Expression analysis of LacZ gene placed in the locus of Cnot7 exhibits its activity in osteoblasts in vivo and in mineralized nodules in vitro.
CCR4-NOT complex 7 (Cnot7) was identified as a regulator of gene expression in yeast and evolutionally conserved in mammals. Cnot7 deficient male mice exhibit abnormality in spermatogenesis. As these mice contained construct to express LacZ, we followed the expression patterning in these animals. LacZ was expressed in osteoblasts located in the primary spongiosa in adult mice. Cellular analysis indicated that LacZ is expressed in osteoblasts but not in osteoclasts. In the mineralized nodules formed in the culture of bone marrow cells obtained from Cnot7 +/- mice, LacZ expression was mainly observed in the cells forming mineralized nodules but not in un-mineralized area scattered around the periphery of the nodules. LacZ blue positive cells were gradually depositing minerals along its time course of the in vitro mineralization assay. Cnot7 expression was enhanced by the treatment with BMP. These data suggest that Cnot7 is expressed in osteoblasts and is associated with mineralization.
Washio-Oikawa
Kaoru
K
Department of Molecular Pharmacology, Medical Research Institute, Tokyo Medical and Dental University, Tokyo 101-0062, Japan.
Nakamura
Takahisa
T
Usui
Michihiko
M
Yoneda
Mitsuhiro
M
Ezura
Youichi
Y
Ishikawa
Isao
I
Nakashima
Kazuhisa
K
Yamamoto
Tadashi
T
Noda
Masaki
M
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D017931:DNA Primers / Q000235:genetics; D005095:Exoribonucleases; D015870:Gene Expression; D006579:Heterozygote; D066298:In Vitro Techniques; D007763:Lac Operon; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008822:Mice, Transgenic; D008903:Minerals / Q000378:metabolism; D010006:Osteoblasts / Q000378:metabolism*; D011506:Proteins / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins; D012260:Ribonucleases
D017931:DNA Primers; D008903:Minerals; D011506:Proteins; D012333:RNA, Messenger; D012097:Repressor Proteins; C489778:Cnot7 protein, mouse; D005095:Exoribonucleases; D012260:Ribonucleases