About the Project

A critical issue for regulation of organ development is the temporal and spatial control of gene and protein expression during progressive developmental stages. To understand morphogenesis and differentiation of specific cell-types, it is necessary to identify the cell-type expression and developmental expression pattern of signaling pathways at a cellular level. Although profiles of gene expression at the mRNA level have been performed in the salivary gland at the mRNA level (sgmap.nidcr.nih.gov), this is the first atlas to profile protein expression in developing salivary glands in developing tissues. With this Atlas, we utilize a novel fluorescent multiplexed immunohistochemistry method that was developed at General Electric’s Global Research Center to profile expression of multiple proteins in the same tissue section.  The multiplexing technology was developed by GE to be used for identifying complex biomarker signatures in human patients. Understanding the development of the salivary gland will provide insight into development of salivary gland cancers since many developmental pathways are reawakened in tumors.

The multiplexed immunohistochemistry technology relies upon antibodies that have been validated for specificity and directly conjugated to a fluorophore. The fluorophore is then inactivated, allowing subsequent rounds of immunohistochemistry to be performed on the same tissue section. The images have been registered to the DAPI stain in the first round and background autofluorescence has been computationally subtracted. Most of the antibodies used in this study are commercially available and information on these antibodies is provided.

This project was funded following peer-review by the American Recovery and Reinvestment Act (ARRA) with funds provided by the National Institutes of Dental Research (NIDCR) at the National Institutes of Health (NIH). It was performed by Dr. Melinda Larsen’s laboratory in collaboration with researchers at GE’s Research and Development laboratory, GE Global Research in Niskayuna, NY, in the Diagnostics and Biomedical Technologies laboratory.

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