Abstract
BackgroundHuman fetal midbrain-derived neural progenitor cells (NPCs) may deliver a tissue source for drug screening and regenerative cell therapy to treat Parkinson’s disease. While glutamate and GABAA receptors play an important role in neurogenesis, the involvement of glycine receptors during human neurogenesis and dopaminergic differentiation as well as their molecular and functional characteristics in NPCs are largely unknown.Methodology/Principal FindingsHere we investigated NPCs in respect to their glycine receptor function and subunit expression using electrophysiology, calcium imaging, immunocytochemistry, and quantitative real-time PCR. Whole-cell recordings demonstrate the ability of NPCs to express functional strychnine-sensitive glycine receptors after differentiation for 3 weeks in vitro. Pharmacological and molecular analyses indicate a predominance of glycine receptor heteromers containing α2β subunits. Intracellular calcium measurements of differentiated NPCs suggest that glycine evokes depolarisations mediated by strychnine-sensitive glycine receptors and not by D-serine-sensitive excitatory glycine receptors. Culturing NPCs with additional glycine, the glycine-receptor antagonist strychnine, or the Na+-K+-Cl− co-transporter 1 (NKCC1)-inhibitor bumetanide did not significantly influence cell proliferation and differentiation in vitro. Conclusions/SignificanceThese data indicate that NPCs derived from human fetal midbrain tissue acquire essential glycine receptor properties during neuronal maturation. However, glycine receptors seem to have a limited functional impact on neurogenesis and dopaminergic differentiation of NPCs in vitro.
Highlights
Glycine is an important inhibitory neurotransmitter in the adult central nervous system acting through ionotropic glycine receptors that are most prominently expressed in the brainstem and spinal cord [1,2,3]
neural progenitor cells (NPCs) differentiated for 3 weeks showed glycine-induced Ca2+ transients that were completely suppressed by the glycine receptor antagonist strychnine but were only slightly affected by D-serine (Fig. 1A)
Calcium imaging and electrophysiological recordings demonstrated the expression of excitatory strychnine-sensitive glycine receptors in differentiated human midbrain-derived NPCs
Summary
Glycine is an important inhibitory neurotransmitter in the adult central nervous system acting through ionotropic glycine receptors that are most prominently expressed in the brainstem and spinal cord [1,2,3]. These receptors belong to the superfamily of Cysloop receptors such as GABAA, nicotinic acetylcholine, and 5HT3 receptors [4]. While glutamate and GABAA receptors play an important role in neurogenesis, the involvement of glycine receptors during human neurogenesis and dopaminergic differentiation as well as their molecular and functional characteristics in NPCs are largely unknown
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