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. 2017 Aug 1;158(11):2222–2232. doi: 10.1097/j.pain.0000000000001027

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Copyright © 2017 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of The International Association for the Study of Pain.

This is an open access article distributed under the Creative Commons Attribution License 4.0 (CCBY), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 1. — Changes in the expression of endocannabinoids (eCBs) and eCB-synthesising enzymes in the brainstem nuclei of the rat and human midbrain during postnatal development. (A and B) Mass spectrometry analysis of anandamide levels in the periaqueductal grey (PAG) and rostroventral medulla (RVM). Expression of anandamide increased during early postnatal development and reached maturity by postnatal day (P)21. (P < 0.05 in the PAG and RVM; 1-way analysis of variance [ANOVA].) (C and D) Mass spectrometry analysis of 2-arachidonoylglycerol (2-AG) levels in the PAG and RVM. No changes in the expression of 2-AG were observed in the PAG (C). Expression of 2-AG increased during early postnatal development and reached maturity by P21 in the RVM (D; P < 0.01; 1-way ANOVA). (E) Taqman real-time (RT) PCR analysis of the eCB-synthesising enzymes NAPE-PLD and DAGLα in the ventral PAG. Expression of NAPE-PLD mRNA increased during early postnatal development and reached maturity by P21 (P < 0.05; 1-way ANOVA). (F) Taqman RT-PCR analysis of NAPE-PLD and DAGLα in the RVM. No significant changes were observed. (G) Expression of NAPE-PLD mRNA was highest in the human infant midbrain compared with all other age groups tested (P < 0.01; 1-way ANOVA). Nine to 11 animals per age group used for mass spectrometry experiments. Three to 4 animals per age group for Taqman RT-PCR and immunohistochemical experiments. Four to 8 human midbrain tissue per age group used for TaqMan RT-PCR. Data shown here represent mean ± SEM. * and ** = P < 0.05 and P < 0.01, respectively, between age comparisons, 1-way ANOVA with Bonferroni multiple comparisons.