Effects of variations in hippocampal slice preparation protocol on the electrophysiological stability, epileptogenicity and graded hypoxia responses of CA1 neurons.
Evoked, extracellularly recorded field potentials and whole-cell current-clamp recordings were used to assay the effects of variations in dissection method and incubation temperature on the electrophysiology of CA1 neurons in hippocampal slices. Slices were cut with either a vibratome or a tissue chopper, and incubated at 28-30 degrees C, room temperature (19-21 degrees C), or in cool solution (13-15 degrees C) which was allowed to passively warm to room temperature while the slices were incubating ('cold-shock', CS). Although no effects of dissection method were observed, it was found that incubation temperature had profound effects on synaptically, but not non-synaptically evoked field potentials. Cold-shocked slices, cut with either a vibratome or a tissue chopper, exhibited epileptiform and spontaneously potentiating orthodromic field potentials. Slices incubated at warmer temperatures demonstrated responses that were larger in amplitude, more stable and much less epileptiform. In response to orthodromic stimulation, CS neurons fired more action potentials than did neurons in slices incubated at room temperature. Further, CS neurons generated smaller inhibitory postsynaptic potentials. Field potential changes resulting from graded hypoxia were not significantly affected by the temperature at which the slices were incubated. These data suggest that the electrophysiology of the hippocampal slice can be altered by the methods used to prepare the tissue. This finding may account for some of the discrepancies that exist between laboratories, and serves to underscore the importance of accurately reporting detailed protocols. Further, CS hippocampal tissue may represent a novel in vitro model of epileptiform activity.
Watson
P L
PL
Department of Physiology, University of Toronto, Ontario, Canada. patricia.watson@uchsc.edu
Weiner
J L
JL
Carlen
P L
PL
D000818:Animals; D015687:Cell Hypoxia / Q000502:physiology*; D002555:Cerebrospinal Fluid / Q000502:physiology; D004558:Electric Stimulation; D004594:Electrophysiology; D004827:Epilepsy / Q000503:physiopathology*; D005071:Evoked Potentials / Q000502:physiology; D005110:Extracellular Space / Q000502:physiology; D006624:Hippocampus / Q000166:cytology / Q000502:physiology*; D066298:In Vitro Techniques; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D009474:Neurons / Q000502:physiology*; D018408:Patch-Clamp Techniques; D051381:Rats; D017208:Rats, Wistar; D013696:Temperature