title: A horizontal slice preparation for examining the functional connectivity of dorsal column fibres in mouse spinal cord.
abstract: In spinal cord injury (SCI) research, axon regeneration across spinal lesions is most often assessed using anatomical methods. It would be extremely advantageous, however, to examine the functional synaptic connectivity of regenerating fibres, using high-resolution electrophysiological methods. We have therefore developed a mouse horizontal spinal cord slice preparation that permits detailed analysis of evoked dorsal column (DCol) synaptic inputs on spinal neurons, using whole-cell patch clamp electrophysiology. This preparation allows us to characterise postsynaptic currents and potentials in response to electrical stimulation of DCol fibres, along with the intrinsic properties of spinal neurons. In addition, we demonstrate that low magnification calcium imaging can be used effectively to survey the spread of excitation from DCol stimulation in horizontal slices. This preparation is a potentially valuable tool for SCI research where confirmation of regenerated, functional synapses across a spinal lesion is critical.
authors: School of Biomedical Sciences and Pharmacy, Faculty of Health and Hunter Medical Research Institute, The University of Newcastle, Callaghan, NSW 2308, Australia. jamie.flynn@newcastle.edu.au
authors: Jamie R
authors: JR
authors: Flynn
authors: Alan M
authors: AM
authors: Brichta
authors: Mary P
authors: MP
authors: Galea
authors: Robert J
authors: RJ
authors: Callister
authors: Brett A
authors: BA
authors: Graham
mesh_terms: D018750:6-Cyano-7-nitroquinoxaline-2,3-dione / Q000494:pharmacology; D000818:Animals; D001640:Bicuculline / Q000494:pharmacology; D001703:Biophysics; D002118:Calcium / Q000378:metabolism; D004558:Electric Stimulation / Q000295:instrumentation* / Q000379:methods; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005260:Female; D058787:GABA-A Receptor Antagonists / Q000494:pharmacology; D018684:Glycine Agents / Q000494:pharmacology; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D009412:Nerve Fibers / Q000187:drug effects / Q000502:physiology*; D009431:Neural Conduction / Q000187:drug effects / Q000502:physiology; D009475:Neurons, Afferent / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques; D011930:Reaction Time / Q000502:physiology; D013116:Spinal Cord / Q000033:anatomy & histology*; D013331:Strychnine / Q000494:pharmacology; D013376:Substantia Gelatinosa / Q000166:cytology* / Q000502:physiology; D009435:Synaptic Transmission / Q000187:drug effects / Q000502:physiology*
chemical_list: D018691:Excitatory Amino Acid Antagonists; D058787:GABA-A Receptor Antagonists; D018684:Glycine Agents; D018750:6-Cyano-7-nitroquinoxaline-2,3-dione; D013331:Strychnine; D002118:Calcium; D001640:Bicuculline