Thermoregulatory Response to Cold at Various Levels of Activation of Peripheral TRPA1 Ion Channel.

The effect of TRPA1-ion channel on thermoregulatory responses depending on the level of its activity was studied in Wistar rats. To activate the TRPA1 ion channel localized in the skin, its agonist allyl isothiocyanate (AITC) was used in different concentrations (0.04, 0.4, 1, and 2.5%). Low concentration of AITC (0.04%) enhanced and high concentrations (1 and 2.5%), on the contrary, inhibited cold-defense responses (decreased their magnitude and led to their later initiation due to an increase in temperature thresholds). With an increase in TRPA1 activation, the increase in temperature thresholds (afferent link) was ahead of the decrease in the magnitude of responses (efferent link), which can attest to different sensitivity of these processes to TRPA1 activation.

Effects of activation of skin ion channels TRPM8, TRPV1, and TRPA1 on the immune response. Comparison with effects of cold and heat exposure.

The effects of pharmacological stimulation of skin ion channels TRPA1, TRPM8, TRPV1 on the immune response are presented. These effects are compared with the effects of different types of temperature exposures - skin cooling, deep cooling, and deep heating. This analysis allows us to clear the differences in the influence on the immune response of thermosensitive ion channels localized in the skin; (2) whether the changes in the immune response under temperature exposures are due to these thermosensitive ion channels. Experiments were performed on Wistar rats. For stimulation of TRPM8 ion channel, an application to the skin of 1% menthol was used, for TRPA1 - 0.04% allylisotiocianate, and for TRPV1 - capsaicin in a concentration of 0.001.The antigen binding in the spleen was two-times stimulated by activation of the cold-sensitive ion channel TRPM8 and much weaker by activation of warm-sensitive TRPV1 (by 15%), and another cold-sensitive ion channel TRPA1 (by 40%). Only the stimulation of TRPA1 significantly (by 140%) increased antibody formation in the spleen, while TRPM8 had practically no effect on this process, and activation of TRPV1 significantly (by 60%) inhibited antibody formation. Stimulation of the TRPM8 ion channel significantly (by 60%) reduced the level of IgG in the blood, which is believed to control of infectious diseases.The obtained results show that pharmacological activation of the skin TRPA1, TRPM8, TRPV1 ion channels can differently affect the immune system. At the epicenter of changes there were the antigen binding and antibody formation in the spleen, as well as the level of IgG in the blood. Exactly stimulation of the TRPM8 ion channel determines the changes in the immune response when only the skin is cooling, while at deep body heating, the changes in the immune response are mostly determined by the activation of the skin TRPV1 ion channel.

Effect of Activation of Peripheral Ion Channel TRPM8 on Gene Expression of Thermosensitive TRP Ion Channels in the Hypothalamus. Comparison with the Effect of Cooling.

Experiments on rats showed that activation of the peripheral ion channel TRPM8 with menthol and rapid cooling (decrease in core temperature by 3°C) led to 1.5-fold activation of the expression of TRPV3 ion channel gene in the posterior hypothalamus, but had no effect on the expression of this gene in the anterior hypothalamus. Neither stimulation of peripheral TRPМ8, nor acute cooling affected the expression of genes for other thermosensitive ion channels (TRPV1, TRPV2, TRPV4, TRPA1, and TRPМ8) in the hypothalamus. Enhanced expression of Trpv3 gene can indicate increased sensitivity of hypothalamic neurons in the range of TRPV3 ion channel functioning (31-39oC). The relationship between the changes in Trpv3 gene expression and the shift of thermoregulatory reaction thresholds is discussed. Our findings attest to the presence of a functional relationship between TRP ion channels of the peripheral nervous system and TRP channels in the central structures of the brain.

Calcium ions facilitate body heat emission response to warming.

Involvement of various areas of the body surface in heat emission response to warming is characterized by a certain succession. The first response preceding the deep body temperature rise is dilation of ear skin vessels. Then, an increase in deep body temperature is counterbalanced by vascular reaction in the tail region, which plays the leading role in up-regulation of heat emission. Calcium ions accelerate the vascular response to warming in both regions, although they produce no effect on the maximum level of heat emission. Our findings confirm the involvement of Ca(2+)-dependent mechanisms in activation of the processes aimed at stabilization of body temperature in warm-blooded animals. The role of heat-sensitive TRPV1 ion channels determining modality of the temperature signal and direction of effector reactions is discussed.

Effects of pharmacological activation of TRPM8 ion channels on the thermoregulatory responses during warming.

Preliminary non-thermal activation of cold-sensitive TRPM8 ion channel facilitates initiation of the heat-defense responses in homoiothermal animals by decreasing temperature threshold of the vasodilatory response. TRPM8 activation leads to earlier heat-initiated increase in oxygen consumption, but reduces its magnitude. Warming inhibits the lipolytic effect of menthol activation of TRPM8 observed under thermoneutral conditions. Thus, modulation of the skin temperature afferent signal by ion channel agonist TRPM8 changes not only cold-defense, but also heat-defense responses of the body.

[Effect of transfer of fragment of chromosome 13, containing gene i16st on parameters of mouse temperature homeostasis].

Mechanisms of maintenance of temperature homeostasis in warm and under effect of cold were studied in mice of AKR strain and of its coherent strain AKR.CBA-D13Mit76 with the changed gene i16st encoding the gp 130 receptor, via which IL-6 performs its action. Under thermoneutral conditions and under action of cold, there were recorded temperature parameters, total oxygen consumption, carbon dioxide release, respiratory coefficient, and electrical muscle activity. Animals of the studied strains demonstrated different reactions to equal cold effect. At cooling, all mice of the AKR strain entered the state of hypothermia by decreasing metabolism. Mice of the AKR.CBA-D13Mit76 line showed 2 different types of reaction: 39 % of the animals of this strain reacted like mice of the AKR strain, but the majority (61 %) resisted actively to the cold action, which was manifested as a marked increase of metabolism. Taking into account the gene penetrance, this can indicate effect of the gene i16st on choice of the active ("regulated") or passive ("dependent") way of the organism reacting to temperature actions.

[Effect of TRPM8 ion channel activation on thermoregulatory response to cooling].

In rats, the effect of activation of the cold- and menthol-sensitive TRPM8 ion channel on different thermoregulatory parameters: total oxygen consumption, carbon dioxide release, respiration coefficient, constriction response of skin blood vessels, muscle activity, was studied. Activation of TRPM8 with menthol even in thermoneutral conditions produces an increase in oxygen consumption and a decrease in respiratory coefficient, which may suggest enhanced non-shivering thermogenesis and lipolysis. Rapid cooling against the background of TRPM8 activation is characterized by a decrease in the temperature thresholds of all thermoregulatory responses without associated changes in sequences of their initiation as well as in enhancement of metabolic component of emergency thermogenesis which leads to improved maintenance of core temperature in conditions when external cold acts on the organism. The obtained data on the effect of TRPM8 activation on metabolic parameters in thermoneutral conditions and under cooling suggest acontinuous involvement of this receptor in regulation of total metabolism and, possibly, in determination of the type of organism's metabolism as well as in determination of organism's response to external cooling.

Modulating effects of calcium on immune response of homoiothermal animal under thermoneutral conditions and during deep cooling.

Preliminary ionophoretic administration of Ca(2+) ions into the skin prevents the inhibitory effects of deep cooling on some processes characterizing the immune response. Differently directed changes in some immune response parameters induced by exogenous calcium and deep cooling suggest that competitive interactions between calcium-dependent processes can serve as mechanisms of functional changes in various physiological systems during the formation of the systemic reaction of a homoiothermal organism to cold.

Effect of rapid slight cooling of the skin in various phases of immunogenesis on the immune response.

Slight cooling had no effect on heat emission and heat production, but modulated the immune response to antigen in animals. Changes in the immune response upon rapid slight cooling of the skin (by 1.5 degrees C) depended on the phase of immunogenesis corresponding to cold exposure. When cooling was performed immediately after immunization, antibody production increased in the spleen and blood, while antigen binding in the spleen remained unchanged. Cold exposure on day 5 after antigen treatment as well as immunization at the peak of cooling did not modulate antibody production, but increased antigen binding in the spleen. Our findings attest to an important role of the temperature factor in the formation of the immune response, which should be taken into account during vaccination.