Whole-cell patch-clamp recordings were made from kidney-related neurons in the intermediolateral cell column (IML) in horizontal slices of thoracolumbar spinal cord from adult rats. Kidney-related neurons were identified in vitro subsequent to inoculation of the kidney with a fluorescent, retrograde, transynaptic pseudorabies viral label (i.e., PRV-152). Kidney-related neurons detected in the IML expressed choline acetyltransferase, characteristic of spinal preganglionic motor neurons. Their mean resting potential was -51±4mV and input resistance was 448±39MΩ. Both spontaneous inhibitory and excitatory post-synaptic currents (i.e., sIPSCs and sEPSCs) were observed in all neurons. The mean frequency for sEPSCs (3.1±1Hz) was approximately 2.5 times that for sIPSCs (1.4±0.3Hz). Application of the glycine and GABAA receptor-linked Cl- channel blocker, picrotoxin (100μM) blocked sIPSCs, while the ionotropic glutamate receptor antagonist, kynurenic acid (1mM) blocked all sEPSCs, indicating they were mediated by GABA/glycine and glutamate receptors, respectively. Thus, using PRV-152 labeling allowed whole-cell patch-clamp recording of neurons in the adult spinal cord, which were kidney-related. Excitatory glutamatergic input dominated synaptic responses in these cells, the membrane characteristics of which resembled those of immature IML neurons. Combined PRV-152 pre-labeling and whole-cell patch-clamp recordings may allow more effective analysis of synaptic plasticity seen in adult models of injury or chronic disease.
|Number of pages||5|
|State||Published - May 2010|
Bibliographical noteFunding Information:
We thank Dr. Lynn Enquist (Princeton University) for the PRV-152. Funded by PVA grant # 2561 (H. Duale), NIHLBI , 1R21HL091293 (A.V. Derbenev), NINDS , R01 NS049901 (A.G. Rabchevsky), NIDDK , R01 DK056132 (B.N. Smith), and NINDS , P30 NS051220 .
- Intermediolateral cell column
- Preganglionic neuron
- Pseudorabies virus
- Whole-cell patch-clamp
ASJC Scopus subject areas
- Neuroscience (all)