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The  Ezh2  gene encodes a histone methyltransferase of the polycomb repressive complex 2 that methylates histone H3 lysine 27. In this study, we investigated whether EZH2 has a role in the development of the pharyngeal apparatus and whether it regulates the expression of the  Tbx1  gene, which encodes a key transcription factor required in pharyngeal development. To these ends, we performed genetic  in vivo  experiments with mouse embryos and used mouse embryonic stem cell (ESC)-based protocols to probe endoderm and cardiogenic mesoderm differentiation. Results showed that EZH2 occupies the  Tbx1  gene locus in mouse embryos, and that suppression of EZH2 was associated with reduced expression of  Tbx1  in differentiated mouse ESCs. Conditional deletion of  Ezh2  in the  Tbx1  expression domain, which includes the pharyngeal endoderm, did not cause cardiac defects but revealed that the gene has an important role in the morphogenesis of the third pharyngeal pouch (PP). We found that in conditionally deleted embryos the third PP was hypoplastic, had reduced expression of  Tbx1 , lacked the expression of  Gcm2 , a gene that marks the parathyroid domain, but expressed  FoxN1 , a gene marking the thymic domain. Consistently, the parathyroids did not develop, and the thymus was hypoplastic. Thus,  Ezh2  is required for parathyroid and thymic development, probably through a function in the pouch endoderm. This discovery also provides a novel interpretational key for the finding of  Ezh2  activating mutations in hyperparathyroidism and parathyroid cancer.

EZH2 is required for parathyroid and thymic development through differentiation of the third pharyngeal pouch endoderm