Murine osteoclasts and spleen cell polykaryons are distinguished by mRNA phenotyping.
To probe osteoclast gene expression, we combined the techniques of cell microisolation and RT-PCR to develop a novel and sensitive method for the isolation and mRNA phenotyping of small numbers of authentic osteoclasts and spleen cell polykaryons. Using this method we report (1) direct evidence for the presence of calcitonin receptor mRNA in osteoclasts, (2) confirmation of the recent finding of osteopontin mRNA in osteoclasts, and (3) demonstration that the specific expression of mRNA for tartrate-resistant acid phosphatase, carbonic anhydrase II, calcitonin receptor, and osteopontin enable one to distinguish the osteoclast from the morphologically similar and developmentally related spleen cell polykaryon. We also show that mRNA associated with the osteoblast phenotype, such as alkaline phosphatase, osteocalcin, and type I collagen, are absent in osteoclasts. This is the first report in which such an approach has been used successfully to distinguish the mRNA expression pattern of an authentic osteoclast from a macrophage polykaryon, and as such it should provide an important new tool for evaluating the results of various cell culture model systems designed to examine the origin and ontogeny of osteoclasts. Our results also indicate that these procedures can be used as an alternative to in situ hybridization methods for the cell-specific localization of specific mRNA in a mixed cell preparation and for colocalization of multiple mRNA species to a single cell type.
Tong
H S
HS
School of Dentistry, University of Southern California, Los Angeles.
Sakai
D D
DD
Sims
S M
SM
Dixon
S J
SJ
Yamin
M
M
Goldring
S R
SR
Snead
M L
ML
Minkin
C
C
D000818:Animals; D001483:Base Sequence; D002469:Cell Separation; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D008264:Macrophages / Q000166:cytology / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010010:Osteoclasts / Q000378:metabolism*; D010641:Phenotype; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D013154:Spleen / Q000166:cytology* / Q000378:metabolism*
D017931:DNA Primers; D012333:RNA, Messenger