Biomedical subjects

M Lafarga

Publications and source records attributed to M Lafarga.

At least 37 records · Page 2Linked to original sources

PubMed1998

Acute osmotic/stress stimuli induce a transient decrease of transcriptional activity in the neurosecretory neurons of supraoptic nuclei.

Administration of hypertonic NaCl solutions by intraperitoneal injection evokes a transient expression of immediate-early genes in the hypothalamic magnocellular neurons of supraoptic nuclei (SON), which is followed by an upregulation of arginine vasopressin synthesis and a general increase in cellular metabolic activity. Here we have analysed the changes that occur in the nucleus of SON neurons during the period of transient Fos expression after injection of hypertonic saline. Within the first 30 minutes after injection, the nuclei become significantly smaller, contain more condensed chromatin and incorporate less 3H-uridine than the controls. By 12 hours these effects are reverting and at 24 hours the nuclei are already more active than the controls. Additionally, we observe an initial decrease in the number of coiled bodies per nucleus within the first 2 hours, followed by a 3-fold increase at 24 hours after injection. As coiled bodies are transcription-dependent subnuclear 'organelles', these results further support the view that injection of hypertonic saline causes a transient inhibition of nuclear activity. Our data show that SON neurons respond to acute osmotic/stress stimuli first with inhibition and then with activation of gene expression. Importantly, inhibition of transcriptional activity occurs simultaneously with maximal accumulation of Fos protein in the nucleus, raising the possibility that activation of c-fos expression may cause repression of target genes.

Animals↗

PubMed1997

Semithin cryosections as a tool to perform high resolution immunofluorescence and in situ hybridization analysis of the nervous tissue: a study in the supraoptic nucleus.

Immunofluorescence and fluorescence in in situ hybridization represent powerful approaches to correlate biochemical and molecular data with the structural organization of cells and tissues. However, the analysis of tissues by fluorescence microscopy is limited by the fact that most methods currently used to preserve the morphological integrity of sectioned samples at high resolution do not allow access of the labeled probes to the target molecules. Here we have made use of semithin cryosections obtained from rat supraoptic nucleus to perform immunofluorescence with antibodies directed against cytoplasmic and nuclear antigens, as well as fluorescence in situ hybridization with antisense oligonucleotide probes complementary to the poly(A) tail of mRNA and to specific mRNAs. In addition, DNA was visualized by incubation of sections with digoxigenin-labeled nucleotides in the presence of Escherichia coli DNA polymerase I. The high resolution of this DNA staining in combination with immunolabeling for nuclear antigens provides a powerful tool to analyze the structural and functional compartmentalization of neuronal cell nuclei. The major conclusion from this study is that performing fluorescence microscopy on 1 micron-thick cryosections provides an important tool to accurately localize proteins, DNA and RNA within nervous tissue in general and particularly in the model of supraoptic nucleus. Moreover, the cryosectioning technique appears particularly suited to the study of the localization of specific mRNA species in the neuronal cytoplasm and represents a useful approach to addressing the functional significance of mRNA localization in protein targeting.

Animals↗

PubMed1997

Insulin-like growth factor I receptor gene expression during postnatal development of rabbit kidney.

BACKGROUND: Insulin-like growth factor-I (IGF-I) is a peptide growth factor whose biological effects are mediated through a specific receptor (IGF-IR). IGF-I and IGF-IR are detected in both fetal and adult kidneys and have both metabolic and growth effects. IGF-IR expression during postnatal kidney development is not well defined and the biological role of this receptor during the postnatal stage is not clearly established. The purpose of the present study was to analyze IGF-IR gene expression during the postnatal development of rabbit kidney to achieve a better understanding of the correlation between growth and differentiation of kidney tissues and IGF-IR expression. METHODS: Using in situ hybridization, we studied changes in IGF-IR expression in the kidneys of newborn rabbits and those up to 35 days old. Evaluation of the stage of kidney development and morphological maturation was made on histological sections stained with hematoxylin-eosin. RESULTS: High levels of IGF-IR gene expression in the rabbit kidney occurred in the last stages of postnatal development and in the adult stages; during the development of the subcapsular metanephrogenic zone, IGF-IR gene expression was not observed. IGF-IR mRNA was expressed by proximal and distal tubules and by collecting ducts after these tissues attained morphological maturation. The appearance of IGF-IR mRNA in these kidney structures followed a precise temporo-spatial sequence. IGF-IR was not expressed by renal corpuscles, Henle's loops, inner medullary collecting ducts, vessels, or interstitial cells at any study stage. CONCLUSIONS: The temporal and spatial patterns of IGF-IR gene expression during postnatal development of the rabbit kidney suggest that IGF-IR and its ligands are relevant for the acquisition of the function, and not for development events, by proximal and distal tubules and collecting ducts. This study also suggests that IGF-IR mRNA localization constitutes a useful marker to determine the functional maturation of these renal structures.

Animals↗

PubMed1997

Fulminant Guillain-Barré syndrome with universal inexcitability of peripheral nerves: a clinicopathological study.

The pathological basis of nerve inexcitability in Guillain-Barré syndrome has not been established with certainty. We report the clinicopathological findings in a 67-year-old patient with fulminant Guillain-Barré syndrome who died 18 days after onset. Three serial electrophysiological studies revealed nerve inexcitability. Antibodies to Campylobacter jejuni were present but there was no antiganglioside reactivity. Spinal root sections revealed extensive and almost pure macrophage-associated demyelination with occasional presence of T lymphocytes and neutrophil leukocytes. Conversely, in femoral, median, and sural nerves the outstanding lesion was axonal degeneration, with some denuded axons remaining. Unmyelinated fibers, posterior root ganglia, and dorsal columns were preserved. Endoneurial postcapillary venules showed plump endothelial cells with loss of their tight junctions. We conclude that both primary demyelination and axonal degeneration secondary to inflammation account for nerve inexcitability. Our findings lend support to the hypothesis of increased endoneurial pressure as the cause of wallerian degeneration in nerve trunks.

Action Potentials↗

PubMed1997

Apoptosis induced by methylazoxymethanol in developing rat cerebellum: organization of the cell nucleus and its relationship to DNA and rRNA degradation.

We present a cytological and biochemical study of the cell death of granule cell precursors in developing rat cerebellum following treatment with the cytotoxic agent methylazoxymethanol (MAM) during the first postnatal week. The density of apoptotic figures per square millimeter progressively increases after 6, 12, 24 and 44 h of treatment, whereas cells immunoreactive for proliferating cell nuclear antigen tend to disappear in the external granular layer (EGL). DNA migration on gel electrophoresis reveals a typical ladder pattern of internucleosomal cleavage following MAM treatment, whereas gel electrophoresis of rRNA shows a conspicuous degradation of both 28S and 18S rRNAs. Ultrastructural analysis has revealed the alterations of structures containing chromatin and ribonucleoprotein (RNP) in dying cells of the EGL. The typical granular beaded configuration of the condensed chromatin changes to a denser, more homogeneous texture suggesting nucleosomal disruption. The reorganization of RNP nuclear domains is reflected by the appearance of dispersed nucleoplasmic RNP particles and the formation of a coiled-body-like structure. However, typical nuclear domains involved in the splicing of RNAs, namely interchromatin granule clusters and typical "coiled bodies", are not found in apoptotic cells. Intranuclear bundles of filaments have also been detected. In the cytoplasm, the presence of dispersed single ribosomes is an initial sign of apoptosis. The massive dispersion and disruption of ribosomes detected after 24 h and 44 h of MAM treatment is reflected by the degradation of both 28S and 18s rRNAs. These results show that MAM treatment provides a useful experimental model for the study of apoptosis in the developing central nervous system. The organization of the cell nucleus in cells undergoing apoptosis clearly reflects a disruption of the nuclear compartments involved in transcription and the processing and transport of RNA and is related to the patterns of DNA and rRNA degradation.

Animals↗

PubMed1996

Schwann cell nuclear remodelling and formation of nuclear and coiled bodies in Guillain-Barré syndrome.

We have examined the reorganization of the cell nucleus in myelin-related Schwann cells (SCs) in a case of acute Guillain-Barré syndrome (GBS). Spinal root samples of the GBS case and human controls were processed for light and electron microscopy. The cytochemical EDTA method for ribonucleoproteins (RNPs) and a specific silver staining technique for nucleolar organizer regions were used on ultrathin sections. In SCs of the GBS case, we observed a significant increase in nuclear size (64.99 +/- 10.47 microns 2 in the GBS vs 35.07 +/- 8.74 microns 2 in the controls, mean +/- SD) accompanying partial decondensation of heterochromatin domains and elaboration of an extensive network of RNP-containing perichromatin fibrils. In addition, the formation of two types of nuclear structures, coiled bodies and nuclear bodies of Bouteille, was induced in SCs of the case of acute GBS. Free coiled bodies were observed in the nucleoplasm and were characteristically stained with both RNP and silver procedures. Typical "simple" and "complex" nuclear bodies were regularly found, sometimes in association with coiled bodies. On the basis of cell nucleus physiology, all of these changes are considered cytological indicators of enhanced transcription and cellular hyperactivity, and they seem to reflect a reactive response of SCs triggered by the constellation of cellular and humoral signals associated with acute GBS.

Aged↗

PubMed1995

Expression of insulin-like growth factor receptor mRNA in rabbit atherosclerotic lesions.

We have found by 'in situ' hybridization a high level of expression of insulin- like growth factor I receptor (IGF-I R) gene in foam cells of atherosclerotic rabbit aortas. By reaction with either anti- rabbit macrophage (RAM11) or anti smooth muscle cell antibodies we also found that most cells expressing increased amounts of IGF-I R mRNA were of smooth muscle cell origin. Thus, increased IGF-I R mRNA levels might be related to the genesis of the atheroma plaque.

Animals↗

PubMed1995

Age-induced hypertrophy of astrocytes in rat supraoptic nucleus: a cytological, morphometric, and immunocytochemical study.

BACKGROUND: In the adult rat, neuron-astroglia interactions in the supraoptic nucleus (SON) are characterized by the structural and functional plasticity of astrocytes in response to several physiological and experimental conditions. This study has analyzed the plasticity of the supraoptic nucleus astrocytes in response to the age-induced changes in neuronal activity. METHODS: The study was performed in 5-, 12-, 18- and 24-month-old rats. The cytology and organization of astrocytes in the SON were examined using glial fibrillary acidic and vimentin immunocytochemistry and ultrastructural and morphometric analysis. RESULTS: No significant age-related variations in the total number of neurons and astrocytes in the SON were detected, although a few degenerating neurons were found in old rats. An age-dependent increase in GFAP immunoreactivity was observed at the ventral glial lamina, perivascularly and between neuronal perikarya. Vimentin overexpression was also detected in ventral lamina astrocytes with advancing age. At the cell nucleus level, we observed an age-associated increase in nuclear size and in the number of coiled bodies, nuclear bodies, and "cleared" nucleoplasmic areas, as well as changes in the nucleolar organization. At the cytoplasmic level, characteristic ultrastructural features in astrocytes of old rats were the hypertrophy of intermediate filament bundles and the formation of an extensive network of Golgi stacks interlinked by tubulovesicular elements. Glial filaments were often associated with the nuclear envelope and polyribosomes. CONCLUSIONS: The increased GFAP and vimentin immunoreactivity and the morphometric and cytological changes in rat SON astrocytes may reflect a sustained upregulation of cellular activity with age, resulting in hypertrophy of glial perikarya and cell processes. Several factors that are known to influence the expression of the astrocytic phenotype, such as signals produced by degenerating neurons and activated microglia, as well as variations in neuronal activity are considered possible causes of the age-associated changes in SON astrocytes.

Aging↗

PubMed1995

Number of nucleoli and coiled bodies and distribution of fibrillar centres in differentiating Purkinje neurons of chick and rat cerebellum.

We used differentiating chick and rat Purkinje cells to investigate in homologous neurons the influence of the number of nucleolar organizer regions (two in the chick and six in the rat) on the behaviour of the nucleolus and coiled bodies. We employed specific silver-staining methods on smear preparations and on semithin and ultrathin sections. In chick Purkinje cells the number of nucleolar silver-staining granules increased from 15.7 +/- 3 (mean +/- SD) at embryonic day 13 to 23.8 +/- 3 at post-hatching day 7. These nucleolar granules were unevenly distributed between the two nucleoli of binucleolated cells. Electron-microscopic cytochemistry showed that nucleolar granules are equivalent to the fibrillar centres with their associated shell of dense fibrillar component. A reduction in the number of nucleoli was found during the differentiation of both chick and rat Purkinje cells, although in mature cells the average number of nucleoli per cell was higher in the chick (1.60) than in the rat (1.07). The number of coiled bodies decreased from 1.33 in newborn rats to 0.47 at postnatal day 90 in the rat. Coiled bodies were not observed in homologous chick Purkinje cells. The dynamic behaviour of nucleoli and coiled bodies during neuronal differentiation and the relationship of these two nuclear organelles with the number of nucleolar organizer regions is discussed.

Animals↗

PubMed1995

Expression of apolipoprotein E in cholesterol-loaded macrophages of extrahepatic tissues during experimental hypercholesterolemia.

To study the expression of extrahepatic apolipoprotein E (apoE) under hypercholesterolemic conditions, apoE mRNA levels were evaluated in 14 tissues of rabbits fed on a cholesterol rich diet and compared to age-matched control animals. In hypercholesterolemic rabbits apoE expression was significantly induced in adipose tissue, adrenals, aorta, lung and spleen. The increase in apoE mRNA levels in lung and spleen was associated with the presence of cholesterol-loaded macrophages. These cells were found to express high levels of apoE mRNA as demonstrated by in situ mRNA hybridization. Our results suggest that extrahepatic tissues would be partially responsible for the rise in serum apoE levels detected under hypercholesterolemic conditions.

Adipose Tissue↗

PubMed1995

Expression of histone H1 (zero) in transcriptionally activated supraoptic neurons.

This study has analysed by immunocytochemistry the expression pattern of histone H1 zero after the osmotically induced activation of transcription in supraoptic nucleus neurons of the rat. In control rats, histone H1 zero was constitutively expressed in neuronal and glial cell nuclei of supraoptic nucleus. After chronic neuronal stimulation by intermittent salt-loading, the majority of neuronal cell nuclei exhibited a marked reduction of immunostaining, which was confirmed by densitometric analysis of immunoreactivity. This effect was reversible, since optical density values returned to control levels when the stimulation of supraoptic neurons was suppressed by rehydration. Ultrastructural immunocytochemistry of histone H1 zero showed that immunogold particles specifically decorated chromatin fibers, with the highest accumulation of particles being on the condensed inactive chromatin. These results indicate that transcriptional activation in supraoptic neurons is accompanied by a depletion of the chromatin-associated histone H1 zero, and also suggest that this transcription-dependent expression of histone H1 zero may be involved in regulating chromatin condensation and gene expression in mature neurons that constitutively express this protein.

Animals↗

PubMed1994

Effects of cycloheximide on the structural organization of the nucleolus and the coiled body in normal and stimulated supraoptic neurons of the rat.

This study was designed to determine the effects of cycloheximide, a protein synthesis inhibitor that interferes with rRNA synthesis and processing, on the nucleoli and coiled bodies of supraoptic nucleus neurons from normally-hydrated and osmotically-stimulated rats. The number of nucleoli and the nucleolar size were estimated on smear preparations of previously silver-impregnated supraoptic nucleus. No significant differences were registered in the mean number of nucleoli per cell in cycloheximide-treated rats. The number of nucleoli per neuron remained constant, at about 1.3, in all animal groups, suggesting that the nucleoli number is strictly regulated in differentiated neurons. By contrast, a significant reduction in the average nucleolar volume of supraoptic nucleus neurons was detected in cycloheximide-treated groups of rats in comparison with their equivalent non-treated groups. By electron microscopy, most nucleoli and coiled bodies of supraoptic nucleus neurons exhibited cycloheximide-induced alterations in their fine structure and configuration. Nucleolar changes included the occurrence of a few large fibrillar centres, the formation of microspherules and small intranucleolar vacuoles or dilated interstices, and the partial segregation of nucleolar components coupled with the transformation of reticulated nucleoli--a nucleolar configuration characteristic of supraoptic nucleus neurons of non-cycloheximide-treated rats--into compact ones. The redistribution of nucleolar components might reflect the interference with rDNA transcription, and also supports the hypothesis that the normal assembly of these components into the nucleolus depends upon ongoing nucleolar transcription. Concerning coiled bodies, most of them revealed ultrastructural alterations, particularly segregation of the amorphous matrix, compactation of coiled threads and formation of coiled body-derived dense bodies of fibrillar nature. Moreover, cycloheximide also induced the formation of smaller dense bodies--here referred to as dense microbodies--which presumably represent a distinct nuclear entity different from coiled bodies. Ultrastructural silver staining of nuclear bodies showed a selective silver reaction on the dense fibrillar component of normal and altered coiled bodies, as well as on the dense microbodies. The possible relationship between the nucleolus and both coiled bodies and dense microbodies is discussed.

Animals↗

PubMed1994

Apolipoprotein E expression in the cerebellum of normal and hypercholesterolemic rabbits.

We have studied the expression of apolipoprotein E (ApoE) mRNA in the cerebella of control and experimental rabbits fed with a cholesterol-rich diet for 8 weeks. Cholesterol-treated rabbits show a dramatic increase in serum cholesterol levels; however, no significant variations in the expression level of cerebellar ApoE mRNA were found in comparison to control rabbits. In addition, no differences were observed between control and hypercholesterolemic rabbits in the in situ hybridization pattern of ApoE mRNA on cerebellar cortex sections. ApoE mRNA was localized in astroglial processes associated with Purkinje cell bodies and dendrites, granule cell clusters, blood vessels and nerve fibers of the white matter. No expression of ApoE mRNA was observed in Purkinje and granule cell neurons. Polarized light examination of cryostat cerebellar sections revealed the absence of cholesterol-rich microglia/macrophage cells induced by the hypercholesterolemia. In this way, neither reactive microglial cells nor perivascular phagocytes were found by ultrastructural analysis in hypercholesterolemic conditions. The pattern of glial fibrillary acidic protein of the astroglial cells of the cerebellar cortex as well as their nuclear size were unchanged following cholesterol treatment, indicating the absence of astroglial activation induced by hypercholesterolemia. Our results suggest that cerebellar ApoE does not contribute to the general cholesterol homeostasis outside of the brain and supports the view that this cerebellar ApoE is involved in paracrine and autocrine functions particularly related with synapse turnover and membrane remodelling of astroglial cells.

Animals↗

PubMed1993

Protein-synthesis inhibition induces perichromatin granule accumulation and intranuclear rodlet formation in osmotically stimulated supraoptic neurons.

The distribution of perichromatin granules (PGs), a storage form of pre-mRNAs, was studied in supraoptic neurons of control and osmotically stimulated rats, and also after treatment with cycloheximide, a protein-synthesis inhibitor. In non-cycloheximide-treated rats, neuronal activation by dehydration significantly decreased the number of PGs. Conversely, PGs were drastically increased in the supraoptic neurons of dehydrated rats treated with cycloheximide for 4 h. This suggests that cycloheximide does not interfere with the transcriptional activation induced by dehydration, but it affects the processing of newly synthesized pre-mRNAs. Moreover, protein-synthesis inhibition was associated with the formation of intranuclear bundles of tubular filaments.

Animals↗

PubMed1993

Nuclear Fos domains in transcriptionally activated supraoptic nucleus neurons.

This study has analysed by light and electron microscopy immunolocalization the nuclear pattern of distribution of Fos-related proteins in supraotic neurons. Two experimental models of transcriptional activation have been used: sustained, global transcriptional activation, at relatively near physiological conditions, by six days of chronic intermittent salt loading; and superinduction of c-fos gene by this salt loading regime plus cycloheximide treatment for 4 h. In the first condition, the ultrastructural analysis showed a distribution of Fos-like immunoreactivity on the reticular network of dispersed chromatin that extends between the nucleolar surface and the nuclear envelope, whereas the Fos-negative adjacent interchromatin spaces appeared rich in interchromatin granules by using a cytochemical staining for ribonucleoproteins. The nucleolus associated heterochromatin, fibrillar centers of the nucleolus and coiled bodies were free of immunoreactivity. This immunoelectron pattern seems to indicate that active genes containing activator protein-1 and cyclic AMP response element recognition sites are extensively distributed in euchromatin regions and suggests that the Fos-positive nuclear domains correspond to the actively transcribing chromatin regions, at least in supraoptic neurons. It also suggests that these Fos-positive transcription domains are complementary to adjacent ribonucleoprotein-rich interchromatin spaces which are involved in the processing and splicing of pre-messenger RNA. Moreover, the absence of immunoreactivity on the fibrillar centers, the sites of pre-ribosomal RNA synthesis, suggests that the Fos protein complexes are not involved in regulating the expression of ribosomal RNA genes. Following superinduction of c-fos gene by osmotic stimulation plus cycloheximide treatment, a conspicuous Fos-like immunoreactivity was detected in dispersed chromatin regions, whereas the heterochromatin masses, nucleoli and coiled bodies showed no immunoreaction. Moreover, this treatment induced the formation of nuclear "dense bodies" of a fibrillar nature which were free of immunolabelling. Since Fos proteins are known to be short-lived, the expression of these nuclear constituents, under conditions of protein synthesis inhibition induced by the cycloheximide, suggests the stabilization of chromatin-bound Fos complexes or, alternatively, a preferential synthesis of Fos proteins.

Animals↗

PubMed1993

Reactive astroglia-neuron relationships in the human cerebellar cortex: a quantitative, morphological and immunocytochemical study in Creutzfeldt-Jakob disease.

In order to investigate the role of neuron-glia interactions in the response of astroglial to a non-invasive cerebellar cortex injury, we have used two cases of the ataxic form of Creutzfeldt-Jakob disease (CJD) with distinct neuronal loss and diffuse astrogliosis. The quantitative study showed no changes in cell density of either Purkinje or Bergmann glial cells in CJ-1, whereas in the more affected CJ-2 a loss of Purkinje cells and an increase of Bergmann glial cells was found. The granular layer in both CJD cases showed a similar loss of granule cells (about 60%) in parallel with the significant increase in GFAP+ reactive astrocytes. GFAP immunostaining revealed greater reactivity of Bergmann glia in CJ-2 than in CJ-1, as indicated by the thicker glial processes and the higher optical density. Granular layer reactive astrocytes were regularly spaced. In both CJD cases there was strict preservation of the spatial arrangement of all astroglial subtypes--Fañanas cells, Bergmann glia and granular layer astrocytes. Reactive Fañanas and Bergmann glial cells and microglia/macrophages expressed vimentin, while only a few vimentin+ reactive astrocytes were detected in the granular layer. Karyometric analysis showed that the increase in nuclear volume in reactive astroglia was directly related with the level of glial hypertrophy. The number of nucleoli per nuclear section was constant in astroglial cells of human controls and CJD, suggesting an absence of polyploidy in reactive astroglia. Ultrastructural analysis revealed junctional complexes formed by the association of macula adherens and gap junctions. In the molecular layer numerous vacant dendritic spines were ensheathed by lamellar processes of reactive Bergmann glia. Our results suggest that quantitative (neuron/astroglia ratio) and qualitative changes in the interaction of neurons with their region-specific astroglial partners play a central role in the astroglial response pattern to the pathogenic agent of CJD.

Astrocytes↗

PubMed1993

Transcriptional response to cAMP in brain: specific distribution and induction of CREM antagonists.

Changes in cAMP levels are often associated with the modulation of neuronal function. The CREM gene encodes both antagonists and activators of the cAMP-dependent transcriptional response by alternative splicing. CREM transcripts in rat brain show a characteristic pattern of expression, being specific for the inner layer of the cerebral cortex, anterior thalamus, hippocampus, and hypothalamus. Strikingly, the CREM transcripts correspond to the antagonist isoforms in these areas, suggesting a down-regulatory role for CREM in brain; in contrast, the expression of CREM tau and CREB activators is more diffuse and generalized. In the supraoptic nucleus, CREM expression is induced after osmotic stimulus. Importantly, this demonstrates physiological inducibility of CREM, which is novel within the CRE/ATF family.

Animals↗

PubMed1993

Nuclear compartmentalization in transcriptionally activated hypothalamic neurons.

Transcription of cell-specific vasopressin and oxytocin genes as well as transcription of those housekeeping genes responsible for general metabolic activation and cellular hypertrophy is induced in supraoptic hypothalamic neurons by rises in plasma osmolarity. In this study, the nuclear volume, the ultrastructure of chromatin and the number and distribution of nuclear particles in the cell nuclei of supraoptic neurons of 3-month-old male Sprague-Dawley rats were analyzed after osmotically induced activation of transcription by periods of acute (1 day) and chronic (6 days) dehydration, and after halting the stimulation by rehydration of animals. The nuclear volume and the ultrastructure of chromatin were assessed on ultrathin sections. The number and distribution of nuclear particles were assessed on freeze-fracture replicas. The initial phase of osmotically induced enhancement of transcription was accompanied by an increase in nuclear volume and by a partial replacement of nuclear particles of large diameter (> 11 nm) by smaller nuclear particles. This latter change affected predominantly the nuclear periphery (0-1,000 nm from the nuclear membrane) and occurred simultaneously with a partial decondensation of chromatin clusters that may be related to chromatin unfolding. In chronically stimulated animals, the decondensation of chromatin and the replacement of large nuclear particles by smaller ones was enhanced in the nuclear periphery and was partially propagated to the interior of the nucleus. After suppression of cellular activation by rehydration of animals, the number of nuclear particles returned to control levels in the nuclear periphery while in the center of the nucleus the number of small particles decreased and the number of large particles increased as compared to control values. These results, together with the observation that in unstimulated cells the nuclear periphery and the nuclear interior differ in their composition of nuclear particles, evidence a structural and functional compartmentalization in the cell nucleus of supraoptic neurons.

Animals↗