Cooperation of Mtmr8 with PI3K Regulates Actin Filament Modeling and Muscle Development in Zebrafish

Background: It has been shown that mutations in at least four myotubularin family genes (MTM1, MTMR1, 2 and 13) are causative for human neuromuscular disorders. However, the pathway and regulative mechanism remain unknown. Methodology/Principal Findings: Here, we reported a new role for Mtmr8 in neuromuscular development of zebrafish. Firstly, we cloned and characterized zebrafish Mtmr8, and revealed the expression pattern predominantly in the eye field and somites during early somitogenesis. Using morpholino knockdown, then, we observed that loss-of-function of Mtmr8 led to defects in somitogenesis. Subsequently, the possible underlying mechanism and signal pathway were examined. We first checked the Akt phosphorylation, and observed an increase of Akt phosphorylation in the morphant embryos. Furthermore, we studied the PH/G domain function within Mtmr8. Although the PH/G domain deletion by itself did not result in embryonic defect, addition of PI3K inhibitor LY294002 did give a defective phenotype in the PH/G deletion morphants, indicating that the PH/G domain was essential for Mtmr8's function. Moreover, we investigated the cooperation of Mtmr8 with PI3K in actin filament modeling and muscle development, and found that both Mtmr8-MO1 and Mtmr8-MO2+LY294002 led to the disorganization of the actin cytoskeleton. In addition, we revealed a possible participation of Mtmr8 in the Hedgehog pathway, and cell transplantation experiments showed that Mtmr8 worked in a non-cell autonomous manner in actin modeling. Conclusion/Significance: The above data indicate that a conserved functional cooperation of Mtmr8 with PI3K regulates actin filament modeling and muscle development in zebrafish, and reveal a possible participation of Mtmr8 in the Hedgehog pathway. Therefore, this work provides a new clue to study the physiological function of MTM family members.

Effect of water temperature on the growth performance and digestive enzyme activities of Chinese longsnout catfish (Leiocassis longirostris Gunther)

The present study was carried out to investigate the influence of water temperature on the growth performance and digestive enzyme (pepsin, trypsin and lipase) activities of Chinese longsnout catfish. Triplicate groups of Chinese longsnout catfish (35.6 +/- 0.48 g, mean +/- SE) were reared at different water temperatures (20, 24, 28 and 32 degrees C). The feeding rate (FR), specific growth rate (SGR) and feed efficiency ratio (FER) were significantly affected by water temperatures and regression relationships between water temperature and FI, SGR as well as FER were expressed as FR=-0.016T2+0.91T-10.88 (n=12, R2=0.8752), SGR=-0.026T2+1.39T-17.29 (n=12, R2=0.7599) and FER=-0.013T2+0.70T-8.43 (n=12, R2=0.7272). Based on these, the optimum temperatures for FR, SGR and FER were 27.66, 26.69 and 26.44 degrees C respectively. The specific activities of digestive enzymes at 24 or 28 degrees C were significantly higher than that at 20 or 32 degrees C. In addition, there was a significant linear regression between FR or SGR and specific activities of pepsin and lipase, which indicated that pepsin and lipase played important roles in regulating growth through nutrient digestion in Chinese longsnout catfish.

ALGAL GROWTH POTENTIAL AND NUTRIENT LIMITATION IN SPRING IN THREE-GORGES RESERVOIR, CHINA

In China, especially in Three-Gorges Reservoir, our knowledge of the algal growth potential and nutrient limitation was still limited. In the spring of 2006, the water column ratios of total nitrogen/total phosphorus were investigated and algal bioassays performed to determine algal growth potential of waters and nutrient limitation of mainstream and Xiangxi Bay of Three-Gorges Reservoir. The results showed sampling sites in mainstream were co-limited by N and P or P-limited alone, and sites in Xiangxi Bay were N-limited alone. Fe likely played an important role in determining the appearance and disappearance of algal blooms of Three-Gorges Reservoir. Native algae, Pseudokirchneriella subcapitata and Cyclotella meneghiniana, had high growth potential in Three-Gorges Reservoir.

Chronic effects of water-borne PFOS exposure on growth, survival and hepatotoxicity in zebrafish: A partial life-cycle test

Perfluorooctane sulfonate (PFOS) is widely distributed and persistent in the environment and wildlife. The main aim of this study was to investigate the impact of long-term exposure to low concentrations of PFOS in zebrafish. Zebrafish fry (F-0, 14d post-fertilization, dpf) were exposed via the water for 70d to 0 (control), 10, 50 and 250 mu g L-1 PFOS, followed by a further 30d to assess recovery in clean water. The effects on survival and growth parameters and liver histopathology were assessed. Although growth suppression (weight and length) was observed in fish treated with high concentrations PFOS during the exposure period, no mortality was observed throughout the 70d experiment. Embryos and larvae (F-1) derived from maternal exposure suffered malformation and mortality. Exposure to 50 and 250 mu g L-1 PFOS could inhibit the growth of the gonads (GSI) in the female zebrafish. Histopathological alterations, primary with lipid droplets accumulation, were most prominently seen in the liver of males and the changes were not reversible, even after the fish were allowed to recover for 30d in clean water. The triiodothyronine (T-3)) levels were not significantly changed in any of the exposure groups. Hepatic vitellogenin (VTG) gene expression was significantly up-regulated in both male and female zebrafish, but the sex ratio was not altered. The overall results suggested that lower concentrations of PFOS in maternal exposure could result in offspring deformation and mortality. (c) 2008 Elsevier Ltd. All rights reserved.

Apo-14 is required for digestive system organogenesis during fish embryogenesis and larval development

Apo-14 is a fish-specific apolipoprotein and its biological function remains unknown. In this study, CagApo-14 was cloned from gibel carp (Carassius auratus gibelio) and its expression pattern was investigated during embryogenesis and early larval development. The CagApo-14 transcript and its protein product were firstly localized in the yolk syncytial layer at a high level during embryogenesis, and then found to be restricted to the digestive system including liver and intestine in later embryos and early larvae. Immunofluorescence staining in larvae and adults indicated that CagApo-14 protein was predominantly synthesized in and excreted from sinusoidal endothelial cells of liver tissue. Morpholino knockdown of CagApo-14 resulted in severe disruption of digestive organs including liver, intestine, pancreas and swim bladder. Moreover, yolk lipid transportation and utilization were severely affected in the CagApo-14 morphants. Overall, this data indicates that CagApo-14 is required for digestive system organogenesis during fish embryogenesis and larval development.

Effects of ammonium on growth, nitrogen and carbohydrate metabolism of Potamogeton maackianus A. Benn

Growth, nitrogen and carbohydrate metabolism in relation to eutrophication were studied for a submerged plant Potamogeton maackianus, a species common in East Asian shallow lakes. The plants were grown in six NH4+-N concentrations (0.05, 0.50, 1.00, 3.50, 5.00 and 10.00 mg/L) for six days. NH4+-N levels in excess of 0.50 mg/L inhibited the plant growth. The relationships between external NH4+-N availability and total nitrogen (TN), protein-N, free amino acid-N (FAA-N) and NH4+-N in plant tissues, respectively, conformed to a logarithmic model suggesting that a feedback inhibition mechanism may exist for ammonium uptake. The response of starch to NH4+-N was fitted with a negative, logarithmic curve. Detailed analysis revealed that the influx NH4+-N had been efficiently incorporated into organic-N and eventually stored as protein at the expense of starch accumulation. These data suggest that this species may be able to tolerate high levels of ammonium when dissolved oxygen is sufficient.

Effect of dietary carbohydrate sources on growth performance and utilization for gibel carp (Carassius auratus gibelio) and Chinese longsnout catfish (Leiocassis longirostris Gunther)

The nutritional function of monosaccharides, disaccharides and polysaccharides for omnivorous gibel carp and carnivorous Chinese longsnout catfish were investigated and the ability of these two species to utilize carbohydrates was compared. For each species, triplicate groups of fish were assigned to each of five groups of isoenergetic and isonitrogenous experimental diets with different carbohydrate sources: glucose, sucrose, dextrin, soluble starch (acid-modified starch) and alpha-cellulose. The carbohydrates were included at 60 g kg(-1) in Chinese longsnout catfish diets and at 200 g kg(-1) in gibel carp diets. A growth trial was carried out in a recirculation system at 27.8 +/- 1.9 degrees C for 8 weeks. The results showed that fish with different food habits showed difference in the utilization of carbohydrate sources. For gibel carp, better specific growth rate (SGR) and feed efficiency (FE) were observed in fish fed diets containing soluble starch and cellulose, but for Chinese longsnout catfish, better SGR and FE were observed in fish fed diets containing dextrin and sucrose. Apparent digestibility coefficient of dry matter (ADC(d)) and apparent digestibility coefficient of energy (ADC(e)) were significantly affected by dietary carbohydrate sources in gibel carp. ADC(d) and ADC(e) significantly decreased as dietary carbohydrate complexity increased in Chinese longsnout catfish except that glucose diet had medium ADC(d) and ADC(e). In both species, no significant difference of apparent digestibility coefficient of protein was observed between different carbohydrate sources. Dietary carbohydrate sources significantly affected body composition, and liver phosphoenolpyruvate carboxykinase (PEPCK), pyruvate kinase (PK), glucose 6-phosphate dehydrogenase (G6PD) and malic enzyme (ME) activities also varied according to dietary carbohydrate complexity. Fish with different food habits showed different abilities to synthesize liver glycogen, and the liver glycogen content in gibel carp was significantly higher than in Chinese longsnout catfish. The influence of carbohydrate source on gluconeogenesis and lipogenesis was also different in the two fish species.

Monitoring and preventing numerical oscillations in 3D simulations with coupled Monte Carlo codes

Previous studies have reported that different schemes for coupling Monte Carlo (MC) neutron transport with burnup and thermal hydraulic feedbacks may potentially be numerically unstable. This issue can be resolved by application of implicit methods, such as the stochastic implicit mid-point (SIMP) methods. In order to assure numerical stability, the new methods do require additional computational effort. The instability issue however, is problem-dependent and does not necessarily occur in all cases. Therefore, blind application of the unconditionally stable coupling schemes, and thus incurring extra computational costs, may not always be necessary. In this paper, we attempt to develop an intelligent diagnostic mechanism, which will monitor numerical stability of the calculations and, if necessary, switch from simple and fast coupling scheme to more computationally expensive but unconditionally stable one. To illustrate this diagnostic mechanism, we performed a coupled burnup and TH analysis of a single BWR fuel assembly. The results indicate that the developed algorithm can be easily implemented in any MC based code for monitoring of numerical instabilities. The proposed monitoring method has negligible impact on the calculation time even for realistic 3D multi-region full core calculations. © 2014 Elsevier Ltd. All rights reserved.

Toxicity of cypermethrin on growth, pigments, and superoxide dismutase of Scenedesmus obliquus

Cypermethrin is a synthetic pyrethroid that is particularly toxic to crustaceans. It is therefore applied as a chemotherapeutant in farms for the treatment of pests. The effective concentrations of cypermethrin on the inhibition of Scenedesmus ohliquus growth at 96h (96h EC50) were determined to be 50, 100, 150, 200, and 250mg/L. Algal growth, pigment fractions, and the activity of superoxide dismutase (SOD) in the algal cells were measured in the exponential phase after exposure to cypermethrin. The results show that higher concentration of cypermethrin is inhibitory for growth and other metabolic activities and the 96h EC50 of cypermethrin to S. ohliquus is 112 +/- 9 mg/L; the potential application of SOD activity in S. ohliquus as a sensitive biomarker for cypermethrin exposure is also discussed. (C) 2004 Elsevier Inc. All rights reserved.

The vertical microdistribution of cyanobacteria and green algae within desert crusts and the development of the algal crusts

Substantial amounts of algal crusts were collected from five different desert experimental sites aged 42, 34, 17, 8 and 4 years, respectively, at Shapotou ( China) and analyzed at a 0.1 mm microscale of depth. It was found that the vertical distribution of cyanobacteria and microalgae in the crusts was distinctly laminated into an inorganic-layer (ca. 0.00 - 0.02 mm, with few algae), an algae-dense-layer ( ca. 0.02 - 1.0 mm) and an algae-sparse-layer ( ca. 1.0 - 5.0 mm). It was interesting to note that in all crusts Scytonema javanicum Born et Flah ( or Nostoc sp., cyanobacterium), Desmococcus olivaceus (Pers ex Ach., green alga) Laundon and Microcoleus vaginatus Gom. ( cyanobacterium) dominated at the depth of 0.02 - 0.05, 0.05 - 0.1 and 0.1 - 1.0 mm, respectively, from the surface. Phormidium tenue Gom. ( or Lyngbya cryptovaginatus Schk., cyanobacterium) and Navicula cryptocephala Kutz.( or Hantzschia amphioxys (Ehr.) Grun. and N. cryptocephala together, diatom) dominated at the depth of 1.0 - 3.0 and 3.5 - 4.0 mm, respectively, of the crusts from the 42 and 34 year old sites. It was apparent that in more developed crusts there were more green algae and the niches of Nostoc sp., Chlorella vulgaris Beij., M. vaginatus, N. cryptocephala and fungi were nearer to the surface. If lichens and mosses accounted for less than 41.5% of the crust surface, algal biovolume was bigger when the crust was older, but the opposite was true when the cryptogams other than algae covered more than 70%. In addition to detailed species composition and biovolume, analyses of soil physicochemical properties, micromorphologies and mineral components were also performed. It was found that the concentration of organic matter and nutrients, electric conductivity, silt, clay, secondary minerals were higher and there were more micro-beddings in the older crusts than the less developed ones. Possible mechanisms for the algal vertical microdistribtion at different stages and the impact of soil topography on crust development are discussed. It is concluded that biomethods ( such as fine species distribution and biovolume) were more precise than mineralogical approaches in judging algal crust development and thus could be a better means to measure the potentiality of algal crusts in desert amelioration.

Spawning areas and early development of long spiky-head carp (Luciobrama macrocephalus) in the Yangtze River and Pearl River, China

The spawning areas and early development of long spiky-head carp, Luciobrama macrocephalus (Lacepede), an endemic fish species in China, were investigated in the Yangtze River and Pearl River of central and southeastern China between 1961 and 1993. The potamodromous fish migrated upstream to spawn between May and July as the floodwater began to rise. The water-hardened eggs drifted down the river, and the embryos and larvae developed in the course of drifting. The spawning areas of the fish were widely found in the upper and middle main channels and large tributaries. Two large dams (Gezhouba dam and Danjiangkou dam) did not significantly impact on the reproduction of the fish. Fifty stages of the early development from one cell to the juvenile with fully formed fins were observed and characterized pictorially. The larvae of long spiky-head carp could be distinguished from the larvae of other co-occurring species by counting the number of somites and comparing the proportion of sizes of eye to otic capsule.

The toxic effects of microcystin-LR on embryo-larval and juvenile development of loach, Misguruns mizolepis Gunthe

Microcystin-LR, a specific and potent hepatotoxin, was tested for its effects oil loach embryo-larval and juvenile development, The results of this study showed that loach embryos were more sensitive when exposed to microcystin-LR at a later than at an earlier stage of development, Juveniles were far less sensitive to MC-LR than were embryos and larvae. Mortality and developmental abnormality were proven to be dose-dependent and to be stage-specific sensitive. Among the abnormal changes noted were: pericardial edema and tubular heart, bradycardia, homeostasis, poor yolk resumption. small head, curved body and tail, and abnormal hatching, Liver and heart were the main targets of microcystin-LR toxicity. Ultrastructural analysis documented a complex set of sublethal effects of microcystin-LR on loach hepatocytes, chiefly including morphological alteration in nuclear and RER of loach liver cells. fit addition, microcystin-LR was lethal to loach juvenile in the subacute (7 days) exposure (LC50) = 593.3 mug/l). (C) 2002 Elsevier Science Ltd. All rights reserved.