Expression of c-kit and Sca-1 and their relationship with multidrug resistance protein 1 in mouse bone marrow mononuclear cells.
P-glycoprotein (Pgp) and multidrug resistance protein 1 (MRP1) are members of the ATP-binding cassette (ABC) family of transporter proteins. Both molecules are membrane-associated, energy-dependent efflux pumps with different substrate selectivity and they may play a role in the activation, differentiation and function of haematopoietic cells. Mouse haematopoietic cells are characterized by the expression of the cell surface molecules c-kit and Sca-1. Herein, the presence and activities of Pgp and MRP1 in mouse bone marrow mononuclear cells (BMMC) and their relationship with the proteins c-kit and Sca-1 were evaluated. Pgp and MRP activities were measured based on the extrusion of rhodamine 123 (for Pgp) and Fluo-3 (for MRP). Cell populations were assessed by cytometry using anti-c-kit and anti-Sca1 antibodies. Pgp activity was present in 5% of BMMC while 50% of BMMC cells showed MRP activity. These findings agreed with the proportion of cells expressing the MRP1 surface molecule (51.3 +/- 4.17%). About 14% of BMMC were positive for c-kit and/or Sca-1 (9.3% c-kit- Sca-1+, 4.2% c-kit+ Sca-1- and 0.9% c-kit+ Sca-1+). Among these subpopulations only c-kit- Sca-1+ cells presented Pgp activity (21.36%). On the other hand, MRP activity was present in all three subpopulations. Most cells (82.5%) of the c-kit+ Sca-1- subpopulation presented MRP1 activity compared to only 54.1% of c-kit+ Sca-1+ and 38.8% of c-kit- Sca-1+. This study demonstrates the expression and activity of MRP1 in BMMC. While only a small proportion of precursor cells had Pgp activity, MRP1 activity was present among different subpopulations of precursor cells. Further studies are necessary to establish the role of these transporters in haematopoietic cells.
Kyle-Cezar
Fernanda
F
Instituto de Bioquímica Médica, UFRJ, Rio de Janeiro, Brazil.
Echevarria-Lima
Juliana
J
dos Santos Goldenberg
Regina Coeli
RC
Rumjanek
Vivian M
VM
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000037:antagonists & inhibitors / Q000378:metabolism* / Q000502:physiology; D000818:Animals; D000950:Antigens, Ly / Q000378:metabolism*; D001693:Biological Transport, Active / Q000502:physiology; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism*; D002454:Cell Differentiation / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D006412:Hematopoietic Stem Cells / Q000166:cytology; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D019009:Proto-Oncogene Proteins c-kit / Q000378:metabolism*
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000950:Antigens, Ly; C074650:Ly6a protein, mouse; D008565:Membrane Proteins; D019009:Proto-Oncogene Proteins c-kit