# Characterization of PDGFR alpha/beta heterodimers > **NIH NIH F32** · UNIVERSITY OF COLORADO DENVER · 2024 · $81,895 ## Abstract Project Summary Craniofacial development is a complex process that requires various signaling pathways to mediate cross-talk between tissues that eventually differentiate into the cartilage and bone of the frontonasal skeleton. Defects in this process result in common craniofacial malformations, such as cleft lip and palate. Signaling through the platelet-derived growth factor receptors (PDGFRs) plays a critical role in both human and mouse craniofacial development. PDGFRa has been shown to play a predominant role in NCC migration, contribute to proliferation of the facial mesenchyme at mid-gestation and promote osteoblast differentiation. Alternatively, PDGFRb primarily regulates proliferation of the facial mesenchyme past mid-gestation. Further, PDGFRa and PDGFRb have been shown to genetically and physically interact in the craniofacial mesenchyme to form functional heterodimers, though the mechanism and function of signaling through these heterodimers remains unknown. We have used an innovative approach, bimolecular fluorescence complementation (BiFC), to explore individual, activated PDGFR dimers, which has revealed preliminary differences in dimer-specific activation, trafficking and downstream signaling dynamics. The goal of this proposal is to fully characterize these dynamics for PDGFRa/b heterodimers in vitro and in vivo, and to identify PDGFR dimer-specific interacting proteins that mediate differential trafficking of the various PDGFR dimers. First, PDGFRa/b heterodimers will be immunoprecipitated using a GFP-Trap nanobody in response to a timecourse of PDGF- BB ligand stimulation to examine the dimerization and autophosphorylation dynamics of PDGFRa/b heterodimers. Then, fluorescence microscopy experiments will be performed to examine co-localization of PDGFRa/b heterodimers with markers of various endosomal compartments in response to a timecourse of PDGF-BB ligand stimulation to examine the trafficking dynamics of PDGFRa/b heterodimers. These findings will be compared to our previous results for the PDGFR homodimers. Second, BiFC-tagged PDGFRa homodimers, PDGFRb homodimers and PDGFRa/b heterodimers will be purified using the GFP-Trap nanobody and analyzed by mass spectrometry to identify PDGFR dimer-specific interacting proteins. Novel proteins with demonstrated roles in receptor trafficking will be both overexpressed and repressed in the relevant PDGFR-BiFC stable cell line in the presence of PDGF ligand, and trafficking of the various PDGFR dimers will be assessed as above. Finally, Venus expression will be analyzed in craniofacial structures of E8.5- E16.5 embryos that are double-homozygous for PdgfraV1 and PdgfrbV2 BiFC knock-in alleles both in whole mount and in coronal frozen sections by fluorescence microscopy to examine the timing and localization of PDGFRa/b heterodimer formation during craniofacial development. The studies proposed here will determine how biological specificity is introduced downstream of individual PDGFR... ## Key facts - **NIH application ID:** 10923825 - **Project number:** 5F32DE032554-02 - **Recipient organization:** UNIVERSITY OF COLORADO DENVER - **Principal Investigator:** Maria B Campana - **Activity code:** F32 (R01, R21, SBIR, etc.) - **Funding institute:** NIH - **Fiscal year:** 2024 - **Award amount:** $81,895 - **Award type:** 5 - **Project period:** 2023-09-01 → 2026-08-31 ## Primary source NIH RePORTER: https://reporter.nih.gov/project-details/10923825 ## Citation > US National Institutes of Health, RePORTER application 10923825, Characterization of PDGFR alpha/beta heterodimers (5F32DE032554-02). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10923825. Licensed CC0. --- *[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*