Effects of electrolytic lesion of hypothalamic paraventricular nucleus and its related areas on the sleep waking cycle in the cat

In order to study putative hypothalamic mechanisms of sleep waking cycle regulation we destroyed, by electrolytic coagulation, a large part of the medial hypothalamus overlapping the paraventricular nucleus in 6 adult cats. We never observed any modification of light slow wave sleep. Three of the six cats presented no paradoxical sleep (PS) impairment, despite an almost total destruction of neurophysin-immunoreactive cells of PVN in two cats and marked signs of diabetes insipidus in the third. Further, in the other three animals a statistically significant decrease of daily quantities of PS and deep slow wave sleep (SWS2) were related to an extensive destruction of the anterior hypothalamic area. These results suggest lack of influence of the PVN in sleep regulation and an involvement of the anterior hypothalamus in the onset of SWS2 and PS.     

Influence of growth room and vessel humidity on the in vitro development of rose plants

An increase of the vapor pressure deficit (VPD) of the growth room atmosphere (from 600 Pa up to 2000 Pa) induced a variation in the air VPD inside the vessels used for rose micropropagation.During the photoperiod, the in vitro plants lost water by evaporation. During the night period, depending upon the VPD of the growth room, plants could take water from the vessel atmosphere.According to the intensity of the transpiration, large changes in the growth and morphology were observed: decrease in multiplication rate, modification of leaf colour and area, reduction of the elongation and changes of the level of axillary buds which grew.     

Cryopreservation of Chrysanthemum cinerariaefolium Vis. cells and its impact on their pyrethrin biosynthesis ability

An efficient protocol for cryogenic storage of high-pyrethrin-producing cell lines of Chrysanthemum cinerariaefolium is described. Optimal survival (92%) was obtained with cells precultured in 1/2 Murashige and Skoog nutrient medium containing 180 g ⋅ l^–1 sucrose for 30 days, then incubated in the same medium in the presence of 5% DMSO for 1 h in an ice bath, cooled slowly to –20°C and immersed for 30 min in liquid nitrogen. After cryopreservation, the cells conserved the same growth pattern, but displayed different biochemical properties. The subculture derived from the thawed cells was characterized by a lower chlorophyll content and a higher pyrethrin biosynthesis ability. Received: 17 February 1997 / Revision received: 27 May 1997 / Accepted: 17 June 1997     

Comparative marker analysis of the ependymocytes of the subcommissural organ in four different mammalian species

Summary The subcommissural organ (SCO), classified as one of the circumventricular organs, is composed mainly of modified ependymal cells, attributable to a glial lineage. Nevertheless, in the rat, these cells do not possess glial markers such as glial fibrillary acidic protein (GFAP), protein S100, or the enzyme glutamine synthetase (GS). They receive a synaptic 5-HT input and show pharmacological properties for uptake of GABA resembling the uptake mechanism of neurons. In this study, we examine the phenotype of several mammalian SCO (cat, mouse, rabbit) and compare them with the corresponding features of the rat SCO. In all these species, the SCO ependymocytes possess vimentin as an intermediate filament, but never express GFAP or neurofilament proteins. They do not contain GS as do glial cells involved in GABA metabolism, and when they contain protein S100 (rabbit, mouse), its rate is low in comparison to classical glial or ependymal cells. Thus, these ependymocytes display characteristics that differentiate them from other types of glial cells (astrocytes, epithelial ependymocytes and tanycytes). Striking interspecies differences in the capacity of SCO-ependymocytes for uptake of GABA might be related to their innervation and suggest a species-dependent plasticity in their function.     

Increase of paradoxical sleep induced by microinjections of ibotenic acid into the ventrolateral part of the posterior hypothalamus in the cat

In order to study putative hypothalamic mechanisms of sleep-waking cycle regulation we injected a neural cell body toxin--ibotenic acid (IBO), 40 to 200 micrograms--into the ventrolateral part of the posterior hypothalamus (HVL). This injection induced a dramatic biphasic and transient hypersomnia immediately after the disappearance of the anesthesia (14 to 24 hours after the injection). The duration of hypersomnia was dose dependent. Its first period was characterized by an increase in paradoxical sleep (PS) (300%). Then, during the second phase, PS disappeared and there was a subsequent increase of slow sleep (SWS) (60%). Finally, on the third day, all cats recovered control level of PS and SWS while, 3 weeks later, the histological analysis revealed the great loss of cell bodies in the HVL in all cats.     

Increase of paradoxical sleep, induced by injection of ibotenic acid in the ventrolateral posterior hypothalamus in the cat

The intratissular injection of ibotenic acid into the ventrolateral part of the posterior hypothalamus induced a dramatic biphasic and transient hypersomnia immediately after disappearance of the anaesthesia (14 to 24 hrs. after injection). The duration of hypersomnia was related to the dose of neurotoxin injected. Its first period was characterized by an increase in paradoxical sleep (PS) (300%). Then, during the second phase, PS disappeared and there was a subsequent increase of slow wave sleep (SWS) (60%). Finally, on the third day, all cats recovered control level of PS and SWS.     

The ultrastructure of micropropagated and greenhouse rose plant stomata

Stomata of leaves from in vitro grown rose plantlets remain opened in the dark. The ultrastructure of their guard cells was studied after a 7 h light and a 7 h dark period, and compared to that of functional stomata from plants which have been acclimatized to greenhouse conditions. Qualitative and quantitative observations concerning the shape of the guard cells, mitochondria, plastids and starch grains, demonstrated the similarity in guard cell ultrastructure. The peculiarity of guard cell ultrastructure of in vitro cultured plants was the inability to close in the dark; vacuolar area was 40% of the whole guard cell area during both light and dark period whereas, in guard cells from greenhouse plants, the vacuolar area was 40% of the whole guard cell area during the light and only 25% during the dark period. These results indicate that stomata from in vitro plants are duly developed and possess an ultrastructure suitable for a typical functioning. The inability to close in the dark results from atypical water relation.     

Paradoxical sleep deprivation increases glutamine synthetase in rat brain.

The modifications of glutamine synthetase (GS) level, an enzyme mainly located in astrocytes, were investigated in rat after paradoxical sleep deprivation and during recovery. An immunotitration method was used to evaluate the relative level of GS in brain tissue. At the end of a 24 hrs. paradoxical sleep deprivation, a significant increase in GS level was observed both in the frontoparietal cortex and the locus coeruleus area. 4 hrs. later, during recovery, the GS level returned to control level in the cortex but was lower in the locus coeruleus area.     

Growth,Stomatal Conductance,Photosynthetic Rate,Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase and Phosphoenolpyruvate Carboxylase Activities during Rooting and Acclimatisation of Rosa Hybrida Plantlets

The rooting of shoots of micropropagated Rosa hybrida cv. Madame Delbard was conducted on MS medium with 30 kg m^–3 sucrose or on hydroponic medium (containing less mineral salts), under higher photosynthetic photon flux density (PPFD) (100 in comparison with 45 µmol m^–2 s^–1) and flushed by ambient air [AC, 340 µmol(CO₂) mol^–1] or by CO₂-enriched air (EC, 2 500 µmol mol^–1) and lower relative humidity (80–90 % vs. 96–99 %). This cultivation led to plantlets with longer roots and adventitious root formation. Net photosynthetic rate and ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) activities, RuBPCO/phosphoenolpyruvate carboxylase activities ratio, and starch accumulation increased under these conditions. After 14 d, plantlets had functional stomata and could be acclimated on open benches without gradual decrease in relative humidity. The percentage of survival was higher when the rooting took place in EC than in AC. However, the advantage acquired during rooting phase by plantlets cultured in liquid medium was not maintained after 4 weeks of acclimatisation.     

Metabolism in orange fruits is driven by photooxidative stress in the leaves

In plants, stress signals propagate to trigger distant responses and thus stress acclimation in non‐exposed organs. We tested here the hypothesis that leaves submitted to photooxidative stress may influence the metabolism of nearby fruits and thus quality criteria. Leaves of orange trees (Citrus sinensis (L.) Osbeck cv. ‘Navelate’) were acclimated to shade for 1 week and then submitted to full (FL) and medium light (ML) conditions. As expected, photoinhibition was detected in leaves of both FL and ML treatments as revealed by stress indicators (F_v/F_m, Performance Index) for at least 99 h after treatments. In the fruits near the stressed leaves, we then determined the activities of enzymes related to oxidative stress, superoxide dismutase, catalase and the enzymes of the ascorbate (AA)/glutathione cycle, as well as the contents in sugars, organic acids and carotenoids. Ascorbate peroxidase and monodehydroascorbate reductase activities in the pulp of fruits were dramatically higher in both treatments when compared to the control. AA and total sugars were not affected by the photooxidative stress. However, the FL treatment resulted in a 16% increase in total organic acids, with succinic acid being the major contributor, a shift towards less glucose + fructose and more sucrose, and a 15% increase in total carotenoids, with cis‐violaxanthin being the major contributor. Our observations strongly suggest the existence of a signal generated in leaves in consequence of photooxidative stress, transmitted to nearby fruits. Exploiting such a signal by agronomic means promises exciting perspectives in managing quality criteria in fruits accumulating carotenoids.