Cell culture of sixteen‐day‐old rat embryo cerebra and associated changes in ganglioside pattern

Schengrund, Cara Lynne; Repman, Mary Ann

doi:10.1111/j.1471-4159.1977.tb10737.x

Cited by 26 publications

(10 citation statements)

References 19 publications

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“…The possibility that the neuraminidase-sensitive component is ganglioside-related is suggested by the following experiments. The presence and accumulation of complex sialoglycolipids during neuronal development in primary cultures of central nervous system neurons is well documented [Schengrund and Repman, 1977;Yavin and Yavin, 19791. The age-in-culture-dependent association of '251-labeled tetanus toxin with the cerebral cells is illustrated in Figure 3 .…”

Section: Sequestration Of Cell Surface-associated Tetanus Toxin Is Nementioning

confidence: 99%

Sequestration of tetanus toxin in developing neuronal cell cultures

Yavin

Kohn

1982

J of Neuroscience Research

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Cultivation of pure population5 of cerebral cells and the unambiguous identification of the neuronal and glial elements is a prerequisite for studying biochemical parameters associated with the developing nervous system under in vitro controlled conditions. We now report on sequestration of tetanus toxin, an exclusibe marker for the neuronal cell surface, into a cellular compartment which prevents its release by neuraminidase. This compartment is associated with cellular elements remaining after detergent extraction of neuronal cells grown in monolayer cultures. Toxin sequestration into this compartmcnt is preceded by a neuraminidase-sensitive receptor mediated binding process which can be correlated with the appearance of lipid-bound sialic acid in devcloping neuronal cultures.

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Section: Sequestration Of Cell Surface-associated Tetanus Toxin Is Nementioning

confidence: 99%

Sequestration of tetanus toxin in developing neuronal cell cultures

Yavin

Kohn

1982

J of Neuroscience Research

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“…Robert et a1 [1979] showed that a morphological differentiation obtained after dibutyryl cyclic-AMP treatment of neonatal Syrian hamster astroblasts (NN) was followed by a marked increase of GM1 and GDla. Schengrund and Repman [1977], then Yavin and Yavin [I9791 have followed the ganglioside composition of 16-day-old rat embryo cerebral cells grown for three weeks in vitro. The high content of GM3 and GMI found by Shengrund and Repman [1977] suggested the presence of glial cells in their cultures within the whole period studied.…”

Section: Discussionmentioning

confidence: 99%

Investigations on myelination in vitro. III. Ultrastructural, biochemical, and immunohistochemical studies in cultures of dissociated brain cells from embryonic mice

Sarliève

Delaunoy

Dierich

et al. 1981

J of Neuroscience Research

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Cerebral hemispheres of 14-to 15-day-old mouse embryos were dissociated mechanically into single cells and cultured in polylysine-coated plastic flasks. The succession of changes and the identification of cell populations have been studied by transmission electron microscopy (TEM). Four distinct cell types can be clearly defined. Neuroblasts, oligodendroblasts, and astroblasts were used to denote embryonic states of neurons, oligodendrocytes, and astrocytes. Cell populations were: 1) neuroblasts; 2) astroblasts, either growing on the surface of the flasks and forming a basal layer of epithelioid cells or scattered at the surface of the bed layer; 3) oligodendroblasts, characterized by a high nucleocytoplasmic ratio and resting on top of the bed layer; 4) macrophages. Incorporation of [3H]thymidine into cells at different days in culture (DIC) delineated at least three periods of DNA synthesis (from 1 to 6, from 10 to 20, and from 25 to 60 DIC). High cell RNA/DNA and protein/DNA ratios indicated an increase in the cytoplasmic mass. Guanylate cyclase (EC 4.6.1.2; GC) and adenylate cyclase (EC 4.6.1 . l ; AC) activities and the ratio AC/GC also agreed with the existence of different periods in culture, but the yin yang hypothesis (postulated by Goldberg et al, 1975) was only partially confirmed. Concerning membrane constituents, the evolution of the pattern of the major gangliosides showed a phase from 1 t o 20 DIC which was characterized during the first half of this period by a high percentage of tri-and tetra-sialogangliosides, thereby confirming the Abbreviations: AC, adenylate cyclase; AChE, acetylcholinesterase; Anti-CAll, anti-carbonic anhydrase isoenzyme 11; Anti-GFAP, anti-glial fibrillary acidic protein; ChAC, choline acetyltransferase; DIC, days in culture; GC, guanylate cyclase; SEM, scanning electron microscopy; TEM, transmission electron microscopy; TLC, thin-layer chromatography.

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“…Neuronal cells from 16-to 17-day-old embryonic rat cortex provide a predom inantly neuronal culture in medium supplemented with fetal calf serum [11,15]. In this study, the neuronal culture was prepared from 17-day-old embryos and incubated in medium containing 3% newborn calf serum to reduce the considerable ef fects of growth factors in calf serum and to retard the proliferation o f nonneuronal cells.…”

Section: Colchicine Inhibition O F Neurite Growth In Primary Culturedmentioning

confidence: 99%

“…In earlier studies [11,15], primary cultures of rat em bryonic neurons were able to synthesize polysialogangliosides, although glial cells also developed and differ entiated in the cultures in the presence of 10-20% fetal calf serum. The recent establishment of chemically de fined medium has enabled neurons to be investigated in the absence of glial cells [16][17][18].…”

Section: Introductionmentioning

confidence: 99%

Effect of Colchicine on Ganglioside Composition of Rat Primary Culture Neurons

Ogiso

Ohta²,

Harada³

et al. 1992

Dev Neurosci

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Developmental changes in gangliosides in the course of neurite outgrowth were examined in dissociated fetal rat cerebral neurons in culture. About a 2-fold increase in ganglioside levels was seen with the progression of neurite formation for up to 24 h in predominantly neuronal cultures. Ganglioside patterns appeared to be unchanged during the first 24 h, subsequently consisted of higher amounts of GD3 and b-series gangliosides (such as GD1b, GT1b, and GQ1b), and lower amounts of a-series gangliosides (GM1 and GD1a). Although the addition of colchicine to the cell growth medium inhibited neurite outgrowth in developing neurons, little if any differences in ganglioside patterns were found between control and colchicine-treated cells. Ganglioside levels decreased slightly in colchicine-treated cells in agreement with the decrease in cell attachment to culture dishes. Although colchicine treatment 8 h after plating caused complete retraction of formed neurites, the ganglioside level of the cells continued to increase during the following 16-hour incubation. Thus, the data suggest that ganglioside synthesis in differentiating neurons does not primarily accompany the expansion in cell surfaces due to neurite formation, and raises the possibility that a large proportion of gangliosides is retained in intracellular compartments.

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Cell culture of sixteen‐day‐old rat embryo cerebra and associated changes in ganglioside pattern

Cited by 26 publications

References 19 publications

Sequestration of tetanus toxin in developing neuronal cell cultures

Sequestration of tetanus toxin in developing neuronal cell cultures

Investigations on myelination in vitro. III. Ultrastructural, biochemical, and immunohistochemical studies in cultures of dissociated brain cells from embryonic mice

Effect of Colchicine on Ganglioside Composition of Rat Primary Culture Neurons

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