title: Transcriptome analysis reveals transmembrane targets on transplantable midbrain dopamine progenitors.
abstract: An important challenge for the continued development of cell therapy for Parkinson's disease (PD) is the establishment of procedures that better standardize cell preparations for use in transplantation. Although cell sorting has been an anticipated strategy, its application has been limited by lack of knowledge regarding transmembrane proteins that can be used to target and isolate progenitors for midbrain dopamine (mDA) neurons. We used a "FACS-array" approach to identify 18 genes for transmembrane proteins with high expression in mDA progenitors and describe the utility of four of these targets (Alcam, Chl1, Gfra1, and Igsf8) for isolating mDA progenitors from rat primary ventral mesencephalon through flow cytometry. Alcam and Chl1 facilitated a significant enrichment of mDA neurons following transplantation, while targeting of Gfra1 allowed for robust separation of dopamine and serotonin neurons. Importantly, we also show that mDA progenitors isolated on the basis of transmembrane proteins are capable of extensive, functional innervation of the host striatum and correction of motor impairment in a unilateral model of PD. These results are highly relevant for current efforts to establish safe and effective stem cell-based procedures for PD, where clinical translation will almost certainly require safety and standardization measures in order to deliver well-characterized cell preparations.
authors.affiliation: Florey Institute for Neuroscience and Mental Health, The University of Melbourne, Parkville, Victoria, Australia 3010; and.
authors.forename: Chris R
authors.identifier: http://orcid.org/0000-0003-1714-5954
authors.initials: CR
authors.lastname: Bye
authors.affiliation: Wallenberg Neuroscience Centre, Department of Experimental Medical Science, Lund University, S-22184 Lund, Sweden.
authors.forename: Marie E
authors.initials: ME
authors.lastname: Jönsson
authors.affiliation: Wallenberg Neuroscience Centre, Department of Experimental Medical Science, Lund University, S-22184 Lund, Sweden anders.bjorklund@med.lu.se lachlant@unimelb.edu.au.
authors.forename: Anders
authors.initials: A
authors.lastname: Björklund
authors.affiliation: Florey Institute for Neuroscience and Mental Health, The University of Melbourne, Parkville, Victoria, Australia 3010; and.
authors.forename: Clare L
authors.initials: CL
authors.lastname: Parish
authors.affiliation: Florey Institute for Neuroscience and Mental Health, The University of Melbourne, Parkville, Victoria, Australia 3010; and anders.bjorklund@med.lu.se lachlant@unimelb.edu.au.
authors.forename: Lachlan H
authors.initials: LH
authors.lastname: Thompson
mesh_terms: D020410:Activated-Leukocyte Cell Adhesion Molecule / Q000235:genetics / Q000378:metabolism; D000818:Animals; D051792:Basic Helix-Loop-Helix Proteins / Q000235:genetics / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D002478:Cells, Cultured; D059290:Dopaminergic Neurons / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D060850:LIM-Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D008636:Mesencephalon / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D008815:Mice, Inbred Strains; D008822:Mice, Transgenic; D018613:Microscopy, Confocal; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism* / Q000637:transplantation; D010300:Parkinson Disease / Q000628:therapy; D017207:Rats, Sprague-Dawley; D033581:Stem Cell Transplantation / Q000379:methods*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D020410:Activated-Leukocyte Cell Adhesion Molecule; D051792:Basic Helix-Loop-Helix Proteins; D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; D049452:Green Fluorescent Proteins; D060850:LIM-Homeodomain Proteins; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D014157:Transcription Factors; C490649:Chl1 protein, mouse; C494988:Gfra1 protein, mouse; C539703:IgSF8 protein, mouse; C489692:Lmx1a protein, mouse; C102938:Neurog2 protein, mouse
keywords: Alcam; Parkinson’s disease; cell sorting; microarray; transplantation