Detection and quantitation of EP3 prostaglandin E2 receptor mRNA along mouse nephron segments by RT-PCR
Taniguchi, S.; Watanabe, T.; Nakao, A.; Seki, G.; Uwatoko, S.; Kurokawa, K.
American Journal of Physiology 266(5 Pt 1): C1453-C1458
1994
ISSN/ISBN: 0002-9513 PMID: 7515568 Document Number: 434347
mRNA of the EP-3 prostaglandin E-2 (PGE-2) receptor was detected and quantitated in microdissected mouse nephron segments by a modified protocol of reverse transcription-polymerase chain reaction (RT-PCR). At the step of RT, a point mutation was introduced in cDNA, which made a new restriction site for the Mbo I enzyme. PCR was performed using a set of primers on the same exon, and genomic DNA was coamplified with cDNA by these primers. PCR products were treated with Mbo I, and signals from genomic DNA and mRNA were separately detected on gel electrophoresis. The relative amount of mRNA per cell was expressed as a ratio of amount of product from mRNA to that from genomic DNA. EP-3 PGE-2 receptor mRNA expression was abundant in thick ascending limb of Henle, present to a lesser extent in distal convoluted tubules and collecting ducts, and undetectable in glomeruli, proximal tubules. and descending thin limb. The results support the notion that PGE-2 modulates water and solute transport through the EP-3 receptor in specific structures of the kidney.