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Evidence for A 1 and A 2B adenosine receptors in baby hamster kidney (BHK) cells
Author(s) -
Mittal Rashmi A.,
Tan CheeHong,
Khoo HoonEng
Publication year - 1998
Publication title -
biofactors
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.204
H-Index - 94
eISSN - 1872-8081
pISSN - 0951-6433
DOI - 10.1002/biof.5520070105
Subject(s) - adenosine , baby hamster kidney cell , adenosine receptor , adenosine a2b receptor , receptor , microbiology and biotechnology , hamster , kidney , adenosine a1 receptor , biology , chemistry , radioligand assay , biochemistry , cell , endocrinology , agonist
Abstract Adenosine is known to produce biphasic effects in the renal tissues via adenosine receptors. However, the presence of more than one subtype of adenosine receptor on a type of kidney cell or tissue has not been conclusively demonstrated. To address this issue, we investigated the presence of A 1 and A 2 adenosine receptors in baby hamster kidney (BHK) cells by use of radioligand binding and the reverse transcription‐polymerase chain reaction. Ligand binding studies with ( 3 H)‐DPCPX revealed a single class of binding site with a K D of 9.2 ± 2.0 nM, a B max of 1.7 ± 0.2 pmol/mg protein and a pharmacological profile characteristic of A 1 adenosine receptor on the BHK cell membrane. As the presence of A 2 adenosine receptors could not be conclusively determined by ligand binding studies, the more sensitive method of RT‐PCR was employed. The presence of A 1 and A 2B adenosine receptors was detected by RT‐PCR with specific primers and the subsequent sequencing of the resultant amplification product. The sequences obtained were 75–90% homologous to the respective adenosine receptor mRNA of rat, mouse and human.