Afferent influences on striatal development in organotypic cocultures.
Organotypic cocultures of striatum, cortex, and ventral mesencephalon were used to study the anatomical and physiological development of striatal neurons in the presence or absence of cortical and nigral (SN/VTA) inputs. Striatum and cortex were dissected from prenatal (E18-E22) or early postnatal (P0-P2) rats, and SN/VTA was dissected from E14-15 fetuses; pieces were maintained up to 3 weeks in static slice culture. Triple cocultures containing SN/VTA exhibited rapid and robust dopamine (DA) innervation of the striatum in a patchy pattern, and homogeneous distribution within the cortical piece, regardless of the orientations of the three pieces. DA fibers within the striatal piece overlapped striatal patch neurons, marked by DARPP-32 immunoreactivity, in striatal cultures prepared from all age rats, but development most analogous to that seen in vivo was observed with the use of late prenatal (E20-E22) striatum. The patch/matrix organization was maintained in cultures prepared from late prenatal striatum in the presence of cortical and nigrostriatal DA afferents. In addition, a more complete transition to a patchy organization was observed in E18/19 striatal cultures in the presence of cortical and DA innervation. Electrophysiological recording demonstrated the presence of both spontaneous and cortically evoked activity in striatal medium spiny neurons; this activity was greatly influenced by the presence of DA innervation. These findings demonstrate the importance of afferent innervation in the maturation of striatal neurons in organotypic cultures.
Snyder-Keller
Abigail
A
David Axelrod Institute, New York State Department of Health, Albany, New York 12208, USA. snykell@wadsworth.org
Tseng
Kuei Y
KY
Lyng
Gregory D
GD
Graber
David J
DJ
O'Donnell
Patricio
P
D000200:Action Potentials / Q000502:physiology; D000344:Afferent Pathways / Q000166:cytology / Q000196:embryology / Q000378:metabolism; D000818:Animals; D002450:Cell Communication / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology*; D002540:Cerebral Cortex / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D018920:Coculture Techniques; D003342:Corpus Striatum / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D004298:Dopamine / Q000378:metabolism*; D051937:Dopamine and cAMP-Regulated Phosphoprotein 32 / Q000378:metabolism; D007150:Immunohistochemistry; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D019581:Neuropil / Q000378:metabolism / Q000648:ultrastructure; D009924:Organ Culture Techniques; D017729:Presynaptic Terminals / Q000378:metabolism / Q000648:ultrastructure; D051381:Rats; D017207:Rats, Sprague-Dawley; D013378:Substantia Nigra / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D009435:Synaptic Transmission / Q000502:physiology
D051937:Dopamine and cAMP-Regulated Phosphoprotein 32; C497930:Ppp1r1b protein, rat; D004298:Dopamine