Skin TRPA1 ion channel participates in thermoregulatory response to cold. Comparison with the effect of TRPM8.

There was no clear evidence of the TRPA1 ion channel involvement in the formation of thermoregulatory responses. The present results convincingly show that the skin TRPA1 ion channel activation has significant influence on the formation of thermoregulatory responses of the body to cooling; it is especially strongly manifested for the metabolic component. At the TRPA1 activation by its agonist AITC (0.04%), an enhancement in thermoregulatory responses is observed: the temperature thresholds for the first phase and the second one of the metabolic response decrease, the values of all components of the metabolic response considerably increase: the increment of oxygen consumption in the first phase increases from 1.8 ± 0.24 in the control to 2.9 ± 0.35 ml/min*kg under AITC, P = 0.04; the increment of oxygen consumption in the second phase increases from 6.2 ± 2.06 to 17.4 ± 1.20 ml/min*kg, P = 0.002, as well as shivering rises from 7.8 ± 1.79 to 15.4 ± 1.87 mV, P = 0.011. In consideration of our previous results on the influence of TRPM8 ion channel activation on thermoregulatory responses (Kozyreva et al., J. Therm.Biol., 2010) it is obvious that the TRPM8 and TRPA1 ion channels have a pronounced, but unequal effects on the values of different phases of the metabolic response to cold. The TRPM8 activation manifests itself in an increase of value only the urgent first phase, this phase is associated with carbohydrate metabolism. As the recent results have shown the influence of the TRPA1 activation is realized predominantly in the clearly marked increase in the second phase of the metabolic response associated with lipid metabolism, as well as in evident shivering gain. The ability to predominantly control different parameters of thermoregulatory responses to cold may indicate the importance of both the TRPM8 and the TRPA1 ion channels in the processes of maintaining temperature homeostasis. The obtained data testify to the joint sequential operation of these thermosensitive ion channels.

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.

Participation of Purinergic P2X Receptors in the Thermoregulatory Response to Cooling.

We studied the role of purinergic P2X receptors in the body response to cooling. In experiments on rats, P2X receptor antagonist PPADS was administered in different modes, which resulted in changes of different characteristics of the thermoregulatory response to cold. Iontophoresis of P2X antagonist into the skin decreased the thermal thresholds of all thermoregulatory responses to cooling, which can attest to a modulating effect of P2X receptors on peripheral thermosensitive afferents. Intraperitoneal administration of P2X antagonist suppressed thermoregulatory activity of skeletal muscles (shivering) developing during cooling without changing the thresholds of thermoregulatory responses. The findings suggest that ATP and P2X receptors play an important role in the formation of the response to cooling.

[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.

The role of alpha1- and beta-adrenoceptors in initiation and development of thermoregulatory reactions during fast and slow cooling.

Ionophoretic application of alpha1- and beta-adrenoceptor blockers into the skin produces no effect on the parameters of thermal homeostasis under thermoneutral conditions. alpha1-Adrenoblocker verapamil inhibits cold shivering during fast and slow cold exposure; it elevates the temperature threshold and moderates the vasoconstrictor response during rapid cooling. These changes are accompanied by a compensatory decrease in the threshold and stimulation of non-shivering thermogenesis. Application of non-selective beta-blocker propranolol had no effect on the temperature thresholds of the thermoregulatory reactions, but augmented the maximum amplitude of shivering during both cooling protocols, thereby compensating the decrease in non-shivering thermogenesis. In the whole organism, block of one type adrenoceptors during cold exposure is accompanied by activation of the compensatory mechanisms mediated by adrenoceptors of the other type.

Effect of substance P on thermoregulation parameters during different cooling modes.

The modulatory effect of ionophoretic application of substance P to the skin on the formation of the cold-triggered thermal protection reactions depends on the rate of cooling. During rapid cooling substance P enhances heat production, while during slow cooling it promotes constriction of skin blood vessels aimed at reduction of heat emission.

Effects of slow and rapid cooling on catecholamine concentration in arterial plasma and the skin.

Norepinephrine (NE) and epinephrine (Epi) concentrations in arterial plasma and in skin tissue were measured chromatographically before and after external cooling. Urethan-anesthetized rats were cooled either slowly (0.004-0.006 degrees C/s) or rapidly (0.03- 0.05 degrees C/s). Blood samples were drawn three times from each animal: 1) before cooling and at a rectal temperature decreased 2) by 0.5 degrees C and 3) by 3-4 degrees C. Skin samples were taken from controls and from rapidly or slowly cooled rats at a rectal temperature lowered by 0.5 degrees C. The resting mean values were 36.7 +/- 0.3 degrees C for rectal temperature, 0.62 +/- 0.079 and 1. 09 +/- 0.203 ng/ml for plasma NE and Epi, and 85.6 +/- 4.1 and 137.6 +/- 34.3 ng/g for skin NE and Epi. A decrease in rectal temperature by 0.5 degrees C at rapid cooling produced a 2.6-fold increase of NE and a 2.8-fold increase of Epi in plasma. Concomitantly, there was a significant decrease in skin NE concentration by 28% and Epi by 86%. At a rectal temperature decreased by 0.5 degrees C after slow cooling, plasma catecholamines did not change; at unaltered skin NE concentration, there was a reduction in skin Epi concentration (60%). When rectal temperature was lowered by 3-4 degrees C, the increase in plasma NE was virtually the same at both cooling rates and only plasma Epi increased more after deep rapid cooling than slow cooling. Thus the sympathoadrenal system may be differently activated depending on cooling rate. Rapid cooling, when the dynamic activity of the skin cold receptors is involved in the cold response, may provide conditions for an earlier activation of the sympathoadrenal system. This may evidence the functional significance of the dynamic activity of the skin cold receptors in the formation of the cold defense responses.

[Characteristics of responses of the sympatho-adrenal system to various types of cooling]. / Osobennosti reaktsii simpatoadrenalovoi sistemy krys pri raznykh tipakh okhlazhdeniia.

Fast cooling involving the dynamic activity of the skin cold receptors seems to establish a condition for changes in catecholamine concentration at a lesser decrease of body temperature as compared with slow cooling.