Prokineticin-1 modulates proliferation and differentiation of enteric neural crest cells

Abstract

Prokineticins (Prok-1 and Prok-2) belong to a newly identified AVIT protein family. They are involved in variety of activities in various tissues, including smooth muscle contraction of the gastrointestinal tract and promoting proliferation of endothelial cells derived from adrenal gland. Importantly, they also act as the survival factors to modulate growth and survival of neurons and hematopoietic stem cells. In this study we demonstrated that Prok-1 (but not Prok-2) protein is expressed in the mucosa and mesenchyme of the mouse embryonic gut during enteric nervous system development. Its receptor, PK-R1 is expressed in the enteric neural crest cells (NCCs). To elucidate the physiological role(s) of Prok-1 in NCCs, we isolated the NCCs from the mouse embryonic gut (E11.5) and cultured them in the form of neurospheres. In an in vitro NCC culture, Prok-1 was able to activate both Akt and MAPK pathways and induce the proliferation and differentiation (but not migration) of NCCs via PK-R1. Knock-down of PK-R1 using siRNA resulted in a complete abolishment of Prok-1 induced proliferation. Taken together, it is the first report demonstrating that Prok-1 acts as a gut mucosa/mesenchyme-derived factor and maintains proliferation and differentiation of enteric NCCs.

Hedgehog/Notch-induced premature gliogenesis represents a new disease mechanism for Hirschsprung disease in mice and humans

“…Digested guts were triturated into single cells and filtered through cell strainers (100-μm strainers and then 40-μm strainers, Falcon, BD Biosciences). Cells were resuspended in neural crest medium, DMEM containing 15% chick embryo extract (SIL), FGF (20 ng/ml, SigmaAldrich), EGF (20 ng/ml, Sigma-Aldrich), Retinoic acid (35 ng/ml, SigmaAldrich), N2 (1%), B27 (2%, Invitrogen), and β-mercaptoethanol (50 mM, Sigma-Aldrich); plated onto poly-d-lysine-coated and fibronectin-coated (20 μg/ml each, Invitrogen) wells; and enriched by multiple replatings as described previously (15,32,33). The ENCCs at passage 3 were harvested for quantitative RT-pCR or seeded at clonal density (1 × 10 5 to 1.…”

Endocrine gland-derived vascular endothelial growth factor in rat pancreas: genetic expression and testosterone regulation

“…The aminoterminal sequence of these proteins is identical; the first four residues are conserved in all of them, giving the name to this family (Soga et al 2002, Kaser et al 2003. EG-VEGF regulates diverse biological functions associated with neuronal survival, angiogenesis, cell proliferation, migration, and fenestration (formation of membrane discontinuities) in capillary endothelial cells , Dorsch et al 2005, Ngan et al 2007a. The human EG-VEGF gene promoter has a binding site for steroidogenic factor 1, which is essential in controlling the expression of all the steroidogenic enzymes and cholesterol transporters required for steroidogenesis (Bakker et al 2001, Morales et al 2006.…”

Genetic Inactivation of Prokineticin Receptor-1 Leads to Heart and Kidney Disorders

“…These 2 secreted proteins from the AVIT secreted protein family are widely distributed in mammalian tissues and human blood cells. 1 In addition to being strong angiogenic factors, 2 prokineticins act as survival/mitogenic factors in various cells, including endothelial cells, 3 neuronal cells, 4 lymphocytes and hematopoietic stem cells, 5 and cardiomyocytes. 6 They exert their biological effects by stimulating 2 closely related receptors, prokineticin receptor-1 (PKR1) and PKR2.…”