Uracil nucleotides stimulate human neural precursor cell proliferation and dopaminergic differentiation: involvement of MEK/ERK signalling

Autor: Herbert Zimmermann, Florian Wegner, Sigrid Schwarz, Ute Roemuss, Javorina Milosevic, Annett Brandt, Johannes Schwarz, Alexander Storch, Antje Arnold
Rok vydání: 2006
Předmět:
MAPK/ERK pathway
Neural precursor cell proliferation
medicine.medical_specialty
P2Y receptor
Patch-Clamp Techniques
Uracil Nucleotides
Dopamine
Cellular differentiation
Blotting
Western
Fluorescent Antibody Technique
Cell Count
Nerve Tissue Proteins
Uridine Triphosphate
Suramin
Biology
Fibroblast growth factor
Biochemistry
Cellular and Molecular Neuroscience
chemistry.chemical_compound
Mesencephalon
Epidermal growth factor
Internal medicine
medicine
Humans
PPADS
Extracellular Signal-Regulated MAP Kinases
Cell Proliferation
Oligonucleotide Array Sequence Analysis
Neurons
Cell Death
Adenine Nucleotides
Receptors
Purinergic P2
Reverse Transcriptase Polymerase Chain Reaction
Stem Cells
Cell Differentiation
Cell biology
Electrophysiology
Endocrinology
chemistry
Pyridoxal Phosphate
RNA
Uracil nucleotide
Zdroj: Journal of Neurochemistry. 99:913-923
ISSN: 1471-4159
0022-3042
DOI: 10.1111/j.1471-4159.2006.04132.x
Popis: Isolation and propagation of neural stem cells derived from human brain tissue uniquely enables the study of human neurogenesis in vitro. In addition, ex vivo-expanded human neural stem/precursor cells (NPCs) may offer novel therapeutic strategies. We investigated the effects of extracellular nucleotides on the proliferation and differentiation of human mesencephalic neural stem/precursor cells (hmNPCs). When combined with the mitogens epidermal growth factor and fibroblast growth factor 2, UTP (1 microm) boosted proliferation of hmNPCs as shown by increased expression of the proliferation marker proliferating cell nuclear antigen (330%). UTP-induced proliferation was abrogated by the preferential P2Y receptor blocker pyridoxal phosphate-6-azophenyl-2',4'-disulphonic acid (PPADS). UTP also stimulated dopaminergic differentiation. Treatment with UTP (100 microm) increased the number of tyrosine hydroxylase (TH)-positive cells and TH protein by 267 and 319% respectively. UTP-stimulated dopaminergic differentiation of hmNPCs was blocked by the P2 receptor antagonists suramin (10 microm) and PPADS (100 microm). In addition, UDP (1 microm) enhanced TH protein expression by 194%. During differentiation, treatment with UTP stimulated the extracellular signal-regulated kinase (ERK) pathway. Both ERK1/2 phosphorylation and dopaminergic differentiation were inhibited by U0126, a selective ERK kinase inhibitor, as well as by suramin. When other P2 receptor agonists (ATP, ADP and adenosine 5'-O-(2-thiophosphate) (ADPbetaS); all 100 microm) were applied, both proliferation and dopaminergic differentiation of NPCs were compromised. We conclude that uracil nucleotides exert specific P2 receptor-mediated effects on midbrain-derived human NPCs, and may be used to enhance both proliferation and dopaminergic differentiation.
Databáze: OpenAIRE
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