Abstract
Microiontophoresis combined with extracellular spike recording is an excellent method for investigating local neuropharmacological effects under in vivo conditions. However, its application has recently become relatively rare in neuroscience research. Now, we aimed to revisit microiontophoresis and demonstrate that it provides valuable data about the pharmacophysiology of neurons and local neuronal networks, in vivo. Extracellular recordings were performed through the central recording channel of multibarrel carbon-fiber microelectrodes in the CA1 pyramidal layer of the hippocampus of anesthetized rats, while N-methyl-D-aspartate (NMDA) was locally administrated by means of microiontophoresis through the surrounding micropipettes of the microelectrode. Various separation procedures were used to distinguish putative pyramidal cells and interneurons. Quality of separation was verified by electrophysiological parameters. After the delivery of NMDA in the vicinity of the examined neurons, firing rate of putative pyramidal cells was increased with a significantly higher grade then that of putative interneurons. The present results in line with previous data indicate that pyramidal cells are more responsive to pharmacological manipulation through NMDA receptors, also confirming the reliability of the separation of different types of neurons in in vivo microiontophoretic experiments.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Alkondon, M., Albuquerque, E. X. (2012) Nicotinic acetylcholine receptor α7 and α4β2 subtypes differentially control GABAergic input to CA1 neurons in rat hippocampus. J. Neurophysiol. 3043–3055.
Avignone, E., Frenguelli, B. G., Irving, A. J. (2005) Differential responses to NMDA receptor activation in rat hippocampal interneurons and pyramidal cells may underlie enhanced pyramidal cell vulnerability. Eur. J. Neurosci. 22, 3077–3090.
Bird, S. J., Aghajanian, G. K. (1976) The cholinergic pharmacology of hippocampal pyramidal cells: a microiontophoretic study. Neuropharmacology 15, 273–282.
Boakes, R. J., Bramwell, G. J., Briggs, I., Candy, J. M., Tempesta, E. (1974) Localization with Pontamine Sky Blue of neurones in the brainstem responding to microiontophoretically applied compounds. Neuropharmacology 13, 475–479.
Budai, D. (2010) Carbon fiber-based microelectrodes and microbiosensors. In: Somerset, V. S. (ed.) Intelligent and Biosensors. InTech, Europa, Rijeka, pp. 269–288. Also available online for open access: https://doi.org/www.intechopen.com
Csicsvári, J., Hirase, H., Czurko, A., Buzsáki, G. (1998) Reliability and state dependence of pyramidal cell-interneuron synapses in the hippocampus: an ensemble approach in the behaving rat. Neuron 21, 179–189.
Czéh, B., Ábrahám, H., Tahtakran, S., Houser, C. R., Seress, L. (2013) Number and regional distribution of GAD65 mRNA-expressing interneurons in the rat hippocampal formation. Acta Biol. Hung. 64, 395–413.
Del Castillo, J., Katz, B. (1955) On the localization of acetylcholine receptors. J. Physiol. 128, 157–181.
Fox, S. E., Ranck, J. B. (1981) Electrophysiological characteristics of hippocampal complex-spike cells and theta cells. Exp. Brain Res. 41, 399–410.
Furue, H., Katafuchi, T., Yoshimura, M. (2007) In vivo patch-clamp technique. In: Walz, W. (ed.) Patch-Clamp Analysis. Humana Press, pp. 229–251.
Gerhardt, G. A., Palmer, M. R. (1987) Characterization of the techniques of pressure ejection and microiontophoresis using in vivo electrochemistry. J. Neurosci. Methods 22, 147–159.
Harris, K. D., Hirase, H., Leinekugel, X., Henze, D. A., Buzsáki, G. (2001) Temporal interaction between single spikes and complex spike bursts in hippocampal pyramidal cells. Neuron 32, 141–149.
Hazan, L., Zugaro, M., Buzsáki, G. (2006) Klusters, NeuroScope, NDManager: a free software suite for neurophysiological data processing and visualization. J. Neurosci. Methods 155, 207–216.
Joëls, M., Urban, I. J. (1982) The effect of microiontophoretically applied vasopressin and oxytocin on single neurones in the septum and dorsal hippocampus of the rat. Neurosci. Lett. 33, 79–84.
Klausberger, T., Somogyi, P. (2008) Neuronal diversity and temporal dynamics: the unity of hippocampal circuit operations. Science 321, 53–57. 21.
Kovács, P., Hernádi, I. (2006) Yohimbine acts as a putative in vivo alpha2A/D-antagonist in the rat prefrontal cortex. Neurosci. Lett. 402, 253–258.
Kovács, P., Dénes, V., Kellényi, L., Hernádi, I. (2005) Microiontophoresis electrode location by neurohistological marking: Comparison of four native dyes applied from current balancing electrode channels. J. Pharmacol. Toxicol. Methods 51, 147–151.
LacKamp, A., Zhang, G.-C., Mao, L.-M., Fibuch, E. E., Wang, J. Q. (2009) Loss of surface N-methyl-D-aspartate receptor proteins in mouse cortical neurones during anaesthesia induced by chloral hydrate in vivo. Br. J. Anaesth. 102, 515–522.
Martina, M., Comas, T., Mealing, G. A. R. (2013) Selective pharmacological modulation of pyramidal neurons and interneurons in the CA1 region of the rat hippocampus. Front. Pharmacol. 4, 24.
Mizuseki, K., Sirota, A., Pastalkova, E., Buzsáki, G. (2009) Theta oscillations provide temporal windows for local circuit computation in the entorhinal-hippocampal loop. Neuron 64, 267–280.
Nyíri, G., Stephenson, F., Freund, T., Somogyi, P. (2003) Large variability in synaptic n-methyl-daspartate receptor density on interneurons and a comparison with pyramidal-cell spines in the rat hippocampus. Neuroscience 119, 347–363.
Pang, K., Rose, G. M. (1987) Differential effects of norepinephrine on hippocampal complex-spike and theta-neurons. Brain Res. 425, 146–158.
Paxinos, G., Watson, C. (1997) The rat brain in stereotaxic coordinates, 3rd Ed. Academic Press, London.
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. Meth. 65, 113–136.
Ranck, J. B. (1973) Studies on single neurons in dorsal hippocampal formation and septum in unrestrained rats. I. Behavioral correlates and firing repertoires. Exp. Neurol. 41, 461–531.
Sah, P., Hestrin, S., Nicoll, R. A. (1990) Properties of excitatory postsynaptic currents recorded in vitro from rat hippocampal interneurones. J. Physiol. 430, 605–616.
Segal, M., Greenberger, V. (1992) Acidic amino acids evoke a smaller [Ca2+]i rise in GABAergic than non-GABAergic hippocampal neurons. Neurosci. Lett. 140, 243–246.
Szegedi, V., Juhász, G., Parsons, C. G., Budai, D. (2010) In vivo evidence for functional NMDA receptor blockade by memantine in rat hippocampal neurons. J. Neural. Transm. 117, 1189–1194.
Vida, I., Frotscher, M. (2000) A hippocampal interneuron associated with the mossy fiber system. Proc. Natl Acad. Sci. USA 97, 1275–1280.
West, M. J., Coleman, P. D., Flood, D. G., Troncoso, J. C. (1994) Differences in the pattern of hippocampal neuronal loss in normal ageing and Alzheimer’s disease. Lancet 344, 769–772.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
About this article
Cite this article
Bali, Z.K., Budai, D. & Hernádi, I. Separation of Electrophysiologically Distinct Neuronal Populations in the Rat Hippocampus for Neuropharmacological Testing under in Vivo Conditions. BIOLOGIA FUTURA 65, 241–251 (2014). https://doi.org/10.1556/ABiol.65.2014.3.1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1556/ABiol.65.2014.3.1