The biochemical identification and immunocytochemical characterization of a cell surface antigen, expressed on globose basal cells (GBCs) of the rodent olfactory epithelium (OE), are described. The monoclonal antibody (MAb) GBC-3 recognizes a surface protein, confirmed by both live cell staining and fluorescence-activated cell sorting. Two-dimensional SDS-PAGE/Western blot followed by tandem mass spectrometry demonstrates that the cell surface GBC-3 antigen is the 40 kDa laminin receptor precursor protein. The MAb GBC-3 labels the vast majority of cells among the GBC population and does not stain either sustentacular cells or horizontal basal cells (HBCs) in the normal rat OE. After epithelial lesion by exposure to methyl bromide, the remaining cells, which are mostly GBCs, are heavily stained by GBC-3, and colabeled with GBC-3 and sustentacular cell or HBC markers. GBC-3 will be a potentially useful tool for identifying and characterizing GBCs.
Indexing terms2D IEF/SDS-PAGE; tandem mass spectrometry; cell surface marker; nonintegrin laminin receptors; tissue stem cells; progenitors The capacity of the olfactory epithelium (OE) for sustained neurogenesis and to recover after injury implies that the adult OE retains a population of tissue stem cells. Based in part on proliferative capacity, basal cells-globose basal cells (GBCs) and horizontal basal cells (HBCs)-have been deemed likely candidates for olfactory stem cells. While still subject to debate, there are ample data to support the notion that among the GBCs are putative stem cells with the capacity to give rise to both neurons and nonneuronal cells during reconstitution of the OE after various types of injury. That GBCs are neuronal progenitors has been well established by studies in unlesioned OE or following bulbectomy including; 1) pulse-chase experiments using 3 H-thymidine (Graziadei, 1973;Graziadei and Monti Graziadei, 1979); 2) increased proliferation of GBCs, but not HBCs, during the accelerated neurogenesis following bulbectomy (Schwartz Levey et al., 1991); and 3) lineage tracing using replication-incompetent, retroviral vectors (Caggiano et al., 1994;Schwob et al., 1994;Huard et al., 1998). However, a broader-than-neuronal potency (that is, multipotency) of
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NIH-PA Author ManuscriptGBCs has been demonstrated in other settings; for example, following MeBr lesion when multiple cell types are destroyed and need replacing. The most direct evidence of broad multipotency derives from experiments using transplantation of FACS-purified cell types into the MeBr-lesioned OE, which is a type of colony-forming unit (CFU) assay in vivo. In aggregate, FACS-sorted GBCs harvested from normal epithelium produce colonies containing olfactory sensory neurons (OSN), sustentacular cells, GBCs, Bowman's gland/ duct (BG/D) cells, and respiratory columnar epithelial cells following transplantation into the MeBr-lesioned epithelium (Chen et al., 2004). Moreover, a subset of GBCs in the normal epithelium has the ...