Abstract
The endogenous cannabinoid (eCB) system is emerging as an important component of female reproductive tract physiology. The eCBs anandamide (AEA), 2-arachidonoyl glycerol (2-AG), and N-arachidonoyl glycine (NAGly) were measured in the rat reproductive tract at five time points in the four-day estrous cycle, in acyclic retired breeders (RB), after ovariectomy (OVX), OVX + estrogen (E2), OVX + progesterone (P4), or OVX with E2+P4. eCBs were measured in the uterus, uterine adipose, ovaries, and ovarian adipose using HPLC/MS/MS. Levels of AEA, 2-AG, and NAGly were highest in the estrus phase of the estrous cycle in the uterus, whereas, only NAGly had differences in production in the ovaries across the cycle. All eCBs were lower in RB ovaries; however, the production of eCBs in the uterus of RB and OVX groups was more varied with NAGly showing the lowest levels of production in these groups. Levels of AEA in uterine fat were significantly higher or equivalent to levels in the uterus. However, levels of 2-AG and NAGly were dramatically lower in uterine fat verses the organ. Ovarian fat had significantly lower levels of all three eCBs. These data provide evidence that the hormonal milieu plays a significant and complex role in the production of eCBs in the female rat reproductive tract.
Highlights
Endocannabinoid ligands were named as the endogenous counterpart of the biologically active phytocannabinoids found in the cannabis plant
The levels measured in the ovaries of all Endocannabinoid ligands (eCB) were significantly lower in the ovarian adipose of retired breeders compared to all days of the cycle (Figures 3A-C)
Comparisons of eCB levels in ovaries and ovarian adipose showed that all eCBs were significantly lower in adipose tissue compared to the ovary with the exception of the levels of arachidonoyl ethanolamine (AEA) in retired breeders, which were equivalent (Figure 4B)
Summary
Endocannabinoid ligands (eCB) were named as the endogenous counterpart of the biologically active phytocannabinoids found in the cannabis plant. We recently showed that eCBs are produced differentially in the brain across the hormonal cycle in the rat [15], supporting the hypothesis that the production of all three eCB ligands is regulated by changes in the hormonal milieu. This hypothesis was suggested earlier when the presence of an estrogen responsive element was identified in the promoter region FAAH [16]. The eCBs AEA, 2-AG, and NAGly were measured in the female rat reproductive tract across the hormonal (estrous) cycle, in anovulatory retired breeders (RB), after ovariectomy (OVX) and with hormone (17β-estradiol and progesterone) replacement. Extractions reflect only those eCBs in tissue and not in luminal fluids
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