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  • 1
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Pregnant rats were treated daily with 1 g/L of caffeine or theophylline in their drinking water during pregnancy and the effect of these methylxanthines on adenosine A1 receptor was assayed using binding and reverse transcription polymerase chain reaction (RT–PCR) assays in brains from both mothers and full-term fetuses. In plasma membranes from pregnant rat brain, caffeine and theophylline caused a significant decrease in total receptor numbers, of the same order in both cases (30%), with no significant changes on receptor affinity. The effect of these adenosine receptor antagonists on plasma membranes from fetal brains was more marked, being detected at approximately 50% of the total receptors detected in control conditions. However, in this tissue, a significant increase in the receptor affinity, of the same order in both cases, was also detected after antagonist administration. No significant variation on the potency of caffeine and theophylline as antagonists was detected after treatments in mothers; however, higher affinities were detected in fetuses. A decrease in the total receptor numbers in fetal brain was associated with an increase in the mRNA coding A1 receptor, as determined by RT–PCR assays, not having detected any mRNA difference in maternal brain. No variation in the levels of mRNA coding A2A receptor was detected in any case. These results suggest that maternal caffeine or theophylline intake modulates adenosine A1 receptor, causing a down-regulation of adenosine A1 receptor in brain in both mothers and fetuses.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Oxford UK : Blackwell Science Ltd.
    Journal of neurochemistry 75 (2000), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Adenosine A1 receptors (A1Rs) have been characterized in primary cultures of neurons from cerebral cortex. The specific adenosine A1 antagonist 8-cyclopentyl-1,3-[3H]dipropylxanthine bound to both membranes and intact cells. When saturation experiments were performed in membranes, a KD value of 0.76 nM and a Bmax of 57 fmol/mg of protein were obtained. Competition assays revealed a pharmacological profile characteristic of A1Rs. The presence of this receptor was further confirmed by RT-PCR analysis. The expression of the receptor showed no significant changes during the period of culture studied, up to 12 days in vitro. A1R agonist inhibited forskolin-stimulated adenylyl cyclase, showing the functional coupling of these receptors with the effector. αGi1,2 protein level, detected by immunoblot, presented an increase during the period of culture. This increase correlated with an increase in the mRNA level of αGi1 but not αGi2. By immunochemical assays, it is shown that these receptors are expressed in both the neuronal cell body and the proximal dendrites. Colocalization of A1Rs with microtubule-associated protein 2 and cell surface adenosine deaminase was shown by confocal microscopy. The high degree of colocalization observed between A1Rs and ectoadenosine deaminase in neurons could suggest an important role of the enzyme in adenosine-mediated neuromodulation.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Science Ltd
    Journal of neurochemistry 88 (2004), S. 0 
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: The status of the adenosine A1 receptor/adenylyl cyclase (A1R/AC) transduction pathway in rat brain was analysed at the end of pregnancy using different approaches. Pregnancy at term caused a significant decrease in the Bmax value obtained by saturation binding assays using [3H]DPCPX as radioligand, suggesting a down-regulation of adenosine A1 receptor. Moreover, A1 receptor immunodetection in pregnant rat membranes and the level of mRNA coding A1 receptor were significantly decreased. This loss of A1 receptor was associated with a significant increase in receptor affinity, since the KD value from the [3H]DPCPX saturation curve and Ki for N6-cyclohexyladenosine (CHA) were decreased in pregnant rats. Surprisingly, CHA-mediated inhibition of adenylyl cyclase was increased, reflecting enhanced receptor responsiveness. On the other hand, immunoblotting of different αGi-protein isoforms revealed a significant increase in αGi3 level in membranes from pregnant rats. Pre-incubation of membranes with anti-αGi3 antibody blocked the guanosine triphosphate (GTP) or CHA inhibitory effect on adenylyl cyclase in both pregnant and non-pregnant rats, pointing to αGi3 as the main isoform involved in the A1 receptor response. These results suggest that, at the end of pregnancy, there is a down-regulation of adenosine A1 receptors counterbalanced with a strengthened functionality, probably due to an increase in both αGi3 protein and receptor affinity.
    Type of Medium: Electronic Resource
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