Abstract
Endocannabinoids are traditionally thought to have an analgesic effect. However, it has been shown that while endocannabinoids can depress nociceptive signaling, they can also enhance non-nociceptive signaling. Therefore, endocannabinoids have the potential to contribute to non-nociceptive sensitization after an injury. Using Hirudo verbana (the medicinal leech), a model of injury-induced sensitization was developed in which a reproducible piercing injury was delivered to the posterior sucker of Hirudo. Injury-induced changes in the non-nociceptive threshold of Hirudo were determined through testing with Von Frey filaments and changes in the response to nociceptive stimuli were tested by measuring the latency to withdraw to a nociceptive thermal stimulus (Hargreaves apparatus). To test the potential role of endocannabinoids in mediating injury-induced sensitization, animals were injected with tetrahydrolipstatin (THL), which inhibits synthesis of the endocannabinoid transmitter 2-arachidonoylglycerol (2-AG). Following injury, a significant decrease in the non-nociceptive response threshold (consistent with non-nociceptive sensitization) and a significant decrease in the response latency to nociceptive stimulation (consistent with nociceptive sensitization) were observed. In animals injected with THL, a decrease in non-nociceptive sensitization in injured animals was observed, but no effect on nociceptive sensitization was observed.
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Abbreviations
- ANOVA:
-
Analysis of variance
- 2-AG:
-
2-Arachidonoylglycerol
- CB:
-
Cannabinoid
- CNS:
-
Central nervous system
- DMSO:
-
Dimethyl sulfoxide
- LPS:
-
Lipopolysaccharide
- N cell:
-
Nociceptive cell
- P cell:
-
Pressure cell
- SUDO:
-
Simplified up and down
- THL:
-
Tetrahydrolipstatin
- T cell:
-
Touch cell
- TRPV:
-
Transient receptor potential vanilloid
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Funding
This work was supported by the University of South Dakota’s Summer Program for Undergraduate Research in Addiction (SPURA), which is funded by the National Institute on Drug Abuse (NIDA) grant R25-DA033674. It was also supported by a National Institute of Neural Disorders and Stroke (NINDS), NIH R01-NS092716 grant.
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MMJ and BDB designed the experiments. MMJ carried out the experiments. MMJ and BDB carried out data analysis. MMJ wrote the first draft of the manuscript in fulfillment of her honors thesis, with revisions made by BDB. BDB obtained funding.
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359_2021_1540_MOESM1_ESM.tif
Supplementary file1 Supplemental Figure 1: Effects of multiple concentrations of THL on response threshold and response latency. (b) 25 and 50 µM THL had no effect on response latency when compared to DMSO-injected animals. 100 µM THL also had no statistically significant effect, although there did appear to be a decrease in response threshold on days 4 and 5. (b) For the initial 4 hour post injury measurement of response latency, 100 µM THL significant reduced response latency compared to the other THL concentrations and the DMSO injected group (*). Data is represented as the average ± the standard error (TIF 6753 kb)
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Jorgensen, M.M., Burrell, B.D. Approaches to studying injury-induced sensitization and the potential role of an endocannabinoid transmitter. J Comp Physiol A 208, 313–323 (2022). https://doi.org/10.1007/s00359-021-01540-z
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DOI: https://doi.org/10.1007/s00359-021-01540-z