Resorption-cycle-dependent polarization of mRNAs for different subunits of V-ATPase in bone-resorbing osteoclasts.
Protein sorting in eukaryotic cells is mainly done by specific targeting of polypeptides. The present evidence from oocytes, neurons, and some other polarized cells suggests that protein sorting can be further facilitated by concentrating mRNAs to their corresponding subcellular areas. However, very little is known about the mechanism(s) involved in mRNA targeting, or how widespread and dynamic such mRNA sorting might be. In this study, we have used an in vitro cell culture system, where large multinucleated osteoclasts undergo continuous structural and functional changes from polarized (resorbing) to a nonpolarized (resting) stage. We demonstrate here, using a nonradioactive in situ hybridization technique and confocal microscopy, that mRNAs for several vacuolar H(+)-ATPase subunits change their localization and polarity in osteoclasts according to the resorption cycle, whereas mRNA for cytoplasmic carbonic anhydrase II is found diffusely located throughout the osteoclast during the whole resorption cycle. Antisense RNA against the 16-kDa or 60-kDa V-ATPase subunit inhibits polarization of the osteoclasts, as determined by cytoskeleton staining. Antisense RNA against carbonic anhydrase II, however, has no such effect.
Laitala-Leinonen
T
T
Department of Anatomy, University of Oulu, Finland.
Howell
M L
ML
Dean
G E
GE
Väänänen
H K
HK
D000818:Animals; D001483:Base Sequence; D001862:Bone Resorption / Q000378:metabolism*; D002256:Carbonic Anhydrases / Q000235:genetics; D002417:Cattle; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003599:Cytoskeleton / Q000378:metabolism; D015870:Gene Expression; D017403:In Situ Hybridization; D051379:Mice; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D010010:Osteoclasts / Q000378:metabolism*; D006180:Proton-Translocating ATPases / Q000235:genetics*; D016372:RNA, Antisense / Q000378:metabolism; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D015183:Restriction Mapping; D025262:Vacuolar Proton-Translocating ATPases
D016372:RNA, Antisense; D012333:RNA, Messenger; D025262:Vacuolar Proton-Translocating ATPases; D006180:Proton-Translocating ATPases; D002256:Carbonic Anhydrases