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Analyze Mouse Knockout Models of UPR Pathway Elements
Chao Zheng1, Cheng Wang2, Qiang Jie3
1Institute of Orthopedic Surgery, Xijing Hospital, Fourth Military Medical University, Xi'an, China.
Abstract:
Evidence from genetic studies in human and mice indicates that defective skeletal development is one of the major phenotypic outcomes for aberrant UPR signaling. Visualization of morphological alterations in whole-mount skeleton and protein secretion and UPR activation on tissue sections is the very first step to investigate skeletal phenotypes of UPR-related mouse models. In this chapter, we introduce the major techniques that have been frequently used in our laboratory to study UPR-induced skeletal disorders with genetically modified mice and provide descriptive directions of mouse genotyping, bone tissue grossing, whole-mount skeletal staining, immunostaining assays of matrix secretion, and UPR activation.
Insights
Aberrant unfolded protein response (UPR) signaling can cause defective skeletal development. This chapter details methods for studying UPR-induced skeletal disorders in genetically modified mice, focusing on skeletal morphology and protein secretion.
Area of Science:
- Molecular Biology
- Genetics
- Developmental Biology
Background:
- Genetic studies in humans and mice link aberrant unfolded protein response (UPR) signaling to defective skeletal development.
- Investigating skeletal phenotypes in UPR-related mouse models requires visualizing morphological alterations and assessing protein secretion and UPR activation.
Purpose of the Study:
- To introduce key techniques for studying UPR-induced skeletal disorders in genetically modified mice.
- To provide detailed protocols for mouse genotyping, bone tissue grossing, whole-mount skeletal staining, and immunostaining assays.
Main Methods:
- Mouse genotyping for genetic modification identification.
- Bone tissue grossing and preparation for analysis.
- Whole-mount skeletal staining for morphological assessment.
- Immunostaining assays to evaluate matrix secretion and UPR activation.
Main Results:
- The chapter outlines established laboratory techniques for analyzing skeletal phenotypes.
- Methods cover the initial steps from genetic validation to detailed tissue analysis.
Conclusions:
- The described methods are crucial for investigating the role of UPR signaling in skeletal disorders.
- This chapter serves as a practical guide for researchers studying UPR-related skeletal phenotypes in mouse models.
