Tissue distribution of the human MDR3 P-glycoprotein.

BACKGROUND

P-glycoproteins (Pgps) belong to a family of well conserved plasma membrane proteins with two members in humans: MDR1 and MDR3. The MDR1 Pgp can transport drugs; the murine homologue of MDR3, mdr2, was recently shown by us to be involved in transport of the phospholipid phosphatidylcholine (lecithin) into bile.

EXPERIMENTAL DESIGN

We have determined the MDR3 mRNA levels in a panel of human tissues by RNase protection. We have also generated polyclonal antibodies specific for the MDR3 Pgp. Detection of the MDR3 Pgp in human tissues with these antibodies was by a streptavidin-ABC procedure.

RESULTS

The RNase protection results show that expression of the MDR3 gene has a more restricted distribution than that of MDR1. A high level of MDR3 mRNA was detected in the liver and in low levels in the adrenal gland, heart, striated muscle, spleen, and tonsil. In all of these tissues, some of the previously described splice variants of MDR3 were abundantly expressed. No indications were found for a tissue-specific regulation of alternative splicing of the MDR3 pre-mRNA. Two MDR3 Pgp-specific antibodies stained the bile canalicular membrane of hepatocytes across the entire liver lobule. No staining was found in the epithelial cells of the bile ductules and gall bladder, indicating that the staining at these sites with C219, a monoclonal antibody that recognizes both MDR1 and MDR3 Pgp, (mainly) represents the MDR1 Pgp. No MDR3 was detected by specific antibodies in the adrenal gland, spleen, and muscle. Since no staining was reported with MDR1-specific antibodies in muscle either, our results indicate that the C219 staining in some fibers of striated muscle represents a cross-reaction with another protein. One of the human MDR3-specific antibodies cross-reacted with the highly homologous mouse mdr2 Pgp. Staining with this antibody showed that the distribution of this protein in mouse liver and striated muscle is very similar to that of MDR3 Pgp in human tissues.

CONCLUSIONS

The highest expression of the MDR3 Pgp was found in liver in the canalicular membranes of hepatocytes. This is in agreement with a role for MDR3 in the transport of phospholipid into bile.