Krotov, V;
Tokhtamysh, A;
Kopach, O;
Dromaretsky, A;
Sheremet, Y;
Belan, P;
Voitenko, N;
(2017)
Functional Characterization of Lamina X Neurons in ex-Vivo Spinal Cord Preparation.
Frontiers in Cellular Neuroscience
, 11
, Article 342. 10.3389/fncel.2017.00342.
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Abstract
Functional properties of lamina X neurons in the spinal cord remain unknown despite the established role of this area for somatosensory integration, visceral nociception, autonomic regulation and motoneuron output modulation. Investigations of neuronal functioning in the lamina X have been hampered by technical challenges. Here we introduce an ex-vivo spinal cord preparation with both dorsal and ventral roots still attached for functional studies of the lamina X neurons and their connectivity using an oblique LED illumination for resolved visualization of lamina X neurons in a thick tissue. With the elaborated approach, we demonstrate electrophysiological characteristics of lamina X neurons by their membrane properties, firing pattern discharge and fiber innervation (either afferent or efferent). The tissue preparation has been also probed using Ca2+ imaging with fluorescent Ca2+ dyes (membrane-impermeable or -permeable) to demonstrate the depolarization-induced changes in intracellular calcium concentration in lamina X neurons. Finally, we performed visualization of subpopulations of lamina X neurons stained by retrograde labeling with aminostilbamidine dye to identify sympathetic preganglionic and projection neurons in the lamina X. Thus, the elaborated approach provides a reliable tool for investigation of functional properties and connectivity in specific neuronal subpopulations, boosting research of lamina X of the spinal cord.
| Type: | Article |
|---|---|
| Title: | Functional Characterization of Lamina X Neurons in ex-Vivo Spinal Cord Preparation |
| Open access status: | An open access version is available from UCL Discovery |
| DOI: | 10.3389/fncel.2017.00342 |
| Publisher version: | http://dx.doi.org/10.3389/fncel.2017.00342 |
| Language: | English |
| Additional information: | © 2017 Krotov, Tokhtamysh, Kopach, Dromaretsky, Sheremet, Belan and Voitenko. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
| Keywords: | Science & Technology, Life Sciences & Biomedicine, Neurosciences, Neurosciences & Neurology, ex-vivo spinal cord preparation, lamina X neurons, electrophysiological recordings, primary afferent stimulation, retrograde labeling, Ca2+ imaging, LOCAL-CIRCUIT NEURONS, NICOTINIC ACETYLCHOLINE-RECEPTORS, SYMPATHETIC PREGANGLIONIC NEURONS, I PROJECTION NEURONS, DORSAL-HORN, SYNAPTIC-TRANSMISSION, POSTNATAL RAT, KROX-24 PROTEINS, C-FOS, EXPRESSION, Ex-vivo spinal cord preparation, lamina X neurons, electrophysiological recordings, primary afferent stimulation, retrograde labeling, Ca2+ imaging |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology > Clinical and Experimental Epilepsy UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Neuro, Physiology and Pharmacology |
| URI: | https://discovery.ucl.ac.uk/id/eprint/10038698 |
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