Olfactory bulbectomy modifies photic entrainment and circadian rhythms of body temperature and locomotor activity in a nocturnal primate

Studies on rodents have emphasized that removal of the olfactory bulbs modulates circadian rhythmicity. Using telemetric recordings of both body temperature (Tb) and locomotor activity (LA) in a male nocturnal primate, the gray mouse lemur, the authors investigated the effects of olfactory bulbectomy on (1) the circadian periods of Tb and LA in constant dim light condition, and (2) photic re-entrainment rates of circadian rhythms following 6-h phase shifts of entrained light-dark cycle (LD 12:12). Under free-running condition, bulbectomized males had significantly shorter circadian periods of Tb and LA rhythms than those of control males. However, the profiles of Tb rhythms, characterized by a phase of hypothermia at the beginning of the subjective day, and Tb parameters were not modified by olfactory bulbectomy. Under a light-dark cycle, olfactory bulbectomy significantly modified the expression of daily hypothermia, especially by an increase in the latency to reach minimal daily Tb, suggesting a delayed response to induction of daily hypothermia by light onset. Reentrainment rates following both a 6-h phase advance and a 6-h phase delay of entrained LD were also delayed in bulbectomized males. Olfactory bulbectomy led to significant fragmentation of locomotor activity and increased locomotor activity levels during the resting period. The shortening of circadian periods in bulbectomized males could partly explain the delayed responses to photic stimuli since in control males, the longer the circadian period, the better the response to light entrainment. This experiment shows for the 1st time that olfactory bulbs can markedly modify the circadian system in a primate.     

The influence of day-length on imaginal diapause in the Red Locust, Nomadacris septemfasciata (Serv.)

The main permanent habitats of the Red Locust (Nomadacris septemfasciata (Serv.)) in tropical Africa lie between latitudes 7° and 9° S and between 10° and 16° N. Breeding is confined to the rainy season, which occurs during the longest days of the year in these areas. Both north and south of the equator, the adults emerge as day-length is diminishing and undergo a prolonged imaginal diapause until the next rainy season.When reared in the laboratory in London, adults which emerge in spring or early summer become sexually mature within two months; those which emerge in late summer and autumn, although kept at the same temperature and humidity, enter diapause and do not copulate or oviposit until the following spring. By controlling the length of the daily photoperiod, it has been shown that the seasonal differences in day-length are responsible for the onset of diapause. Constant short photoperiods usually induce a rather shorter diapause than photoperiods diminishing so as to simulate autumn day-length at this latitude (51.5° N). The critical constant photoperiod for the establishment of diapause lies between 12 and 13 hours which corresponds approximately to maximum and minimum day-lengths at 7° N or S. It is therefore possible that day-length is responsible for the establishment of the natural diapause.

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Resume On a déjà constaté que, quand on élève le criquet rouge, Nomadacris septemfasciata (Serv.), au laboratoire à Londres, les adultes, malgré le fait qu'on les retient dans une ambiance très constante, montrent de grandes différences saisonnières dans la durie du développement qui conduit à la maturité sexuelle. Les adultes qui émergent à partir du mois d'avril jusque'au mois de juillet deviennent matures sexuellement sans diapause en 11/2 à 2 mois; tandis que ceux qui émergent au début de l'automne, à partir de la mi-août approximativement jusqu'à la fin de septembre, ont une diapause prolongée de 7 à 8 mois; et ceux qui émergent encore plus tard dans l'année, du mois d'octobre jusqu'à mi-décembre, ont une diapause plus courte de 41/2 à 51/2 mois. Les périodes de maturation des adultes qui émergent à partir de janvier et plus tard, diminuent progressivement, en se fondant dans les groupes des non-diapause, qui commencent à émerger au début d'avril. Les expériences exposées dans cette thèse indiquent que les différences saisonnières de la longueur des jours, sont responsables des temps de maturation.Certains groupes de criquets, émergeant comme adultes vers la fin de l'été ou en automne, ont été soumis à des journées de longueur artificielle qui correspondait à la durée des jours de printems qui n'exerce son influence que sur la vie adulte, suffit à produire ce dans les délais semblables à ceux des groupes qui émergent au printemps. La longueur des jours de printemps qui n'excerce son influence que sur la vie adulte, suffit à produire ce résultat.Quelques larves étaient élevées simultanément dans les conditions d'une durée artificielle des jours du printemps, d'une durée artificielle des jours d'automne, et d'une lumière naturelle des jours d'été à Londres. Les adultes qui avaient émergé au mois de juin étaient ou maintenus dans le même régime de longueur de journée que les larves, ou transférés dans une des autres conditions. Tous les temps de maturation étaient appropriés aux saisons qui correspondaient au régime de durée du jour auquel les adultes étaient retenus, et il n'y avait nulle évidence qu'ils étaient influencés par la durée des jours pendant leur vie larvaire.Les groupes diapause, quand ils étaient soumis à la lumière normale du laboratoire, sont devenus matures, au printemps, au moment où les jours s'allongeaient. Pourtant la maturation n'est pas due à la stimulation causée par un accroissement de la longueur du jour, car on à observé qu'elle arrive en même temps si les criquets ne la subissent pas. Ceci est vrai également à l'égard des deux groupes, celui de longue diapause (qui émerge au début de l'automne) et celui de courte diapause (qui émerge vers la fin de l'automne); il s'ensuit que la diapause plus courte du dernier groupe n'est pas attribuable, comme il semblait probable, au fait qu'on l'avait exposé à la longueur des jours du printemps plus tôt dans la vie adulte. On a trouvé que ces groupes de la dernière partie de l'automne étaient devenus matures dans le même temps s'ils étaient retenus aux courtes photo-périodes constantes de 8 ou 11 heures, ou à la durée changeante des jours d'hiver. On a conclu qu'ils répondent comme à la durée de constants jours courts.Les groupes intermédaires (qui émergent à partir de janvier jusqu'a mars) d'autre part, sont exposés aux jours plus longs du printemps et de l'été encore plus tôt dans la vie adulte, et dans leur cas la diapause est empêchée, ou au moins abrégée, par les jours plus longs. Quand ces groupes sont retenus aux constantes photo-périodes courtes, ils deviennent matures plus lentement que s'ils étaient maintenus à la lumière du jour, et ils ont une diapause de la même longueur que les groupes de courte-diapause.On a conclu de ces résultats que, quand les groupes sont retenus dans conditions de jours-courts, les processus de maturation (manifestés par un accroissement du poids du corps, et par le développement de l'ovaire) sont repris à un certain stade indépendamment de la longueur du jour et sous l'action de stimuli internes. L'exposition aux conditions de jours longs n'est effective que pour accélérer la maturation si elle a lieu avant que ce stade soit atteint.L'observation que les adultes émergeant au début de l'automne ont une diapause plus longue que ceux qui émergent vers la fin de l'automne, reste sans explication. Ceux-là sont exposés à la durée de jours qui décroissent rapidement pendant la vie larvaire et du commencement de la vie adulte, et il semble probable que le changement propre des jours longs aux jours courts puisse amener une diapause plus longue que ne le feraient des jours courts dès le début. A présent, on étudie les effets de la variation de durée des jours.Les adultes retenus dans l'obscurité constante sont devenus matures sans diapause.La grande fluctuation de la longueur des jours, caractéristique de la latitude de Londres, n'est pas nécessaire pour induire les deux types de maturation diapause ou non-diapause. Les plus grandes différences de réponse aux photo-périodes constantes ont eu lieu entre les moyennes correspondant à peu près avec les minimum et maximum de longueurs de jour (y compris les périodes de crépuscule) dans les habitats tropicaux du criquet.Il est discuté ici de la probabilité que le cours de la diapause dans l'habitat tropical naturel soit influencé par la faible fluctuation des longueurs de jour de cet habitat.

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The author is Mrs O. W. Richards     

Abi1 regulates the activity of N-WASP and WAVE in distinct actin-based processes

Neural Wiskott-Aldrich syndrome protein (N-WASP) and WAVE are members of a family of proteins that use the Arp2/3 complex to stimulate actin assembly in actin-based motile processes. By entering into distinct macromolecular complexes, they act as convergent nodes of different signalling pathways. The role of WAVE in generating lamellipodial protrusion during cell migration is well established. Conversely, the precise cellular functions of N-WASP have remained elusive. Here, we report that Abi1, an essential component of the WAVE protein complex, also has a critical role in regulating N-WASP-dependent function. Consistently, Abi1 binds to N-WASP with nanomolar affinity and, cooperating with Cdc42, potently induces N-WASP activity in vitro. Molecular genetic approaches demonstrate that Abi1 and WAVE, but not N-WASP, are essential for Rac-dependent membrane protrusion and macropinocytosis. Conversely, Abi1 and N-WASP, but not WAVE, regulate actin-based vesicular transport, epidermal growth factor receptor (EGFR) endocytosis, and EGFR and transferrin receptor (TfR) cell-surface distribution. Thus, Abi1 is a dual regulator of WAVE and N-WASP activities in specific processes that are dependent on actin dynamics.     

Undersulfation of heparan sulfate restricts differentiation potential of mouse embryonic stem cells

Heparan sulfate proteoglycans, present on cell surfaces and in the extracellular matrix, interact with growth factors and morphogens to influence growth and differentiation of cells. The sulfation pattern of the heparan sulfate chains formed during biosynthesis in the Golgi compartment will determine the interaction potential of the proteoglycan. The glucosaminyl N-deacetylase/N-sulfotransferase (NDST) enzymes have a key role during biosynthesis, greatly influencing total sulfation of the heparan sulfate chains. The differentiation potential of mouse embryonic stem cells lacking both NDST1 and NDST2 was studied using in vitro differentiation protocols, expression of differentiation markers, and assessment of the ability of the cells to respond to growth factors. The results show that NDST1 and NDST2 are dispensable for mesodermal differentiation into osteoblasts but necessary for induction of adipocytes and neural cells. Gene expression analysis suggested a differentiation block at the primitive ectoderm stage. Also, GATA4, a primitive endoderm marker, was expressed by these cells. The addition of FGF4 or FGF2 together with heparin rescued the differentiation potential to neural progenitors and further to mature neurons and glia. Our results suggest that the embryonic stem cells lacking both NDST1 and NDST2, expressing a very low sulfated heparan sulfate, can take the initial step toward differentiation into all three germ layers. Except for their potential for mesodermal differentiation into osteoblasts, the cells are then arrested in a primitive ectoderm and/or endoderm stage.     

New insights into the organisation and intracellular localisation of the two subunits of glucose-6-phosphatase

Glucose-6 phosphatase (G6Pase), a key enzyme of glucose homeostasis, catalyses the hydrolysis of glucose-6 phosphate (G6P) to glucose and inorganic phosphate. A deficiency in G6Pase activity causes type 1 glycogen storage disease (GSD-1), mainly characterised by hypoglycaemia. Genetic analyses of the two forms of this rare disease have shown that the G6Pase system consists of two proteins, a catalytic subunit (G6PC) responsible for GSD-1a, and a G6P translocase (G6PT), responsible for GSD-1b. However, since their identification, few investigations concerning their structural relationship have been made. In this study, we investigated the localisation and membrane organisation of the G6Pase complex. To this aim, we developed chimera proteins by adding a fluorescent protein to the C-terminal ends of both subunits. The G6PC and G6PT fluorescent chimeras were both addressed to perinuclear membranes as previously suggested, but also to vesicles throughout the cytoplasm. We demonstrated that both proteins strongly colocalised in perinuclear membranes. Then, we studied G6PT organisation in the membrane. We highlighted FRET between the labelled C and N termini of G6PT. The intramolecular FRET of this G6PT chimera was 27%. The coexpression of unlabelled G6PC did not modify this FRET intensity. Finally, the chimera constructs generated in this work enabled us for the first time to analyze the relationship between GSD-1 mutations and the intracellular localisation of both G6Pase subunits. We showed that GSD1 mutations did neither alter the G6PC or G6PT chimera localisation, nor the interaction between G6PT termini. In conclusion, our results provide novel information on the intracellular distribution and organisation of the G6Pase complex.     

Copper redistribution in murine macrophages in response to Salmonella infection

The movement of key transition metal ions is recognized to be of critical importance in the interaction between macrophages and intracellular pathogens. The present study investigated the role of copper in mouse macrophage responses to Salmonella enterica sv. Typhimurium. The copper chelator BCS (bathocuproinedisulfonic acid, disodium salt) increased intracellular survival of S. Typhimurium within primary mouse BMM (bone-marrow-derived macrophages) at 24 h post-infection, implying that copper contributed to effective host defence against this pathogen. Infection of BMM with S. Typhimurium or treatment with the TLR (Toll-like receptor) 4 ligand LPS (lipopolysaccharide) induced the expression of several genes encoding proteins involved in copper transport [Ctr (copper transporter) 1, Ctr2 and Atp7a (copper-transporting ATPase 1)], as well as the multi-copper oxidase Cp (caeruloplasmin). Both LPS and infection with S. Typhimurium triggered copper accumulation within punctate intracellular vesicles (copper 'hot spots') in BMM as indicated by the fluorescent reporter CS1 (copper sensor 1). These copper hot spots peaked in their accumulation at approximately 18 h post-stimulation and were dependent on copper uptake into cells. Localization studies indicated that the copper hot spots were in discrete vesicles distinct from Salmonella containing vacuoles and lysosomes. We propose that copper hot spot formation contributes to antimicrobial responses against professional intracellular bacterial pathogens.     

Genetics of canine olfaction and receptor diversity

Olfaction is a particularly important sense in the dog. Humans selected for this capacity during the domestication process, and selection has continued to be employed to enhance this ability. In this review we first describe the different olfactory systems that exist and the different odorant receptors that are expressed in those systems. We then focus on the dog olfactory receptors by describing the olfactory receptor gene repertoire and its polymorphisms. Finally, we discuss the different uses of dog olfaction and the questions that still need to be studied.     

Control of pollen-mediated gene flow in transgenic trees

Pollen elimination provides an effective containment method to reduce direct gene flow from transgenic trees to their wild relatives. Until now, only limited success has been achieved in controlling pollen production in trees. A pine (Pinus radiata) male cone-specific promoter, PrMC2, was used to drive modified barnase coding sequences (barnaseH102E, barnaseK27A, and barnaseE73G) in order to determine their effectiveness in pollen ablation. The expression cassette PrMC2-barnaseH102E was found to efficiently ablate pollen in tobacco (Nicotiana tabacum), pine, and Eucalyptus (spp.). Large-scale and multiple-year field tests demonstrated that complete prevention of pollen production was achieved in greater than 95% of independently transformed lines of pine and Eucalyptus (spp.) that contained the PrMC2-barnaseH102E expression cassette. A complete pollen control phenotype was achieved in transgenic lines and expressed stably over multiple years, multiple test locations, and when the PrMC2-barnaseH102E cassette was flanked by different genes. The PrMC2-barnaseH102E transgenic pine and Eucalyptus (spp.) trees grew similarly to control trees in all observed attributes except the pollenless phenotype. The ability to achieve the complete control of pollen production in field-grown trees is likely the result of a unique combination of three factors: the male cone/anther specificity of the PrMC2 promoter, the reduced RNase activity of barnaseH102E, and unique features associated with a polyploid tapetum. The field performance of the PrMC2-barnaseH102E in representative angiosperm and gymnosperm trees indicates that this gene can be used to mitigate pollen-mediated gene flow associated with large-scale deployment of transgenic trees.