Related Experiment Video
Updated: Feb 24, 2026
Isolation of Whole Cell Protein Lysates from Mouse Facial Processes and Cultured Palatal Mesenchyme Cells for Phosphoprotein Analysis
Published on: April 1, 2022
Differential expression of transforming growth factor-beta1, connective tissue growth factor, phosphorylated-SMAD2/3
Shubo Li1, Yihuai Pan2,3
1The Institute of Stomatology, School and Hospital of Stomatology, Wenzhou Medical University, Wenzhou, Zhejiang, People's Republic of China.
Abstract:
Connective tissue growth factor (CTGF) is a downstream mediator of transforming growth factor-beta 1 (TGF-β1) and TGF-β1-induced CTGF expression is regulated through SMAD and mitogen-activated protein kinase (MAPK) signaling pathways. The fine modulation of TGF-β1 signaling is very important to the process of tooth development. However, little is known about the localization of CTGF, MAPK and SMAD in the context of TGF-β1 signaling during odontogenesis. Hence, we aimed to investigate the expression of TGF-β1, CTGF, phosphorylated-SMAD2/3 (p-SMAD2/3) and phosphorylated-ERK1/2 (p-ERK1/2). ICR mice heads of embryonic (E) day 13.5, E14.5, E16.5, postnatal (PN) day 0.5 and PN3.5 were processed for immunohistochemistry. Results revealed that at E13.5, TGF-β1 and CTGF were strongly expressed in dental epithelium (DE) and dental mesenchyme (DM), while p-SMAD2/3 was intensely expressed in the internal side of DE. p-ERK1/2 was not present in DE or DM. At E14.5 and E16.5, strong staining for TGF-β1 and CTGF was detected in enamel knot (EK) and dental papilla (DPL). DPL was intensely stained for p-ERK1/2 but negatively stained for p-SMAD2/3. There was no staining for p-SMAD2/3 and p-ERK1/2 in EK. At PN0.5 and PN3.5, moderate to intense staining for TGF-β1 and CTGF was evident in preameloblasts (PA), secretary ameloblasts (SA) and dental pulp (DP). p-SMAD2/3 was strongly expressed in SA and DP but sparsely localized in PA. p-ERK1/2 was intensely expressed in DP, although negative staining was observed in PA and SA. These data demonstrate that TGF-β1 and CTGF show an identical expression pattern, while p-SMAD2/3 and p-ERK1/2 exhibit differential expression, and indicate that p-SMAD2/3 and p-ERK1/2 might play a regulatory role in TGF-β1 induced CTGF expression during tooth development.
Insights
Transforming growth factor-beta 1 (TGF-β1) and connective tissue growth factor (CTGF) share similar expression patterns during tooth development. Phosphorylated SMAD2/3 and ERK1/2 signaling pathways show differential localization, suggesting regulatory roles in TGF-β1-induced CTGF expression.
Area of Science:
- Developmental Biology
- Cell Signaling
- Molecular Biology
Background:
- Connective tissue growth factor (CTGF) mediates transforming growth factor-beta 1 (TGF-β1) effects.
- TGF-β1 signaling, involving SMAD and MAPK pathways, is crucial for tooth development.
- The precise localization of CTGF, MAPK, and SMAD during odontogenesis remains unclear.
Purpose of the Study:
- To investigate the spatiotemporal expression of TGF-β1, CTGF, phosphorylated-SMAD2/3 (p-SMAD2/3), and phosphorylated-ERK1/2 (p-ERK1/2) during mouse tooth development.
Main Methods:
- Immunohistochemistry was performed on ICR mouse heads at embryonic days 13.5, 14.5, 16.5, and postnatal days 0.5, 3.5.
Main Results:
- TGF-β1 and CTGF exhibited consistent expression patterns throughout tooth development.
- p-SMAD2/3 and p-ERK1/2 displayed distinct expression patterns in various tooth germ compartments.
- Differential expression of p-SMAD2/3 and p-ERK1/2 was observed in the dental epithelium, mesenchyme, enamel knot, dental papilla, and ameloblasts.
Conclusions:
- TGF-β1 and CTGF expression patterns are similar during odontogenesis.
- p-SMAD2/3 and p-ERK1/2 signaling pathways exhibit differential localization, indicating distinct roles.
- These findings suggest that p-SMAD2/3 and p-ERK1/2 may regulate TGF-β1-induced CTGF expression during tooth development.
