title: Islet expression of Rhombotin and Isl-1 suggests cell type specific exposure of LIM-domain epitopes.
abstract: The homeodomain protein Isl-1 and the proto-oncogene Rhombotin (a LIM-only protein), share a double zinc-binding LIM domain and have both been implicated in neural and possibly endocrine development. Isl-1 is expressed in all endocrine cell-types of the islet of Langerhans while Rhombotin mRNA expression was reported in rat insulinoma cells. We have cloned and sequenced Rhombotin cDNA from rat insulinoma (99.4% identical to human and mouse sequences) and demonstrate that it is expressed in normal islets, intestinal tissue, and testis, in addition to the brain; but absent in all other organs tested. Rhombotin mRNA is expressed in phenotypically distinct islet tumours (α-, β, and δ-tumours) at levels comparable to that of normal islets. Antisera raised against two distinct epitopes contained within a short synthetic peptide representing part of the N-terminal LIM domain of Rhombotin surprisingly stain α- and δ-cells, respectively, on sections of rat pancreas. Rhombotin is undetectable by immunocytochemistry using LIM-domain antisera on intact monolayer islet tumor cells or transfected fibroblasts while readily detectable when equipped with a FLAG epitope, as detected with FLAG antiserum. In contrast, recombinant FLAG-Rhombotin is efficiently recognised by Western blotting or immunoprecipitation with all LIM-specific antisera. Almost identical results were obtained with LIM-specific versus homeodomain/C-terminal Isl-1 antisera staining α-cell cytoplasm or all islet nuclei, respectively. We conclude that Rhombotin in addition to Isl-1 is expressed in the islet of Langerhans and propose that the differential staining patterns obtained with antisera towards the LIM domains versus flanking epitopes of both proteins reflect (1) cell-specific protein-protein interactions of these domains or, alternatively, (2) islet cell type specific expression of novel homologous LIM domain proteins.
authors.affiliation: Hagedorn Research Institute, Niels Steensensvej 6, DK2820, Gentofte, Denmark.
authors.forename: K
authors.initials: K
authors.lastname: Lund
authors.forename: J S
authors.initials: JS
authors.lastname: Petersen
authors.forename: J
authors.initials: J
authors.lastname: Jensen
authors.forename: N
authors.initials: N
authors.lastname: Blume
authors.forename: T
authors.initials: T
authors.lastname: Edlund
authors.forename: S
authors.initials: S
authors.lastname: Thor
authors.forename: O D
authors.initials: OD
authors.lastname: Madsen