Developmental expression of adenosine deaminase in the upper alimentary tract of mice

Chinsky, Jeffrey M.; Ramamurthy, V.; Fanslow, William C.; Ingolia, Diane E.; Blackburn, Michael R.; Shaffer, Kathryn T.; Higley, Howard R.; Trentin, John J.; Rudolph, Frederick B.; Knudsen, Thomas B.; Kellems, Rodney E.

doi:10.1111/j.1432-0436.1990.tb00759.x

Cited by 67 publications

(45 citation statements)

References 52 publications

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“…3) corresponding in size to the 3.9 kb rat 5'-NT mRNA reported (Misumi et al, 1990); the mouse ADA cRNA probe recognized a major band (Fig. 4) corresponding in size to the 1.7 kb murine ADA mRNA re- ported (Chinsky et al, 1990). Results from developmental Northern blots (Figs.…”

Section: The Patterns Of Y-nt and Ada Expression Differ In The Antimementioning

confidence: 89%

“…The mouse ADA probe, provided to us by Dr. R.E. Kellems (Department of Biochemistry and Molecular Biology, Baylor College of Medicine), consisted of the 5'(310 bp) PstI fragment from murine ADA cDNA in pGEM-2 (Chinsky et al, 1990). Plasmid linearized with EcoRI and incubated with T7 RNA polymerase yielded antisense cRNA; that linearized with Hind111 and incubated with SP6 RNA polymerase yielded sense cRNA.…”

Section: Northern Blot Analysismentioning

confidence: 99%

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Adenosine levels in the postimplantation mouse uterus: Quantitation by HPLC‐fluorometric detection and spatiotemporal regulation by 5′‐nucleotidase and adenosine deaminase

Blackburn

Gao

Airhart

et al. 1992

Developmental Dynamics

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Extracellular adenosine has the potential to influence many aspects of target cell metabolism. The present study has determined the endogenous levels of adenosine in the pregnant mouse uterus and developing embryodecidual unit with respect to the expression of two key enzymes of adenosine metabolism, 5'-nucleotidase (5'-NT; EC 3.1.3.5) and adenosine deaminase (ADA; EC 3.5.4.4). To measure adenosine levels, nucleoside extracts were etheno-derivatized and quantitated by high-performance liquid chromatography-fluorescence detection (0.03 pmollmg protein sensitivity). Adenosine levels were determined to be 0.18 nmol/mg protein in the nonpregnant uterus; however, two statistically significant changes were identified in the pregnant uterus: (1) a periimplantation surge between day 3 (0.24 nmoVmg protein) and day 5 (0.59 nmoYmg protein) of gestation (plug day 0; implantation day 4); and (2) an early postimplantation decline between day 6 (0.54 nmollmg protein) and day 7 (0.10 nmollmg protein). The periimplantation adenosine surge coincided with uterine expression of 5'-NT, an enzyme which catalyzes the irreversible dephosphorylation of 5'-AMP to adenosine. 5'-NT expression was shown by Northern blot analysis to peak in the embryo-decidual unit on day 5 of gestation and then to decline through day 9; transcripts remained elevated in the placenta between day 9 and day 13 (the latest day examined in this study). By use of specific enzyme histochemistry, most 5'-NT activity was localized to the primary decidual :zone on day 5. This expression subsequently declined during regression of the primary decidua; however, 5'-NT appeared on giant trophoblast ((days 7-13) and the metrial gland (days 11-13).Other purine catabolic enzymes degrading AMP (adenylate deaminase) or generating adenosine (S-adenosylhomocysteine hydrolase) were not detected in the embryo-decidual unit suggesting that the net flux of utero-placental AMP catabolism proceeds with adenosine as an intermediate, this being the major pathway of adenosine formation.The sharp drop in adenosine levels between day 6 and day 7 coincided with a rise in the activity and mRNA expression of ADA, an enzyme which catalyzes the irreversible deamination of adenosine to inosine. ADA was previously localized to the secondary decidual zone (days 6 1 l), secondary giant cells (days 7-13), and spongiotrophoblasts (days 8-13) in the mouse (Knudsen et al., 1991). Results of developmental Northern blot analysis demonstrated a direct correlation of relative 5'-NT/ADA mRNA band intensity to adenosine content between day 4 and day 9 of gestation, suggesting that the local availability of adenosine in the antimesometrium is dependent upon the distribution of these enzymatic activities. Purine nucleoside phosphorylase and xanthine oxidase, which are two catabolic enzymes acting subsequent to 5'-NT and ADA in the sequential degradation of AMP to xanthine, remained low and constant in the tissues examined suggesting that the catabolic pathway is geared toward regulation of adenosine le...

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Section: The Patterns Of Y-nt and Ada Expression Differ In The Antimementioning

confidence: 89%

Section: Northern Blot Analysismentioning

confidence: 99%

Adenosine levels in the postimplantation mouse uterus: Quantitation by HPLC‐fluorometric detection and spatiotemporal regulation by 5′‐nucleotidase and adenosine deaminase

Blackburn

Gao

Airhart

et al. 1992

Developmental Dynamics

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“…Preliminary observations suggests that rescued ADA deficient animals exhibit lymphopenia and immune deficiency. 2 ADA is expressed at high levels at three different places and times during the murine life cycle: first in the trophoblasts of the placenta during prenatal development (6,7), next in the gastrointestinal epithelium of the adult (16), and then in the deciduum of the pregnant uterus (6). The lack of ADA in the first of these places, the placenta, results in a phenotype that precludes our ability to investigate its importance in adult tissues.…”

Section: Figmentioning

confidence: 99%

“…B, Southern blot analysis of the Ada locus and ADA minigene locus from an intercrossing between animals hemizygous for the ADA minigene locus and heterozygous for the null Ada allele. Genomic DNA was isolated from tails at weaning, digested with BamHI, and hybridized with the internal 3.5-kb probe shown in Fig creases at birth (16). The high levels of ADA normally found in the placenta, together with placental replacement and subsequent rescue of ada m1 /ada m1 fetuses, suggest that placental ADA plays an important role in murine fetal development.…”

Section: Fig 2 Detection Of the Ada Minigene Locus In Ada-deficientmentioning

confidence: 99%

Tissue-specific Rescue Suggests That Placental Adenosine Deaminase Is Important for Fetal Development in Mice

Blackburn¹,

Wakamiya

Caskey

et al. 1995

Journal of Biological Chemistry

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Adenosine deaminase (ADA, EC 3.5.4.4) is an essential enzyme of purine metabolism that is expressed at very high levels in the murine placenta where it accounts for over 95% of the ADA present at the fetal gestation site. We have recently shown that ADA-deficient fetuses, which also lack ADA in their adjoining placentas, die during late fetal development in association with profound purine metabolic disturbances and hepatocellular impairment. We have now investigated the potential importance of placental ADA by genetically restoring the enzyme to placentas of ADA-deficient fetuses. This genetic engineering strategy corrected most of the purine metabolic disturbances, prevented serious fetal liver damage, and rescued the fetuses from perinatal lethality. Our findings suggest that placental ADA is important for murine fetal development and illustrate a general strategy for the tissue specific correction of phenotypes associated with null mutations in mice.During mammalian development the first differentiation event gives rise to the trophectoderm, which in turn provides lineages for specialized extraembryonic cells known as trophoblasts. These cells make the physical connection between the embryo and the maternal environment and play important roles in the implantation process and placental function (1). Inadequate placental development is associated with a high incidence of early embryonic mortality (2-4) and serious pregnancy disorders such as preeclampsia (5). Although the formation of a functional placenta is essential for mammalian embryogenisis and fetal development, relatively little is known about the molecular mechanisms that govern trophoblast differentiation and subsequent function. Determining what proteins are produced in trophoblasts and elucidating their physiological roles is an important part of understanding how this cell lineage contributes to the formation of a functional placenta.One such protein that is highly abundant in the murine placenta is adenosine deaminase (ADA).1 ADA is an essential enzyme of purine metabolism that is expressed at very high levels in trophoblasts of the murine placenta (6, 7). The physiological importance of ADA in trophoblasts of the placenta is not known. However, recent evidence suggests that ADA is essential during fetal stages of development. ADA-deficient fetuses, which also lacked ADA in trophoblasts of their adjoining placentas, died perinatally in association with profound purine metabolic disturbances and hepatocellular impairment (8, 9). Considering that greater than 95% of ADA enzymatic activity found in the fetal gestation site resides in trophoblasts of the placenta, it is likely that placental ADA plays an essential role during fetal development. Here we show that genetically restoring ADA to placentas of ADA-deficient fetuses rescued them from perinatal lethality, thereby providing compelling evidence for the importance of placental ADA for fetal development. MATERIALS AND METHODS ADA Minigene Construction and Transgenic Mouse Generation-Theplasmid, Ϫ...

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“…As for tissues, immunohistochemical localization of ADA studies have been performed extensively using tissues from the brain [13,22] and digestive tract [7,25]. However, immunohistochemical localization of ADA has not been performed with human tissues, except for the thymus [4].…”

Section: Introductionmentioning

confidence: 99%

Immunohistochemical Demonstration of Adenosine Deaminase (ADA1) in Human Tissues

Moriwaki

Takahashi

Tsutsumi

et al. 2004

Acta Histochem. Cytochem

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Developmental expression of adenosine deaminase in the upper alimentary tract of mice

Cited by 67 publications

References 52 publications

Adenosine levels in the postimplantation mouse uterus: Quantitation by HPLC‐fluorometric detection and spatiotemporal regulation by 5′‐nucleotidase and adenosine deaminase

Adenosine levels in the postimplantation mouse uterus: Quantitation by HPLC‐fluorometric detection and spatiotemporal regulation by 5′‐nucleotidase and adenosine deaminase

Tissue-specific Rescue Suggests That Placental Adenosine Deaminase Is Important for Fetal Development in Mice

Immunohistochemical Demonstration of Adenosine Deaminase (ADA1) in Human Tissues

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