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D Gros

Publications and source records attributed to D Gros.

At least 55 records · Page 3Linked to original sources

Atrial natriuretic factor messenger ribonucleic acid and peptide in the human heart during ontogenic development.

We have investigated the level of expression of the atrial natriuretic factor (ANF) gene in the human heart during ontogenic development by determining the concentrations of ANF messenger ribonucleic acid (ANF mRNA), of immunoreactive ANF (IR ANF) and of receptor reactive ANF (RR ANF), in myocardial samples of the various heart chambers. We found the level was high and almost identical in the left and right ventricles in utero. It gradually decreased during ontogenic development to reach the low adult levels, with a more rapid decrease in the right than in the left ventricle after birth. In the atria, ANF gene expression was high as early as the 13th week of gestation, was higher in the right than in the left atrium, and appeared little affected by ontogenic development.

Aging↗

Cross-linking of cardiac gap junction connexons by thiol/disulfide exchanges.

SDS-polyacrylamide gel electrophoresis and immunoblotting were used to investigate inter- and intramolecular disulfide bonds to connexin 43 (the cardiac gap junctional protein) in isolated rat heart gap junctions and in whole heart fractions. In gap junctions isolated in the absence of alkylating agent, connexin 43 molecules are cross-linked by disulfide bonds. The use of iodoacetamide (100 mM) for the first steps of isolation procedure prevents the formation of these artifactual linkages. Investigation of connexin 43 in whole heart fractions by means of antibodies confirms the results obtained with isolated gap junctions; that is, connexin 43 molecules are not interconnected with disulfide bridges. In whole heart fractions treated with alkylating agents, a 38 kD protein, immunologically related to connexin 43, and containing intramolecular disulfide bonds is detected. It is hypothesized that this protein might be a folded form of connexin 43, a precursory form of the molecules embedded in the gap junctions.

Animals↗

Tissue-specific transcriptional control of alpha- and beta-tropomyosins in chicken muscle development.

During muscle maturation, isoform switching of contractile proteins to attain the adult phenotype involves both stage-specific and muscle-specific regulatory mechanisms. Chicken pectoralis major (PM) provides an interesting model to study the latter since a specific pattern of tropomyosin (TM) with repression of the beta TM isoform is displayed by the adult PM. The developmental pattern of alpha and beta fast skeletal muscle tropomyosins' (alpha f and beta TM) RNAs was investigated with 3' untranslated region specific probes. In PM, the beta TM messenger ceased to accumulate after hatching through a transcriptional control, as shown by run-on assays, so that, at Day 8 ex ovo, no beta TM mRNA was detected. In this same muscle, in parallel with the disappearance of the beta TM mRNA, there was a boost in the accumulation of the alpha f TM mRNA. In the leg muscles, following hatching, there was only a moderate increase in the level of the alpha f TM mRNA, together with a slight decrease in the accumulation of the beta TM mRNA. Taken together, these results show that chicken muscle maturation involves tissue-specific transcriptional control of tropomyosin genes and could suggest a possible coordinate regulation of the two genes.

Animals↗

Immunological characterization of rat cardiac gap junctions: presence of common antigenic determinants in heart of other vertebrate species and in various organs.

Antibodies to the following synthetic peptide, SALGKLLDKVQAY, were purified by affinity chromatography and characterized by ELISA and immunoblotting. These antibodies, shown to be specific to the major protein constituent of isolated rat heart junctions: connexin 43, cross-reacted with a homologous protein in immunoreplicas of whole heart fractions of trout, frog, chicken, guinea pig, mouse and rat, suggesting a phylogenic conservation of connexin 43 in vertebrates. By immunoblotting of whole organ fractions it was also demonstrated that these antibodies cross-reacted with major proteins of Mr 32 and 22 kD in rat and mouse liver, of Mr 41 kD in rat cerebellum, of Mr 43 kD in uterus, stomach and kidney of rat, of Mr 46 and 70 kD in rat lens, suggesting that these proteins share common or related epitopes with the synthetic peptide and connexin 43.

Amino Acid Sequence↗

[Regulation of the expression of alpha and beta tropomyosin genes during development of the pectoral muscle in the chicken].

Accumulation of mRNAs coding for alpha and beta skeletal tropomyosins was investigated using specific probes and normalized to muscle creatine kinase (M-CK) mRNA by slot-blot assays. In developing pectoralis muscle, the ratio of alpha TM messenger/M-CK remained constant until hatching, at which time the messenger disappeared within a week. However, in the leg, this ratio remained constant until 8 days after hatching, whereafter it decreased progressively to reach 30% in the adult. The alpha TM/M-CK ratios were almost the same in the embryonic leg and pectoralis muscle. After hatching, there was a large increase in pectoralis muscle (x 3 at day +4, x 0 at day +21) whereas, the increase was less pronounced and more progressive in the leg (x 3 at day 21). Run-on assays showed that nuclei isolated from 15-day in ovo leg and pectoralis muscles had similar patterns of muscle specific gene transcription whereas post-hatched pectoralis muscle nuclei were shown to have a higher rate of alpha to beta tropomyosin gene transcription. These data are in accordance with the results obtained for protein analysis of leg and pectoralis muscles and support the notion that changes in the protein pattern of developing muscle can be relevant to coordinate regulation of gene transcription.

Animals↗

[Expression of MMTV proviruses in lymphopoietic organs of C3H Bi mice].

We show that transcription of MMTV proviruses takes place in lymphopoietic tissues (spleen, thymus) of C3H Bi mice, a strain with high incidence of tumors. This transcription is found in the spleen of 21 day old mice but is no longer detectable in the spleen of 4 month old mice. In the mammary tumor, the 3.8 kb RNA (24S) which encodes the viral envelope proteins is well transcribed whereas the 1.7 kb RNA (13 S) is barely detectable. In contrast, the 1.7 kb (13 S) is the major species present in the spleen of 21 day old mice. It appears that the 7.8 kb genomic RNA (35 S) is differentially spliced into a mature RNA of either 3.8 kb (24 S) or 1.7 kb (13 S). The function of the latter is still unknown, though it could encode a protein involved in the induction of carcinogenesis.

Animals↗

Evolution of the surface of mouse myocardial cells during ontogenesis. II. Changes of fluorescent lectin-binding sites.

Fluorescein isothiocyanate-conjugated lectins (FTC-lectins) have been used to study the occurrence and the distribution of their receptors in the coating of mouse myocardial cells throughout their differentiation (from 10 days post-coitum to the adult stage). FTC-Con A, -WGA and -PNA specifically inhibited by alpha-mannosyl, beta-N-acetylglucosaminyl and beta-galactosyl residues, respectively, bind to the cell surface whatever the stage of differentiation. FTC-SBA and -HPA, specifically inhibited by alpha-N-acetylgalactosaminyl residues, bind to the coating of adult cells; their receptor sites appear only at day 18 of development. FTC-UeA, specifically inhibited by fucosyl residues also binds to adult cells. In sections of embryonic hearts, FTC-UeA is trapped by a lectin-like component; this UeA binding can be prevented with mannosyl-substituted serum albumin. Isolated embryonic myocardial cells do not bind FTC-UeA. FTC-BS I-A4 and -BS I-B4, specifically inhibited by alpha-N-acetylgalactosaminyl and alpha-galactosyl residues, respectively, bind only to the capillaries of adult heart. These results show that the differentiation of myocardial cells is accompanied by the sequential acquisition of lectin-binding sites. These changes express the maturation of cell coating glycoconjugates.

Age Factors↗

The three cortical membranes of the gregarines. I. Ultrastructural organization of Gregarina blaberae.

Gregarines, parasitic protozoa of invertebrates, possess a highly differentiated cell surface, with three cortical membranes and associated structures. Transmission electron microscopy and freeze-fracture reveal the presence of two cytomembranes lying uniformly under the plasma membrane. The density and the distribution of the intramembraneous particles (IMPs) in the plasma membrane of Gregarina blaberae are similar to those reported for other eukaryotic cells. The IMP density is lower in the cytomembranes than in the plasma membrane. The distribution of IMPs in the different fracture faces of the two cytomembranes suggests that they are in topological continuity, forming either side of a flattened vesicle or cisterna. The sizes of the cytomembrane IMPs show a high variability. The nature of the IMPs, both for the plasma membrane and the cytomembrane, is discussed with regard to the integral proteins and glycoproteins of the ghost. The cell surface of G. blaberae exhibits numerous longitudinal folds with three types of cortical membrane-associated structures: 12 nm filaments, an internal lamina, and homogeneous structures described as 'rippled dense structures'. The 12 nm filaments, running under the cytomembranes along the longitudinal axis of each fold, exhibit the properties of intermediate filaments. Their distribution in mature cells and during the growth process suggests a participation in cell surface morphogenesis. The internal lamina, also localized under the cytomembranes, would stabilize each fold and assure a scaffolding function between the numerous folds. The rippled dense structures, settled on the external cytomembrane, show a regular distribution at the top of each fold. The membrane-associated structures are discussed with regard to the gliding movement mechanism.

Apicomplexa↗

Further studies on the junctional complex in the intestine of Sagitta setosa. Freeze-fracture of the pleated septate junction.

The intramembrane structures of the pleated septate junction which occur in the junctional complex of the intestine of the chaetognath Sagitta setosa have been investigated. The pleated septate junction is made up of linear rows of irregularly shaped and sized particles, often fused into short rods, and pits which can be fused into furrows. The distribution of these structures on E and P faces depends upon the preparative methods used. Many of the morphological characteristics are the same as those of the "lower invertebrate pleated septate junction type" defined by Green (1981 a). The physiological significance of this junction is obscure. On the basis of the presence of septate junctions (both of the paired septate junction and pleated septate junction types) which have mainly morphological characteristics of the "lower invertebrate pleated septate junction" we can add to the hypothesis that chaetognaths are not related to the molluscs and arthropods.

Animals↗

Ultrastructural localization of concanavalin A and wheat germ agglutinin binding sites in adult and embryonic mouse myocardium.

The lectins, concanavalin A (Con A) and wheat germ agglutinin (WGA), have been used to localize with precision glycosyl residues in adult and embryonic mouse myocardium. They were detected by means of an affinity method using peroxidase and chitobiosylperoxidase, respectively, which then were revealed with 3,3'-diaminobenzidine and H2O2. Exhaustive controls have shown that the binding of Con A and WGA is reversible when experiments are performed with adult specimens (tissue blocks or ultrathin sections of glycol methacrylate-embedded material) or with isolated embryonic cells. Experiments carried out with tissue blocks from embryonic hearts have shown peroxidase binding. This finding is discussed on the basis of the presence of the endogenous lectin-like components in embryonic hearts. Results show that the surface of adult and embryonic myocardial cells specifically bind both Con A and WGA, thus indicating the presence of glycosyl residues similar to alpha-methyl-D-mannoside and N-acetyl-D-glucosamine. In adult heart the transverse tubular system was also labeled. The absence of Con A and WGA receptor sites in the gap junction regions was demonstrated by means of an electron microscope postembedding staining method.

Animals↗

The caveolae in rabbit sinus node and atrium.

Caveolae or membrane vesicles are commonly observed in smooth and skeletal muscle as well as in working heart muscle. Using sections of fixed tissue and replicas of freeze-cleaved material, we show in this study that caveolae are also very numerous in sinus node cells of the rabbit, and to a lesser degree, in the atrial cells. Caveolae increase the plasma membrane surface area by 115% in the leading sinus node, and by 56% in the atrial cells. In these two cell types, the membrane of the caveolae contains four times fewer intramembranous particles than the rest of the plasma membrane, and this difference applies to both PF and EF faces. The role of the caveolae is still unclear, but it does not seem that they have a pinocytotic function.

Animals↗

Ultrastructural studies of the junctional complex in the musculature of the arrow-worm (Sagitta setosa) (Chaetognatha).

In the A fibres of the primary musculature of Sagitta, the junctional complex is made up of three kinds of junctions. From the apex to the base they occur in the following order: an apical zonula adherens, a columnar zonula then columnar maculae intermingled with gap junction. Each columnar junction joins two intracellular filament networks in adjacent cells; this cytoskeleton is largely developed around the nucleus of the A fibres and in close relation with the contractile apparatus, especially at the I band level. The B fibres, which never reach the general cavity, lack zonula adherens and columnar zonula. The columnar junction constitutes a new type of junction which seems to belong to the adherens kind. At their level fibrous columns cross the extracellular space, joining the membranes. Each column faces two cytoplasmic densities localized against the cytoplasmic leaflets of the membranes. A cytoskeleton composed of bunldes of cytoplasmic filaments is in close contact with these cytoplasmic densities. The great number of columnar junctions and associated cytoskeleton assure the cohesion of the tissue and the distribution of contractile forces in the absence of connective tissue. The abundance of gap junctions can account for the metabolic and ionic coupling of the fibres.

Animals↗

The junctional complex in the intestine of Sagitta setosa (Chaetognatha): the paired septate junction.

The junctional complex of the intestine of Sagitta setosa has been studied in tissues stained with uranyl acetate or after lanthanum impregnation, and by freeze-cleavage. All types of junctions have been characterized in both perpendicular and tangential planes. From the apex to the base of the cell the following junctions occur in this order: a zonula adhaerens; a septate junction where the septa occur in pairs; a pleated sheet septate junction; and numerous gap junctions of the A-type. From the upper part of the cells inwards to the septate junction, the membranes follow a relatively straight path. In the lower part of the cells the membranes are deeply interdigitating. At the intersection between 3 cells a very different junction is to be observed where small units, periodically disposed, bind the membranes of the 3 adjoining cells. Each unit is composed of 3 short segments which bind the cell membranes to a central ring 16.6 +/- 2.3 nm in outer diameter. The paired septate junction constitutes a new type. Its main features are that the septa are paired and occur in 2 formations, one the 'loose formation', with elements between the septa of each pair, and the other, a 'tight formation'. After lanthanum impregnation, the thickness of each septum is seen to be about 3 nm and the undulation period 12.6 +/- 1.6 nm. On freeze-fractures 10-nm particles are found on crests on the PF face and in furrows on the EF face. The possible significance of this type of junction is discussed. The junctional complex described is analogous to those found in various invertebrate epithelia.

Animals↗

The myocardial plasma membrane during development: influence of glutaraldehyde fixation on the density and size of intramembranous particles.

The ultrastructure of the plasma membrane of mouse myocardial cells has been studied during development (from 10 days post-coïtum to the adult stage) using the freeze-cleave method, and the effect of glutaraldehyde-fixation on this structure has also been evaluated. Blebs, free of intramembranous particles (IMP), were found to be dependent upon the developmental stage and upon fixation with glutaraldehyde. On P and E fracture faces, the presence of IMP-free areas was age-dependent and fixation-independent. No difference was demonstrated in the IMP density during heart development. Fixation with glutaraldehyde retains in the plasma membrane approximately 40% of IMP, probably by preventing an alteration induced by glycerol. Whatever the developmental stage, E face-associated particles are significantly larger than P face-associated particles. The diameter of both types of particle increases slightly but irregularly as myocardial cells mature. Fixation reduces the particle size on both fracture faces in such a way that no significant difference between them can be demonstrated.

Aldehydes↗