Effect of a Ketogenic Diet on Acute Inflammatory Pain Induced by Complete Freund’s Adjuvant (CFA) in Male Mice

Isabella C. Sturdevant ’20, David N. Ruskin, PhD, Susan A. Masino, PhD

3 thoughts on “Effect of a Ketogenic Diet on Acute Inflammatory Pain Induced by Complete Freund’s Adjuvant (CFA) in Male Mice

  1. Hi Izzy,

    Nice job. Here’s a fun question: Using pharmacology, what could you do (assuming all options for all kinds of experiments are on the table) to assess whether there is indeed no adenosine A1R signaling in your KO mice? Because one way to interpret your findings is that there is indeed functional A1R signaling in the KO mice, thereby explaining why they look similar to WT animals whether they are on the control or ketogenic diet. So imagine you want to challenge your assumption that these animals are indeed successful KOs. I’ll start you in the direction I’m thinking about by posing a potential experiment: Imagine that you cultured some neurons from your KO and WT mice. You grow these cells up in a dish, and then you start applying some pharmacological agents to them. Your dependent measure is going to be a cAMP assay. How would you expect cAMP levels in your KO vs. WT cultures to respond to application of CPA vs. DPCPX, assuming that your KO animals don’t have functional A1R signaling?

  2. Thank you for your comment and interesting proposal for a possible experiment. Activation of A1 receptors inhibits adenylyl cyclase activity and thus decreases cAMP levels. Since CPA is an A1 receptor agonist, I would expect it to increase the inhibitory effects of the A1 receptor on cAMP levels in WT mice, resulting in less cAMP. DPCPX is an A1 receptor antagonist, so it should reduce A1 receptor activity, leading to less inhibition of adenylyl cyclase and an increase in cAMP levels in WT mice. Assuming the KO mice had insufficient adenosine signaling due to reduced A1 receptors, I would expect them to have higher cAMP levels than the WT mice. CPA and DPCPX act on the A1 receptors, so the knockout mice would lack the ability to respond sufficiently to these applications. If the KO cultures are not true A1 receptor knockouts, then I would expect them to exhibit similar results to the WT mice and have normal adenosine signaling in response to CPA and DPCPX applications.

    • Hi Izzy,

      Excellent answer.

      A somewhat different approach to begin to assess whether the KO is really a KO is just to see if you detect any A1 receptor expression (using a western blot or IHC) in the KO mice. If both types of experiments were combined, and if they played out in the way you would expect for KO vs. WT mice, then you could have more confidence that your KOs do not have functional A1Rs.

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