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Studies on gene expression in the kidneys of P2Y2 receptor knockout mice
Author(s) -
Tuttle Jonathon D.,
Kishore Bellamkonda K.
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.970.7
Subject(s) - gene isoform , gene expression , microbiology and biotechnology , knockout mouse , gene knockout , receptor , gene , biology , complementary dna , prostanoid , real time polymerase chain reaction , chemistry , biochemistry
Research conducted in the host laboratory showed that P2Y2 receptor (P2Y2‐R) plays a key role in the maintenance of water homeostasis in the renal medullary collecting duct by interacting with vasopressin and prostanoid systems. Based on these observations, the aim of this study is to examine the gene expression of key molecules of prostanoid biosynthesis and NOS isoforms in P2Y2‐R knockout (KO) and wild type (WT) mice. RNA was extracted from the cortex and medullary tissue samples of KO and WT mice (N = 5 per group). Reverse transcription (RT) was performed on the RNA samples using SuperScript III Super Mix. Quantitative PCR was performed on the cDNA samples using gene specific primers for COX‐1, COX‐2, mPEGS‐1, nNOS, iNOS, eNOS and β‐actin and TaKaRa Taq R‐PCR Version in the Smart Cycler II System. SYBR Green was used for detection. Relative expression values were computed by normalizing the expression of target genes to that of β‐actin. Statistical analysis showed that expression of none of the molecules studied was significantly altered in the KO mice as compared to WT mice, although, numerically higher or lower values were observed for some of the molecules. These data indicate that genetic deletion of P2Y2‐R may not significantly alter the expression of molecules of prostanoid biosynthetic pathway and NOS isoforms at least under basal conditions. It is possible that when challenged by subjecting the KO and WT mice to experimental conditions that are known to alter the prostanoid biosynthesis or NOS isoforms, significant differences between the KO and WT mice could be observed. Work supported by the Dept. of Veterans Affairs and NKF of Utah and Idaho.