Functional Implication of Nucleolin in the Mouse First Molar Development

Xie, Ming; Kobayashi, Ieyoshi; Kiyoshima, Tamotsu; Yamaza, Haruyoshi; Honda, Jun; Takahashi, Keiko; Enoki, Norio; Akamine, Akifumi; Sakai, Hidetaka

doi:10.1074/jbc.m610779200

Cited by 17 publications

(29 citation statements)

References 45 publications

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“…This is the first report to show a possible relationship between the strong expression of Rpl21 mRNA and the morphological development of the tooth germ. The predictable roles of Rpl21 in the tooth germ development are also supported by previous studies of the in situ signals of Tb4, Pgk1, Hsc73, Hsj2, Hsp86, nucleolin and Set-a mRNAs, because the strong signal pattern of Rpl21 was similar to patterns of those genes in the thickened epithelium-bell stages of the tooth germ development (Akhter et al 2005;Honda et al 2008;Wada et al 2002;Xie et al 2007;Yamaza et al 2001a, b). In addition, an inhibitory assay for nucleolin expression with antisense sulfur-substituted oligodeoxynucleotide (AS-S-ODN) probe apparently suppressed the morphogenesis of the tooth germ.…”

Section: Discussionsupporting

confidence: 62%

“…In situ hybridization for Rpl21 mRNA In situ hybridization using the antisense RNA probe was performed according to our previous studies (Akhter et al 2005;Honda et al 2008;Kobayashi et al 2006;Wada et al 2002;Xie et al 2007;Yamaza et al 2001a, b). The specific day old embryos were fixed in 4% paraformaldehyde (PFA) in diethylpyrocarbonate (DEPC)-treated phosphate buffered saline (PBS, pH 7.4) for 12 h. Heads dissected from the fixed embryos were embedded in OCT compound (Sakura Finetechnical Co. Ltd, Tokyo, Japan).…”

Section: Methodsmentioning

confidence: 99%

“…In addition, 47 highly expressed positive clones were also obtained from the E12.0 mandible (Yamaza et al 2001a). Minute in situ expression patterns of the thymosin beta 4 (Tb4), Pgk1, Hsc73, Hsj2, Hsp86, nucleolin and Set-a genes were observed in the developing lower first molar tooth germs (Akhter et al 2005;Honda et al 2008;Wada et al 2002;Xie et al 2007;Yamaza et al 2001a, b).…”

Section: Introductionmentioning

confidence: 99%

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In situ expression of ribosomal protein L21 in developing tooth germ of the mouse lower first molar

et al. 2009

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We previously performed cDNA subtraction between the mouse mandibles at embryonic day 10.5 (E10.5) in the pre-initiation stage of the odontogenesis and E12.0 in the late initiation stage to investigate the key regulator genes in odontogenesis. Ribosomal protein L21 (Rpl21) is one of differentially expressed genes in the E12.0 mandible. This study examined the precise expression pattern of Rpl21 mRNA in the mouse mandibular first molar by in situ hybridization. Rpl21 mRNA was expressed in the presumptive dental epithelium and the underlying mesenchyme at E10.5, and in the thickened dental epithelium at E12.0. Strong in situ signals were observed in the epithelial bud at E14.0, and in the enamel organ at E15.0. However, either no (E14.0) or only a weak (E15.0) in situ signal was found in the primary enamel knot at these gestational days. Rpl21 was strongly expressed in the inner enamel epithelium, cervical loop and dental lamina from E16.0 to E18.0. In addition, Rpl21 mRNA was also demonstrated in various developing cranio-facial organs. These results suggest that Rpl21 participates in the synthesis of various polypeptides which might be related to the initiation and the development of such tooth germ, and also in the synthesis of enamel components in the presecretory stage of the ameloblast. Rpl21 for protein synthesis might also be related to the morphogenesis of the developing cranio-facial organs.

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Section: Discussionsupporting

confidence: 62%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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In situ expression of ribosomal protein L21 in developing tooth germ of the mouse lower first molar

et al. 2009

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“…The culture period of both the E11.0 mandibles and E15.0 tooth germs was determined based on previous studies (Kobayashi et al, 2006;Xie et al, 2007;Ookuma et al, 2013). The protocols for organ culture were identical to those used in these studies.…”

Section: Inhibition Assay For Tb10 Using Sirna On Organ Culturementioning

confidence: 99%

“…Paraffin-embedded explants were cut into 5-mm thick sections in the anteroposterior direction, and HE staining was performed. The explants were prepared for the histological analysis as previously described (Kobayashi et al, 2006;Xie et al, 2007;Ookuma et al, 2013).…”

Section: Histological Analysis Of the Cultured Explantsmentioning

confidence: 99%

The expression and function of thymosin beta 10 in tooth germ development

Shiotsuka¹,

Wada²,

Kiyoshima³

et al. 2013

Int. J. Dev. Biol.

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This study presents the expression pattern and functions of thymosin beta 10 (Tb10), a Tb4 homologue during the development of mouse lower first molars. An in situ signal of Tb10 was detected on embryonic day 10.5 (E10.5)-E15.5 mainly in dental mesenchymal cells as well as in dental epithelial cells, while Tb4 was expressed in dental epithelial cells. In the late bell stage, preodontoblasts with strong Tb10 expression and preameloblasts with strong Tb4 expression exhibited face-to-face localization, suggesting that an intimate cell-cell interaction might exist between preodontoblasts and preameloblasts to form dentin and enamel matrices. A strong Tb10 signal was found in odontoblasts in the lateral side of the dental pulp and in Hertwig's epithelial root sheath, thus suggesting that Tb10 participates in the formation of the outline of the tooth root. An inhibition assay using Tb10-siRNA in E11.0 mandibles showed significant growth inhibition in the tooth germ. The Tb10-siRNA-treated E15.0 tooth germ also showed significant developmental arrest. The number of Ki67-positive cells significantly decreased in the Tb10-siRNA-treated mandibles. The cellular proliferative activity was also significantly suppressed in Tb10-siRNA-treated cultured mouse dental pulpal and epithelial cells. These results indicate that developmental arrest of the tooth germ might be caused by a reduction in cell proliferative activity. The stage-specific temporal and spatial expression pattern of Tb10 in the developing tooth germ is indicative of multiple functions of Tb10 in the developmental course from initiation to root formation of the tooth germ.

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Calpain‐truncated CRMP‐3 and ‐4 contribute to potassium deprivation‐induced apoptosis of cerebellar granule neurons

et al. 2009

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Increasing evidence shows that calpain-mediated proteolytic processing of a selective number of proteins plays an important role in neuronal apoptosis. Study of calpain-mediated cleavage events and related functions may contribute to a better understanding of neuronal apoptosis and neurodegenerative diseases. We, therefore, investigated the role of calpain substrates in potassium deprivation-induced apoptosis of cerebellar granule neurons (CGNs). Twelve previously known and seven novel candidates of calpain substrates were identified by 2-D DIGE and MALDI-TOF/TOF MS analysis. Further, the identified novel calpain substrates were validated by Western blot analysis. Moreover, we focused on the collapsin response mediator proteins (CRMP-1, -2, -3 and -4 isoforms) and found that CRMPs were proteolytically processed by calpain but not by caspase, both in vivo and in vitro. To clarify the properties of the calpain-mediated proteolysis of CRMPs, we constructed the deletion mutants of CRMPs for additional biochemical studies. In vitro cleavage assays revealed that CRMP-1, -2 and -4 were truncated by calpain at the C-terminus, whereas CRMP-3 was cleaved at the N-terminus. Finally, we assessed the role of CRMPs in the process of potassium deprivation-triggered neuronal apoptosis by overexpressing the truncated CRMPs in CGNs. Our data clearly showed that the truncated CRMP-3 and -4, but not CRMP-1 and -2, significantly induced neuronal apoptosis. These findings demonstrated that calpain-truncated CRMP-3 and -4 act as pro-apoptotic players when CGNs undergo apoptosis.

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Functional Implication of Nucleolin in the Mouse First Molar Development

Cited by 17 publications

References 45 publications

In situ expression of ribosomal protein L21 in developing tooth germ of the mouse lower first molar

In situ expression of ribosomal protein L21 in developing tooth germ of the mouse lower first molar

The expression and function of thymosin beta 10 in tooth germ development

Calpain‐truncated CRMP‐3 and ‐4 contribute to potassium deprivation‐induced apoptosis of cerebellar granule neurons

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