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Use of Hyperosmolar Stress to Measure Stress-Activated Protein Kinase Activation and Function in Human HTR Cells and Mouse Trophoblast Stem Cells

Department of Anatomy and Cell Biology Wayne State University School of Medicine, Detroit, Michigan

Yufen Xie, MD

CS Mott Center for Human Growth and Development Wayne State University School of Medicine, Detroit, Michigan

Yingchun Wang, MD

CS Mott Center for Human Growth and Development Wayne State University School of Medicine, Detroit, Michigan

Jennifer Lewis, MS

CS Mott Center for Human Growth and Development Wayne State University School of Medicine, Detroit, Michigan

Anna Trostinskaia, MD

CS Mott Center for Human Growth and Development Wayne State University School of Medicine, Detroit, Michigan

Fangfei Wang, MD

CS Mott Center for Human Growth and Development Wayne State University School of Medicine, Detroit, Michigan

Elizabeth E. Puscheck, MD, MS

Department of Obstetrics/Gynecology, Hutzel Hospital Wayne State University School of Medicine, Detroit, Michigan

Daniel Allen Rappolee, PhD

Department of Anatomy and Cell Biology Wayne State University School of Medicine, Detroit, Michigan, drappole{at}med.wayne.edu, CS Mott Center for Human Growth and Development Wayne State University School of Medicine, Detroit, Michigan, Department of Obstetrics/Gynecology, Hutzel Hospital Wayne State University School of Medicine, Detroit, Michigan, Institute for Environmental Health and Safety Wayne State University School of Medicine, Detroit, Michigan, Department of Reproductive Sciences, Department of Physiology and Obstetrics/ Gynecology Wayne State University School of Medicine, Detroit, Michigan

Embryo growth is inversely correlated with hyperosmolar stress-induced stress-activated protein kinase/jun kinase (SAPK/JNK) induction. To examine whether stress has similar effects in stem cells derived from the embryo, the authors test trophoblast stem cells. The stress response of human placental and mouse trophoblast stem cell lines are tested here. Peak phosphorylated SAPK/JNK was induced by 400 mM sorbitol at 0.5 hours. At this dose, there is an SAPK/JNK-dependent decrease in mitogenic, phosphorylated cMyc at 0.5 hours preceding an SAPK/JNK-dependent decrease in cell cycle entrance at 24 hours. At 0.5 hours, SAPK/JNK decreases terminal deoxynucleotidyltransferase dUTP nick end labeling/apoptosis at sorbitol doses from 50 mM to 400 mM and induces phosphorylated cJun prior to an SAPK/JNK-dependent, approximate 8-fold increase in apoptosis by 24 hours at 400 mM. SAPK/JNK phosphorylation peaked at 0.5 to 4 hours and largely subsided by 12 hours. Thus, total SAPK/JNK exists before stress and mediates rapid, homeostatic molecular responses that become biologic consequences after phosphorylated SAPK/JNK ends. This suggests continuity in the homeostatic mechanisms and functions of SAPK/JNK in placental lineage cells during implantation, in which SAPK/JNK is completely responsible for cell cycle arrest and largely responsible for apoptosis.

Key Words: HTR human placental cells • trophoblast stem cells • SAPK/JNK-terminal protein kinase • TUNEL/apoptosis • cell cycle arrest.

Reproductive Sciences, Vol. 14, No. 6, 534-547 (2007)
DOI: 10.1177/1933719107307182


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