Biomedical subjects

Ludovic Collin

Publications and source records attributed to Ludovic Collin.

3 recordsLinked to original sources

PubMed2006

Nucleofection of primary neurons.

Efficient gene transfer is an important tool for the study of neuronal function and biology. This has proved difficult and inefficient with traditional transfection strategies, which can also be fairly toxic, whereas viral-mediated gene transfer, although highly efficient, is often time-consuming. The recently developed Amaxa Nucleofector technology, based on electroporation with preset parameters in a cell-type-specific solution, enables direct delivery of DNA, small interfering (si)RNA oligonucleotides and siRNA vectors into the cell nucleus. This strategy results in reproducible, rapid, and efficient transfection of a broad range of cells, including primary neurons. Nucleofected neurons survive for up to 3 weeks and remain functional. We are currently using this transfection method to examine the contribution of Rho GTPase signaling pathways in the establishment of neuronal polarity, neuronal migration, and neurite outgrowth. Here, we describe three protocols to efficiently nucleofect rat cerebellar granule, cortical, and hippocampal neurons.

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PubMed2003

Motor training compensates for cerebellar dysfunctions caused by oligodendrocyte ablation.

The role played by oligodendrocytes (OLs), the myelinating cells of the CNS, during brain development has not been fully explored. We have addressed this question by inducing a temporal and reversible ablation of OLs on postnatal CNS development. OL ablation in newborn mice leads to a profound alteration in the structure of the cerebellar cortex, which can be progressively rescued by newly generated cells, leading to a delayed myelination. Nevertheless, the temporal shift of the OL proliferation and myelinating program cannot completely compensate for developmental defects, resulting in impaired motor functions in the adult. Strikingly, we show that, despite these abnormalities, epigenetic factors, such as motor training, are able to fully rescue cerebellar-directed motor skills.

Animals↗

PubMed2003

Oligodendrocyte ablation impairs cerebellum development.

Oligodendrocytes (OLs) are the glial cells of the central nervous system and are classically known to form myelin sheaths around most axons of higher vertebrates. Whether these cells might have other roles, in particular during development, has not been studied. Taking advantage of a transgenic mouse model in which OLs can be selectively killed in a desired time-frame, we have investigated the impact of OL ablation on cerebellar development. OL ablation was induced during the first 3 postnatal weeks, a time at which cerebellum development is ongoing. Strikingly, OL ablation triggers a profound perturbation of the known cerebellum developmental program, characterized by the disorganization of the cortical layers, abnormal foliation and a complete alteration of Purkinje cell dendritic arborization and axonal fasciculation. This phenotype is accompanied by decreased granule cell density, a disorganized Bergmann glia network and impaired migration of interneurons in the molecular layer. These results demonstrate a previously ignored role of OLs in the formation of the cerebellar cytoarchitecture.

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