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E Dicou

Publications and source records attributed to E Dicou.

At least 37 records · Page 2Linked to original sources
PubMed1991

Expression of the beta-nerve growth factor gene in male sex organs of the mouse, rat, and guinea pig.

Steady-state nerve growth factor (NGF) mRNA levels were estimated in male sex organs of the mouse, rat, and guinea pig by RNA blot hybridization analysis. The abundance of NGF mRNAs was in the order vas deferens greater than epididymis greater than or equal to seminal vesicles much greater than testis. NGF mRNA levels in these organs were compared with those estimated for other rat peripheral tissues and were found to correlate with the density of their sympathetic innervation, with the exception of guinea pig prostate. Castration had no significant effect on NGF mRNA levels in the guinea pig prostate, suggesting that NGF synthesis in this tissue is not under direct androgen control. NGF-like and proNGF-like immunoreactivities were localized by immunohistochemical techniques in the secretory cells of the glandular epithelium of the guinea pig prostate and in germ cells in the seminiferous tubules of the mouse testis.

Animals↗
PubMed1991

Naturally occurring antibodies against nerve growth factor in human and rabbit sera: comparison between control and herpes simplex virus-infected patients.

Antibodies against nerve growth factor (NGF) in sera were detected by enzyme-linked immunosorbent assays (ELISA), by their isolation after passage of sera through NGF immunoadsorbent columns and by their specificity to bind and immunoprecipitate mouse NGF as well as to stain by immunohistochemical methods cellular sites of NGF synthesis. Increased levels of anti-NGF antibodies were found in sera of herpes simplex virus (HSV)-infected patients but not in HSV-inoculated rabbits. As HSV latency is known to be promoted by NGF in vitro, these results may suggest that anti-NGF antibodies modulate the cytokine function of NGF and thus might play a role in HSV infection. The biological function of circulating antibodies against NGF, in general, is now open to future investigation.

Adult↗
PubMed1990

Nerve growth factor (NGF), proNGF and NGF receptor-like immunoreactivity in BB rat retina.

Nerve growth factor (NGF) and NGF precursor (proNGF)-like material were localized immunohistochemically in diabetic and non-diabetic adult rat retina using affinity-purified immunoglobulins to mature NGF and to synthetic peptides that reproduce sequences of the proNGF. Immunoreactivity for NGF and proNGF was detected in retinal ganglion cells, Müller cells, retinal pigment epithelium and areas adjacent to the internal limiting membrane. Immunolocalization of the NGF receptor using a monoclonal antibody (MC192) revealed positive staining of retinal pigment epithelium and Müller cells but not of ganglion cells. The staining intensity and distribution of NGF, proNGF, and NGF receptor-like immunoreactivity were similar in retinas of diabetic rats. These results suggest that NGF is synthesized endogenously in the retina by both neuronal and non-neuronal cell constituents, and that diabetes has no detectable effect on NGF/NGF receptor distribution and possibly on NGF biosynthesis.

Animals↗
PubMed1990

Neuronal localization of the nerve growth factor precursor-like immunoreactivity in the rat brain.

The distribution of the nerve growth factor precursor(proNGF)-like immunoreactivity was examined in the adult rat brain with affinity-purified antisera directed against synthetic peptides that reproduce sequences of the precursor protein. Immunoreactivity was localized in defined areas of the neocortex, hippocampal formation, thalamus/hypothalamus, striatum, olfactory bulb, pons and spinal cord, which are regions previously reported to contain NGF mRNA. Interestingly, immunoreactivity was also observed in the septum and diagonal band of Broca known to contain very low NGF mRNA levels. Using immunohistochemical methods combined with the retrograde transport of a protein gold complex we demonstrate that proNGF-like immunoreactivity is localized within neuronal cell bodies, in the cortex, hippocampus and septum. These results suggest that the immunological approach may not only identify NGF-synthesizing cells, but also cells which may accumulate proNGF or some cleavage products by an uptake mechanism.

Animals↗
PubMed1989

Interaction of antibodies to synthetic peptides of proNGF with in vitro synthesized NGF precursors.

Sera raised against three synthetic peptides that reproduce sequences of the pro-nerve growth factor (proNGF) protein were tested in immunoprecipitation experiments using in vitro translation products of SP6-directed NGF mRNA in a rabbit reticulocyte lysate. The interaction of these antibodies with bacterially synthesized chimeric preproNGF was also examined. Digestion of the translation products by the gamma-subunit generated the 22 and 18 kDa intermediates. A predominant 13 kDa intermediate was obtained after digestion of translation products in wheat germ extract. This is shown to be the N-terminal peptide by immunoprecipitation with an anti-peptide serum. These antibodies may be used to detect NGF precursor cleavage products in vivo.

Animals↗
PubMed1989

Synthesis of chimeric mouse nerve growth factor precursor and human beta-nerve growth factor in Escherichia coli: immunological properties.

The complete mouse prepro-nerve growth factor (NGF) DNA was fused to the carboxyl terminus of the beta-galactosidase (lac-z) gene of Escherichia coli. Similarly, a genomic fragment encoding the human NGF comprising codons 11 to 106 (from a total of 118) was fused to the fifth codon of the amino terminus of beta-galactosidase. Both bacterial vectors produce high amounts of the chimeric proteins. After cell lysis most of the chimeric mouse preproNGF protein is insoluble and appears in the pellet, whereas the majority of the chimeric human beta-NGF remains in the supernatant. Purification of the fusion proteins from the soluble fraction was achieved by affinity chromatography to p-aminophenyl beta-D-thio-galactoside Sepharose. Yields of the purified chimeric proteins were increased threefold to fourfold by the addition of protease inhibitors in the lysis and chromatography buffers. Their antigenic similarity to the preproNGF and mouse beta-NGF was examined by their interaction to sera raised against synthetic peptides which reproduce sequences of the precursor protein and to sera directed against native and denatured mouse beta-NGF using enzyme-linked immunoabsorbent assay (ELISA) techniques. Antibodies to the peptide N2 (-163 to -139) interacted with high affinity with the chimeric mouse preproNGF protein. Antisera to native and denatured mouse beta-NGF interacted with both chimeric proteins but with a variable degree of affinity. These results provide direct evidence that certain antisera to mouse beta-NGF can cross-react with the human beta-NGF molecule.

Animals↗
PubMed1988

Co-localization of the nerve growth factor precursor protein and mRNA in the mouse submandibular gland.

Indirect immunofluorescence on frozen sections of the mouse submandibular gland (MSG) and affinity-purified sera directed against synthetic peptides that reproduce sequences of the nerve growth factor (NGF) precursor protein permitted its localization in the basal parts of the cells forming the secretory tubules. In situ hybridization experiments employing 35S-labeled NGF cDNA probe localized the NGF mRNA in the same region. Conversion of proNGF to mature NGF results in an altered localization of the cleaved peptide throughout the cytoplasm of the tubular cells with a preferential concentration at their apical pole.

Animals↗
PubMed1987

Retinoic acid increases the expression of NGF gene in mouse L cells.

Retinoic acid (RA), the acid form of vitamin A, is shown to enhance the synthesis of nerve growth factor (NGF) in cultures of mouse L cells. Maximal stimulation was observed in cells growing in a serum-free medium supplemented with 10(-6)M RA during 48 h. The drug increased both the level of NGF mRNA and the amount of mature NGF protein secreted by the cells. RA was previously reported to increase the number of NGF receptors on some neuroblastoma cells (Haskell et al., 1987 Cell and Tiss. Res., 247, 67-73). It seems, therefore, that RA may influence nerve cell differentiation by promoting both the synthesis of the neurotrophic factor and the responsiveness of target cells.

Animals↗
PubMed1986

Synthesis and secretion of beta-nerve growth factor by mouse teratocarcinoma cell lines.

Using a cDNA probe and a two-site enzyme immunoassay, beta-nerve growth factor (beta NGF) synthesis was monitored in several mouse teratocarcinoma cell lines. Trace amounts of NGF mRNA were detected in the embryonal carcinoma (EC) PCC4, F9 and 1003 clones, whereas the myocardial (PCD1), myogenic (1168) and adipogenic (1246) clones contained significantly higher levels of NGF mRNA and secreted mature beta NGF peptide in the culture medium. The 1003, 1168 and 1246 strains were derived from the same teratocarcinoma cell line and their ability or inability to synthesize the neurotrophic factor may reflect a developmental decision for divergent differentiation programs. Induction of NGF mRNA and protein synthesis was observed in a differentiated derivative of an SV40-transformed F9 clone which expresses the viral T antigen. Southern blot analysis of the genomic DNAs revealed no structural alterations of the NGF locus between teratocarcinoma cells that express the NGF gene and those that do not. Similar analysis of the DNA methylation pattern in C-C-G-G sequences using the Hpa II and Msp I isoschizomers indicated no methylation changes of the NGF gene in the teratocarcinoma DNAs. At least two, and probably all four, of the already mapped Msp I sites within the NGF gene are methylated in all teratocarcinoma DNAs examined, as well as in the male mouse submaxillary gland DNA, the organ richest in this factor.

Animals↗
PubMed1986

Synthesis of nerve growth factor mRNA and precursor protein in the thyroid and parathyroid glands of the rat.

Nerve growth factor (NGF) mRNAs were detected in the thyroid and parathyroid glands by hybridization to a preproNGF and a NGF-beta cDNA probe. The thyroid NGF transcript was identical in size to that found in the mouse submaxillary gland. Mature NGF-beta was not detected in the thyroid tissue either by immunoprecipitation or by immunohistochemical methods. In contrast, an antiserum directed against the predicted precursor protein sequence immunoprecipitated a protein of Mr 30,000-31,000 in thyroid extracts. A protein of identical molecular weight was also detected by this serum in the mouse submaxillary gland extracts. In vitro translation of hybrid-selected NGF RNAs detected two peptides of Mrs 35,000 and 28,000. These peptides closely correspond to the proposed translation products of the preproNGF mRNA sequence, suggesting that two different initiation codons can be utilized, at least in vitro. The proNGF antiserum also cross-reacted with the in vitro synthesized Mr 35,000 peptide. The antigenic determinant recognized by the precursor-specific antibody was localized to the convoluted granular tubules of the mouse submaxillary gland, known to be the site of synthesis of the NGF-beta. We present evidence for the localization of unprocessed proNGF in the parathyroid gland and in some parafollicular cells of the thyroid.

Animals↗
PubMed1985

Serum and thyroid hormones T3 and T4 regulate nerve growth factor mRNA levels in mouse L cells.

Mouse L cells synthesize and secrete a neurotrophic factor related to the beta subunit of the submaxillary gland nerve growth factor (NGF) of male mice. Use of a cDNA probe which encodes the beta-NGF mRNA demonstrated that L cells produce a transcript identical in size to that of the submaxillary gland. Moreover, target sites of restriction enzymes EcoRI, PstI and BamHI were not significantly rearranged in the beta-NGF gene locus of these cells. The abundance of the beta-NGF transcript was found to depend on culture conditions. Removal of serum depressed the cellular content of polyadenylated RNA by a factor of 1.7, and decreased specifically the pool of beta-NGF transcript by an additional factor of 4. The presence of 10(-7) M testosterone in the serum-free medium did not modify the level of beta-NGF mRNA, while addition of 10(-7) M T3 (or T4) increased this level by a factor of 1.5. These data provide the first evidence that the beta-NGF mRNA of L cells is subjected to regulation, but in a way apparently different from that described for the submaxillary gland.

Animals↗
PubMed1984

Precursors of the nerve growth factor gamma subunit and renin bind to microtubules.

Translation products of a reticulocyte lysate reaction, programmed with poly(A)-rich RNAs from the male mouse submaxillary gland, were subjected to affinity chromatography on a tubulin-Sepharose column. Analysis of the bound proteins in sodium dodecylsulfate/polyacrylamide gels revealed two polypeptides of Mr 27 000 and 45 000, that were shown to bind to tubulin in a specific manner. These polypeptides were absent from the translation products coded by poly(A)-rich RNAs from the female mouse. They were eluted from the tubulin-Sepharose resin under conditions similar to those employed for the dissociation of immune complexes. The Mr-27 000 and Mr-45 000 proteins were identified by immunoprecipitation with specific antisera as the precursors of the gamma subunit of the nerve growth factor (NGF) and renin respectively. These two precursors as well as a third, unidentified polypeptide of Mr 38 000, probably unrelated to the beta subunit of NGF, bound also to microtubules. The mature form of renin, purified from the submaximillary gland, also displayed an affinity for the microtubules. In contrast, the mature form of the gamma subunit of NGF did not bind to the microtubules. The possible involvement of the microtubules (tubulin) in the biosynthesis of these two secretory proteins is discussed.

Animals↗
PubMed1984

Synthesis and partial maturation of the alpha- and gamma-subunits of the mouse submaxillary gland nerve growth factor in Xenopus laevis oocytes.

Sera raised against the alpha-, beta- and gamma-subunits of the mouse 7 S NGF were used to characterize translation products coded by submaxillary gland mRNAs microinjected into Xenopus oocytes. Anti-beta NGF sera did not cross-react with any material. In contrast, the precursors of the alpha- and gamma-subunits, as well as that of renin were identified. Use of tunicamycin, and a comparison of the translation products obtained in oocytes or in the reticulocyte lysate indicated that oocytes achieved the cleavage of signal sequences, the glycosylation of the alpha- and gamma-precursors, and the subsequent secretion of the 3 proteins. In the submaxillary gland, however, the mature forms of alpha NGF, gamma NGF and renin are composed of peptides of smaller size than those produced by the oocytes. These latter appear to lack specific proteases involved in the terminal processing of the submaxillary gland proteins.

Animals↗
PubMed1984

L cells potentiate the effect of the extracellular NGF activity in co-cultures with PC12 pheochromocytoma cells.

Two mouse cell lines, 3T3 and L, reported to secrete an NGF-like activity in the culture medium were co-cultured with the pheochromocytoma cell line PC12 which responds to NGF in vitro. In these co-cultures mitomycin-treated L or 3T3 cells were employed at low cell density (1 000 cells/cm2). L cells, but not 3T3, promoted efficiently neurite outgrowth of PC12. The response of the PC12 cells was blocked by an antiserum to male mouse submaxillary gland beta NGF. The NGF secreted by the L cells and immunoprecipitated by this antiserum co-migrated with the submaxillary gland beta NGF monomer in SDS-polyacrylamide gels. Surprisingly the neurite-promoting activity of media conditioned by L or by L-PC12 co-cultures was at most one-tenth of that expected on the basis of the response of PC12 cells in the co-cultures. This was not due to proteolytic degradation of the NGF-like factor or to losses by manipulation of the media. It seems therefore that co-cultures provide conditions which enhance the effect of the factor. Possible mechanisms responsible for this effect are discussed.

Animals↗
PubMed1983

[The neurotrophic factor nerve growth factor (NGF) synthetized by murine L-929 cells is not associated with alpha and gamma subunits].

L-929 cells secrete a neurotrophic factor which is immunologically related to the beta NGF present in the male Mouse submaxillary gland. In order to determine whether L-929 cells also synthesize the alpha and gamma subunits which are associated with the beta NGF in the gland, immunoprecipitations were performed with appropriate antisera. Our results indicate the absence of alpha and gamma subunits in the extracellular medium or in the cell extracts and suggest that the synthesis and secretion of the beta NGF by the L cells does not require these two proteins.

Animals↗
PubMed1982

An intracellular modulator of the 3'5'-cGMP hydrolysis in growing Dictyostelium discoideum amoebae.

A heat-stable and acid-stable macromolecular factor present in the cytosol of growing Dictyostelium discoideum amoebae affects specifically the intracellular cGMP phosphodiesterase. It decreases the V of the enzyme but does not alter its Km. It has no effect on the cAMP or cGMP hydrolysis catalyzed by the intracellular cAMP-cGMP phosphodiesterases or by the extracellular phosphodiesterase. It is also expressed in a mutant (HPX235), defective in the synthesis of the cAMP-cGMP phosphodiesterases but capable of intracellular transduction of the chemotactic signal. This factor is resistant to several nucleases, proteases and phospholipases, and has an apparent molecular weight between 3500-10000. In contrast, the protein phosphodiesterase inhibitor secreted by the amoebae exerts an opposite inhibition on the intracellular phosphodiesterases. These two inhibitory factors may regulate intracellular cGMP hydrolysis during the chemotactic response.

3',5'-Cyclic-GMP Phosphodiesterases↗