Cyclooxygenase 1 mRNA expression is undetectable in Madin Darby Canine Kidney cells.

Guillaume Pelletier,Bhaja K Padhi

doi:10.1186/s13104-015-1049-4

Abstract

BackgroundMadin Darby Canine Kidney (MDCK) cells form polarized epithelium in vitro and are routinely used in research fields ranging from protein trafficking to influenza. However, the canine origin of these cells also means that compared to man or mouse, genomic resources are more limited and performance of commercially available antibodies often untested. The synthesis of pro-inflammatory prostaglandins in the kidney is mediated by the constitutively expressed cyclooxygenase 1 and the inducible cyclooxygenase 2 (COX-1 and COX-2, respectively). There are conflicting reports on the expression of COX-1 and COX-2 in MDCK cells and this lingering uncertainty about such important pharmacological targets may affect the interpretation of results obtained from this cell line.ResultsIn order to definitively settle the issue of cyclooxygenase expression in MDCK cells, we designed PCR primers based on dog genomic sequences to probe COX-1 and COX-2 mRNA expression in MDCK cells and dog kidney. We report that while COX-1 and COX-2 genes are both expressed in dog kidney, COX-1 expression is undetectable in MDCK cells.ConclusionsBy improving the characterization of cyclooxygenase expression in MDCK cells, this study will contribute to a better understanding of the properties of this cell line and lead to improved experimental designs and data interpretations.

Highlights

Madin Darby Canine Kidney (MDCK) cells form polarized epithelium in vitro and are routinely used in research fields ranging from protein trafficking to influenza
Cyclooxygenases are important pharmacological targets, as they are involved in the synthesis of pro-inflammatory prostaglandins [1]
In addition to the constitutively expressed cyclooxygenase 1 (COX-1), the inducible cyclooxygenase 2 (COX-2) is robustly expressed in the kidney where it plays a role in cellular adaptation to osmotic stress [2,3]

Summary

Introduction

Madin Darby Canine Kidney (MDCK) cells form polarized epithelium in vitro and are routinely used in research fields ranging from protein trafficking to influenza. In the decade or so between identification of the inducible COX-2 gene [6] and sequencing of dog cyclooxygenases [7,8,9,10], studies on their expression in MDCK cells assessed COX-1 and COX-2 protein levels by Western blot [2,11,12,13,14] and mRNA levels by Northern blot or RT-PCR [11,13,15,16], see Table 1 These studies which were based on antibodies raised against cyclooxygenases from other organisms or on orthologous nucleotide sequences from different species reached conflicting conclusions

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Conclusion