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D Verrier

Publications and source records attributed to D Verrier.

15 recordsLinked to original sources

Assessing a relationship between bone microstructure and growth rate: a fluorescent labelling study in the king penguin chick (Aptenodytes patagonicus).

Microstructure-function relationships remain poorly understood in primary bone tissues. The relationship between bone growth rate and bone tissue type, although documented in some species by previous works, remains somewhat unclear and controversial. We assessed this relationship in a species with extreme adaptations, the king penguin (Aptenodytes patagonicus). These birds have a peculiar growth, interrupted 3 months after hatching by the austral winter. Before this interruption, chicks undergo extremely rapid statural and ponderal growth. We recorded experimentally (by means of fluorescent labelling) the growth rate of bone tissue in four long bones (humerus, radius, femur and tibiotarsus) of four king penguin chicks during their fastest phase of growth (3-5 weeks after hatching) and identified the associated bone tissue types ('laminar', 'longitudinal', 'reticular' or 'radial' fibro-lamellar bone tissue). We found the highest bone tissue growth rate known to date, up to 171 microm day(-1) (mean 55 microm day(-1)). There was a highly significant relationship between bone tissue type and growth rate (P<10(-6)). Highest rates were obtained with the radial microarchitecture of fibro-lamellar bone, where cavities in the woven network are aligned radially. This result supports the heuristic value of a relationship between growth rate and bone primary microstructure. However, we also found that growth rates of bone tissue types vary according to the long bone considered (P<10(-5)) (e.g. growth rates were 38% lower in the radius than in the other long bones), a result that puts some restriction on the applicability of absolute growth rate values (e.g. to fossil species). The biomechanical disadvantages of accelerated bone growth are discussed in relation to the locomotor behaviour of the chicks during their first month of life.

Analysis of Variance↗

The enzyme indoleamine 2,3-dioxygenase is induced in the mouse brain in response to peripheral administration of lipopolysaccharide and superantigen.

The essential amino-acid, L-tryptophan, is the precursor of serotonin. Its availability in the brain is controlled by indoleamine 2,3-dioxygenase (IDO). This enzyme is inducible by cytokines such as interferon-gamma (IFN-gamma) and is the first and rate-limiting enzyme of the catabolism pathway of tryptophan. Since induction of IDO has been proposed to mediate the influence of cytokines on mood in patients with various somatic disorders, the present study aimed at analyzing the relationships between changes in brain IDO activity and serum IFN-gamma levels in response to peripheral immune stimulation by lipopolysaccharide (LPS) and superantigen in mice. Each of these treatments induced an increase in serum IFN-gamma at 6 h post-treatment followed 24 h later by a two-fold increase in IDO activity in the brain. These results support the involvement of peripheral IFN-gamma in the control of L-tryptophan catabolism in the brain.

Animals↗

Characterization of interleukin-1 receptor antagonist isoform expression in the brain of lipopolysaccharide-treated rats.

The endogenous interleukin-1 receptor antagonist is the natural inhibitor of the biological effects of interleukin-1 during inflammation. Interleukin-1 receptor antagonist refers to three isoforms: one secreted and two intracellular forms (types I and II). The objective of the present study was to investigate the expression of interleukin-1 receptor antagonist isoforms in the rat brain in vivo in response to an i.p. injection of lipopolysaccharide. The interleukin-1 receptor antagonist was studied at the messenger and protein levels by reverse transcription-polymerase chain reaction and western blot analysis, respectively. Interleukin-1 receptor antagonist messenger RNA was constitutively expressed in the brain and its expression increased in response to lipopolysaccharide. The three interleukin-1 receptor antagonist protein isoforms were up-regulated after lipopolysaccharide treatment in a time-dependent manner. Their relative expression differed according to the isoform and brain region studied. Double immunofluorescence staining revealed interleukin-1 receptor antagonist positive neurons and microglia in hippocampus 24h after lipopolysaccharide stimulation. These results demonstrate for the first time that brain cells are able to produce interleukin-1 receptor antagonist isoforms in response to a peripheral immune challenge with a predominance of the secreted over intracellular forms.

Blotting, Western↗

Rat microglial cells secrete predominantly the precursor of interleukin-1beta in response to lipopolysaccharide.

Little is known on the forms of interleukin-1beta (IL-1beta) that are produced by microglial cells in the nervous system. Mixed glial cell cultures of rats produced IL-1beta in response to lipopolysaccharide (LPS). Using Western blot, pro-IL-1beta was found to be localized both intracellularly and in the supernatant, whereas mature IL-1beta was found only in the supernatant but in lower quantities than pro-IL-1beta. Immunocytochemistry confirmed that microglial cells are the exclusive source of IL-1beta. Blockade of the IL-1beta-converting enzyme (ICE) by Tyr-Val-Ala-Asp-aldehyde (YVAD-CHO) decreased the levels of mature IL-1beta but had no effect on pro-IL-1beta. Release of pro-IL-1beta was not associated with cell death nor with the extracellular release of ICE. Using gelatin zymography, glial cells were found to express constitutive matrix metalloproteinases (MMP) in the form of MMP-2. Exposure to LPS induced MMP-9 expression in a time-dependent manner similar to the pro-IL-1beta expression profile. MMP activation and inhibition experiments indicated a possible role of MMPs in the cleavage of pro-IL-1beta but not in the generation of mature IL-1beta. Microglial cells share with macrophages the ability to release large amounts of pro-IL-1beta of which the extracellular role remains to be determined.

Animals↗

Production of interleukin-1 receptor antagonist isoforms by microglia in mixed rat glial cells stimulated by lipopolysaccharide.

Although the natural interleukin-1 receptor antagonist (IL-1Ra) has been shown to be produced by microglial cells in response to immune stimuli, nothing was known about the ability of these cells in primary culture to produce the different isoforms of IL-1Ra. Using RT-PCR, we first confirmed that mixed glial cell cultures from newborn rats respond to the cytokine inducer, lipopolysaccharide, by synthesizing IL-1Ra mRNA. Using double immunostaining, we showed that IL-1Ra was detected in microglia but not in astrocytes. Using Western blotting, we finally demonstrated that the IL-1Ra1 isoform was secreted in the supernatant of mixed glial cell cultures, and its production increased in response to lipopolysaccharide. The three different IL-1Ra isoforms were constitutively expressed in cell lysates and their levels increased after lipopolysaccharide treatment, except for IL-1Ra3. These results point to the ability of microglial cells in primary culture to produce the different isoforms of IL-1Ra.

Animals↗

Reproducibility of 1H-NMR integrals: a collaborative study.

The quantitative use of NMR spectroscopy was investigated by a reproducibility study of 1H-NMR integrals involving five laboratories. A significant laboratory effect was found confirming the difficulty to obtain very precise data by integration of complex signals. The reproducibility of any NMR assay measurement, which requires a high precision should be validated by an interlaboratory study.

Evaluation Studies as Topic↗

Apoptosis occurs independently of the release of interleukin-1 beta in the anterior pituitary of end-lactating rats.

The mechanism of Interleukin-1 beta (IL-1 beta) release remains unknown. Because of the absence of typical peptide signal on the precursor, IL-1 beta is not secreted through the classical pathway. The aim of this study was to determine whether IL-1 beta is released during apoptosis, as has been reported for activated macrophages. We chose anterior pituitary cells of end-lactating rats because of their capacity to produce IL-1 beta spontaneously and because this organ undergoes cellular degeneration. The combination of two techniques, reverse haemolytic plaque assay (RHPA) and terminal transferase dUTP nick end labelling (TUNEL), allowed us to observe simultaneously the release of IL-1 beta and apoptosis. Our results show that in these conditions apoptosis is not the mechanism of IL-1 beta release.

Animals↗

EGF release by rat gonadotroph cells: characteristics and effects of LHRH.

The ability of anterior pituitary cells of immature female rats to secrete epidermal growth factor (EGF) was studied using the reverse hemolytic plaque assay. An average of 22% of cells (22.12 +/- 0.49%) spontaneously released low amount of EGF, as measured by the small area of plaques surrounding cells (1230 +/- 67 micron2). To determine the cellular origin of EGF from the rat anterior pituitary gland, a combination of immunocytochemistry and reverse hemolytic plaque assay was used. Most of the EGF-secreting cells were identified as luteinizing hormone (LH) containing cells (72.14 +/- 2.97%). Treatment with luteinizing hormone releasing hormone (LHRH; 10 nM) significantly increased the plaque area formed around EGF-secreting cells (2438 +/- 114 micron2) without altering the number of EGF-secreting cells. These results indicate that EGF is mostly secreted by gonadotrophs and that this EGF release is enhanced by LHRH.

Animals↗

Identification of epidermal growth factor-secreting cells in the anterior pituitary of lactating female rats.

Epidermal growth factor (EGF) is synthesized and secreted by mammalian anterior pituitary cells. It stimulates GH and prolactin (PRL) secretion, but the cellular origin of EGF is relatively unexplored. The objective of this study was to characterize the cells that secrete EGF in the anterior pituitary of lactating rats. An EGF reverse haemolytic plaque assay (RHPA) was used to identify EGF-secreting cells and this RHPA was combined with immunofluorescence using antibodies to the six major adenohypophysial hormones (i.e. PRL, GH, LH, FSH, TSH and ACTH). Approximately 20% (20.33 +/- 2.96%) of the cells in the pituitary of lactating rats secrete EGF. The EGF-secreting cell population was composed of the following labelled cells: PRL (27%), GH (20%), LH (18%), FSH (14%), TSH (14%) and ACTH (5%). The present study showed that EGF is released by a subpopulation of anterior pituitary cells composed of all the classic hormone-containing cells.

Animals↗

Evidence that TRH controls prolactin release from rat lactotrophs by stimulating a calcium influx.

Prolactin (PRL) release and intracellular free calcium concentration [Ca2+]i were measured in two populations of normal rat lactotrophs (light and heavy fractions) in culture. Spontaneous PRL release of heavy fraction cells was more sensitive to dihydropyridines (DHPs; Bay K 8644 and nifedipine) when compared to the light fraction lactotrophs. The stimulatory effect of thyrotropin-releasing hormone (TRH) on PRL release from heavy fraction cells was inhibited by Cd2+ and mimicked by Bay K 8644. Indo-1 experiments revealed that TRH-increased [Ca2+]i was reversibly inhibited by Cd2+. In a Ca(2+)-free EGTA-containing medium, TRH did not modify [Ca2+]i.

3-Pyridinecarboxylic acid, 1,4-dihydro-2,6-dimethy↗

Lipopolysaccharide induces sickness behaviour in rats by a vagal mediated mechanism.

To assess the possibility that lipopolysaccharide (LPS) induces sickness behaviour by activating primary afferent nerves, the effects of LPS (1.25 mg kg-1, intraperitoneally) were compared in vagotomized and sham-operated rats. Subdiaphragmatic vagotomy blocked the LPS-induced depression of social investigation but had no effect on LPS-induced increases in levels of IL-1 beta in plasma and peritoneal macrophages and on LPS-induced changes in dehydrogenase activity of peritoneal macrophages.

Animals↗

Different expression of the two dopaminergic D2 receptors, D2415 and D2444, in two types of lactotroph each characterised by their response to dopamine, and modification of expression by sex steroids.

Dopamine inhibits prolactin liberation acting via the D2 type receptor. Two different electrophysiological responses to dopamine have been shown to characterise two types of lactotroph isolated from the lactating female rat. It is now known that differential splicing of the pre-messenger RNA coding for the D2 receptor leads to the production of two D2 subtypes, D2(415) and D2(444). These subtypes differ in the region which is believed to be responsible for the binding of G proteins, and could thus lead to the activation of different intracellular second messenger systems. Here we show that the pre-messenger RNA for the D2 receptor is differentially spliced in such a way that the ratio D2(415)/D2(444) is significantly different (2.91 +/- 0.6 vs 1.29 +/- 0.14) between two populations of lactotrophs, each enriched in cells showing one type of response to DA. We further show that the ratio D2(415)/D2(444) can be changed by treatment of prolactin cells in primary culture with progesterone or testosterone. Estrogen did not change the ratio, but diminished the total amount of D2 cDNA. Regulation of differential splicing by sex steroids could provide a mechanism for modifying lactotroph responsiveness to DA in different physiological situations.

Animals↗

Evidence for a relationship between lactotroph heterogeneity and physiological context.

Two lactotroph subpopulations differing in their functional properties have previously been isolated from lactating female rats. It was found that adult female rats at proestrus similarly yield two subpopulations of lactotrophs, whereas those from adult female rats at metestrus show different properties and in males only one functional population is found. As the period of lactation and proestrus are characterised by high circulating estrogen levels, estrogen is considered to modify lactotroph properties in the female, and as the functional differences concern the lactotroph responsiveness to TRH, it is supposed that lactotroph heterogeneity provides a means for particular secretory patterns during lactation and at proestrus.

Animals↗

[Histamine and prolactin liberation in the rhesus monkey].

The effect of intra venous (i.v.) or intra cerebroventriculaire (i.c.v.) administration of histamine (HA) on plasma prolactin (PRL) levels was investigated in ovariectomized Rhesus Monkeys. Intra venous injection of 50 micrograms/kg HA increased the plasma PRL concentration but icv administration of 10 and 50 micrograms decreased PRL plasma levels. Intra venous injection of 2-thiazolyl-éthylamine, a H1 receptor agonist, rapidly stimulated PRL release (peak PRL concentration at 5 min) suggesting a direct effect on the pituitary. In contrast intra venous administration of the H2 receptor agonist, impromidine, inhibited PRL release at low doses. High doses of impromidine increased PRL concentrations but this effect was delayed (PRL peak values were reached at 20 minutes). Our results show that HA may influence PRL release in the primate via H1 and H2 receptors located at both pituitary and central levels.

Animals↗

Angiotensin II stimulates prolactin release in the rhesus monkey.

The effect of angiotensin II (AII) on plasma prolactin (PRL) concentrations was evaluated in ovariectomized rhesus monkeys which had been conditioned to chair restraining. Intracerebro-ventricular (i.v.t.) microinjections of AII increased PRL plasma levels in a dose-dependent manner. PRL values peaked at 10 min after injection and declined rapidly thereafter. Peak levels of PRL after 50 micrograms AII were approximately 7 times higher than those of baseline controls and were significantly reduced when 500 micrograms saralasin were injected via the ventricular route 15 min before AII injections. Measurements of vasopressin (AVP) plasma levels in some experiments showed that AVP release occurred following any dose of AII, irrespective of PRL liberation. In 2 animals, central injections of AII induced a dose-related increase in blood pressure. This study shows that, in primates, AII is able to influence PRL secretion via specific AII receptors. This action of AII is independent of AVP release and may be indirectly mediated through vasoactive effects of the octapeptide.

Angiotensin II↗