Comparative proteomic analysis between fifth‐instar nymphs and adults of Asian citrus psyllid Diaphorina citri

Asian citrus psyllid Diaphorina citri Kuwayama is extremely problematic worldwide, particularly where Huanglongbing (HLB) disease, the most serious and devastating of citrus diseases, is found. The threat is a result of its ability to transmit the causal agent of HLB, Candidatus Liberibacter asiaticus (CLas) bacterium. Improvements in proteomics, mass spectrometry, bioinformatics tools and gene ontology annotation facilitate the mapping and large‐scale identification and quantification of proteins. To date, only a few comparative proteomic studies report the developmental proteomic changes of hemimetabolous and plant–disease vector insects. Two‐dimensional gel electrophoresis analysis of D. citri total protein is able to detect qualitative and quantitative developmental differences. Liquid chromatography‐tandem mass spectrometry identifies 89 protein spots. Most proteins are metabolism and bioenergetics‐related. Nineteen protein spots are found to be implicated in stress/defence/immunity; 7 in development regulation; 9 in nervous system functions; 4 in the reproductive system; 23 in cytoskeleton and muscle organization; and 4 in movement, flight and other processes. Significant increases in the level of proteins related to structural constitution of the skeleton, stress/defence/immunity, reproduction system, muscles, locomotion and flight are found in adults, consistent with the fact that D. citri is a hemimetabolous insect, whereas proteins involved in developmental regulation are higher in the nymphal stage. The identification of these variably expressed proteins between the nymph and adult stages, linked with the basis of their physiological roles, will lead to a better understanding of the factors influencing development in D. citri and the regulation of some crucial metabolic pathways. It may also help to identify targets for genetic manipulation using RNA interference or other techniques to disrupt Asian citrus psyllid development, lifespan or its ability to transmit CLas.     

Etude au microscope électronique de la neurohypophyse de Rana esculenta L.

Résumé L'éminence médiane et la pars nervosa de Rana esculenta diffèrent du point de vue de leur structure.L'éminence médiane se compose de 2 zones différentes: la zone externe placée près du lobe distal et la zone interne située sous l'épendyme. Dans la zone externe, on distingue, d'après la taille et la forme des grains de sécrétion, 5 types de terminaisons.1. des terminaisons avec de fins granules sphériques denses de 800 à 1000 Å de diamètre; 2. des terminaisons avec des granules de 1000 à 1200 Å de diamètre; 3. des terminaisons avec des grains de forme irrégulière de diamètre supérieur aux précédents (1200 à 1600 Å); 4. des terminaisons avec de volumineux grains denses sphériques d'environ 1200 à 1800 Å de diamètre; 5. un petit nombre de terminaisons ne contenant que des vésicules. Les terminaisons des catégories 3 et 4 sont probablement du type neurosécrétoire.La zone interne contient de nombreuses fibres neurosécrétrices. Elles sont de 2 types, l'une avec de gros granules (1600–2400 Å), l'autre avec des granules moins volumineux (1300–2000 Å). Des fibres non neurosécrétrices ont également été observées.Dans la pars nervosa, on rencontre deux types principaux de fibres neurosécrétrices, l'une avec des grains denses de 1600 à 2400 Å de diamètre, l'autre avec des grains moins denses d'environ 1300 à 2000 Å de diamètre. Dans la zone externe bordant la pars intermedia des fibres aminergiques avec de fines granulations ont été observées.

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Electron microscopic study of the neurohypophysis of Rana esculenta L.

Summary The median eminence and the pars nervosa of Rana esculenta have a different structure.The median eminence has 2 different zones: the outer zone situated near the pars distalis and the inner zone under the ependyme. In the outer zone there are, according to the size and the shape of the granules, 5 types of nerve terminals.1. Endings containing spherical fine dense granules of 800 to 1000 Å in diameter; 2. Endings with spherical granules from 1000 to 1200 Å in diameter; 3. Endings with granules of irregular shape which are bigger than the former (1200 to 1600 Å); 4. Endings with spherical dense granules of about 1200 to 1800 Å in diameter; 5. A few endings containing only clear vesicles. Type 3 and type 4 endings are probably neurosecretory.The inner zone contains numerous neurosecretory fibres. They are of two types: one with big granules (1600–2400 Å), the second with smaller granules (1300–2000 Å). Non-neurosecretory fibres have also been observed.The pars nervosa contains two principal types of neurosecretory fibres: one with dense granules of 1600 to 2400 Å in diameter, the other with lighter granules of about 1300 to 2000 Å. In the external zone lining the pars intermedia, aminergic fibres with fine granules have been observed.

Je tiens à exprimer mes vifs remerciements, à Monsieur le Professeur E. Follenius pour l'intérêt constant qu'il prend à ce travail. Je remercie également Madame R. O. Clauss, collaboratrice technique et Madame Schwoerer, photographe, pour leur aide précieuse.     

Amphibian Pleurodeles waltl Fibronectin: cDNA Cloning and Developmental Expression of Spliced Variants

Partial cDNA clones encoding approximately the carboxy terminal half of Pleurodeles fibronectin (FN) were isolated. They account for 4.7 Kbp of the 3′ region of the FN mRNA. The cDNA nucleotide sequence comprises all three alternatively spliced segments designated EIIIA, EIIIB and V-segment, respectively. All three segments are included in FN mRNA synthesized during early embryogenesis whereas, from the tailbud stage onward the V-region was partially excluded. The isolation of Pleurodeles cDNA clones including the three different spliced EIIIA, EIIIB and V segment raises new possibilities for the study of the precise role of specific regions of FN in early amphibian development.     

Ultraviolet radiation and Vitamin D3 in amphibian health, behaviour, diet and conservation

Amphibians are currently suffering a period of mass extinction with approximately 20% of species under severe threat and more than 120 species already extinct. In light of this crisis there is an urgency to establish viable ex situ populations and also find the causes of in situ declines. The role of ultraviolet radiation and Vitamin D₃ in amphibian health directly influences both ex situ and in situ populations. Vitamin D₃ can be photosynthesised endogenously via UV-B radiation (UV-B), or acquired through the diet, and then metabolised to calcitriol the biologically active hormonal form. Although, there is a lack of literature concerning Vitamin D₃ requirements and calcitriol synthesis in amphibians, amphibians are likely to have similar Vitamin D₃ requirements and metabolic processes as other vertebrates due to the phylogenetically conservative nature of calcitriol biosynthesis. Deficiencies in calcitriol in amphibians result in nutritional metabolic bone disease (NMBD) and could compromise reproduction and immunity. However, excess biologically active UV radiation has also proven detrimental across all three amphibian life stages and therefore could impact both in situ and ex situ populations. Here we review the role and necessity of UV-B and calcitriol in amphibians and the potential for negative impacts due to excessive exposure to UV radiation. We also identify priorities for research that could provide critical information for maintaining healthy in ex situ and in situ populations of amphibians.     

Desiccation resistance in megalopae of the terrestrial hermit crab Coenobita compressus: water loss and the role of the shell

Abstract. The terrestrial hermit crab Coenobita compressus H. M ilne E dwards undergoes larval development in the sea and then moves to land as a megalopa, where it metamorphoses and remains for the rest of its life. As a small organism (generally <3 mg), in a body adapted for pelagic life, the megalopa must avoid desiccating in air to make a successful sea-to-land transition. In this study, I measured rates of water loss in 1 to 26-day-old megalopae without mollusk shells to determine if there is an improvement in desiccation resistance with age. I also exposed 26-day-old megalopae with or without shells to different relative-humidity conditions for 1 h to determine if shells worn by megalopae allow them to function in air that is not fully saturated. Megalopae without shells did not survive exposure to a relative humidity (RH) of less than 99%, while those with shells survived 52% RH. Older megalopae lost water more slowly than younger ones. However, the amount of body water explained more of the variation in water-loss rate than age; individuals with smaller body-water masses showed lower rates of absolute water loss. Though megalopae of C. compressus become less water permeable as they approach metamorphosis on land, most of their ability to avoid desiccation comes from the shell. Shell-wearing can be considered a pre-adaptation to a terrestrial life-style because shell-wearing behavior predates land invasion in hermit crabs, and desiccation-proofing in air is a novel function for shells.     

Neurogenesis during caudal spinal cord regeneration in adult newts

 After tail amputation in urodele amphibians, dramatic changes appear in the spinal cord rostral to the amputation level. Transection induces a proliferation response in cells lining the ependymal canal, giving rise to an ependymal tube in which neurogenesis occurs. Using the thymidine analog bromodeoxyuridine (BrdU) in short- and long-term labeling of cells undergoing DNA synthesis (S phase of the cell cycle), specific cell markers, and cell cultures, we show that neurons derive from the proliferative ependymal layer of the ependymal tube. Received: 30 November 1998 / Accepted: 22 December 1998     

Reproductive strategy of an isopod Onisocryptus ovalis,parasitizing a bioluminescent myodocope ostracod Vargula hilgendorfii

The functional morphology and the reproductive strategy of a parasitic isopod Onisocryptus ovalis in a bioluminescent ostracod Vargula hilgendorfii as its final host were studied based on video and SEM observations. During its lifetime, Onisocryptus ovalis dramatically metamorphoses several times, changing sex from male to female in the final host's carapace. At nearly the last ontogenetical stage, the parasite anchors its body with a pair of thoracopods to the posterodorsal region of the host ostracod's trunk and loses all the other appendages and thus its mobility as well. Thereafter, the parasite reverses bodily orientation during the final moulting so as to locate its mouth in the midst of the host eggs, and finally consumes them, leaving only the egg membrane. Such a mode of feeding of the parasite following the fixation of the body is interpreted in terms of the adaptation to escape elimination from the ostracod carapace by the host's cleaning appendages (the seventh limbs) and to obtain as much space as possible for the parasite's own eggs/embryos at the sacrifice of the mother's mobility. The synchronization between the timing of metamorphosis of the parasite and the reproductive cycle of the host animal can be expected to guarantee the parasite the opportunity to exploit sufficient nutrition from the eggs of the host.     

Teratogenic effects of tetrabromobisphenol A on Xenopus tropicalis embryos

Tetrabromobisphenol A (TBBPA) is the most widely used brominated flame retardant and a known thyroid disruptor. We reported exposing Xenopus tropicalis embryos (NF10) to 0.01, 0.1 or 1 mg/L of TBBPA with or without 70 µg/L triiodothyronine (T₃). Compared with the controls, 1 mg/L of TBBPA significantly reduced the body length of embryos after 24, 36 and 48 h of exposure. Embryos treated with TBBPA showed multiple malformations, including: abnormal eyes, skin hypopigmentation, enlarged proctodaeum, narrow fins and pericardial edemas. The effect of abnormal eyes manifested itself in the loss of pigmentation, reduction in size, or absence of external eyes. The degree of eye malformations was quantified with the index of eye malformations (IEM) with 0 being normal and 3 being severe. In the 1 mg/L TBBPA treatment groups, the incidence of total malformations (ITM) was 68–93%, and IEM was 0.8–0.9. T₃ showed no teratogenic effects on embryos, but it significantly enhanced TBBPA-induced teratogenic effects. In the T₃ + 1 mg/L TBBPA treatment groups, ITM was 91–99%, and IEM was 1.8–1.9. Histological observations showed that the retinas were generally smaller, and the lenses were underdeveloped or even absent. These results indicate that TBBPA at relatively high concentration has teratogenic effects on X. tropicalis embryos. The results also suggest that thyroid hormone signaling might be involved in TBBPA induced-teratogenicity.     

Distribution patterns of flora and fauna in southern Chilean Coastal rain forests: Integrating Natural History and GIS

Knowledge of species richness centers is necessary for the design of conservation areas. In this study, we present a GIS analysis of two years of field data on animal and plant diversity distributions in evergreen, coastal rain forests of southern Chile (39°30′–41°25′ S). Despite their high endemism, these forests have remained largely unprotected. Field records were complemented with data from museum collections and scientific literature. We used selected environmental variables (evapotranspiration, altitude) and, in some cases, forest types as predictors of species distributions. Our study focused on the distribution of forest bryophytes, vascular plants, soil invertebrates, amphibians and birds. We generated distributional maps for each taxa based on their field records in the study area, complemented by natural history information, except in the case of bryophytes and soil invertebrates. In general, species richness was lower at 600 m elevation or above for all the taxa studied. Species richness tends to increase in the northern sector of the study area. We observed a greater richness of vascular plants near rivers and streams, and noted important floristic differences between west and east-facing slopes of the Coastal Range, with more species in the oriental side. Because species in high altitude forests are not a subset of those found at lower elevations, we propose that conservation strategies should prioritize the protection of the entire altitudinal gradient of the southern Coastal Range, especially in the more diverse oriental and northern sectors.