The Mouse Cerebellar Cortex in Organotypic Slice Cultures: An In Vitro Model to Analyze the Consequences of Mutations and Pathologies on Neuronal Survival, Development, and Function

Abstract

Thin acute slices and dissociated cell cultures taken from different parts of the brain have been widely used to examine the function of the nervous system, neuron-specific interactions, and neuronal development (specifically, neurobiology, neuropharmacology, and neurotoxicology studies). Here, we focus on an alternative in vitro model: brain-slice cultures in roller tubes, initially introduced by Beat Gähwiler for studies with rats, that we have recently adapted for studies of mouse cerebellum. Cultured cerebellar slices afford many of the advantages of dissociated cultures of neurons and thin acute slices. Organotypic slice cultures were established from newborn or 10-15-day-old mice. After 3-4 weeks in culture, the slices flattened to form a cell monolayer. The main types of cerebellar neurons could be identified with immunostaining techniques, while their electrophysiological properties could be easily characterized with the patch-clamp recording technique. When slices were taken from newborn mice and cultured for 3 weeks, aspects of the cerebellar development were displayed. A functional neuronal network was established despite the absence of mossy and climbing fibers, which are the two excitatory afferent projections to the cerebellum. When slices were made from 10-15-day-old mice, which are at a developmental stage when cerebellum organization is almost established, the structure and neuronal pathways were intact after 3-4 weeks in culture. These unique characteristics make organotypic slice cultures of mouse cerebellar cortex a valuable model for analyzing the consequences of gene mutations that profoundly alter neuronal function and compromise postnatal survival.

Cell death and proliferation in acute slices and organotypic cultures of mammalian CNS

“…the axons of the PNs, may be spared in slices containing the cerebellar nuclei that represent the main synaptic target of these neurons. Roller tube cultures obtained from P0 to P1 mice share many of the features of the adult mouse cerebellum in vivo (Dupont et al, 2006;Lonchamp et al, 2006). In cultures obtained with the interface method, the maturation of PNs was used by some authors as a means to study culture vitality and architecture.…”

Transcriptome Analysis Identifies Key Metabolic Changes in the Hooded Seal (Cystophora cristata) Brain in Response to Hypoxia and Reoxygenation

“…Firstly and as already mentioned, transcriptome studies do not allow the detection of changes that occur on the posttranscriptional level. Secondly, although the brain slice technique is well established in animal models [24, 56, 57], it is unknown whether the responses in the slices, in fact, reflect the situation in vivo . Thirdly, our data only reflect the global changes and do not allow discriminating between the expression of genes in neurons and glia cells; in fact, there is ample immunohistological evidence that changes in the interplay between neurons and astrocytes is an important component in the adaptation of the hooded seal brain to hypoxia [27–29].…”

Regenerating cortical connections in a dish: the entorhino-hippocampal organotypic slice co-culture as tool for pharmacological screening of molecules promoting axon regeneration

“…For most purposes, slices of developing brain, termed organotypic slice cultures (OSC), preserve a high degree of cellular differentiation and tissue organization. OSC have been prepared from a variety of brain regions, including hippocampus, neocortex, striatum, spinal cord, hypothalamus and cerebellum [3][4][5][6][7][8][9][10] . As OSC obviate the need for extensive animal surgery and equipment, their use in basic and applied research has increased over the years 1 .…”