Silencing of the Tandem Pore Domain Halothane-inhibited K+ Channel 2 (THIK2) Relies on Combined Intracellular Retention and Low Intrinsic Activity at the Plasma Membrane

The tandem pore domain halothane-inhibited K⁺ channel 1 (THIK1) produces background K⁺ currents. Despite 62% amino acid identity with THIK1, THIK2 is not active upon heterologous expression. Here, we show that this apparent lack of activity is due to a unique combination of retention in the endoplasmic reticulum and low intrinsic channel activity at the plasma membrane. A THIK2 mutant containing a proline residue (THIK2-A155P) in its second inner helix (M2) produces K⁺-selective currents with properties similar to THIK1, including inhibition by halothane and insensitivity to extracellular pH variations. Another mutation in the M2 helix (I158D) further increases channel activity and affects current kinetics. We also show that the cytoplasmic amino-terminal region of THIK2 (Nt-THIK2) contains an arginine-rich motif (RRSRRR) that acts as a retention/retrieval signal. Mutation of this motif in THIK2 induces a relocation of the channel to the plasma membrane, resulting in measurable currents, even in the absence of mutations in the M2 helix. Cell surface delivery of a Nt-THIK2-CD161 chimera is increased by mutating the arginines of the retention motif but also by converting the serine embedded in this motif to aspartate, suggesting a phosphorylation-dependent regulation of THIK2 trafficking.     

Mise en evidence de Campanien continental (facies provencal) a Salavas (Ardeche,S-E de la France): Description,sedimentologie, paleogeographie

The continental sediments in the Northern Alès Basin are dated as Campanian age based on gastropod Melania praelonga MATHERON. The lithology is made up by red shales, conglomerates and pedologic limestones (crusts and nodules) with microcodiums. The conglomerates are heterometric and made up by limestone (Urgonian facies) and standstone boulders of Cenomanian and Turonian age. Also sandy limestones of coniacian age are found, all belonging to the local substratum. This discovery helps to understand the evolution of the Upper Cretaceous paleogeography.     

Selective Acylation of Plasma Membrane Proteins of Mycoplasma agalactiae: The Causal Agent of Agalactia

Revealed by in vivo labeling with ¹⁴C-palmitic acid, about 15 acylated proteins were identified in the plasma membrane of Mycoplasma agalactiae (type strain PG2), including the major component p40. Triton X-114 phase partitioning and Western blotting demonstrated the amphiphilic properties of the acyl proteins and showed that they were also antigenic components. Chemical analyses of fatty acids bound to proteins revealed the following selectivity order within acylation: stearic acid (18:0) > linoleic acid (18:2c) ≈ palmitic acid (16:0) > oleic acid (18:1c) > myristic acid (14:0), with 16:0 and 18:1c preferred for the O-acylation and 18:0 for the N-acylation. The ratio [O-ester- + amide-bound acyl chains]/O-ester-linked chains being close to 1.4 as well as the presence of S-glycerylcysteine suggest that acyl proteins in M. agalactiae are true lipoproteins containing N-acyl diacyl glycerylcysteine, probably processed by a mechanism analogous to that described for Gram-negative eubacteria. Received: 22 June 1999 / Accepted: 2 August 1999     

Contrasting effects of peroxisome-proliferator-activated receptor (PPAR)gamma agonists on membrane-associated prostaglandin E2 synthase-1 in IL-1beta-stimulated rat chondrocytes: evidence for PPARgamma-independent inhibition by 15-deoxy-Delta12,14prostaglandin J2

Microsomal prostaglandin E synthase (mPGES)-1 is a newly identified inducible enzyme of the arachidonic acid cascade with a key function in prostaglandin (PG)E2 synthesis. We investigated the kinetics of inducible cyclo-oxygenase (COX)-2 and mPGES-1 expression with respect to the production of 6-keto-PGF1alpha and PGE2 in rat chondrocytes stimulated with 10 ng/ml IL-1beta, and compared their modulation by peroxisome-proliferator-activated receptor (PPAR)gamma agonists. Real-time PCR analysis showed that IL-1beta induced COX-2 expression maximally (37-fold) at 12 hours and mPGES-1 expression maximally (68-fold) at 24 hours. Levels of 6-keto-PGF1alpha and PGE2 peaked 24 hours after stimulation with IL-1beta; the induction of PGE2 was greater (11-fold versus 70-fold, respectively). The cyclopentenone 15-deoxy-Delta12,14prostaglandin J2 (15d-PGJ2) decreased prostaglandin synthesis in a dose-dependent manner (0.1 to 10 microM), with more potency on PGE2 level than on 6-keto-PGF1alpha level (-90% versus -66% at 10 microM). A high dose of 15d-PGJ2 partly decreased COX-2 expression but decreased mPGES-1 expression almost completely at both the mRNA and protein levels. Rosiglitazone was poorly effective on these parameters even at 10 microM. Inhibitory effects of 10 microM 15d-PGJ2 were neither reduced by PPARgamma blockade with GW-9662 nor enhanced by PPARgamma overexpression, supporting a PPARgamma-independent mechanism. EMSA and TransAM analyses demonstrated that mutated IkappaBalpha almost completely suppressed the stimulating effect of IL-1beta on mPGES-1 expression and PGE2 production, whereas 15d-PGJ2 inhibited NF-kappaB transactivation. These data demonstrate the following in IL-1-stimulated rat chondrocytes: first, mPGES-1 is rate limiting for PGE2 synthesis; second, activation of the prostaglandin cascade requires NF-kappaB activation; third, 15d-PGJ2 strongly inhibits the synthesis of prostaglandins, in contrast with rosiglitazone; fourth, inhibition by 15d-PGJ2 occurs independently of PPARgamma through inhibition of the NF-kappaB pathway; fifth, mPGES-1 is the main target of 15d-PGJ2.     

Comparison of fertility and embryo mortality following artificial insemination of common duck females (Anas Platyrhynchos) with semen from common or Muscovy (Cairina Moschata) drakes

The purpose of this study was to compare fertility and early embryo mortality rates (< or = 5 days of incubation) following artificial insemination (AI) of common duck females (Anas Platyrhynchos) with semen from either common or Muscovy (Cairina Moschata) drakes at various periods of the reproductive season (Period I, 27-35 weeks; Period II, 39-43 weeks and Period III, 49-56 weeks). Based on observations performed by stereomicroscopy on eggs laid from Days 2 to 10 after AI, we confirmed that fertility was significantly lower in the interbred compared to the purebred cross at each of the periods tested (purebred 58.1, 61.2 and 54.2 versus crossbred 31.0, 40.4 and 39.5 at Periods I, II and II, respectively; 0.01 < P < 0.001). In a complementary experiment, we demonstrated that the number of perivitelline spermatozoa (NPS) was markedly lower in mule (crossbred) eggs compared to common (purebred) eggs, a strong indication that initial sperm selection occurring in the lower oviduct is probably more intense after crossbred compared to purebred insemination. Comparison of early embryo mortality (EEM) between mule and common duck eggs indicated that increased levels of EEM in mule embryos corresponded to Stages II-IV of the Eyal-Giladi and Kochav classification (EGK). While a similar age-dependent increase in early embryo mortality was observed in eggs from both genetic origins during the latter periods of the reproductive season, it was also established that embryo mortality due to parental age was related rather to Stages X-XIV of the EGK classification in eggs from both genetic origins. It is concluded that the relative subfertility of mule compared to common duck eggs is probably the consequence of a more intense rate of selection of heterologous than homologous spermatozoa occurring in the vaginal portion of the oviduct while the causal origins of EEM in mule duck eggs can at least in part be identified on the basis of precise staging (by stereomicroscopy) of dead embryos.     

Tunic morphogenesis in the sea peach (Halocynthia papillosa,Ascidiacea, rochordata): Cellular events,the initiation of twisted fibrous arrangements and spine formation

Summary— The adult tunic of the sea peach (Halocynthia papillosa) shows a high degree of organisation. Tunic morphogenesis was monitored from the onset of tunic secretion until juveniles reached the age of 3 months. While some characteristics of the adult tunic are still missing, like certain types of intratunical cells and striated bodies, its main features have already developed by this time. Crucial events take place at or soon after the onset of metamorphosis (stage M 0). Cuticular spines cover the external surface of the juvenile. At least two types of intratunical cells enter the tunic and the fibrous material adopts a three-dimensional twisted helicoidal architecture. The initiation of this helicoidal arrangement of fibrils directly after stage M 0 is discussed regarding accompanying developmental events. The existence of cells that penetrate the outer compartment of the tunic at the end of larval life is reported for the first time.     

Feeding rates of the woodlouse Armadillidium vulgare on herb litters produced at two levels of atmospheric CO2

The consumption and assimilation rates of the woodlouse Armadillidium vulgare were measured on leaf litters from five herb species grown and naturally senesced at 350 and 700 µl l^-1 CO₂. Each type of litter was tested separately after 12, 30 and 45 days of decomposition at 18°C. The effects of elevated CO₂ differed depending on the plant species. In Medicago minima (Fabaceae), the CO₂ treatment had no significant effect on consumption and assimilation. In Tyrimnus leucographus (Asteraceae), the CO₂ treatment had no significant effect on consumption, but the elevated CO₂ litter was assimilated at a lower rate than the ambient CO₂ litter after 30 days of decomposition. In the three other species, Galactites tomentosa (Asteraceae), Trifolium angustifolium (Fabaceae) and Lolium rigidum (Poaceae), the elevated CO₂ litter was consumed and/or assimilated at a higher rate than the ambient CO₂ litter. Examination of the nitrogen contents in these three species of litter did not support the hypothesis of compensatory feeding, i.e. an increase in woodlouse consumption to compensate for low nitrogen content of the food. Rather, the results suggest that in herbs that were unpalatable at the start of the experiment (Galactites, Trifolium and Lolium), more of the the litter produced at 700 µl l^-1 CO₂ was consumed than of that produced at 350 µl l^-1 because inhibitory factors were eliminated faster during decomposition.     

Litter decomposition, climate and liter quality

Litter decomposition is controlled by three main factors: climate, litter quality and the nature and abundance of the decomposing organisms. Climate is the dominant factor in areas subjected to unfavourable weather conditions, whereas litter quality largely prevails as the regulator under favourable conditions. Litter quality remains important until the late decomposition stages through its effects on humus formation. Interest in the role of litter decomposition in the global carbon cycle has increased recently since (1) increased atmospheric carbon dioxide will probably affect the chemical quality of litter (especially nitrogen content), and (2) global warming may enhance decomposition rates.     

Searching for an indicator of N evolution during organic matter decomposition based on amino acids behaviour: a study on litter layers of pine forests

Soil microflora can directly take up the amino acids (Aas) released by decomposing plants and use them, together with de novo synthesized Aas, as building blocks for their own structures, which are expected to have an Aa composition that differs from plant-derived structures. The degree of change in the Aa composition during OM decomposition should reflect the degree of N evolution. Here we apply this concept to the study of three litter decomposition sequences in European forests: boreal (Pinus sylvestris L., Jädraås, Sweden), cool Atlantic (Pinus sylvestris L., La Gileppe, Belgium), and warm Atlantic (Pinus pinaster L., Montemor, Portugal). Litter was sorted into six decomposition stages according to morphological features: from intact, light-brown needles to dark needle fragments with faunal perforations. At each stage, the Aa composition of the litter was studied by acid hydrolysis plus liquid chromatography of the hydrolysates. Both the Aa content of the litter and the unhydrolyzable/total OC ratio increased with decomposition, but this was not the case with the unhydrolyzable/total N ratio. There is no sole pattern of Aa change with decomposition: in Jädraås and Montemor most of the changes occur in the initial and final steps, while in La Gileppe the changes seem quite evenly spread throughout the decomposition. The fact that the Aa composition of the litter did not converge with decomposition suggests the existence of site-specific biotas with contrasted Aa fingerprints. The abundance of a given Aa relative to the total differed among Aas and sites; but for some Aas it was possible to detect common patterns of decomposition behaviour (either a consistent increase or a consistent decrease). The aa/total ratio for Gly, Ala and Thr consistently increased with decomposition. These Aas were used to search for a numerical index, the Aa signature (Aa_SIG), which would reflect the changes in Aa composition from fresh debris to highly decomposed organic materials. The change in the proposed Aa_SIG was found to correlate well with the mathematical distance from the starting point (calculated as the euclidean distance); this suggests that it could be potentially useful as an indicator of N evolution during litter decomposition, at least in its first stages.