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G Gargiulo

Publications and source records attributed to G Gargiulo.

At least 91 records · Page 5Linked to original sources
PubMed1983

Analysis of the chromatin assembled in germinal vesicles of Xenopus oocytes.

We have injected circular DNA, labeled with 32P at a single restriction site, into germinal vesicles of Xenopus laevis oocytes in order to study the nucleosome arrangement on the assembled minichromosomes. Two types of genes were used in these studies, the somatic 5 S RNA gene unit of Xenopus borealis and the histone gene unit of Drosophila melanogaster. We find that injections of labeled DNA alone, at 1 ng DNA per oocyte, results in irregularly spaced nucleosomes and partially supercoiled DNA molecules. However, perfectly spaced nucleosomes are assembled and fully supercoiled DNA is recovered if 5 to 20 nanograms of cold vector DNA is coinjected with the labeled DNA. At the optimum chromatin assembly conditions, the nucleosomes are perfectly spaced with a 180 base-pair periodicity, but they are randomly positioned on the DNA. The assembly of a periodic chromatin structure is accompanied by a dramatic enhancement in the expression of the injected 5 S RNA gene.

Animals↗
PubMed1983

Escherichia coli single-strand binding protein stabilizes specific denatured sites in superhelical DNA.

Escherichia coli single-strand binding protein relaxes supercoiled DNA molecules containing the Drosophila melanogaster histone gene repeat unit, by stabilizing denaturation bubbles that map near the boundaries of the genes, at sites that in native chromatin have been shown to be hypersensitive to nucleases. A similar process may contribute to the propagation of such hypersensitive sites after their induction on the activation of gene expression.

Bacterial Proteins↗
PubMed1983

Chromatin fine structure of the histone gene complex of Drosophila melanogaster.

We have used salt extractions of nuclei and long agarose gels to dissect the chromatin fine structure of the histone gene repeat of Drosophila melanogaster. Extraction of nuclei with 0.35 M KCl removes many non-histone chromosomal proteins but does not significantly disturb the overall nucleosome arrangement of the repeat unit. After extraction of nuclei with 0.55 M KCl, which also removes histone Hl, the basic arrangement of nucleosome core particles in the repeat unit is not greatly disturbed and the exposed DNA segments near the 5' ends of the histone genes are also retained. Extraction of nuclei with 0.75 M or higher KCl concentrations causes extensive nucleosome sliding and rearrangement with accompanying changes in the nucleoprotein organization of the histone gene complex and loss of the 5' hypersensitive sites. Our results indicate that the histone gene repeat displays a highly organized chromatin structure in vivo.

Animals↗
PubMed1983

A simple procedure for parallel sequence analysis of both strands of 5'-labeled DNA.

Ligation of a 5'-labeled DNA restriction fragment results in a circular DNA molecule carrying the two 32Ps at the reformed restriction site. Double digestions of the circular DNA with the original enzyme and a second restriction enzyme cleavage near the labeled site allows direct chemical sequencing of one 5'-labeled DNA strand. Similar double digestions, using an isoschizomer that cleaves differently at the 32P-labeled site, allows direct sequencing of the now 3'-labeled complementary DNA strand. It is possible to directly sequence both strands of cloned DNA inserts by using the above protocol and a multiple cloning site vector that provides the necessary restriction sites. The simultaneous and parallel visualization of both DNA strands eliminates sequence ambiguities. In addition, the labeled circular molecules are particularly useful for single-hit DNA cleavage studies and DNA footprint analysis. As an example, we show here an analysis of the micrococcal nuclease-induced breaks on the two strands of the somatic 5S RNA gene of Xenopus borealis, which suggests that the enzyme may recognize and cleave small AT-containing palindromes along the DNA helix.

Animals↗
PubMed1982

Analogous cleavage of DNA by micrococcal nuclease and a 1-10-phenanthroline-cuprous complex.

We have examined the DNA cleavage specificity of a 1,10-phenanthroline-cuprous complex and find this agent to recognize the same sites and cleave with the same relative preferences as micrococcal nuclease. In contrast, DNase I and bleomycin-ferrous complex cleave the same 5000 bp D. melanogaster histone gene DNA with different specificities, although some of the sites appear to be recognized and cleaved by all four reagents. Our results suggest that the reagents used probably detect discrete conformational perturbations along the DNA double helix.

Base Sequence↗
PubMed1980

Comparison between rDNA magnification and bb lethal mutation frequencies in Drosophila melanogaster.

Unequal mitotic sister strand crossing over has been evoked to explain the occurrence of phenotypically bb+ males in the progeny of phenotypically bobbed males during magnification. If this is the case, complementary bbl loci should be obtained together with the bb+. To test this hypothesis we compared the frequency of bb lethal mutations in the sperms of bb males with the percentages of phenotypically bb+ males obtained during magnification of these bb males. We then compared these values with those occurring in phenotypically bb+ control males. We found that, while the number of bb+ males obtained during magnification, though variable, is high, the bb lethal mutation occurs at a very low frequency in all the genetic conditions, whatever the phenotype of the parental male.

Animals↗
PubMed1979

Different behaviour of Xbb+ and Xbb chromosomes in D. melanogaster with only one nucleolus organizer.

We have examined the rDNA content of male and female adult flies having only one nucleolus organizer (NO), using X chromosomes carrying wild or partially deleted bobbed loci (Xbb+/O, Xbb+/XNO- and Xbb/O, Xbb/XNO-). The results show that in Xbb+/O and Xbb+/XNO- flies, where only somatic gene compensation is supposed to occur, the rDNA increase, althought less pronounced than previously reported, is directly proportional to the number of rRNA genes initially present in the nucleolus organizer. In Xbb/O and in Xbb/Xbb/XNO- flies the rDNA increase is relatively much higher than that observed in flies carrying bb+ instead of bb. It is suggested that this may be due to rDNA premagnification and somatic gene compensation occurring simultaneously in the former flies.

Animals↗
PubMed1979

Effects of intraventricular and intra-aquaductal 6-hydroxydopamine on monoamine containing neurons of the fowl paleostriatum, diencephalon and mesencephalon.

The effects of a single intraventricular or intra-aquaductal infusion of 6-hydroxydopamine (6-OHDA) on monoamine fluorescence in the paleostriatum augmentatum, diencephalon and mesencephalon were studied. Present experiments have enable us to better characterize the distribution of catecholaminegic axons in the paleostriatum augmentatum, preoptic area, hypothalamus, median eminence, periventricular areas as well as of cell-bodies and axons exbiting green fluorescence in the n. mesencephalicus properties and other mesencephalic nuclei.

Animals↗
PubMed1978

Effect of central nervous system-acting drugs after selective destruction by neurotoxins of 5-hydroxytryptamine fibers in the brain.

Some behavioral effects of CNS-acting drugs have been studied in two animal species after functional impairment of central serotonergic activity. In rabbits, pretreatment with p-CPA or with 5,6-DHT counteracted the hyperthermia induced by d-amphetamine or apomorphine; the same pretreatments were ineffective in modifying pyrogen-induced fever. These data indicate a modulatory role of the 5-HT system in the hyperthermic response to dopaminergic agonists, such as d-amphetamine and apomorphine, and rule out 5-HT involvement in the hyperpyrexia induced by pyrogen. A previous intraventricular injection of 5,6-DHT significantly potentiated the increase in body temperature induced by 5-HTP in combination with a MAO inhibitor. Pretreatment with p-CPA, instead, strongly inhibited the hyperthermic response to 5-HTP. In unanesthetized fowl, at the time of the most dramatic degenerative signs of indoleaminergic neurons induced by 5,6-DHT, as documented by histofluorescence pictures, intraventricular infusion of 5-HT induced more intense behavioral, electrocortical, and body temperature modifications than in control animals. These results suggest the existence of receptor supersensitivity after 5-HT nerve ending degeneration but not after brain 5-HT depletion. The hyperthermic effect of a presynaptically acting drug, fenfluramine, was counteracted in rabbits and in fowl pretreated with either p-CPA or 5,6-DHT. Since identical behavioral, electrocortical, and body temperature effects have been observed after 5-HT or 5,6-DHT infusion into the third ventricle of fowl, it may be concluded that 5,6-DHT behaves also as a central 5-HT receptor agonist.

5-Hydroxytryptophan↗
PubMed1978

Time course of 5,6-dihydroxytryptamine neurotoxic effects on fowl diencephalon and upper brain stem monoaminergic pathways.

In adult fowls the time-course of the neurotoxic effects of 5,6-DHT on indolaminergic neurons (in comparison with controls and with animals pre-treated with mebanazine, a MAO-inhibitor) was studied. It was shown that 5,6-DHT given into the third cerebral ventricle produced a rather selective degeneration of indolaminergic neurons which was most dramatic 4 and 7 days after the injection. Injured axons characterized by a pile-up of strongly yellow fluorescent material were seen in areas bordering the third ventricle, and in the posterior, medium and anterior hypothalamus, preoptic area, median eminence, n. tuberalis and paleostriatum augmentatum. A decrease in yellow fluorescence in areas distal to the injured axons was only detectable after 19 days. A certain regrowth of some terminal networks was observed 30 days after 5,6-DHT administration. Present data indicate that, in fowls, ascending indolamine neurons mainly originate in tegmental cell bodies and are organized in a large tract which runs along the medial forebrain bundle, sending terminals innervating several diencephalic and other subcortical areas.

5,6-Dihydroxytryptamine↗
PubMed1976

Acute central effects of 5,6-dihydroxytryptamine in fowl.

In adult hens (Gallus domesticus) infusion into the IIIrd cerebral ventricle of 5,6-DHT (50, 75 and 100 mug) produced, after 5-10 min behavioural and electrocortical sedation and sleep lasting about 6-8 hr,and a monophasic or biphasic increase in body temperature for about 8 hr, Two and a half hr after 5,6-DHT administration, the histochemical picture showed a sustained decrease in yellow and green fluorescence in areas adjacent to the IIIrd ventricle, in the anterior hypothalamus and in the preoptic area, whereas no significant changes were detected in fluorescence of the tegmental cell-bodies. The present experiments suggest that behavioural and electrocortical effects evoked by 5,6-DHT seem to be due to a synergistic action of 5-HT and catecholamines displaced and released by this compound, whereas hyperthermic effects seem to be due to a more sustained release of 5-HT and/or to a direct action on 5-HT receptors and/or inhibition of 5-HT reuptake.

5,6-Dihydroxytryptamine↗