Rabbit Anti-Human ERK1/ERK2 (phospho Tyr204) monoclonal antibody, clone C.853.6

In mammals, human chorionic gonadotropin (hCG), a luteinizing hormone (LH) analogue, induces MAPK3/1 phosphorylation in the granulosa cells and this event is largely dependent on epidermal growth factor receptor (EGFR) activity. However, whether this mechanism also works in other vertebrates such as fish remains unknown. Here, we showed that treatment of cultured zebrafish ovarian follicle cells with hCG also resulted in MAPK3/1 phosphorylation without affecting the total protein level of MAPK3/1. The phosphorylation level peaked at 5 min and then declined to the basal level after 40 min of hCG treatment. Further experiment showed that H89 (a PICA inhibitor) could abolish hCG-stimulated MAPK3/1 phosphorylation, but had no effect on EGF-induced phosphorylation, suggesting a mediating role for cAMP/PKA in hCG activation of MAPK3/1. On the other hand, AG1478 (an EGFR inhibitor) completely blocked EGF-stimulated MAPK3/1 phosphorylation, but had no effect on the hCG-induced MAPK3/1 phosphorylation. These data indicate that similar to its action in mammals, hCG/LH also stimulated MAPK3/1 phosphorylation in the zebrafish ovarian follicle cells; however, unlike the situation in the mammalian ovary, the hCG-stimulated MAPK3/1 phosphorylation in cultured zebrafish ovarian follicle cells was independent of EGFR. (C) 2013 Elsevier Inc. All rights reserved.

Hepcidin (HAMP) synthesis is suppressed by erythropoiesis to increase iron availability for red blood cell production. This effect is thought to result from factors secreted by erythroid precursors. Growth differentiation factor 11 (GDF11) expression was recently shown to increase in erythroid cells of beta-thalassaemia, and decrease with improvement in anaemia. Whether GDF11 regulates hepatic HAMP production has never been experimentally studied. Here, we explore GDF11 function during erythropoiesis-triggered HAMP suppression. Our results confirm that exogenous erythropoietin significantly increases Gdf11 as well as Erfe (erythroferrone) expression, and Gdf11 is also increased, albeit at a lower degree than Erfe, in phlebotomized wild type and beta-thalassaemic mice. GDF11 is expressed predominantly in erythroid burst forming unit- and erythroid colony-forming unit- cells during erythropoiesis. Exogeneous GDF11 administration results in HAMP suppression in vivo and in vitro. Furthermore, exogenous GDF11 decreases BMP-SMAD signalling, enhances SMAD ubiquitin regulatory factor 1 (SMURF1) expression and induces ERK1/2 (MAPK3/1) signalling. ERK1/2 signalling activation is required for GDF11 or SMURF1-mediated suppression in BMP-SMAD signalling and HAMP expression. This research newly characterizes GDF11 in erythropoiesis-mediated HAMP suppression, in addition to ERFE.