The developmental loss of the ability of Purkinje cells to regenerate their axons occurs in the absence of myelin: an in vitro model to prevent myelination.
Axonal regeneration in the mammalian CNS is a property of immature neurons that is lost during development. Using organotypic culture of cerebellum, we have shown that in vitro Purkinje cells lose their regenerative capacity in parallel with the process of myelination. We have investigated whether myelination is involved in the age-dependent loss of regeneration of these neurons. By applying a high dose of bromodeoxyuridine in the culture medium of newborn cerebellar slices during the first 3 d in vitro, we have succeeded in obtaining cultures with oligodendrocyte depletion, together with a lack of ameboid microglia and enhancement of Purkinje cell survival. These cultures, after 14 d in vitro, are completely devoid of myelin. We have compared the ability of Purkinje cells to regenerate their axons in the presence or absence of myelin. Purkinje cells in cerebellar explants taken at birth, treated with bromodeoxyuridine and axotomized after 7 d in vitro, survive better than similar neurons in untreated cultures. However, despite the lack of myelin and the enhanced survival, Purkinje cells do not regenerate, whereas they do regenerate when the axotomy is done at postnatal day 0. Thus, the Purkinje cell developmental switch from axonal regeneration to lack of regeneration does not appear to be regulated by myelin.
Bouslama-Oueghlani
Lamia
L
Institut National de la Santé et de la Recherche Médicale U106, Hôpital de la Salpêtrière, 75651 Paris, France.
Wehrlé
Rosine
R
Sotelo
Constantino
C
Dusart
Isabelle
I
D000367:Age Factors; D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000187:drug effects; D001369:Axons / Q000187:drug effects / Q000502:physiology*; D019771:Axotomy; D001973:Bromodeoxyuridine / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D066298:In Vitro Techniques; D051379:Mice; D018345:Mice, Knockout; D017628:Microglia / Q000187:drug effects; D009186:Myelin Sheath / Q000187:drug effects / Q000378:metabolism*; D009416:Nerve Regeneration / Q000187:drug effects / Q000502:physiology*; D009836:Oligodendroglia / Q000187:drug effects; D011689:Purkinje Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D013997:Time Factors
D001973:Bromodeoxyuridine