RGD Reference Report - The ecto-enzymes CD73 and adenosine deaminase modulate 5'-AMP-derived adenosine in myofibroblasts of the rat small intestine. - Rat Genome Database

RGD Reference Report - The ecto-enzymes CD73 and adenosine deaminase modulate 5'-AMP-derived adenosine in myofibroblasts of the rat small intestine. - Rat Genome Database

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The ecto-enzymes CD73 and adenosine deaminase modulate 5'-AMP-derived adenosine in myofibroblasts of the rat small intestine.
Authors:	Bin, Anna Caputi, Valentina Bistoletti, Michela Montopoli, Monica Colucci, Rocchina Antonioli, Luca De Martin, Sara Castagliuolo, Ignazio Orso, Genny Giaroni, Cristina Debetto, Patrizia Giron, Maria Cecilia
Citation:	Bin A, etal., Purinergic Signal. 2018 Dec;14(4):409-421. doi: 10.1007/s11302-018-9623-6. Epub 2018 Sep 29.
RGD ID:	152995394
Pubmed:	PMID:30269308 (View Abstract at PubMed)
PMCID:	PMC6298927 (View Article at PubMed Central)
DOI:	DOI:10.1007/s11302-018-9623-6 (Journal Full-text)
Adenosine is a versatile signaling molecule recognized to physiologically influence gut motor functions. Both the duration and magnitude of adenosine signaling in enteric neuromuscular function depend on its availability, which is regulated by the ecto-enzymes ecto-5'-nucleotidase (CD73), alkaline phosphatase (AP), and ecto-adenosine deaminase (ADA) and by dipyridamole-sensitive equilibrative transporters (ENTs). Our purpose was to assess the involvement of CD73, APs, ecto-ADA in the formation of AMP-derived adenosine in primary cultures of ileal myofibroblasts (IMFs). IMFs were isolated from rat ileum longitudinal muscle segments by means of primary explant technique and identified by immunofluorescence staining for vimentin and α-smooth muscle actin. IMFs confluent monolayers were exposed to exogenous 5'-AMP in the presence or absence of CD73, APs, ecto-ADA, or ENTs inhibitors. The formation of adenosine and its metabolites in the IMFs medium was monitored by high-performance liquid chromatography. The distribution of CD73 and ADA in IMFs was detected by confocal immunocytochemistry and qRT-PCR. Exogenous 5'-AMP was rapidly cleared being almost undetectable after 60-min incubation, while adenosine levels significantly increased. Treatment of IMFs with CD73 inhibitors markedly reduced 5'-AMP clearance whereas ADA blockade or inhibition of both ADA and ENTs prevented adenosine catabolism. By contrast, inhibition of APs did not affect 5'-AMP metabolism. Immunofluorescence staining and qRT-PCR analysis confirmed the expression of CD73 and ADA in IMFs. Overall, our data show that in IMFs an extracellular AMP-adenosine pathway is functionally active and among the different enzymatic pathways regulating extracellular adenosine levels, CD73 and ecto-ADA represent the critical catabolic pathway.

Biological Process

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Object Symbol	Species	Term	Qualifier	Evidence	With	Notes	Source	Original Reference(s)
Nt5e	Rat	adenosine biosynthetic process		IMP			RGD
Ada	Rat	adenosine catabolic process		IMP			RGD
Nt5e	Rat	AMP catabolic process		IMP			RGD

Genes (Rattus norvegicus)


Ada (adenosine deaminase)	Nt5e (5' nucleotidase, ecto)