PubMed2003
Three sub-populations of sensory neurons may be distinguished based on responses to a decrease in temperature: one has a relatively low threshold for activation (cool fibers), a second has a high threshold for activation (cold nociceptors), and the third is unresponsive to a decrease in temperature. Results from several recent studies suggest that the ability to detect a decrease in temperature reflects an intrinsic property(ies) of sensory neurons and therefore may be characterized via the study of the sensory neuron cell body in vitro. However, while three unique ionic mechanisms of cold transduction have recently been identified (i.e. activation of the transient receptor potential channel M8 [TRPM8] or an epithelial Na(+) channel [ENaC] or inhibition of two pore K(+) channel [TREK-1]), the possibility that these "mechanisms" may be differentially distributed among sensory neurons in a manner consistent with predictions based on in vivo observations has not been investigated. To investigate this possibility, we have characterized the influence of cooling on isolated trigeminal ganglion (TG) neurons from adult rats in vitro with Ca(2+) microfluorimetry in combination with a series of pharmacological interventions. We report that neurons responded to a decrease in temperature from approximately 34 degrees C to approximately 12 degrees C in one of two ways: 1) with a low threshold (30.1+/-0.6 degrees C) for activation demonstrating an increase in fluorescence with a minimal decrease in bath temperature (12.3%); 2) with a high threshold for activation (21.5+/-0.6 degrees C), demonstrating an increase in fluorescence only after a substantial decrease in bath temperature (13.3%); 74.4% did not respond to a decrease in temperature with an increase [Ca(2+)](i). These responses also were distinguishable on the basis of their rate of activation and degree of desensitization in response to prolonged application of a cold stimulus: low threshold responses were associated with a rapid (tau=12.0+/-5.7 s) increase in [Ca(2+)](i) and a time constant of desensitization of 85.8+/-20.7 s while high threshold responses were associated with a slow (tau=38.1+/-8.2 s) increase in [Ca(2+)](i) and demonstrated little desensitization over 4 min of stimulation. We refer to low threshold and high threshold cold responsive TG neurons as LT(cool) and HT(cool) neurons, respectively. LT(cool) and HT(cool) neurons were distributed among two distinct subpopulations of TG neurons distinguishable on the basis of cell body size and isolectin B4 staining. Both ENaC and TRPM8 appear to contribute to cold transduction, but neither is sufficient to account for all aspects of cold transduction in either population of TG neurons. Furthermore, inhibition of Ba(2+) and/or Gd(3+) sensitive two-pore K(+) channels (i.e. TREK-1 and TRAAK) was insufficient to account for cold transduction in HT(cool) or LT(cool) neurons. Our results suggest that cold transduction in sensory neurons is a complex process involving the activation and inhibition of several different ion channels. In addition, there appear to be both similarities and differences between mechanisms underlying cold transduction in LT(cool) and HT(cool) neurons. Identification of specific mechanisms underlying cold transduction in LT(cool) and HT(cool) neurons may enable the development of novel therapeutic interventions for the treatment of pathological conditions such as cold allodynia.