title: Adenosine receptor expression and gene reference evaluation in human leukocytes.
abstract: BACKGROUND
  Peripheral blood mononuclear cells and isolated polymorphonuclear neutrophilis were used to evaluate gene expression studies. Unfortunately, there are many methodological problems related to these cellular models, limiting their use. The aim was to evaluate a fast and easy procedure for the extraction of total RNA from leukocytes obtained from human whole blood (WB) < 10 mL; to determine adenosine receptor (AR) mRNA expression in WB samples of normal subjects and to establish the most stable reference genes for data normalization.
  
  
  METHODS
  mRNA expression was performed by Real-Time PCR.
  
  
  RESULTS
  The most stably expressed genes were TPT1, EEF1A, and RPL13A. Similar levels of mRNA expression were observed for A2aR, A2bR, and A3R while lower levels were measured for A1R (p = 0.02 A1R vs. A2aR; p = 0.04 A1R vs. A3R).
  
  
  CONCLUSIONS
  Our study represents an important and useful starting point for future investigations devoted to evaluate the expression of ARs in human diseases.
authors.affiliation: CNR Institute of Clinical Physiology, Laboratory of Cardiovascular Biochemistry, Pisa, Italy.
authors.forename: Laura
authors.initials: L
authors.lastname: Sabatino
authors.forename: Manuela
authors.initials: M
authors.lastname: Cabiati
authors.forename: Chiara
authors.initials: C
authors.lastname: Caselli
authors.forename: Silvia
authors.initials: S
authors.lastname: Del Ry
mesh_terms: D000328:Adult; D000704:Analysis of Variance; D000818:Animals; D004587:Electrophoresis, Agar Gel; D005819:Genetic Markers / Q000235:genetics; D006801:Humans; D007962:Leukocytes / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D051379:Mice; D012333:RNA, Messenger / Q000096:biosynthesis / Q000097:blood* / Q000737:chemistry / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D018047:Receptors, Purinergic P1 / Q000096:biosynthesis / Q000097:blood / Q000235:genetics*; D012016:Reference Values; D000090324:Tumor Protein, Translationally-Controlled 1
chemical_list: D005819:Genetic Markers; D012333:RNA, Messenger; D018047:Receptors, Purinergic P1; C000715647:TPT1 protein, human; C000715648:Tpt1 protein, mouse; D000090324:Tumor Protein, Translationally-Controlled 1