Maintenance of immunocytologically identified Purkinje cells from mouse cerebellum in monolayer culture

Weber, Andrée; Schachner, Melitta

doi:10.1016/0006-8993(84)91404-5

Cited by 73 publications

(51 citation statements)

References 31 publications

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“…1). These neurons were identified as PNs based the following: (1) the specificity of the antibody for PNs in the mature cultures, (2) the specificity of the antibody for PNs in histological sections from mature and immature cerebella (Lohmann et al, 1981;Levitt et al, 1984), and (3) the similarity of the cultured PNs to published photographs and descriptions of immature PNs in histological sections from cerebella of young animals (Ramon y Cajal, 1960;Meller and Glees, 1969;Woodward et al, 1969a;Altman, 1972;Crepe1 et al, 1980;Hendleman and Aggerwal, 1980;Laxson and King, 1983) and in other types of cerebellar culture preparations (Hendleman and Aggerwal, 1980;Weber and Schachner, 1984). Staining for cGPK was light for PNs in 4 d cultures and increased in intensity with culture age and neuronal maturation.…”

Section: Resultsmentioning

confidence: 99%

Morphological and physiological differentiation of Purkinje neurons in cultures of rat cerebellum

Gruol¹,

Franklin²

1987

J. Neurosci.

115

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During ontogeny, vertebrate CNS neurons differentiate from relatively simple stem cells to complex units that express unique morphological and electrophysiological characteristics. We have examined several aspects of this developmental process in an identified CNS neuronal type, the Purkinje neuron (PN) of the cerebellum. Our approach has included the use of a tissue culture preparation and immunohistochemical and electrophysiological techniques. Using immunohistochemical techniques, we have identified immature PNs in culture and examined their morphological and synaptic development. These studies have shown that PNs undergo extensive morphological and synaptic development in culture, the morphological characteristics of the immature PNs in culture and the developmental sequence and time course are reflective of that described for PNs in vivo, synapse formation is initiated at an early stage of PN development in culture and proceeds concurrently with the morphological development, and the main period of synapse formation is associated with the main period of dendritic development, reflecting the preferential location of synaptic sites at the dendritic region of mature PN. Using electrophysiological techniques, we have examined the physiological development of PNs in culture and have correlated the stages in physiological, morphological, and synaptic development. Results from these studies show the following. Mature PNs in culture exhibit complex electrophysiological properties, including the ability to generate 2 types of spike events, simple and complex spikes, and endogenously generated activity. Expression of electrophysiological properties begins at an early stage in PN development, when the PNs consist of little more than a soma with a few fine perisomatic processes. The earliest physiological characteristics to be expressed by the PN include sensitivity to transmitters, the ability to respond to synaptic input, and the ability to generate simple spikes. Synaptic input produces spontaneous activity in young PNs, but the patterns of activity change during development as mechanisms underlying endogenously generated activity and complex spike generation are expressed, and synapse formation proceeds.(ABSTRACT TRUNCATED AT 400 WORDS)

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Section: Resultsmentioning

confidence: 99%

Morphological and physiological differentiation of Purkinje neurons in cultures of rat cerebellum

Gruol¹,

Franklin²

1987

J. Neurosci.

115

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“…The cultures have been reported to contain ϳ95% small interneurons, predominantly granule neurons (Thangnipon et al, 1983), and 4 -6% GABAergic neurons (Resink et al, 1994). This is achieved in part by the huge abundance of granule neurons in the cerebellum and in part by the isolation procedure that does not allow the survival of larger, earlier differentiating neurons such as Purkinje cells (Weber and Schachner, 1984). Culturing in serum-containing medium supplemented with the antimitotic agent arabinosyl cytosine is more effective than culturing without serum at reducing the proportion of contaminating cells.…”

Section: Methodsmentioning

confidence: 99%

Neurotrophins Protect Cultured Cerebellar Granule Neurons against the Early Phase of Cell Death by a TwoComponent Mechanism

1997

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Cerebellar granule neurons cultured with serum develop a mature neuronal phenotype, including stimulus-coupled release of glutamate, and depend on elevated potassium for survival. We find that cells cultured with serum undergo two phases of cell death. By 6 d in vitro, 30 -50% of the cells present are dead; after this time the remaining cells die. Elevated potassium prevents only this later phase of death, whereas neurotrophins protect these cells against the early phase of death. Factors that bind p75 NTR or TNF-R, members of the same receptor family, exhibit voltage-sensitive calcium channel-dependent protection, whereas ligands of expressed Trk receptors show additional calcium channel-independent protection. The cells express TrkB protein and show elevated c-Fos and c-Jun levels in response to BDNF. No TrkA is detected, although p75 NTR protein is expressed and NGF induces depolarizationdependent elevation of c-Jun levels. In the presence of the protein kinase C inhibitor bisindolylmaleimide, BDNF-induced survival promotion is reduced partially, whereas NGF-induced death is unmasked. Basal survival mechanisms are insensitive to inhibition of PK-C or PI-3 kinase. We conclude that BDNF promotes survival in part via its TrkB receptor, whereas there is an additional pathway promoting survival and elevating c-Jun evoked by both NGF and BDNF via a non-Trk receptor.

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“…C erebellar Purkinje cells were cultured on several coverslips coated with poly-D-lysine (P-6407; Sigma, St. L ouis, MO) as reported previously (Weber and Schachner, 1984;Hirano et al, 1986;Kataoka and Ohmori, 1996). We used Purkinje cells after 3-5 weeks in culture for the experiments reported here.…”

Section: Methodsmentioning

confidence: 99%

A Postsynaptic Excitatory Amino Acid Transporter with Chloride Conductance Functionally Regulated by Neuronal Activity in Cerebellar Purkinje Cells

Kataoka

Morii²,

Watanabe³

et al. 1997

J. Neurosci.

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Excitatory amino acid (EAA) neurotransmitters induce postsynaptic depolarization by activating receptor-mediated cation conductances, a process known to underlie changes in synaptic efficacy. Using a patch-clamp method, we demonstrate here an EAA-dependent postsynaptic anion conductance mediated by EAA transporters present on cerebellar Purkinje cell bodies and dendrites in culture. This transporter-mediated current was modulated by neuronal activity: it exhibited facilitation for Ͼ20 min after transient depolarization accompanied by Ca 2ϩ influx.Evidence is presented suggesting that the transporter facilitation is mediated by arachidonate release after Ca 2ϩ -dependent activation of phospholipase A 2 , which exists in Purkinje cells. This postsynaptic reuptake system may represent a novel modulatory mechanism of synaptic transmission as well as prevent neuronal excitotoxicity.

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Maintenance of immunocytologically identified Purkinje cells from mouse cerebellum in monolayer culture

Cited by 73 publications

References 31 publications

Morphological and physiological differentiation of Purkinje neurons in cultures of rat cerebellum

Morphological and physiological differentiation of Purkinje neurons in cultures of rat cerebellum

Neurotrophins Protect Cultured Cerebellar Granule Neurons against the Early Phase of Cell Death by a TwoComponent Mechanism

A Postsynaptic Excitatory Amino Acid Transporter with Chloride Conductance Functionally Regulated by Neuronal Activity in Cerebellar Purkinje Cells

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