Microarray analysis of thapsigargin - Induced stress to the endoplasmic reticulum of mouse osteoblasts

Kazunori Hamamura, Yunlong Liu, Hiroki Yokota

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Activating transcription factor 4 (ATF4) protein has a dual role in osteoblasts. It functions as a responder to stress to the endoplasmic reticulum (ER) as well as a transcription factor for bone formation. Little is known about molecular pathways that can potentially lead to stress-induced apoptosis or homeostasis of extracellular matrix (ECM) molecules. Based on microarray-derived mRNA expression data for mouse osteoblasts (MC3T3 E1 cells, clone 4), we analyzed the ER-stress responses in the presence of 10 nM Thapsigargin using two computational approaches: "Gene Set Enrichment Analysis (GSEA)" and "Ingenuity Pathways Analysis (IPA)." GSEA presented a strong linkage to an expression pattern observed in the responses to hypoxia, and IPA identified two molecular pathways: ATF4-unlinked connective tissue development and ATF4-linked organ morphology. Real-time polymerase chain reaction (PCR) and Western blot analyses validated eIF2α-driven translational regulation as well as ATF4-linked transcriptional activation of transcription factors and growth factors including FOS, FGF-9, and BMP-2. Consistent with the role of p38 MAPK in hypoxia, phosphorylation of p38 MAPK was activated in nonapoptotic osteoblasts under surviving ER stress. Furthermore, the level of phosphorylated PERK was elevated. These results support cross-talk between p38 MAPK and ER kinase, presenting a similarity to the responses to hypoxia as well as a pathway toward connective tissue development and organ morphology.

Original languageEnglish
Pages (from-to)231-240
Number of pages10
JournalJournal of Bone and Mineral Metabolism
Volume26
Issue number3
DOIs
StatePublished - 2008

Fingerprint

Activating Transcription Factor 4
Endoplasmic Reticulum Stress
Thapsigargin
Microarray Analysis
Osteoblasts
p38 Mitogen-Activated Protein Kinases
Connective Tissue
Growth Differentiation Factor 2
Transcription Factors
Osteogenesis
Endoplasmic Reticulum
Transcriptional Activation
Genes
Extracellular Matrix
Real-Time Polymerase Chain Reaction
Intercellular Signaling Peptides and Proteins
Homeostasis
Phosphotransferases
Clone Cells
Western Blotting

Keywords

  • Endoplasmic reticulum
  • Microarray
  • Osteoblasts p38
  • PERK
  • Stress

ASJC Scopus subject areas

  • Endocrinology

Cite this

Microarray analysis of thapsigargin - Induced stress to the endoplasmic reticulum of mouse osteoblasts. / Hamamura, Kazunori; Liu, Yunlong; Yokota, Hiroki.

In: Journal of Bone and Mineral Metabolism, Vol. 26, No. 3, 2008, p. 231-240.

Research output: Contribution to journalArticle

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