- TWO WAY ANOVA FOR MICE BEHAVIOR EXPIREMENT GRAPHPAD PRISM 6 SKIN
- TWO WAY ANOVA FOR MICE BEHAVIOR EXPIREMENT GRAPHPAD PRISM 6 FREE
The protocols for induction of either histaminergic or non-histaminergic itch were performed according to our previous work. When compounds were delivered subcutaneously (s.c.), a standard low volume of 20 μl was injected.
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The pruritogens chloroquine diphosphate (Sigma-Aldrich) and histamine (Sigma-Aldrich), as well as ethosuximide (Sigma-Aldrich), were dissolved in PBS solution.
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All experiments were carried out between 9 am and 3 pm. The housing room was kept at a temperature of 23 ± 1 ☌ and under a 12-h light/dark cycle (lights on at 7 am).
TWO WAY ANOVA FOR MICE BEHAVIOR EXPIREMENT GRAPHPAD PRISM 6 FREE
Mice were housed at five per cage (30 × 20 × 15 cm) with free access to food and water. Here, we report that ethosuximide potently inhibits scratching responses associated with subcutaneous delivery of histamine or chloroquine in both male and female mice.Įxperiments were carried out on adult (7–11 weeks old) male or female C57BL/6J mice purchased from Jackson laboratories following approval by the institutional Animal Care Committee. Therefore, as a first step, we wanted to obtain preclinical evidence of possible anti pruritonergic effects of locally and systemically delivered ethosuximide in mouse models of acute itch. Given that ethosuximide is generally well tolerated and thought to have few drug interactions, we surmised that this molecule could perhaps be used to treat itch. These data collectively suggested that Cav3.2 channels mediate an important role in itch responses in the skin, presumably by controlling action potential initiation in peripheral nerve endings.Įthosuximide (Zarontin) is an archetypal T-type calcium channel therapeutic that is used clinically to treat absence epilepsy, particularly in children. Furthermore, subcutaneous injection of an experimental small molecule inhibitor of Cav3.2 channels significantly attenuated scratching responses to injection of histamine or chloroquine in wild type mice. We found that Cav3.2 null mice were almost completely resistant to the actions of both histamine and chloroquine. We have recently explored the putative role of Cav3.2 channels in the transmission of histamine and chloroquine-dependent itch. The mammalian genome encodes three distinct T-type calcium channel isoforms, Cav3.1, Cav3.2 and Cav3.3, with Cav3.2 having been prominently associated with peripheral pain transmission. Based on their biophysical properties, T-type calcium channels are uniquely suited towards regulating neuronal excitability. Distinct but perhaps partially overlapping populations of neurons are involved in transmitting pain and itch related information, however, there is accumulating evidence that both pain and itch transmitting neurons express T-type calcium channels. Similar to painful stimuli, pruritogens activate peripheral sensory neurons that then communicate in the spinal cord with second order neurons that project to brain regions where itch is perceived as an unpleasant sensation.
TWO WAY ANOVA FOR MICE BEHAVIOR EXPIREMENT GRAPHPAD PRISM 6 SKIN
Pruritus significantly influences quality of life measures, such as mood, the ability to concentrate, and sleep, and impacts quality of life more than many other skin conditions.
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Our findings also suggest that ethosuximide could be explored further as a possible therapeutic for the treatment of itch.
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Overall, our results are consistent with an important role of Cav3.2 T-type calcium channels in modulating histamine-dependent and histamine-independent itch transmission in the primary sensory pathway. When co-delivered subcutaneously together with either pruritogenic agent ethosuximide was also effective in inhibiting scratching responses in both male and female animals. When delivered intraperitoneally ethosuximide significantly reduced scratching behavior of mice of both sexes in response to subcutaneous injection of either histamine or chloroquine. Here, we evaluated the effect of the clinically used anti-seizure medication ethosuximide, a well known inhibitor of T-type calcium channels, on male and female mice subjected to histaminergic- and non-histaminergic itch. We have recently reported that the Cav3.2 T-type calcium channel which is well known for its key role in pain signalling, also mediates a critical function in the transmission of itch/pruritus.