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Neuroscience Bulletin logoLink to Neuroscience Bulletin
. 2015 Feb 4;31(2):265–270. doi: 10.1007/s12264-014-1497-1

Preserving GABAergic interneurons in acute brain slices of mice using the N-methyl-D-glucamine-based artificial cerebrospinal fluid method

Geng Pan 1, Yue Li 1, Hong-Yan Geng 1, Jian-Ming Yang 1, Ke-Xin Li 1, Xiao-Ming Li 1,2,
PMCID: PMC5562650  PMID: 25648546

Abstract

Defects in the function and development of GABAergic interneurons have been linked to psychiatric disorders, so preservation of these interneurons in brain slices is important for successful electrophysiological recording in various ex vivo methods. However, it is difficult to maintain the activity and morphology of neurons in slices from mice of >30 days old. Here we evaluated the N-methyl-D-glucamine (NMDG)-based artificial cerebrospinal fluid (aCSF) method for the preservation of interneurons in slices from mice of up to ∼6 months old and discussed the steps that may affect their quality during slicing. We found that the NMDG-aCSF method rescued more cells than sucrose-aCSF and successfully preserved different types of interneurons including parvalbumin- and somatostatin-positive interneurons. In addition, both the chemical and electrical synaptic signaling of interneurons were maintained. These results demonstrate that the NMDG-aCSF method is suitable for the preservation of interneurons, especially in studies of gap junctions.

Keywords: artificial cerebrospinal fluid, acute brain slice, electrophysiology, N-methyl-D-glucamine, parvalbumin, somatostatin

References

  • [1].Aghajanian GK, Rasmussen K. Intracellular studies in the facial nucleus illustrating a simple new method for obtaining viable motoneurons in adult rat brain slices. Synapse. 1989;3:331–338. doi: 10.1002/syn.890030406. [DOI] [PubMed] [Google Scholar]
  • [2].Nashmi R, Velumian AA, Chung I, Zhang L, Agrawal SK, Fehlings MG. Patch-clamp recordings from white matter glia in thin longitudinal slices of adult rat spinal cord. J Neurosci Methods. 2002;117:159–166. doi: 10.1016/S0165-0270(02)00096-1. [DOI] [PubMed] [Google Scholar]
  • [3].Peca J, Feliciano C, Ting JT, Wang W, Wells MF, Venkatraman TN, et al. Shank3 mutant mice display autisticlike behaviours and striatal dysfunction. Nature. 2011;472:437–442. doi: 10.1038/nature09965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [4].Zhao S, Ting JT, Atallah HE, Qiu L, Tan J, Gloss B, et al. Cell type-specific channelrhodopsin-2 transgenic mice for optogenetic dissection of neural circuitry function. Nat Methods. 2011;8:745–752. doi: 10.1038/nmeth.1668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [5].Ye JH, Zhang J, Xiao C, Kong JQ. Patch-clamp studies in the CNS illustrate a simple new method for obtaining viable neurons in rat brain slices: glycerol replacement of NaCl protects CNS neurons. J Neurosci Methods. 2006;158:251–259. doi: 10.1016/j.jneumeth.2006.06.006. [DOI] [PubMed] [Google Scholar]
  • [6].Robillard JM, Gordon GR, Choi HB, Christie BR, MacVicar BA. Glutathione restores the mechanism of synaptic plasticity in aged mice to that of the adult. PLoS One. 2011;6:e20676. doi: 10.1371/journal.pone.0020676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [7].Brahma B, Forman RE, Stewart EE, Nicholson C, Rice ME. Ascorbate inhibits edema in brain slices. J Neurochem. 2000;74:1263–1270. doi: 10.1046/j.1471-4159.2000.741263.x. [DOI] [PubMed] [Google Scholar]
  • [8].Espanol MT, Xu Y, Litt L, Chang LH, James TL, Weinstein PR, et al. Modulation of edema by dizocilpine, kynurenate, and NBQX in respiring brain slices after exposure to glutamate. Acta Neurochir Suppl (Wien) 1994;60:58–61. doi: 10.1007/978-3-7091-9334-1_15. [DOI] [PubMed] [Google Scholar]
  • [9].Werth JL, Park TS, Silbergeld DL, Rothman SM. Excitotoxic swelling occurs in oxygen and glucose deprived human cortical slices. Brain Res. 1998;782:248–254. doi: 10.1016/S0006-8993(97)01286-9. [DOI] [PubMed] [Google Scholar]
  • [10].Chesler M, Chen JC, Kraig RP. Determination of extracellular bicarbonate and carbon dioxide concentrations in brain slices using carbonate and pH-selective microelectrodes. J Neurosci Methods. 1994;53:129–136. doi: 10.1016/0165-0270(94)90169-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [11].Desagher S, Glowinski J, Prémont J. Pyruvate protects neurons against hydrogen peroxide-induced toxicity. J Neurosci. 1997;17:9060–9067. doi: 10.1523/JNEUROSCI.17-23-09060.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • [12].Casalino E, Sblano C, Landriscina C. A possible mechanism for initiation of lipid peroxidation by ascorbate in rat liver microsomes. Int J Biochem Cell Biol. 1996;28:137–149. doi: 10.1016/1357-2725(95)00131-X. [DOI] [PubMed] [Google Scholar]
  • [13].MacGregor DG, Chesler M, Rice ME. HEPES prevents edema in rat brain slices. Neurosci Lett. 2001;303:141–144. doi: 10.1016/S0304-3940(01)01690-1. [DOI] [PubMed] [Google Scholar]
  • [14].Aoyama K, Suh SW, Hamby AM, Liu J, Chan WY, Chen Y, et al. Neuronal glutathione deficiency and age-dependent neurodegeneration in the EAAC1 deficient mouse. Nat Neurosci. 2006;9:119–126. doi: 10.1038/nn1609. [DOI] [PubMed] [Google Scholar]
  • [15].Steullet P, Neijt HC, Cuénod M, Do KQ. Synaptic plasticity impairment and hypofunction of NMDA receptors induced by glutathione deficit: relevance to schizophrenia. Neuroscience. 2006;137:807–819. doi: 10.1016/j.neuroscience.2005.10.014. [DOI] [PubMed] [Google Scholar]
  • [16].Lippiat JD, Standen NB, Davies NW. Block of cloned BKCa channels (rSlo) expressed in HEK 293 cells by N-methyl D-glucamine. Pflugers Arch. 1998;436:810–812. doi: 10.1007/s004240050708. [DOI] [PubMed] [Google Scholar]
  • [17].Terasawa K, Nakajima T, Iida H, Iwasawa K, Oonuma H, Jo T, et al. Nonselective cation currents regulate membrane potential of rabbit coronary arterial cell: modulation by lysophosphatidylcholine. Circulation. 2002;106:3111–3119. doi: 10.1161/01.CIR.0000039345.00481.1D. [DOI] [PubMed] [Google Scholar]
  • [18].Geiger JRP, Bischofberger J, Vida I, Fröbe U, Pfitzinger S, Weber HJ, et al. Patch-clamp recording in brain slices with improved slicer technology. Pflugers Arch. 2002;443:491–501. doi: 10.1007/s00424-001-0735-3. [DOI] [PubMed] [Google Scholar]

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