Abstract
This chapter summarizes the extracellular recording and juxtacellular labeling method and its application to the characterization of cardiorespiratory brainstem neurons, although this technique could be applied to any neuron. The combination of this method with immunohistochemical and in situ hybridization techniques is emphasized in detail.
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References
Pinault D (1996) A novel single-cell staining procedure performed in vivo under electrophysiological control: morpho-functional features of juxtacellularly labeled thalamic cells and other central neurons with biocytin or Neurobiotin. J Neurosci Methods 65:113–136
Guyenet PG, Stornetta RL, Weston MC, McQuiston T, Simmons JR (2004) Detection of amino acid and peptide transmitters in physiologically identified brainstem cardiorespiratory neurons. Auton Neurosci 114:1–10
Duque A, Tepper JM, Detari L, Ascoli GA, Zaborszky L (2007) Morphological characterization of electrophysiologically and immunohistochemically identified basal forebrain cholinergic and neuropeptide Y-containing neurons. Brain Struct Funct 212:55–73
Manns ID, Alonso A, Jones BE (2000) Discharge profiles of juxtacellularly labeled and immunohistochemically identified GABAergic basal forebrain neurons recorded in association with the electroencephalogram in anesthetized rats. J Neurosci 20:9252–9263
Tang X, Neckel ND, Schramm LP (2003) Locations and morphologies of sympathetically correlated neurons in the T10 spinal segment of the rat. Brain Res 976:185–193
Mulkey DK, Stornetta RL, Weston MC, Simmons JR, Parker A, Bayliss DA, Guyenet PG (2004) Respiratory control by ventral surface chemoreceptor neurons in rats. Nat Neurosci 7:1360–1369
St-John WM, Stornetta RL, Guyenet PG, Paton JF (2009) Location and properties of respiratory neurones with putative intrinsic bursting properties in the rat in situ. J Physiol 587:3175–3188
Neumann E, Kakorin S, Toensing K (1999) Fundamentals of electroporative delivery of drugs and genes. Bioelectrochem Bioenerg 48:3–16
Boudes M, Pieraut S, Valmier J, Carroll P, Scamps F (2008) Single-cell electroporation of adult sensory neurons for gene screening with RNA interference mechanism. J Neurosci Methods 170:204–211
Nevian T, Helmchen F (2007) Calcium indicator loading of neurons using single-cell electroporation. Pflugers Arch 454:675–688
Nagayama S, Zeng S, Xiong W, Fletcher ML, Masurkar AV, Davis DJ, Pieribone VA, Chen WR (2007) In vivo simultaneous tracing and Ca(2+) imaging of local neuronal circuits. Neuron 53:789–803
Brown DL, Guyenet PG (1985) Electrophysiological study of cardiovascular neurons in the rostral ventrolateral medulla in rats. Circ Res 56:359–369
Stornetta RL, Moreira TS, Takakura AC, Kang BJ, Chang DA, West GH, Brunet JF, Mulkey DK, Bayliss DA, Guyenet PG (2006) Expression of Phox2b by brainstem neurons involved in chemosensory integration in the adult rat. J Neurosci 26:10305–10314
Lazarenko RM, Milner TA, Depuy SD, Stornetta RL, West GH, Kievits JA, Bayliss DA, Guyenet PG (2009) Acid sensitivity and ultrastructure of the retrotrapezoid nucleus in Phox2b-EGFP transgenic mice. J Comp Neurol 517:69–86
Fortuna MG, West GH, Stornetta RL, Guyenet PG (2008) Botzinger expiratory-augmenting neurons and the parafacial respiratory group. J Neurosci 28:2506–2515
Stornetta RL, Spirovski D, Moreira TS, Takakura AC, West GH, Gwilt JM, Pilowsky PM, Guyenet PG (2009) Galanin is a selective marker of the retrotrapezoid nucleus in rats. J Comp Neurol 512:373–383
Dutschmann M, Wilson RJA, Paton JFR (2000) Respiratory activity in neonatal rats. Auton Neurosci 84:19–29
Acknowledgments
This work was supported by National Institutes of Health Grants HL74011 and HL28785.
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Stornetta, R.L. (2012). Juxtacellular Labeling in Combination with Other Histological Techniques to Determine Phenotype of Physiologically Identified Neurons. In: Badoer, E. (eds) Visualization Techniques. Neuromethods, vol 70. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-897-9_9
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DOI: https://doi.org/10.1007/978-1-61779-897-9_9
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