title: Nuclear transfer embryonic stem cells provide an in vitro culture model for Parkinson's disease.
abstract: Somatic cell nuclear transfer enables the generation of embryonic stem cells (ESCs) that genetically match the donor and can be used to treat disease through cell replacement therapies or to recapitulate patient-specific disease via in vitro differentiation. We performed a "proof-of-principle" study using tail tip fibroblasts from a mouse model of Parkinson's disease (Aphakia) as the donor cell nuclei for nuclear transfer and derived "customized" ESCs for in vitro analysis. Aphakia mice contain deletions in the pitx3 gene and show selective loss of dopamine neurons of the substantia nigra, specifically the neuron population susceptible to degeneration in Parkinson's disease. Using electrofusion nuclear transfer, we produced cloned Aphakia oocytes at rates similar to those for control, cloned oocytes. Aphakia ESCs were isolated and live mice were generated using tetraploid embryo complementation. In vitro differentiation of Aphakia ESCs to dopaminergic neurons revealed significantly fewer TH+ neurons that expressed MAP2, DAT, synaptophysin, VMAT2, and AHD2 compared to control nuclear transfer ESC cultures, supporting a role for Pitx3 in mesodiencephalic dopamine neuron maturation. Taken together, our studies define a customized in vitro ESC culture system used to analyze gene-specific contribution to dopamine neuron generation, maturation, and susceptibility to degeneration.
authors.affiliation: Center for Regenerative Biology, University of Connecticut, Storrs, USA.
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Amano
authors.forename: Theodora
authors.initials: T
authors.lastname: Papanikolaou
authors.forename: Li-Ying
authors.initials: LY
authors.lastname: Sung
authors.forename: Jessica
authors.initials: J
authors.lastname: Lennington
authors.forename: Joanne
authors.initials: J
authors.lastname: Conover
authors.forename: Xiangzhong
authors.initials: X
authors.lastname: Yang
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D004195:Disease Models, Animal; D050483:Dopamine Plasma Membrane Transport Proteins / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000378:metabolism* / Q000473:pathology; D018398:Homeodomain Proteins / Q000235:genetics; D051379:Mice; D008817:Mice, Mutant Strains; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D055495:Neurogenesis; D009474:Neurons / Q000378:metabolism* / Q000473:pathology; D053652:Nuclear Transfer Techniques; D010300:Parkinson Disease / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D050697:Retinal Dehydrogenase / Q000378:metabolism; D016708:Synaptophysin / Q000378:metabolism; D014157:Transcription Factors / Q000235:genetics; D050493:Vesicular Monoamine Transport Proteins / Q000378:metabolism
chemical_list: D050483:Dopamine Plasma Membrane Transport Proteins; D018398:Homeodomain Proteins; D008869:Microtubule-Associated Proteins; C079240:Mtap2 protein, mouse; C493453:Slc18a2 protein, mouse; D016708:Synaptophysin; D014157:Transcription Factors; D050493:Vesicular Monoamine Transport Proteins; C433629:homeobox protein PITX3; D050697:Retinal Dehydrogenase