title: Extensive clear zone and defective ruffled border formation in osteoclasts of osteopetrotic (ia/ia) rats: implications for secretory function.
abstract: The cellular distribution of osteoclast integrin subunits alpha(v) and beta(3), the tissue distribution, and level of the apparent ligand osteopontin (OPN) as well as of the putative regulatory enzyme tartrate-resistant acid phosphatase (TRAP) were studied along with the intracellular distribution of the activation marker c-src in osteopetrotic ia/ia (incisors-absent) mutant rats and their normal littermates. In ia/ia rats, the osteoclasts are incapable of bone matrix resorption. Ultrastructurally the cells exhibit extended clear zones at the expense of ordinary ruffled borders. A secretory dysfunction in the mutant is strongly suggested by the absence of detectable extracellular TRAP, concomitant with an accumulation of the enzyme in abundant small cytoplasmic vesicles. Moreover, TRAP mRNA, protein content, as well as enzymatic activity were elevated. Furthermore, increased levels of integrin subunits alpha(v) and beta(3) were detected at the clear zone of mutant osteoclasts. OPN mRNA levels were elevated in long bones from mutants. In ia/ia rats, immunolabeling for OPN was homogeneously distributed at the surface facing osteoclasts, while in normal littermates it was concentrated at the clear zone area and barely detectable at ruffled borders. The absence of OPN labeling in the abundant, putative intracellular secretory vesicles in mutant osteoclasts suggests that these cells do not produce OPN. The osteoclasts of ia/ia rats appeared to produce and translocate the c-src protein to the cell membrane. In ia/ia a defect ruffled border-formation is observed along with extensive clear zone formation and decreased secretory function. The lesion may be due to a signaling defect, but in that case the defect seems to be located downstream to or not involving the c-src pathway. Our results illustrate the close relationship between secretory function and ruffled border formation in osteoclasts, a relationship that appears to be necessary for proper resorptive function.
authors.affiliation: Department of Immunology, Microbiology, Pathology and Infectious Diseases, Karolinska Institutet, Huddinge, S-141 86, Sweden. f.p.reinholt@labmed.uio.no
authors.forename: F P
authors.initials: FP
authors.lastname: Reinholt
authors.forename: K
authors.initials: K
authors.lastname: Hultenby
authors.forename: D
authors.initials: D
authors.lastname: Heinegård
authors.forename: S C
authors.initials: SC
authors.lastname: Marks
authors.forename: M
authors.initials: M
authors.lastname: Norgård
authors.forename: G
authors.initials: G
authors.lastname: Anderson
mesh_terms: D000135:Acid Phosphatase / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D015703:Antigens, CD / Q000302:isolation & purification; D001862:Bone Resorption / Q000503:physiopathology*; D002448:Cell Adhesion; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D039564:Integrin alphaV; D039661:Integrin beta3; D007527:Isoenzymes / Q000096:biosynthesis / Q000235:genetics; D010010:Osteoclasts / Q000473:pathology* / Q000648:ultrastructure; D010022:Osteopetrosis / Q000503:physiopathology*; D053495:Osteopontin; D010980:Platelet Membrane Glycoproteins / Q000302:isolation & purification; D016392:Proto-Oncogene Proteins pp60(c-src) / Q000096:biosynthesis / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011922:Rats, Mutant Strains; D012795:Sialoglycoproteins / Q000096:biosynthesis / Q000235:genetics; D015398:Signal Transduction; D000071681:Tartrate-Resistant Acid Phosphatase
chemical_list: D015703:Antigens, CD; D039564:Integrin alphaV; D039661:Integrin beta3; D007527:Isoenzymes; D010980:Platelet Membrane Glycoproteins; D012333:RNA, Messenger; D012795:Sialoglycoproteins; C506897:Spp1 protein, rat; D053495:Osteopontin; D016392:Proto-Oncogene Proteins pp60(c-src); D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase