title: Cathepsin K mRNA detection is restricted to osteoclasts during fetal mouse development.
abstract: We recently identified a novel cysteine protease, cathepsin K, by random sequencing of an osteoclast cDNA library, and in situ hybridization studies in adult human tissues demonstrated high and specific expression in osteoclasts. To determine whether the expression of cathepsin K mRNA during mouse embryogenesis was more widespread, cryostat sections of early (day 11-13) and late (day 15-17) mouse fetuses were analyzed by in situ hybridization. Serial cross-sections were collected through each fetus, and co-reacted for tartrate-resistant acid phosphatase (TRAP) and nonspecific esterase (NSE), selective markers for the osteoclast, and precursor cells derived from the macrophage/monocyte lineage, respectively. In the 11-13 day fetuses, cathepsin K mRNA was not expressed in any extraskeletal tissue; at this stage of embryogenesis, no osteoclasts are present. However, in the 15-17 day fetuses, a distinctive, developmental stage-dependent pattern of cathepsin K expression was observed in osteoclasts and preosteoclasts at sites of cartilage and bone modeling. Cathepsin K positive osteoclasts differentiated within a peripheral zone of the osteogenic stacked cell layer of the cartilage rudiments (prior to ossification), migrated and/or resorbed the bone collar, and invaded the cartilage core. Furthermore, following the invasive penetration of vasculature into the degenerating cartilage core, the calcified cartilage was resorbed by cathepsin K positive mononuclear osteoclast precursors (NSE+ve, negligible TRAP); cells positive for both enzymes were identified indicative of osteoclast differentiation. The deposition of bone by osteoblasts onto the cartilage remnants is followed by mononucleated and multinucleated osteoclastic resorption; these osteoclasts demonstrated intense cathepsin K expression. Similar expression patterns were observed at sites of intramembranous ossification. No expression was observed in chondrocytes, osteoblasts, marrow, or in any other nonskeletal tissue at these time points. These data indicated that cathepsin K expression during embryogenesis occurred only following the onset of osteoclast differentiation.
authors.affiliation: SmithKline Beecham Pharmaceuticals, Department of Cellular Biochemistry, King of Prussia, Pennsylvania 19406, USA.
authors.forename: R A
authors.initials: RA
authors.lastname: Dodds
authors.forename: J R
authors.initials: JR
authors.lastname: Connor
authors.forename: F
authors.initials: F
authors.lastname: Drake
authors.forename: J
authors.initials: J
authors.lastname: Feild
authors.forename: M
authors.initials: M
authors.lastname: Gowen
mesh_terms: D000135:Acid Phosphatase / Q000032:analysis; D000818:Animals; D001846:Bone Development / Q000502:physiology*; D016723:Bone Remodeling / Q000502:physiology; D002113:Calcification, Physiologic; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000032:analysis; D002356:Cartilage / Q000378:metabolism; D056657:Cathepsin K; D002403:Cathepsins / Q000032:analysis* / Q000235:genetics; D002454:Cell Differentiation; D004396:Coloring Agents; D005314:Embryonic and Fetal Development / Q000502:physiology*; D005260:Female; D005865:Gestational Age; D017403:In Situ Hybridization; D007527:Isoenzymes / Q000032:analysis; D051379:Mice; D008807:Mice, Inbred BALB C; D010010:Osteoclasts / Q000378:metabolism*; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis*; D000071681:Tartrate-Resistant Acid Phosphatase
chemical_list: D004396:Coloring Agents; D007527:Isoenzymes; D012333:RNA, Messenger; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; D000135:Acid Phosphatase; C000605511:Acp5 protein, mouse; D000071681:Tartrate-Resistant Acid Phosphatase; D002403:Cathepsins; C534130:CTSK protein, human; D056657:Cathepsin K; C534132:Ctsk protein, mouse