Enzymatic Synthesis of Anandamide, an Endogenous Cannabinoid Receptor Ligand, through N-Acylphosphatidylethanolamine Pathway in Testis: Involvement of Ca2+-Dependent Transacylase and Phosphodiesterase Activities

Sugiura, Takayuki; Kondo, Sachiko; Sukagawa, Akihiro; Tonegawa, Takashi; Nakane, Shinji; Yamashita, Atsushi; Waku, Keizo

doi:10.1006/bbrc.1996.0020

Cited by 193 publications

(105 citation statements)

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“…In other organs, such as rat skin and spleen, and human heart and spleen, anandamide levels were found to be below 20 pmol͞g tissue (26). Rat testes were recently shown to contain both NAEs and NAPEs with anandamide amounting to 6 pmol͞g tissue (28). Mouse uterus therefore contains by far the highest levels of anandamide detected in any mammalian tissue.…”

Section: Resultsmentioning

confidence: 98%

Changes in anandamide levels in mouse uterus are associated with uterine receptivity for embryo implantation

Schmid

Paria

Krebsbach

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

237

166

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Anandamide (N-arachidonoylethanolamine) is an endogenous ligand for both the brain-type (CB1-R) and spleen-type (CB2-R) cannabinoid receptors. This investigation demonstrates that the periimplantation mouse uterus contains the highest levels of anandamide (142-1345 pmol͞ mol lipid P; 1-7 g͞g wet weight) yet discovered in a mammalian tissue. The levels f luctuate with the state of pregnancy; down-regulation of anandamide levels is associated with uterine receptivity, while up-regulation is correlated with uterine refractoriness to embryo implantation. Anandamide levels are highest during the nonreceptive phase in the pseudopregnant uterus and in the interimplantation sites, and lowest at the site of embryo implantation. The lower levels of uterine anandamide at the implantation sites may be a mechanism by which implanting embryos protect themselves from the detrimental effects of this endogenous ligand. We also observed a reduced rate of zona-hatching of blastocysts in vitro in the presence of anandamide, and inhibition of implantation by systemic administration of a synthetic cannabinoid agonist CP 55,940. These adverse effects were reversed by SR141716A, a specific CB1-R antagonist. Taken together, the results suggest that an aberrant synthesis of anandamide and͞or expression of the cannabinoid receptors in the uterus͞embryo may account for early pregnancy failure or female infertility.

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Section: Resultsmentioning

confidence: 98%

Changes in anandamide levels in mouse uterus are associated with uterine receptivity for embryo implantation

Schmid

Paria

Krebsbach

et al. 1997

Proc. Natl. Acad. Sci. U.S.A.

237

166

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“…So far, NAT has been studied with crude or partially purified enzyme preparations from the brains (16,21,24) and testes (17) of rats and the brains (23) and hearts (20,22) of dogs. The enzyme catalyzes the transfer of an O-acyl chain from the sn-1 position of various glycerophospholipids, including PC, PE, lyso-PC, and cardiolipin (16 -24), to the amino group of PE and lyso-PE (16 -24) and exhibits a broad specificity regarding the transferred fatty acyl species (24).…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies showed the presence of Ca 2ϩ -dependent, membrane-bound NAT in rat testis (17,24,30). Hence, we were interested in seeing how Ca 2ϩ -dependent NAT and RLP-1 share the transacylation activity in rat testis (Fig.…”

Section: And 1-palmitoyl-2-[mentioning

confidence: 99%

Discovery and Characterization of a Ca2+-independent Phosphatidylethanolamine N-Acyltransferase Generating the Anandamide Precursor and Its Congeners

Jin

Okamoto

Morishita

et al. 2007

Journal of Biological Chemistry

132

105

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N-Acylphosphatidylethanolamines (NAPEs) are precursors of bioactive N-acylethanolamines, including the endocannabinoid anandamide. In animal tissues, NAPE is formed by transfer of a fatty acyl chain at the sn-1 position of glycerophospholipids to the amino group of phosphatidylethanolamine (PE), and this reaction is believed to be the principal rate-limiting step in N-acylethanolamine synthesis. However, the Ca 2؉ -dependent, membrane-associated N-acyltransferase (NAT) responsible for this reaction has not yet been cloned. In this study, on the basis of the functional similarity of NAT to lecithin-retinol acyltransferase (LRAT), we examined a possible PE N-acylation activity in two rat LRAT homologous proteins. Upon overexpression in COS-7 cells, one protein, named rat LRAT-like protein (RLP)-1, catalyzed transfer of a radioactive acyl group from phosphatidylcholine (PC) to PE, resulting in the formation of radioactive NAPE. However, the RLP-1 activity was detected mainly in the cytosolic rather than membrane fraction and was little stimulated by Ca 2؉ . Moreover, RLP-1 did not show selectivity with respect to the sn-1 and sn-2 positions of PC as an acyl donor and therefore could generate N-arachidonoyl-PE (anandamide precursor) from 2-arachidonoyl-PC and PE. In contrast, under the same assay conditions, partially purified NAT from rat brain was highly Ca 2؉ -dependent, membrane-associated, and specific for the sn-1-acyl group of PC. RLP-1 mRNA was expressed predominantly in testis among various rat tissues, and the testis cytosol exhibited an RLP-1-like activity. These results reveal that RLP-1 can function as a PE N-acyltransferase, catalytically distinguishable from the known Ca 2؉ -dependent NAT.

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“…By hydrolysis of the sn ‐1 and sn ‐2 bonds of NAPE the resulting glycerophospho‐NAE species can be determined either by LC–MS 9, 21, GC–MS 22, 23 or HPLC coupled to fluorescence detection 24. However, this approach groups all species with the same N ‐acyl chain together without the possibility for individual detection of intact NAPE molecular species.…”

Section: Introductionmentioning

confidence: 99%

Quantitative analysis of N‐acylphosphatidylethanolamine molecular species in rat brain using solid‐phase extraction combined with reversed‐phase chromatography and tandem mass spectrometry

Triebl

Weissengruber

Trötzmüller

et al. 2016

J of Separation Science

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A novel method for the sensitive and selective identification and quantification of N‐acylphosphatidylethanolamine molecular species was developed. Samples were prepared using a combination of liquid–liquid and solid‐phase extraction, and intact N‐acylphosphatidylethanolamine species were determined by reversed‐phase high‐performance liquid chromatography coupled to positive electrospray tandem mass spectrometry. As a result of their biological functions as precursors for N‐acylethanolamines and as signaling molecules, tissue concentrations of N‐acylphosphatidylethanolamines are very low, and their analysis is additionally hindered by the vast excess of other sample components. Our sample preparation methods are able to selectively separate the analytes of interest from any expected biological interferences. Finally, the highest selectivity is achieved by coupling chromatographic separation and two N‐acyl chain specific selected reaction monitoring scans per analyte, enabling identification of both the N‐acyl chain and the phosphatidylethanolamine moiety. The validated method is suitable for the reliable quantification of N‐acylphosphatidylethanolamine species from rat brain with a lower limit of quantification of 10 pmol/g and a linear range up to 2300 pmol/g. In total, 41 N‐acylphosphatidylethanolamine molecular species with six different N‐acyl chains, amounting to a total concentration of 3 nmol/g, were quantified.

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Enzymatic Synthesis of Anandamide, an Endogenous Cannabinoid Receptor Ligand, through N-Acylphosphatidylethanolamine Pathway in Testis: Involvement of Ca2+-Dependent Transacylase and Phosphodiesterase Activities

Cited by 193 publications

References 0 publications

Changes in anandamide levels in mouse uterus are associated with uterine receptivity for embryo implantation

Changes in anandamide levels in mouse uterus are associated with uterine receptivity for embryo implantation

Discovery and Characterization of a Ca2+-independent Phosphatidylethanolamine N-Acyltransferase Generating the Anandamide Precursor and Its Congeners

Quantitative analysis of N‐acylphosphatidylethanolamine molecular species in rat brain using solid‐phase extraction combined with reversed‐phase chromatography and tandem mass spectrometry

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