Glial cells promote dendrite formation and the reception of synaptic input in Purkinje cells from postnatal mice.
Previous studies suggest that glial cells contribute to synaptogenesis in specific neurons from the postnatal CNS. Here, we studied whether this is true for Purkinje cells (PCs), which represent a unique neuronal cell type due to their large size, massive synaptic input, and high vulnerability. Using new glia-free cultures enriched in PCs from postnatal mice we show that these neurons survived and grew, but displayed only low levels of excitatory and inhibitory synaptic activity. Coculture with glial cells strongly enhanced the frequency and size of spontaneous and miniature excitatory synaptic currents as well as neurite growth and branching. Immunocytochemical staining for microtubule-associated protein 2- (MAP2-) positive neurites revealed impaired dendrite formation in PCs under glia-free conditions, which can explain the absence of synaptic activity. Glial signals strongly enhanced dendritogenesis in PCs and thus their ability to receive excitatory synaptic input from granule cells (GCs). The enhancement of dendrite formation was mimicked by glia-conditioned medium (GCM), whereas the increase in synaptic activity required physical presence of glia. This indicated that dendrite development is necessary but not sufficient for PCs to receive excitatory synaptic input and that synaptogenesis requires additional signals. The level of inhibitory synaptic activity was low even in cocultures due to a low incidence of inhibitory interneurons. Taken together, our results reinforce the idea that glial cells promote synaptogenesis in specific neuronal cell types.
Buard
Isabelle
I
CNRS UPR 3212, University of Strasbourg, Institute for Cellular and Integrative Neurosciences (INCI), 5, rue Blaise Pascal, F-67084 Strasbourg Cedex, France.
Steinmetz
Celine C
CC
Claudepierre
Thomas
T
Pfrieger
Frank W
FW
D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology; D018920:Coculture Techniques / Q000379:methods; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003712:Dendrites / Q000187:drug effects / Q000502:physiology*; D019706:Excitatory Postsynaptic Potentials / Q000187:drug effects / Q000502:physiology; D005968:Glutamate Decarboxylase / Q000378:metabolism; D039841:Leukocyte L1 Antigen Complex / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009457:Neuroglia / Q000737:chemistry / Q000502:physiology*; D018408:Patch-Clamp Techniques / Q000379:methods; D011689:Purkinje Cells / Q000166:cytology*; D013569:Synapses / Q000187:drug effects / Q000502:physiology*; D018800:Thy-1 Antigens / Q000378:metabolism
D017077:Culture Media, Conditioned; D039841:Leukocyte L1 Antigen Complex; D008869:Microtubule-Associated Proteins; C079240:Mtap2 protein, mouse; D018800:Thy-1 Antigens; D005968:Glutamate Decarboxylase