PubMed Health⌕ Search

Biomedical subjects

Lorella Vecchio

Publications and source records attributed to Lorella Vecchio.

5 recordsLinked to original sources

Chromatin domains and the interchromatin compartment form structurally defined and functionally interacting nuclear networks.

In spite of strong evidence that the nucleus is a highly organized organelle, a consensus on basic principles of the global nuclear architecture has not so far been achieved. The chromosome territory-interchromatin compartment (CT-IC) model postulates an IC which expands between chromatin domains both in the interior and the periphery of CT. Other models, however, dispute the existence of the IC and claim that numerous chromatin loops expand between and within CTs. The present study was undertaken to resolve these conflicting views. (1) We demonstrate that most chromatin exists in the form of higher-order chromatin domains with a compaction level at least 10 times above the level of extended 30 nm chromatin fibers. A similar compaction level was obtained in a detailed analysis of a particularly gene-dense chromosome region on HSA 11, which often expanded from its CT as a finger-like chromatin protrusion. (2) We further applied an approach which allows the experimental manipulation of both chromatin condensation and the width of IC channels in a fully reversible manner. These experiments, together with electron microscopic observations, demonstrate the existence of the IC as a dynamic, structurally distinct nuclear compartment, which is functionally linked with the chromatin compartment.

Animals↗

The effect of the enkephalin DADLE on transcription does not depend on opioid receptors.

[D-Ala(2),D-Leu(5)] enkephalin (DADLE) is a synthetic peptide capable of inducing a hibernation-like state in mammals in vivo and in cultured cells in vitro. The effects of DADLE seem to be due to its binding to opioid receptors; however, it inhibits the growth of LNCaP cells, devoid of opioid receptors. We have investigated the effects of DADLE on this cell line using transmission electron microscopy, immunocytochemistry and cytometry, in order to elucidate the general mechanism(s) by which this enkephalin affects cell metabolism. We demonstrated that, similar to cell lines provided with opioid receptors, in LNCaP cells DADLE induces structural modifications of cytoplasmic and nuclear constituents, as well as a decrease in transcription and proliferation. However, DADLE does not provoke an increase in apoptotic or necrotic cell fraction, and, after removing the enkephalin from the culture medium, all effects disappear. We also demonstrated that DADLE molecules enter the cytoplasm and the nucleus of LNCaP cells, mostly binding to perichromatin fibrils and dense fibrillar component, where transcription and early splicing of pre-mRNAs and pre-rRNAs occur. In conclusion, our data demonstrate that the effect of DADLE on transcription and on cultured cells does not depend on opioid receptors. DADLE can, therefore, be envisaged as an extremely promising molecule to be used for inducing a reversible hypometabolic state in various cultured cells, without provoking cell damage or death.

Cell Death↗

DADLE induces a reversible hibernation-like state in HeLa cells.

[D-Ala(2)-D-Leu(5)-Enkephalin] (DADLE) can induce hibernation when injected into ground squirrels in summer and is able to increase the survival time of explanted organs such as liver and lung. Since cell metabolism is a target of this peptide, we have treated HeLa cells with DADLE and investigated its possible effect on transcription and proliferation as well as the resumption of metabolic activity after treatment. The labelling for Pol I, Pol II and for splicing factors such as snRNPs and SC-35 decreased after treatment as did the nucleolar labelling for UBF. In treated cells, several spherical nuclear bodies were found to be labelled for hnRNPs. In parallel, the number of proliferating cells decreased after treatment with DADLE. After recovery, there was a gradual resumption of cell function: transcription and splicing factors had a distribution similar to that of controls; proliferation resumed; nuclear bodies, representing storage sites for RNPs, disappeared.

Antimetabolites↗

Ultrastructural and immunocytochemical analyses of opioid treatment effects on PC3 prostatic cancer cells.

Some opioid peptides are able to inhibit the growth of human prostatic cancer cells; in particular, the [D-Ala(2),D-Leu(5)] enkephalin (DADLE) reduces PC3 cell growth. In order to understand how DADLE decreases cell proliferation, we investigated, by electron microscopy, its effects on PC3 cellular components. PC3 cells were incubated with DADLE and processed for both ultrastructural morphology and immunoelectron microscopy. Some cells were incubated with BrU to determine the transcriptional rate. BrU and DADLE molecules were detected by immunogold techniques and the labeling was quantitatively evaluated. Modifications of some cytoplasmic and nuclear components were observed in DADLE-treated cells. Moreover, treated cells incorporated lower amounts of BrU than control cells. DADLE molecules were located in the cytoplasm and in the nucleus, especially on mRNA transcription and early splicing sites. Our data suggest that DADLE is able to slow down the synthetic activity of PC3 cells, perhaps interfering with nuclear functions.

Cell Division↗

Rearrangement of nuclear ribonucleoprotein (RNP)-containing structures during apoptosis and transcriptional arrest.

The aim of this paper is to review the data in the literature concerning ribonucleoprotein components during apoptosis, where a major rearrangement of RNPs takes place. In parallel with chromatin changes, the nucleoplasmic constituents (perichromatin fibrils; perichromatin granules; interchromatin granules and nuclear bodies) as well as the nucleoli aggregate into heterogeneous clusters called HERDS, in the interchromatin space. Later, these RNP-containing structures are extruded from the nucleus and leave the cell within cytoplasmic blebs. We propose also a role for HERDS as markers of irreversible transcriptional arrest.

Animals↗