Biotechnology and Bioprocess Engineering

, Volume 19, Issue 6, pp 1069–1076 | Cite as

In-situ detection of neurotransmitter release from PC12 cells using Surface Enhanced Raman Spectroscopy

Research Paper
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Abstract

Rat pheochromocytoma PC12 cells have frequently been used as a dopaminergic neuron model due to their various functions, including the synthesis, storage, and secretion of catecholamines. Furthermore, PC12 cells release a measurable amount of dopamine (DA) in response to some chemicals. PC12 cells are thus considered to be one of the most common invitro models for studying neurotransmitter release. Here, we applied Surface-enhanced Raman Spectroscopy (SERS) to determine with high sensitivity the in-situ short-time effects of cisplatin (cisdiamine- dichloroplatinum), bisphenol-A, and cyclophosphamide on the extracellular DA level released from PC12 cells. In addition, using the SERS technique, changes in the biochemical composition of the PC12 cell lysates were investigated to determine the intracellular DA level. Gold nano-patterned substrates were fabricated based on electrochemical deposition of Au nanorods onto ITO substrates; these substrates were then used as SERS-active surfaces. The Raman spectroscopy results demonstrated that the changes in the Raman spectra depending on the treatment agent were in agreement with the HPLC results on the extracellular DA level. Therefore, the SERS technique can overcome the limitations of other detection techniques, and can be used with cellular nanoarrays to study the effect of a wide range of chemicals.

Keywords

Raman spectroscopy neurotransmitter PC12 cell gold nanorods nanobiochip drug discovery 
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References

  1. 1.
    Taylor, S. C., E. Carpenter, M. L. Roberts, and C. Peers (1999) Potentiation of quantal catecholamine secretion by glibenclamide: Evidence for a novel role of sulphonylurea receptors in regulating the Ca(2+) sensitivity of exocytosis. J. Neurosci. 19: 5741–5749.Google Scholar
  2. 2.
    Pothos, E., M. Desmond, and D. Sulzer (1996) l–3,4-Dihydroxyphenylalanine increases the quantal size of exocytotic dopamine release in vitro. J. Neurochem. 66: 629–636.CrossRefGoogle Scholar
  3. 3.
    Zachor, D. A., J. F. Moore, C. Brezausek, A. Theibert, and A. K. Percy (2000) Cocaine inhibits NGF-induced PC12 cells differentiation through D1-type dopamine receptors. Brain Res. 869: 85–97.CrossRefGoogle Scholar
  4. 4.
    Presse, F., B. Cardona, L. Borsu, and J.-L. Nahon (1997) Lithium increases melanin-concentrating hormone mRNA stability and inhibits tyrosine hydroxylase gene expression in PC12 cells. Mol. Brain Res. 52: 270–283.CrossRefGoogle Scholar
  5. 5.
    Tischler, A. S., R. L. Perlman, G. M. Morse, and B. E. Sheard (1983) Glucocorticoids increase catecholamine synthesis and storage in PC12 pheochromocytoma cell cultures. J Neurochem. 40: 364–370.CrossRefGoogle Scholar
  6. 6.
    Brownell, A. L., B. G. Jenkins, and O. Isacson (1999) Dopamine imaging markers and predictive mathematical models for progressive degeneration in Parkinson’s disease. Biomed. Pharmacother. 53: 131–140.CrossRefGoogle Scholar
  7. 7.
    Carr, D. B., P. O’Donnell, J. P. Card, and S. R. Sesack (1999) Dopamine terminals in the rat prefrontal cortex synapse on pyramidal cells that project to the nucleus accumbens. The J. Neurosci. 19: 11049–11060.Google Scholar
  8. 8.
    Shi, B., W. Huang, and J. Cheng (2007) Determination of neurotransmitters in PC 12 cells by microchip electrophoresis with fluorescence detection. Electrophoresis 28: 1595–1600.CrossRefGoogle Scholar
  9. 9.
    Steiner, J. P., T. M. Dawson, M. Fotuhi, and S. H. Snyder (1996) Immunophilin regulation of neurotransmitter release. Mol. Med. 2: 325–333.Google Scholar
  10. 10.
    Vo, T. D. L., J.-S. Ko, S. H. Lee, S. J. Park and S. H. Hong (2013) Overexpression of Neurospora crassa OR74A glutamate decarboxylase in Escherichia coli for efficient GABA production. Biotechnol. Bioproc. Eng. 18: 1062–1066.CrossRefGoogle Scholar
  11. 11.
    Zuriani, R., S. Vigneswari, M. N. M. Azizan, M. I. A. Majid and A. A. Amirul (2013) A high throughput nile red fluorescence method for rapid quantification of intracellular bacterial polyhydroxyalkanoates. Biotechnol. Bioproc. Eng. 18: 472–478.CrossRefGoogle Scholar
  12. 12.
    Park, J. K., Z.-H. Kim, C. G. Lee, A. Synytsya, H. S. Jo, S. O. Kim, J. W. Park, and Y. I. Park (2011) Characterization and immunostimulating activity of a water-soluble polysaccharide isolated from Haematococcus lacustris. Biotechnol. Bioproc. Eng. 16: 1090–1098.CrossRefGoogle Scholar
  13. 13.
    Schulze, H. G., L. S. Greek, B. B. Gorzalka, A. V. Bree, M. W. Blades, and R. F. B. Turner (1995) Artificial neural network and classical least-squares methods for neurotransmitter mixture analysis. J. Neurosci. Methods 56: 155–167.CrossRefGoogle Scholar
  14. 14.
    El-Said, W. A., J.-H. Lee, B.-K. Oh, and J.-W. Choi (2011) Electrochemical sensor to detect neurotransmitter using gold nano-island coated ITO electrode. J. Nanosci. Nanotechnol. 11: 6539–6543.CrossRefGoogle Scholar
  15. 15.
    El-Said, W. A., T. H. Kim, C. H. Yea, H. Kim, and J. W. Choi (2011) Fabrication of gold nanoparticle modified ITO substrate to detect beta-amyloid using surface-enhanced Raman scattering. J. Nanosci. Nanotechnol. 11: 768–772.CrossRefGoogle Scholar
  16. 16.
    El-Said, W. A., T.-H. Kim, H. Kim, and J.-W. Choi (2010) Detection of effect of chemotherapeutic agents to cancer cells on gold nanoflower patterned substrate using surface-enhanced Raman scattering and cyclic voltammetry. Biosens. Bioelectron. 26: 1486–1492.CrossRefGoogle Scholar
  17. 17.
    Chen, J., J. Jiang, X. Gao, G. Liu, G. Shen, and R. Yu (2008) A new aptameric biosensor for cocaine based on surface-enhanced Raman scattering spectroscopy. Chem. (Weinheim an der Bergstrasse, Germany). 14: 8374–8382.Google Scholar
  18. 18.
    Talley, C. E., L. Jusinski, C. W. Hollars, S. M. Lane, and T. Huser (2004) Intracellular pH sensors based on surface-enhanced Raman scattering. Anal. Chem. 76: 7064–7068.CrossRefGoogle Scholar
  19. 19.
    Wang, Z., A. Bonoiu, M. Samoc, Y. Cui, and P. N. Prasad (2008) Biological pH sensing based on surface enhanced Raman scattering through a 2-aminothiophenol-silver probe. Biosens. Bioelectron. 23: 886–891.CrossRefGoogle Scholar
  20. 20.
    Nowak-Lovato, K. L. and K. D. Rector (2012) Live cells as dynamic laboratories: Time lapse raman spectral microscopy of nanoparticles with both IgE targeting and pH-sensing functions. Int. J. Anal. Chem. 2012: 390182.CrossRefGoogle Scholar
  21. 21.
    El-Said, W. A., T. H. Kim, H. Kim, and J. W. Choi (2011) Analysis of intracellular state based on controlled 3D nanostructures mediated surface enhanced Raman scattering. PloS one. 6: e15836.CrossRefGoogle Scholar
  22. 22.
    Orendorff, C. J., L. Gearheart, N. R. Jana, and C. J. Murphy (2006) Aspect ratio dependence on surface enhanced Raman scattering using silver and gold nanorod substrates. Physic. Chem. Chem. Phys. 8: 165–170.CrossRefGoogle Scholar
  23. 23.
    Kelly, K. L., E. Coronado, L. L. Zhao, and G. C. Schatz (2002) The optical properties of metal nanoparticles: The influence of size, shape, and dielectric environment. J. Phys. Chem. B. 107: 668–677.CrossRefGoogle Scholar
  24. 24.
    Niesen, B., B. P. Rand, P. Van Dorpe, H. Shen, B. Maes, J. Genoe, and P. Heremans (2010) Excitation of multiple dipole surface plasmon resonances in spherical silver nanoparticles. Optics Exp. 18: 19032–19038.CrossRefGoogle Scholar
  25. 25.
    Huang, H., L. Zhu, B. R. Reid, G. P. Drobny, and P. B. Hopkins (1995) Solution structure of a cisplatin-induced DNA interstrand cross-link. Science 270: 1842–1845.CrossRefGoogle Scholar
  26. 26.
    Ta, L. E., L. Espeset, J. Podratz, and A. J. Windebank (2006) Neurotoxicity of oxaliplatin and cisplatin for dorsal root ganglion neurons correlates with platinum–DNA binding. Neurotoxicol. 27: 992–1002.CrossRefGoogle Scholar
  27. 27.
    Kasabdji, D., V. Shanmugam, and A. Rathinavelu (1996) Effect of cisplatin on dopamine release from PC12 cells. Life Sci. 59: 1793–1801.CrossRefGoogle Scholar
  28. 28.
    Peng, S., J. M. McMahon, G. C. Schatz, S. K. Gray, and Y. Sun (2010) Reversing the size-dependence of surface plasmon resonances. Proc. Natl. Acad. Sci. U.S.A. 107: 14530–14534.CrossRefGoogle Scholar
  29. 29.
    Kim, J., J. Lee, S. Kwon, and S. Jeong (2009) Preparation of biodegradable polymer/silver nanoparticles composite and its antibacterial efficacy. J. Nanosci. Nanotechnol. 9: 1098–1102.CrossRefGoogle Scholar

Copyright information

© The Korean Society for Biotechnology and Bioengineering and Springer-Verlag Berlin Heidelberg 2014

Authors and Affiliations

  1. 1.Interdisciplinary program of Integrated BiotechnologySogang UniversitySeoulKorea
  2. 2.Department of Chemical & Biomolecular EngineeringSogang UniversitySeoulKorea
  3. 3.Department of Chemistry, Faculty of ScienceAssiut UniversityAssiutEgypt

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