. Martinac B (2004) Mechanosensitive ion channels: molecules of mechanotransduction. J Cell Sci 117:2449–2460. doi:10.1242/jcs.01232 CrossRefGoogle Scholar
  23. Mehic E, Xu JM, Caler CJ, Coulson NK, Moritz CT, Mourad PD (2014) Increased anatomical specificity of neuromodulation via modulated focused ultrasound. PLoS One 9:e86939. doi:10.1371/journal.pone.0086939 CrossRefGoogle Scholar
  24. Mihran RT, Barnes FS, Wachtel H (1990a) Temporally-specific modification of myelinated axon excitability in vitro following a single ultrasound pulse. Ultrasound Med Biol 16:297–309CrossRefGoogle Scholar
  25. Mihran RT, Barnes FS, Wachtel H (1990b) Transient modification of nerve excitability in vitro by single ultrasound pulses. Biomed Sci Instrum 26:235–246Google Scholar
  26. Miller DL, Smith NB, Bailey MR, Czarnota GJ, Hynynen K, Makin IR (2012) Overview of therapeutic ultrasound applications and safety considerations. J Ultrasound Med 31:623–634CrossRefGoogle Scholar
  27. Morin FO, Takamura Y, Tamiya E (2005) Investigating neuronal activity with planar microelectrode arrays: achievements and new perspectives. J Biosci Bioeng 100:17–24CrossRefGoogle Scholar
  28. Philips WB (2002) In vitro modification of nerve excitability via high frequency ultrasound pulses. Dissertation, Arizona State University, TempeGoogle Scholar
  29. Phillips WB, Larson PJ, Towe BC (2004) Ultrasonically-assisted intracortical microstimulation of the rat. Conf Proc IEEE Eng Med Biol Soc 6:4217–4220Google Scholar
  30. Potter SM, DeMarse TB (2001) A new approach to neural cell culture for long-term studies. J Neurosci Methods 110:17–24CrossRefGoogle Scholar
  31. Sachs F, Morris CE (1998) Mechanosensitive ion channels in nonspecialized cells. Rev Physiol Biochem Pharmacol 132:1–77CrossRefGoogle Scholar
  32. Stephenson J (2005) Fetal ultrasound safety, vol 293. doi:10.1001/jama.293.3.286-c
  33. Sutton Y, Shaw A, Zeqiri B (2003) Measurement of ultrasonic power using an acoustically absorbing well. Ultrasound Med Biol 29:1507–1513CrossRefGoogle Scholar
  34. Takagi SF, Higashino S, Shibuya T, Osawa N (1960) The actions of ultrasound on the myelinated nerve, the spinal cord and the brain. Jpn J Physiol 10:183–193CrossRefGoogle Scholar
  35. Tarantal AF, O’Brien WD, Hendrickx AG (1993) Evaluation of the bioeffects of prenatal ultrasound exposure in the cynomolgus macaque (Macaca fascicularis): III. Dev Hematol Studies Teratol 47:159–170Google Scholar
  36. Treeby BE, Jaros J, Rendell AP, Cox BT (2012) Modeling nonlinear ultrasound propagation in heterogeneous media with power law absorption using a k-space pseudospectral method. J Acoust Soc Am 131:4324–4336. doi:10.1121/1.4712021 CrossRefGoogle Scholar
  37. Tufail Y et al (2010) Transcranial pulsed ultrasound stimulates intact brain circuits. Neuron 66:681–694. doi:10.1016/j.neuron.2010.05.008 CrossRefGoogle Scholar
  38. Tyler WJ, Tufail Y, Finsterwald M, Tauchmann ML, Olson EJ, Majestic C (2008) Remote excitation of neuronal circuits using low-intensity, low-frequency ultrasound. PLoS One 3:e3511CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  • Massoud L. Khraiche
    • 1
    • 2
  • William B. Phillips
    • 1
  • Nathan Jackson
    • 1
    • 3
  • Jit Muthuswamy
    • 1
  1. 1.Harrington Department of Bioengineering ECG 334Arizona State UniversityTempeUSA
  2. 2.Department of OphthalmologyUniversity of California San DiegoLa JollaUSA
  3. 3.Tyndall National InstituteUniversity College CorkCorkIreland

Personalised recommendations