anisomycin and biocytin

Deep dorsal horn neurons are involved in the processing of nociceptive information in the spinal cord. Previous studies revealed a role of the intrinsic bioelectrical properties (plateau potentials) of deep dorsal horn neuron in neuronal hyperexcitability, indicating their function in pain sensitization. These properties were considered to rely on L-type calcium currents. Two different isotypes of L-type calcium channel alpha 1 subunit have been cloned (Ca(V)1.2 and Ca(V)1.3). Both are known to be expressed in the spinal cord. However, no data were available on their subcellular localization. Moreover, possible changes in Ca(V)1.2 and Ca(V)1.3 expression had never been investigated in nerve injury models. Our study provides evidence for a differential expression of Ca(V)1.2 and Ca(V)1.3 subunits in the somato-dendritic compartment of deep dorsal horn neurons. Ca(V)1.2 immunoreactivity is restricted to the soma and proximal dendrites whereas Ca(V)1.3 immunoreactivity is found in the whole somato-dendritic compartment, up to distal dendritic segments. Moreover, these specific immunoreactive patterns are also found in electrophysiologically identified deep dorsal horn neurons expressing plateau potentials. After nerve injury, namely total axotomy or partial nerve ligation, Ca(V)1.2 and Ca(V)1.3 expression undergo differential changes, showing up- and down-regulation, respectively, both at the protein and at the mRNA levels. Taken together, our data support the role of L-type calcium channels in the control of intrinsic biolectrical regenerative properties. Furthermore, Ca(V)1.2 and Ca(V)1.3 subunits may have distinct and specific roles in sensory processing in the dorsal horn of the spinal cord, the former being most likely involved in long-term changes after nerve injury.

Topics: Animals; Anisomycin; Antibodies; Axotomy; Blotting, Western; Calcium Channels; Calcium Channels, L-Type; Cycloheximide; Immunohistochemistry; In Situ Hybridization; In Vitro Techniques; Ligation; Lysine; Male; Membrane Potentials; Microtubule-Associated Proteins; Posterior Horn Cells; Protein Synthesis Inhibitors; Rats; Rats, Wistar; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Sciatic Neuropathy; Spinal Cord; Time Factors