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D N Currie

Publications and source records attributed to D N Currie.

11 recordsLinked to original sources
PubMed1982

Development of Thy-1 antigen on cerebellar neurons in culture.

The Thy-1.1 cell surface antigen was demonstrated by indirect immunofluorescence on two types of neurons in dissociated cell cultures of developing rat cerebellum. In cultures from postnatal rats, small cells predominated. They bound tetanus toxin, a neuronal marker, and did not have the capacity to take up gamma-aminobutyric acid (GABA) as measured by autoradiography. From these properties, we conclude that they are granule cells. These neurons began to express the surface antigen Thy-1.1 by 2 weeks in culture, and by 4 weeks, the antigen was detected on about 40% of all of the surviving neurons. The second type of neuron, which may be a heterogeneous mixture of cerebellar neurons, was numerous only in cultures prepared from embryonic rats. They were larger than granule cells and expressed Thy-1.1 antigen after a few days in culture. Such cells bound tetanus toxin and were negative for the glial markers galactocerebroside and glial fibrillary acidic protein. Thy-1-positive cells and cells with high GABA uptake were most frequent in embryonic day 19 cultures, where 40 to 50% of all of the neurons were positive for either property. They survived about 1 week in culture. The size and frequency of Thy-1 cells and the frequency of similar cells with high GABA uptake suggest that the embryonic rat cultures included many Purkinje cells, which express Thy-1.

Animals↗
PubMed1981

Potassium-stimulated, calcium-dependent release of [3H]GABA from neuron- and glia-enriched cultures of cells dissociated from rat cerebellum.

The release of [3H]GABA from cultures of cells dissociated from the rat cerebellum was investigated. The culture system contained a population of neurons (stellate and basket cells) which were capable of accumulating [3H]GABA and releasing it in a calcium-dependent manner in response to 50 mM potassium. In addition, the release of [3H]GABA for cultured astrocytes was found to be insensitive to potassium depolarization.

Animals↗
PubMed1981

An improved method for the bulk isolation of viable perikarya from postnatal cerebellum.

A modified method for the isolation of large numbers of viable perikarya from postnatally developing rat cerebellum is described in comparison with an earlier published version. Advantages of the modified preparation include greater cell yields which emphasize the preservation of the larger sized perikarya (inhibitory interneurons and Purkinje cells). In addition, the use of cytotoxic EDTA is eliminated, and by combining several steps and using a simpler buffer system, the preparation time is reduced by half to less than 1 h.

Animals↗
PubMed1980

[3H]GABA uptake as a marker for cell type in primary cultures of cerebellum and olfactory bulb.

Uptake of [3H]GABA into cell cultures of rat cerebellum and olfactory bulb was studied by autoradiography, using beta-alanine and aminocyclohexane carboxylic acid to distinguish neuronal-specific and glial-specific uptake. Neurons and astrocytes were also labeled by tetanus toxin and anti-GFAP respectively. This combination of markers allowed identification and quantification of several cell types. Cerebellar cultures were found to contain 77% granule neurons, 7.5% inhibitory neurons (probably stellate and basket cells) and 15% astrocytes. Olfactory bulb cultures were over 50% in small neurons which accumulated GABA, the olfactory bulb granule neuron being GABAergic in vivo.

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PubMed1979

Monolayer cultures of perikarya isolated from postnatal rat cerebellum.

Viable cerebellar perikarya of mixed cell type obtained from 7--9-day-old rats were maintained in monolayer cutlure for up to 12 days. During this time extensive neurite formation and outgrowth occurred. The large majority of the cells developing in culture were tentatively identified as granule neurons. This identification was based on the large number isolated from the starting tissue, and the cells' general morphological features in culture such as perikaryal and nuclear size, the bipolar nature of neurite extension, and their migratory behaviour.

Animals↗
PubMed1978

Separation of cell types from the developing cerebellum.

The heterogenous population of perikarya (cells) obtained by dissociating cerebellar tissue of developing postnatal rats was separated by sedimentation at unit gravity. Peak fractions, defined by monitoring the distribution of different size classes with a particle analyzer, were enriched in ultrastructurally well-preserved and metabolically competent specific cell types. These fractions included the peak of rapidly sedimenting cells comprising large neurones, such as Purkinje cells, which accounted for about 50% of the cells (vs. 2% in the initial cell suspension) and for a much greater proportion of the total cell mass (over 80%). More slowly sedimenting peak fractions contained 2-5-fold enrichments in replicating cells (assessed in terms of [3H]thymidine incorporation into DNA or by the frequency of mitotic cells), astroglia-like cells and external and differentiating granule cells, respectively. Separated, replicating cells continued synthesizing DNA in vitro; the [3H]thymidine incorporation rate was about 5-fold greater in the fraction enriched in proliferating cells than in the total cell suspension. Besides their structural integrity, the viability of the cells was also indicated by the finding that the proportion of trypan blue-excluding cells in all fractions exceeded 80%. Moreover, protein synthesis, in terms of incorporation of labelled amino acids, continued in the separated cells at a linear rate for a relatively long time. The rate per cell was highest in the large neuronal fraction, and lowest in the astroglia-like fraction.

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