Estimation of turgor pressure through comparison between single plant cell and pressurized shell mechanics.

While plant growth is well known to rely on turgor pressure, it is challenging to quantify the contribution of turgor pressure to plant cell rheology. Here we used a custom-made micro-rheometer to quantify the viscoelastic behavior of isolated plant cells while varying their internal turgor pressure. To get insight into how plant cells adapt their internal pressure to the osmolarity of their medium, we compared the mechanical behavior of single plant cells to that of a simple, passive, pressurized shell: a soccer ball. While both systems exhibited the same qualitative behavior, a simple mechanical model allowed us to quantify turgor pressure regulation at the single cell scale.

Effects of daytime and nighttime stress on Fos-like immunoreactivity in the paraventricular nucleus of the hypothalamus, the habenula, and the posterior paraventricular nucleus of the thalamus.

Circadian effects on basal and stress-induced Fos-like immunoreactivity (IR) in the paraventricular nucleus of the hypothalamus (PVN), the habenula (Hab) and the posterior paraventricular nucleus of the thalamus (PVN-Thal) were examined. Stress induced a significant increase in the number of Fos-like IR cells within all 3 brain regions. In the Hab, expression was localized specifically to the medial region of the lateral Hab. No differences between the effects of daytime vs nighttime stress on numbers of Fos-like IR cells in the PVN and PVP-Thal were observed. Significantly fewer Fos-like IR cells were observed, however, in the lateral habenula of nighttime vs daytime non-stressed controls, resulting in a significantly greater percentage increase in Fos-like IR in the lateral habenula following nighttime vs daytime stress.

Immobilisation stress induces a paradoxical sleep rebound in rat.

An immobilisation stress (IS) of 2 h applied to rats at the beginning of the dark period (12 h), i.e. when the animals are more active, induces during the 10 consecutive h a significant rebound (+92%) of paradoxical sleep (PS) while slow-wave sleep state (SWS) is poorly affected. Two h of sleep deprivation, also applied at the beginning of the dark period and realized either by the platform technique or by maintaining the animals awake with gentle handling, do not affect significantly subsequent SWS and PS. Finally, when repetitive IS are inflicted to the animals (one IS of 2 h every 3 days) an attenuation of the PS rebound is observed. These data suggest that a qualitative aspect of the waking state as in an intense stressful situation might be the source of a hormonal process inducing a PS excess.

Detection of vasopressin mRNA in the neurointermediate lobe of the rat pituitary.

A messenger ribonucleic acid (mRNA) homologous to the transcript that encodes vasopressin (VP) was detected in the neurointermediate lobe (NIL) of the rat pituitary. The abundance of this transcript is approximately 1/100th the amount detected in the hypothalamus. In rats drinking 2% NaCl-water for 0,2,4, or 10 days, or for 10 days and then tap water for 14 days, the levels of VP mRNA in the NIL were altered in a fashion that paralleled changes in the hypothalamus.

A monoclonal antibody directed against CLIP (ACTH 18-39). Anatomical distribution of immunoreactivity in the rat brain and hypophysis with quantification of the hypothalamic cell group.

After the recent demonstration of the facilitatory effect exerted by corticotropin-like intermediate lobe peptide (CLIP or adrenocorticotropic hormone (ACTH) 18-39) on paradoxical sleep in the rat (Chastrette and Cespuglio, 1985), we undertook the production of monoclonal antibodies against this peptide. Wistar rats were immunized against CLIP and their spleen cells fused with mouse myeloma cells. After recloning, 25 supernatants were found to give positive immunohistochemical reactions in the rat brain. In immunohistochemical tests performed by preabsorption, the 25 supernatants presented similar properties, i.e. recognized CLIP, ACTH (1-39) and ACTH (25-39), but not ACTH (1-24) and the C-terminal fragment (34-39). We assume that the epitope(s) recognized by the 25 supernatants is (are) located between the amino-acids Asn25 and Ala34 of the CLIP molecule. The immunoreactivity observed in the rat brain and hypophysis with this antibody was distributed with a pattern quite similar to that described for anti-ACTH antibodies. A main group of immunoreactive cell bodies was located in the mediobasal hypothalamus and a small group in the nucleus of the solitary tract. Immunoreactive fibres were distributed from the olfactory nucleus to the spinal cord and formed particularly rich networks in the hypothalamus and preoptic area. Among other locations, immunoreactive axons were also present in the brainstem centres involved in the control of the sleep-waking cycle, which is in accordance with the influence of CLIP on paradoxical sleep. Using Abercrombie's formula, the number of immunoreactive cells in the mediobasal hypothalamus was estimated at about 3000 neurons. We conclude that our monoclonal anti-CLIP antibody can be considered as a good marker of proopiomelanocortin neurons.

Proopiomelanocortin (POMC)-derived peptides and sleep in the rat. Part 1--Hypnogenic properties of ACTH derivatives.

The sleep-wake effects of the proopiomelanocortin (POMC)-derived peptides, i.c.v. injected, are reported. Adrenocorticotropic hormone (ACTH, 1 microgram) induces an awakening effect, while its two derivatives, desacetyl-alpha-MSH (des-alpha-MSH, 1ng) and corticotropin-like intermediate lobe peptide (CLIP, 10 ng), are respectively able to increase slow wave sleep (SWS) and paradoxical sleep (PS); the hypnogenic effect of CLIP is also observed in hypophysectomized rats. Furthermore, two hypothalamic factors known to be involved in the control of POMC derivatives were also injected; MSH inhibiting Factor (MIF) does not influence the vigilance states, while Corticotropin Releasing Factor (CRF, 1 microgram) increases the waking state. Finally, some preliminary results, obtained with a restraint stress and suggesting a possible interrelation between stress, sleep and POMC derivatives, are discussed.

Proopiomelanocortin (POMC)-derived peptides and sleep in the rat. Part 2--Aminergic regulatory processes.

Apomorphine (Apo), a D1/D2 Dopamine (DA) agonist, at high doses (500 micrograms/kg) induces a short-lasting insomnia, antagonized by a secondary injection of corticotropin-like intermediate lobe peptide (CLIP, 10 ng); these effects are also observed with hypophysectomized (hypoX) rats. The administration of the serotonin (5-HT) agonist 8-hydroxy-2-di-n-propylamino-tetralin (8-OHDPAT, 0.3 mg/kg) induces also an insomnia which, unlike Apo, is followed by a significant PS rebound. CLIP, again, antagonizes the 8-OHDPAT-induced insomnia. Finally, Bromocriptine (5 mg/kg), an agonist for both DA and 5-HT, induces first an insomnia (antagonized by CLIP), followed by a PS rebound; these effects persist in hypoX rats.

Voltammetric detection of the release of 5-hydroxyindole compounds throughout the sleep-waking cycle of the rat.

In the present work, differential pulse voltammetry (DPV) measurements of the extracellular fraction of 5-hydroxyindole compounds were performed in rats under long-term chronic conditions. In the nucleus Raphe Dorsalis (n.RD), the voltammetric signal measured at +300 mv (peak 3) disappeared completely 70 to 90 min after injection of Clorgyline (10 mg/kg), a monoamine oxidase inhibitor type A (MAOI-A); the signal measured in such conditions is thus dependent upon extracellular 5-hydroxyindoleacetic acid (5-HIAA peak 3). Deprenyl, an MAOI type B, at the same dose, induced only a slight increase in peak 3 height; according to the fact that MAO-B is selectively located in the 5-HT neurons and since their inhibition does not decrease 5-HIAA peak 3 nor the endogenous 5-HIAA content as measured with High Performance Liquid Chromatography (HPLC), 5-HIAA measured with DPV in the extracellular fluid of untreated animals might come from 5-HT released and metabolized by MAO-A outside the 5-HT neurons. In animals implanted for measurements of both voltammetric and polygraphic parameters, the 5-HIAA peak 3 measured mainly in the anterior and ventral part of the n.RD exhibited large increases in its height during slow-wave sleep (SWS: +39%) and paradoxical sleep (PS = +71%) as compared to the waking state (W = 100%); these variations could reflect the dendritic release of 5-HT. In the Caudate nucleus (n.Cd) the same voltammetric signal presented reverse fluctuations, i.e. an increase during W and a decrease during SWS and PS.(ABSTRACT TRUNCATED AT 250 WORDS)

[Opposite variations of extracellular concentrations of 5-hydroxyindoleacetic acid (5-HIAA) measured by voltammetry of axonal terminals and cell bodies of the dorsal raphe nucleus through the sleep-wake cycle]. / Variations opposées des concentrations extracellulaires de l'acide 5-hydroxyindolacétique (5-HIAA) mesurées par la voltamétrie au niveau des terminaisons axonales et des corps cellulaires du noyau raphe dorsalis, à travers le cycle veille-sommeil.

Differential pulse voltammetry was used for 5-hydroxyindoleacetic acid (5-HIAA) detection in the rat caudate (n. Cd) and Raphe Dorsalis (n. RD) nuclei, in chronic experimental conditions. In the anterior and ventral part of n. RD, large increases in the extracellular concentrations of 5-HIAA were reported during slow wave sleep (SWS) and paradoxical sleep (PS) whereas a decrease occurred during waking. These variations could reflect the dendritic release of serotonin. In n. Cd, opposite variations of the extracellular concentrations of 5-HIAA were observed i.e. increase during waking state and decrease during SWS and PS.

Differential pulse voltammetry in vivo with working carbon fiber electrodes: 5-hydroxyindole compounds or uric acid detection?

Differential pulse voltammetry was performed in rats chronically implanted with carbon fiber electrodes in the caudate (n.Cd) and raphe dorsalis (n.RD) nuclei. The electrochemical signal obtained at the +300 mV potential (peak 3) in animals implanted for more than one week (long term chronic conditions, greater than 7 days) could be dependent upon the extracellular fraction of 5-hydroxyindolacetic acid (5-HIAA) since a single injection of Pargyline is sufficient to suppress it in n.Cd and n.RD. This result was obtained despite the tendency of Pargyline to increase n.Cd and n.RD endogenous concentrations of Uric Acid (UA) measured by High Performance Liquid Chromatography (HPLC). In contrast, in animals implanted for less than one week (short term chronic conditions, less than 7 days) peak 3 recorded in the same structure could be dependent upon extracellular fractions of 5-HIAA and UA since consecutive injections of Pargyline and Allopurinol are necessary to suppress this signal. The source of extracellular UA measured in brain by voltammetry, in such short term chronic conditions, might result from surgical trauma.

Influence of proopiomelanocortin-derived peptides on the sleep-waking cycle of the rat.

The effects of i.c.v. injections of different proopiomelanocortin-derived peptides were tested on the rat sleep-waking cycle: adrenocorticotropic hormone (ACTH) had no effect. In contrast, administration of two ACTH-derived peptides was followed by a selective and significant increase of each sleep state: desacetyl-alpha-melanocyte-stimulating hormone induced an increase of slow-wave sleep, while corticotropin-like intermediate lobe peptide was followed by an increase of paradoxical sleep. Both alpha-melanocyte-stimulating hormone in its acetylated form and alpha-endorphin were inactive.

[Differential pulse voltammetry: focus on the measurement of 5-hydroxyindole compounds and uric acid in the brain]. / Voltamétrie impulsionnelle différentielle: mise au point concernant la mesure des composés 5-hydroxyindols et de l'acide urique au niveau du cerveau.

The electrochemical signal obtained at the +300 mV potential (peak 3) in rats implanted for more than one week could be dependent upon extracellular fraction of 5-hydroxyindolacetic acid (5-HIAA) since a single injection of Pargyline is sufficient to suppress it in caudate and raphe dorsalis nuclei. In contrast, in rats implanted for less than one week, this signal could be dependent upon extracellular fractions of 5-HIAA and uric acid since consecutive injections of Pargyline and Allopurinol are necessary to suppress it.

[Hypnogenic effects of des-acetyl-alpha-MSH and CLIP (ACTH 18-39) in the rat]. / Effets hypnogènes de la des-acetyl-alpha-MSH et du CLIP (ACTH 18-39) chez le rat.

In the rat, the intraventricular administration of ACTH (adrenocorticotropic hormone) has no effect upon the sleep-waking cycle. However, administration of two ACTH-derived peptides is followed by a selective and significant increase of each sleep state: Des-alpha-MSH (Des-acetyl-alpha-melanocyte stimulating hormone) administration induces an increase of slow wave sleep, while CLIP (corticotropin-like intermediate lobe peptide) is followed by an increase of paradoxical sleep.