Heat shock protein 1 and the mitogen-activated protein kinase 14 pathway are important for mouse trophoblast stem cell differentiation.

Differentiation of trophoblast cells is a critical process for the proper establishment of the placenta and is, therefore, necessary to maintain embryonic development. Trophoblast stem (TS) cells grown in culture can differentiate into different trophoblast subtypes in vitro mimicking normal trophoblast cell differentiation. Therefore, TS cells are a valuable model system that can be used to elucidate genetic factors that regulate trophoblast cell differentiation. Several transcription factors, when analyzed by targeted gene mutation in mice, have resulted in embryonic lethality due to placental defects and, more specifically, defects of the trophoblast lineages. These studies have helped improve our knowledge about trophoblast cell differentiation, but much is still unknown about the specific mechanisms involved. This study uses TS cell culture to detect proteins with differential expression in proliferating and differentiating TS cells in order to identify proteins with potential roles in the differentiation process. We identified four proteins with differential expression: dimethylarginine dimethylaminohydrolase1 (DDAH1), keratin 8, keratin 18, and HSPB1 (also known as heat shock protein 25, HSP25). Further investigation confirmed the presence of HSPB1 protein during in vitro TS cell differentiation. In addition, we confirmed that phosphorylation of HSPB1 and MAP kinase-activated protein kinase 2 (MAPKAPK2) increased in TS cells during differentiation. Inhibition of MAPK14 (also known as p38 MAPK) resulted in a reduction of HSPB1 phosphorylation and an increase in cell death during TS cell differentiation. These results suggest that HSPB1 and the MAPK14 pathway are important during TS cell differentiation.