scholarly journals Rapid, sequential changes in surface morphology of PC12 pheochromocytoma cells in response to nerve growth factor

1979 ◽  
Vol 82 (3) ◽  
pp. 820-827 ◽  
Author(s):  
JL Connolly ◽  
LA Greene ◽  
RR Viscarello ◽  
WD Riley
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cell Surface ◽  
Rna Synthesis ◽  
Dorsal Surface ◽  
Synthesis Inhibitor ◽  
Nerve Growth ◽  
Pheochromocytoma Cells ◽  
Phase Microscopy ◽  
Ngf Receptor

The effect of nerve growth factor (NGF), a substance that promotes the differentiation and maintenance of certain neurons, was studied via scanning electron microscopy utilizing the PC12 clonal NGF-responsive pheochromocytoma cell line. After 2-4 d of exposure to NGF, these cells acquire many of the properties of normal sympathic neurons. However, by phase microscopy, no changes are discernible within the first 12-18 h. Since the primary NGF receptor appears to be a membrane receptor, it seemed likely that some of the initial responses to the factor may be surface related. PC12 cells maintained without NGF are round to ovoid and have numerous microvilli and small blebs. After the addition of NGF, there is a rapidly initiated sequential change in the cell surface. Ruffles appear over the dorsal surface of the cells with 1 min, become prominent by 3 min, and almost disappear by 7 min. Microvilli, conversely, disappear as the dorsal ruffles become prominent. Ruffles are seen at the the periphery of cell at 3 min, are prominent on most of the cells by 7 min and are gone by 15 min. The surface remains smooth from 15 min until 45 min when large blebs appear. The large blebs are present on most cells at 2 h and are gone by 4 h. The surface remains relatively smooth until 6-7 h of NGF treatment, when microvilli reappear as small knobs. These microvilli increase in both number and length to cover the cell surface by 10 h. These changes were not observed with other basic proteins, with α-bungarotoxin (which binds specifically to PC12 membranes), and were not affected by an RNA synthesis inhibitor that blocks initiation of neurite outgrowth. Changes in the cell surface architecture appear to be among the earlist NGF responses yet detected and may represent or reflect primary events in the mechanism of the factor's action.

scholarly journals Pit formation and rapid changes in surface morphology of sympathetic neurons in response to nerve growth factor.

1981 ◽  
Vol 90 (1) ◽  
pp. 176-180 ◽  
Author(s):  
J L Connolly ◽  
S A Green ◽  
L A Greene
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cell Surface ◽  
Sympathetic Neurons ◽  
Cell Types ◽  
Coated Pits ◽  
Nerve Growth ◽  
Pheochromocytoma Cells ◽  
Pit Formation ◽  
Transmission Electron

Scanning and transmission electron microscope studies were carried out on the rapid cell surface responses of cultured newborn rat sympathetic neurons to nerve growth factor (NGF), a substance that promotes their survival and differentiation. The somas of sympathetic neurons continuously exposed to NGF or deprived of the factor for 4-5 h have a very smooth surface. After readdition of NGF to the latter type of cultures, there is rapidly initiated a transient, sequential change in the cell surface. Microvilli and small ruffles appear within 30 s and are most prominent by 1 min. By 3 min of exposure, the microvilli and ruffles decrease in prominence, and by 7 min the somal surface is again smooth. By 30 s after NGF readdition, as increase in the number of 60- tp 130-nm coated pits is also detectable. This increase reaches a maximum of about threefold from 0.5 to 3 min and then gradually decreases. Alterations in the surface did not occur on the nonneuronal cell types present in the cultures and were not observed in response to another basic protein (cytochrome c) or to physical manipulation. Changes in cell surface architecture induced by NGF in normal sympathetic neurons and, as previously described, in PC12 pheochromocytoma cells indicate that such responses may present or reflect primary events in the mechanism of the factor's action.

scholarly journals PC12 cell mutants that possess low- but not high-affinity nerve growth factor receptors neither respond to nor internalize nerve growth factor.

1986 ◽  
Vol 102 (3) ◽  
pp. 830-843 ◽  
Author(s):  
S H Green ◽  
R E Rydel ◽  
J L Connolly ◽  
L A Greene
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Cell Surface ◽  
Pc12 Cells ◽  
Selection Procedure ◽  
Division Rate ◽  
Nerve Growth ◽  
High Affinity ◽  
Ngf Receptor ◽  
Epidermal Growth

Four mutant PC12 pheochromocytoma cell lines that are nerve growth factor (NGF)-nonresponsive (PC12nnr) have been selected from chemically mutagenized cultures by a double selection procedure: failure both to grow neurites in the presence of NGF and to survive in NGF-supplemented serum-free medium. The PC12nnr cells were deficient in all additional NGF responses surveyed: abatement of cell proliferation, changes in glycoprotein composition, induction of ornithine decarboxylase, rapid changes in protein phosphorylation, and cell surface ruffling. However, PC12nnr cells closely resembled non-NGF-treated PC12 cells in most properties tested: cell size and shape; division rate; protein, phosphoprotein, and glycoprotein composition; and cell surface morphology. All four PC12nnr lines differed from PC12 cells in three ways in addition to failure of NGF response: PC12nnr cells failed to internalize bound NGF by the normal, saturable, high-affinity mechanism present in PC12 cells. The PC12nnr cells bound NGF but entirely, or nearly entirely, at low-affinity sites only, whereas PC12 cells possess both high- and low-affinity NGF binding sites. The responses to dibutyryl cyclic AMP that were tested appeared to be enhanced or altered in the PC12nnr cells compared to PC12 cells. Internalization of, and responses to, epidermal growth factor were normal in the PC12nnr cells ruling out a generalized defect in hormonal binding, uptake, or response mechanisms. These findings are consistent with a causal association between the presence of high-affinity NGF receptors and of NGF responsiveness and internalization. A possible relationship is also suggested between regulation of cAMP responses and regulation of NGF responses or NGF receptor affinity.

Nerve growth factor (NGF)-dependent kinase activities in PC12 pheochromocytoma cells

1988 ◽  
Vol 117 (4_Suppl) ◽  
pp. S51
Author(s):  
ANKE-PEGGY HOLTORF ◽  
K. UNSICKER ◽  
H.-D. HOFMANN
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Nerve Growth ◽  
Pheochromocytoma Cells ◽  
Pc12 Pheochromocytoma Cells

scholarly journals Expression of beta-nerve growth factor and its receptor in rat seminiferous epithelium: specific function at the onset of meiosis.

1992 ◽  
Vol 117 (3) ◽  
pp. 629-641 ◽  
Author(s):  
M Parvinen ◽  
M Pelto-Huikko ◽  
O Söder ◽  
R Schultz ◽  
A Kaipia ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Sertoli Cells ◽  
Seminiferous Epithelium ◽  
Receptor Protein ◽  
Tyrosine Kinase Receptor ◽  
Specific Expression ◽  
Nerve Growth ◽  
Ngf Receptor

beta-Nerve growth factor (NGF) is expressed in spermatogenic cells and has testosterone-downregulated low-affinity receptors on Sertoli cells suggesting a paracrine role in the regulation of spermatogenesis. An analysis of the stage-specific expression of NGF and its low affinity receptor during the cycle of the seminiferous epithelium in the rat revealed NGF mRNA and protein at all stages of the cycle. Tyrosine kinase receptor (trk) mRNA encoding an essential component of the high-affinity NGF receptor was also present at all stages. In contrast, expression of low affinity NGF receptor mRNA was only found in stages VIIcd and VIII of the cycle, the sites of onset of meiosis. The low-affinity NGF receptor protein was present in the plasma membrane of the apical Sertoli cell processes as well as in the basal plasma membrane of these cells at stages VIIcd to XI. NGF was shown to stimulate in vitro DNA synthesis of seminiferous tubule segments with preleptotene spermatocytes at the onset of meiosis while other segments remained nonresponsive. We conclude that NGF is a meiotic growth factor that acts through Sertoli cells.

CRK protein binds to two guanine nucleotide-releasing proteins for the Ras family and modulates nerve growth factor-induced activation of Ras in PC12 cells

1994 ◽  
Vol 14 (8) ◽  
pp. 5495-5500
Author(s):  
M Matsuda ◽  
Y Hashimoto ◽  
K Muroya ◽  
H Hasegawa ◽  
T Kurata ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Pc12 Cells ◽  
Sh2 Domain ◽  
Single Amino Acid ◽  
Guanine Nucleotide ◽  
Nerve Growth ◽  
Tyrosine Residues ◽  
Ngf Receptor ◽  
Ras Family

It has been reported that growth factors activate Ras through a complex of an adaptor type SH2-containing molecule, Grb2, and a Ras guanine nucleotide-releasing protein (GNRP), mSos. We report on the involvement of another adaptor molecule, CRK, in the activation of Ras. Overexpression of wild-type CRK proteins CRK-I and CRK-II enhanced the nerve growth factor (NGF)-induced activation of Ras in PC12 cells, although the basal level of GTP-bound active Ras was not altered. In contrast, mutants with a single amino acid substitution in either the SH2 or SH3 domain of the CRK-I protein inhibited the NGF-induced activation of Ras. Two GNRPs for the Ras family, mSos and C3G, were coimmunoprecipitated with the endogenous Crk proteins in PC12 cells. The association between C3G and the CRK mutants was dependent upon the presence of intact SH3. The SH2 domain of CRK bound to the SHC protein phosphorylated on tyrosine residues by NGF stimulation. The results demonstrate that, in addition to Grb2, CRK participates in signaling from the NGF receptor and that two GNRPs appear to transmit signals from these adaptor molecules to Ras.

Expression of nerve growth factor and neurotrophin receptors in testicular cells suggest novel roles for neurotrophins outside the nervous system

1996 ◽  
Vol 8 (7) ◽  
pp. 1075 ◽  
Author(s):  
K Seidl ◽  
A Buchberger ◽  
C Erck
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Germ Cells ◽  
Sertoli Cells ◽  
Leydig Cells ◽  
Expression Patterns ◽  
Myoid Cells ◽  
Nerve Growth ◽  
High Affinity ◽  
Ngf Receptor

The present study was designed to clarify the non-neurotrophic role for neurotrophins in mouse testis. By means of SI nuclease protection assay we could demonstrate that the gene coding for the low-affinity nerve growth factor (NGF) receptor p75NGFR is transiently expressed during germ cell development. Gene expression for p75NGFR was detected in late-meiotic spermatocytes and early spermatids and was found to be co-expressed with trkB and trkC, two tyrosine kinase receptors, commonly regarded as the high-affinity receptors for brain-derived neurotrophic factor and neurotrophin-3. Gene transcripts for the high-affinity NGF receptor trkA were found exclusively in non-germ cells. Isolated Leydig cells, peritubular myoid cells and Sertoli cells, but not germ cells, could be identified as potential testicular NGF sources. Non-germ cells respond after incubation for several days with a sharp induction in NGF synthesis, which is accompanied by a loss of phenotypic expression patterns. The fact that p75NGFR mRNA expression was induced in cultured Sertoli cells and peritubular myoid cells suggests an autocrine mode of NGF action in these cells. Induction of NGF synthesis in cultured Leydig cells could be prevented by the glucocorticoid dexamethasone. Results indicate different roles for the individual neurotrophins in distinct testicular compartments and suggest that these neurotrophins might support testicular functions by signalling between individual cell types in an autocrine and paracrine manner.

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