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  • Itm2a expression in the developing mouse first lower molar, and the subcellular localization of Itm2a in mouse dental epithelial cells.

Itm2a expression in the developing mouse first lower molar, and the subcellular localization of Itm2a in mouse dental epithelial cells.

PloS one (2014-08-01)
Makiko Kihara, Tamotsu Kiyoshima, Kengo Nagata, Hiroko Wada, Hiroaki Fujiwara, Kana Hasegawa, Hirotaka Someya, Ichiro Takahashi, Hidetaka Sakai
ABSTRACT

Itm2a is a type II transmembrane protein with a BRICHOS domain. We investigated the temporospatial mRNA and protein expression patterns of Itm2a in the developing lower first molar, and examined the subcellular localization of Itm2a in murine dental epithelial (mDE6) cells. From the initiation to the bud stage, the in situ and protein signals of Itm2a were not detected in either the dental epithelial or mesenchymal cells surrounding the tooth bud. However, at the bell stage, these signals of Itm2a were primarily observed in the inner enamel epithelium of the enamel organ. After the initiation of the matrix formation, strong signals were detected in ameloblasts and odontoblasts. Itm2a showed a punctate pattern in the cytoplasm of the mDE6 cells. The perinuclear-localized Itm2a displayed a frequent overlap with the Golgi apparatus marker, GM130. A tiny amount of Itm2a was colocalized with lysosomes and endoplasmic reticulum. Minimal or no overlap between the Itm2a-EGFP signals with the other organelle markers for endoplasmic reticulum, lysosome and mitochondria used in this study noted in the cytoplasm. These findings suggest that Itm2a may play a role in cell differentiation during odontogenesis, rather than during the initiation of tooth germ formation, and may be related to the targeting of proteins associated with enamel and dentin matrices in the secretory pathway.

MATERIALS
Product Number
Brand
Product Description

Supelco
Digoxigenin, analytical standard
Sigma-Aldrich
Anti-GM130 (C-terminal) antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Digoxigenin, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
Anti-Mitofusin-2 (N-Terminal) antibody produced in rabbit, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
Anti-LAMP2 antibody produced in rabbit