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.
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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.
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.
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.
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.
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.
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.
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.
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.
Steiner, J. P., T. M. Dawson, M. Fotuhi, and S. H. Snyder (1996) Immunophilin regulation of neurotransmitter release. Mol. Med. 2: 325–333.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Kasabdji, D., V. Shanmugam, and A. Rathinavelu (1996) Effect of cisplatin on dopamine release from PC12 cells. Life Sci. 59: 1793–1801.
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.
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.
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El-Said, W.A., Choi, JW. In-situ detection of neurotransmitter release from PC12 cells using Surface Enhanced Raman Spectroscopy. Biotechnol Bioproc E 19, 1069–1076 (2014). https://doi.org/10.1007/s12257-014-0092-7
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DOI: https://doi.org/10.1007/s12257-014-0092-7