Separation of dorsal and ventral dopaminergic neurons from embryonic rat mesencephalon by buoyant density fractionation: disassembling pattern in the ventral midbrain.
The dopaminergic neurons of the ventral mesencephalon, though physically mixed with non-dopamine neurons, are organized into dorsal and ventral 'tiers' with regard to their ontogeny, efferent projections and their relative position in the various mesencephalic sub-nuclei. We have employed buoyant density fractionation to separate the dopaminergic neurons of the two compartments and compare their subsequent phenotype development with respect to their expression of the gene encoding tyrosine hydroxylase, the rate-limiting enzyme in the catecholamine biosynthetic pathway. Using immunocytochemistry, separately and combined with in situ hybridization, we demonstrate here that sedimentation of cell suspensions from E19 rat ventral mesencephalon on 5-step Percoll gradients produces cell fractions enriched in ventral and dorsal tier DA neurons, respectively.
Silverman
W F
WF
Department of Morphology, Zlotowski Center for Neuroscience, Ben-Gurion University of the Negev, Beer Sheva, Israel. szeev@bgumail.bgu.ac.il
Alfahel-Kakunda
A
A
Dori
A
A
Barker
J L
JL
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D001973:Bromodeoxyuridine; D064032:Calbindin 2; D002469:Cell Separation / Q000379:methods; D002501:Centrifugation, Zonal / Q000379:methods; D004525:Efferent Pathways / Q000166:cytology / Q000196:embryology; D004622:Embryo, Mammalian; D008636:Mesencephalon / Q000166:cytology* / Q000196:embryology / Q000378:metabolism; D009474:Neurons / Q000145:classification / Q000166:cytology* / Q000378:metabolism; D010320:Parvalbumins / Q000032:analysis; D011205:Povidone; D051381:Rats; D017207:Rats, Sprague-Dawley; D064030:S100 Calcium Binding Protein G / Q000032:analysis; D012822:Silicon Dioxide; D014158:Transcription, Genetic; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics; D014741:Video Recording
D015415:Biomarkers; D064032:Calbindin 2; D010320:Parvalbumins; D064030:S100 Calcium Binding Protein G; C016039:Percoll; D012822:Silicon Dioxide; D014446:Tyrosine 3-Monooxygenase; D011205:Povidone; D001973:Bromodeoxyuridine