Immunohistochemically-detected differential localization of CXCL14 in mouse cerebellum suggesting the presence of neuronal and glial forms of CXCL14

Abstract

In this study we compared staining profiles in mouse cerebellum, using two anti-CXCL14 antibodies, which we designated α and β. The α anti-CXCL14 antibody labeled Purkinje neurons; this was confirmed by double-immunofluorescence staining with calbindin D-28k, a marker for Purkinje neurons. The β anti-CXCL14 antibody marked Bergmann glia; this was confirmed by combinational staining with glial fibrillary acidic protein, a marker for Bergmann glia. Both staining profiles were abolished by preabsorption of each antibody with recombinant human CXCL14. Western blot analyses using these two antibodies indicated that they both recognized recombinant human and mouse CXCL14s, which migrated at approximately a 12 kDa position. Furthermore, the α anti-CXCL14 antibody labeled major bands at approximately 15 kDa and 21 kDa, and a minor band at approximately 19 kDa from a Triton-X fraction of mouse cerebellum. However, there were no immunostaining bands from a Tris-HCl fraction. In contrast, the β anti-CXCL14 antibody labeled a band of approximately 18 kDa from a Tris-HCl fraction of cerebellum, but no immunoreactive bands were detected from a Triton-X fraction. These results suggest that mouse cerebellum has at least two forms of CXCL14; presumably a neuronal form and a glial form.