Identification of granulocyte-macrophage colony stimulating factor receptor mRNA by non-isotopic in situ hybridization in bone marrow biopsies

Abstract

BACKGROUND. In the last few years many studies have been published on GM-CSF receptors, focusing on molecular structure, function and distribution. Nevertheless, protocols for detecting GM-CSF receptors on formalin-fixed paraffin-embedded histological sections, to our knowledge, have not been described. METHODS. A method based on non-isotopic in situ hybridization (ISH) using a 21-base antisense DNA oligoprobe whose 3'-end was labeled with digoxigenin 11-dUTP was devised. The probe was applied on 20 routinely processed bone marrow trephine biopsies which were considered as positive controls. RESULTS. The hybridization signal was seen in myeloid cells, erythroid progenitors and rare megakaryocytes. CONCLUSIONS. Non-isotopic ISH represents an alternative to current methodologies for the assessment of GM-CSF receptor expression; since it is suitable for routinely processed samples, it can be regarded as a helpful tool for diagnostic determination of GM-CSF receptors in tumors from patients receiving GM-CSF and for retrospective studies on archival material.