CD44 isoform expression in periodontal tissues: cell-type specific regulation of alternative splicing.
CD44 functions as a receptor for various extracellular matrices and plays crucial roles in homotypic and heterotypic cell-cell interactions. Recently, the molecular structure of CD44 has been extensively analyzed and multiple isoforms produced by alternative splicing of messenger RNA have been identified. In this study, we examined the expression of CD44 isoforms on different cell types isolated from periodontal tissue. In order to examine tissue differences in CD44 isoform expression, we established in vitro cell culture of human gingival fibroblasts (HGF), human periodontal ligament cells (HPDL) and human gingival epithelial cells (HGEC). These cells all expressed CD44 protein and messenger RNA. However, immunoprecipitation and Northern blot analysis revealed that HGEC expressed larger CD44 isoforms than HGF and HPDL. Reverse transcription-polymerase chain reaction with primers flanking the insertion site of alternatively spliced exons was used to study details of the heterogeneity. All cells examined expressed a major band in the absence of alternatively spliced exons and additional larger bands. In particular, HGEC contained more abundant high molecular mass species. In vitro stimulation by IL-1 beta, TNF alpha or phorbol 12-myristate 13-acetate induced an increase in total CD44 messenger RNA in HGF but not change in overall patterns of CD44 isoform expression. However, the isoform expression of HGEC was sensitive to cell density. The amount of larger isoform was decreased by culturing cells beyond confluence. These findings suggest that CD44 isoform expression is cell type-specifically regulated in periodontium and altered according to growth phase of HGEC.
Hirano
F
F
Department of Periodontology and Endodontology, Osaka University Faculty of Dentistry, Japan.
Hirano
H
H
Hino
E
E
Takayama
S
S
Saito
K
K
Kusumoto
Y
Y
Shimabukuro
Y
Y
Murakami
S
S
Okada
H
H
D017398:Alternative Splicing; D015152:Blotting, Northern; D002450:Cell Communication; D002452:Cell Count; D002453:Cell Cycle / Q000235:genetics; D002478:Cells, Cultured; D017931:DNA Primers; D004847:Epithelial Cells / Q000166:cytology / Q000276:immunology; D005091:Exons / Q000235:genetics; D005109:Extracellular Matrix / Q000276:immunology; D005347:Fibroblasts / Q000166:cytology / Q000276:immunology; D005786:Gene Expression Regulation; D005881:Gingiva / Q000166:cytology / Q000276:immunology*; D006801:Humans; D018960:Hyaluronan Receptors / Q000032:analysis / Q000145:classification / Q000235:genetics*; D007375:Interleukin-1 / Q000494:pharmacology; D010513:Periodontal Ligament / Q000166:cytology / Q000276:immunology*; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D012333:RNA, Messenger / Q000235:genetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D017931:DNA Primers; D018960:Hyaluronan Receptors; D007375:Interleukin-1; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D013755:Tetradecanoylphorbol Acetate