Identification of nerve growth factor receptors in primary cultures of chick neural crest cells

End, David W.; Pevzner, Leonid Z.; Lloyd, Aaron; Guroff, Gordon

doi:10.1016/0165-3806(83)90169-4

Cited by 16 publications

(4 citation statements)

References 14 publications

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“…Thus, the most straightforward explanation for the lack of a response to NGF by CA+ cells is that they lack NGF receptors. It should be noted, however, that other cells in these neural crest cultures do possess NGF receptors (End et al, 1983;Bemd, 1985Bemd, , 1986bGreiner et al, 1985. Recent data by Bemd (1986a) suggest that when neural crest cultures are grown in the presence of NGF, some TH+ cells with NGF receptors are present.…”

Section: Ca Uptake and Storage Propertiesmentioning

confidence: 94%

Phenotypic properties of catecholamine-positive cells that differentiate in avian neural crest cultures

1987

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We have investigated several phenotypic features of the catecholamine-positive (CA+) cell population that develops in quail neural crest cultures. The number, spatial distribution, and morphology of CA+ and tyrosine hydroxylase-positive (TH+) cells are similar at all ages examined, suggesting that these 2 cell classes are identical. Neither CA+ nor TH+ cell bodies or processes were stained using antisera that recognize the 70 or 160 kDa subunits of chicken neurofilament protein. Other cell bodies and fibers in the cultures (which were CA- and TH-) were stained with these neurofilament antisera. The uptake and storage of 3H-norepinephrine by neural crest cultures containing CA+ cells were inhibited in the presence of desmethylimipramine and by incubation at 0 degrees C, but were unaffected by normetanephrine. Overnight treatment with reserpine eliminated histochemically detectable CA fluorescence from the cultures. Chronic reserpine treatment from day 2 to 7 in vitro prevented the appearance of CA+ cells, while normal numbers of TH+ and somatostatin-like immunoreactive (SLI) cells developed. The number and light-microscopic morphology of the CA+ cells that developed in these cultures were not dramatically altered by either exogenous NGF or 6-hydroxydopamine. Using the method of Grillo et al. (1974), we have demonstrated that the CA+ cells observed in the light microscope corresponded to cells containing abundant cytoplasmic granular vesicles (GV) characteristic of catecholamine storage granules observed in other systems. The GV diameters were quite similar in cells examined after 5, 7, 14, and 21 d in vitro. Most GV were 50-200 nm in diameter and were distributed in a unimodal manner, with the observed modal values in the range of 85-115 nm at the ages examined. The number of GV/micron2 of cytoplasmic area remained quite constant at all ages examined. These data, taken together with other available information, suggest that the CA+ cells that differentiate in our neural crest cultures resemble, in many respects, the small, intensely fluorescent cells found in autonomic ganglia and extra-adrenal chromaffin tissue of many species. At present, we do not know if the CA+ cells that differentiate in our neural crest cultures are a stable endpoint of development or whether they are a developmental intermediate in adrenergic differentiation that is normally observed only transiently during the development of avian sympathetic ganglia in vivo, but that can persist under our tissue culture conditions.

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Section: Ca Uptake and Storage Propertiesmentioning

confidence: 94%

Phenotypic properties of catecholamine-positive cells that differentiate in avian neural crest cultures

1987

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“…Bars, 25 pm. Bernd Previous studies performed on neural crest cultures grown in the absence of NGF revealed no effects of NGF on morphology or production of catecholamines (Norr, 1973;Cohen and Konigsberg, 1975;Fauquet et al, 198 1;End et al, 1983). NGF, however, influenced ornithine decarboxylase levels (End et al, 1983).…”

Section: Discussionmentioning

confidence: 94%

“…These neurons express both high-and low-affinity forms of the receptor (Frazier et al, 1974;Andres et al, 1977;Sutter et al, 1979;Olender and Stach, 1980;Riopelle et al, 1980;Olender et al, 1981;Lyons et al, 1983). The binding of iodinated NGF (1251-NGF) to cultured neural crest cells is specific and saturable (End et al, 1983;Bernd, 1985Bernd, , 1986, but only the low-affinity form of the NGF receptor appears to be present on a subpopulation of cells in this system (Bernd, 1986).…”

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confidence: 99%

Selective expression of high-affinity nerve growth factor receptors on tyrosine hydroxylase-containing neuron-like cells in neural crest cultures

Bernd

1988

J. Neurosci.

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A subpopulation of cultured neural crest cells undergoing differentiation have receptors for nerve growth factor (NGF) that exhibit a binding constant similar to that of the low-affinity NGF binding site (3.2 nM). Recent studies have shown that NGF receptors are not present on neuron-like cells immunoreactive for tyrosine hydroxylase (TH), serotonin, or vasoactive intestinal polypeptide. Since tissues innervated by sympathetic neurons in vivo produce NGF, we sought to determine whether NGF deficits in the tissue culture microenvironment may be one parameter preventing the expression of NGF receptors on TH-containing neuron-like cells. Neural crest cultures were therefore grown in complete tissue culture medium (15% fetal bovine serum and 5% chicken embryo extract), with or without exogenous NGF (50 ng/ml). Examination of light-microscopic radioautographs following incubation with 125I-NGF revealed that, if the cultures were supplemented with NGF for 7 d, approximately 33% of neuron-like cells exhibiting TH-like immunoreactivity possessed NGF receptors. There were no obvious morphological differences between TH-containing cells that did or did not have NGF receptors. Scatchard analysis of cultures grown under these conditions again demonstrated the sole presence of the low-affinity form of the NGF receptor (Kd, 3.4 nM). Embryonic catecholaminergic sympathetic neurons exhibit both high- and low-affinity forms of the NGF receptor, raising the possibility that the Scatchard analysis may not have been sensitive enough to detect the high-affinity form of the receptor on a relatively small population of cells. Therefore we used a morphological approach that took advantage of the different dissociation rates of the 2 receptor types.(ABSTRACT TRUNCATED AT 250 WORDS)

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“…5,17 Thus, NGF may influence the size of the neuronal pool during the generative phase either by a direct effect on mitosis or, indirectly, by acting as a survival factor for rapidly dividing neuroblasts. The presence of neurotrophin receptors on mitotic neural crest cells and differentiating sympathoblasts 4,20,21,29,58,59 indicates that these cells have the potential to respond to NGF or its homologues during this early developmental phase.…”

Section: J Saltis and R A Rushmentioning

confidence: 99%

Effects of nerve growtfactor on sympathetic neuron development in normal and limbless chick embryos

Saltis

Rush

1995

Intl J of Devlp Neuroscience

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Wing bud removal in chick embryos has been shown to affect the generation of sympathetic neurons prior to the normal period of limb innervation [Saltis and Rush (1995) J. auton. nerv. Sys., 51, 117-127.]. Pyknotic activity occurred earlier within the peripherally deprived ganglion, suggesting that a precocious cell death of dividing sympathoblasts led to the reduced neuronal population. We have now sought to test whether the effect of limb bud extirpation can be overcome by the administration of nerve growth factor (NGF). Specifically, the peripherally deprived ganglion has been examined for mitotic activity and total neuronal numbers. In brachial ganglia from the operated side, neuron numbers decreased by 67% by embryonic day (E) 13, but by only 28% when NGF was administered from E9. Ganglia on the unoperated side were unaffected by the NGF treatment. In contrast, in embryos receiving NGF from E5 to E9, neuron numbers in the ganglia increased by more than 100%, on both the intact and operated side. This increase was accompanied by a greater proportion of 3H-thymidine-labelled neurons. We therefore conclude that NGF, in addition to its previously described role of preventing naturally occurring neuron death, can also affect the generation of sympathetic neurons. This ability of NGF to affect gangliogenesis is most likely achieved by increasing the survival of dividing neuroblasts, although a direct effect on mitosis has not been excluded.

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Identification of nerve growth factor receptors in primary cultures of chick neural crest cells

Cited by 16 publications

References 14 publications

Phenotypic properties of catecholamine-positive cells that differentiate in avian neural crest cultures

Phenotypic properties of catecholamine-positive cells that differentiate in avian neural crest cultures

Selective expression of high-affinity nerve growth factor receptors on tyrosine hydroxylase-containing neuron-like cells in neural crest cultures

Effects of nerve growtfactor on sympathetic neuron development in normal and limbless chick embryos

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