Kidney Week

Abstract: FR-OR046

Mass Spectrometric Analysis of the Extracellular Matrix During Renal Development

Session Information

• Development and Stem Cells
  November 08, 2019 | Location: 152, Walter E. Washington Convention Center
  Abstract Time: 05:30 PM - 05:42 PM

Category: Development, Stem Cells, and Regenerative Medicine

• 500 Development, Stem Cells, and Regenerative Medicine

Authors

• Lipp, Sarah N., Purdue University, West Lafayette, Indiana, United States

Sarah N. Lipp



Sarah Lipp is in the Calve lab at Purdue University, studying extracellular matrix changes with development. She is a 2nd-year biomedical engineering graduate student in the Indiana University School of Medicine medical scientist training program.

• Jacobson, Kathryn R., Purdue University, West Lafayette, Indiana, United States

Kathryn R. Jacobson,

• Hains, David S., Indiana University, Zionsville, Indiana, United States

David S. Hains,

• Schwaderer, Andrew L., Indiana University, Zionsville, Indiana, United States

Andrew L. Schwaderer,

• Calve, Sarah, Purdue University, West Lafayette, Indiana, United States

Sarah Calve,

Background

The extracellular matrix (ECM) is a network of macromolecules that interacts with renal cells to regulate development, physiology, and pathology. Defects in specific ECM associated genes affect metanephric kidney initiation, branching, tubulogenesis, terminal differentiation of intercalated cells, and integrity of the glomerular basement membrane. However, global changes in the composition and turnover of kidney ECM as a function of development are unknown.

Methods

We used mass spectrometry (MS) to analyze embryonic (E18.5) and adult murine kidney proteins fractionated by differential solubilities and decellularization techniques for ECM 3D visualization using confocal microscopy

Results

We resolved increased expression of COL4A3, COL4A4, COL4A5, LAMA5, and LAMB2, which are critical for the integrity of the glomerular basement membrane in the adult. Additionally, FREM1, FREM2, and FRAS1, which are necessary for metanephric induction, were selectively upregulated at E18.5 (Fig 1a). The ECM of decellularized kidneys from different developmental stages could be visualized in 3D using confocal microscopy (Fig 1b).

Conclusion

Together, this work shows that the ECM changes during development can be resolved by MS. We aim to observe how global protein abundance determined by MS correlates to 3D spatiotemporal organization in ECM proteins in the kidney. This information can be used to tailor the scaffold environment in regenerative medicine applications.

Figure 1: (a) Volcano plot comparing ECM of E18.5 and adult murine kidneys. Proteins that are different between time points (>2-fold difference with a false discovery rate of 1%) are indicated with a circle. (b) Decellularization enables the 3D visualization of E13.5-P20 kidneys stained with wheat germ agglutinin to visualize the ECM. Color portrays depth (blue to yellow), scale bar=100 µm.

Funding

• Other NIH Support