Deficiency of manganese superoxide dismutase in hepatocytes disrupts zonated gene expression in mouse liver

The liver acinus displays a physiological periportal to perivenous oxygen gradient. This gradient was implicated to use reactive oxygen species (ROS) as mediators for the zonal gene expression. Mitochondria use oxygen and produce ROS, therefore they may contribute to the zonation of gene expression. To further elucidate this, we used the Cre-loxP system to generate a hepatocyte-specific null mutation of the mitochondrial antioxidant enzyme manganese superoxide dismutase (MnSOD) in mice. We found that ROS levels were enhanced in livers of MnSOD(-/-) mice which were reduced in size and displayed signs of liver failure such as intracellular protein droplets, increased apoptotic bodies and Bax levels as well as multinuclear hepatocytes. Further, the zonation of glutamine synthetase, glucokinase and phosphoenolpyruvate carboxykinase was no longer preserved. We conclude that deficiency of mitochondrial MnSOD initiates a dysregulation of zonated gene expression in liver.     

Fluff deposition on intertidal sediments: effects on benthic biota, ammonium fluxes and nitrification rates

The effects of fluff deposit on benthic biota,NH₄ ⁺ fluxes and nitrification was studied in thelaboratory using waterlogged and reflooded intertidal sediments fromMarennes-Oléron Bay, France. The fluff deposit was enriched inNH₄ ⁺ compared to underlying sediments, and promotedchanges of the sediment pH, E_h, C:N ratio, C:chla ratio and the NH₄ ⁺ efflux tooverlying water. Statistical analysis showed that pore waterNH₄ ⁺ concentrations were strongly influenced byinteractions between fluff, drying, depth and bioturbation. The fluff depositresulted in anoxia in the top sediments and moved the nitrification zone tosurface layers in fluff. However, the NH₄ ⁺ enrichment influff did not significantly change actual nitrification rates (range 0–1mmol m^–2 d^–1) or potentialnitrification rates (range 3–11 mmolNO₃ ^– m^–2d^–1).     

海桑属红树植物次生木质部解剖特征极其对潮间带生境的适应

通过光学显微镜和扫描电子显微镜详细观察了相似生境条件下生长的海桑属(Sonneratia)植物以及低、高潮位生长的海桑(S. caseolaris)和杯萼海桑(S. alba)次生木质部的形态特征,应用Lasersharp软件测量了其次生木质部的数量特征.结果表明:海桑属植物次生木质部形态特征的特化是与潮间带生境相适应的,能在水分胁迫的生境中,有效地协调水分输导的有效性和安全性.其特化结构主要包括:1)宽、窄导管并存,2)管孔密度较大,复孔率高,3)存在纤维状导管、形状不规则的导管和少量环管管胞,4)螺旋雕纹、附物纹孔、管壁具疣等许多导管壁的微观结构,有利于水分输导的安全性,5)射线细胞和分隔木纤维内的淀粉粒是渗透调节的物质基础,有利于促进水分上升,6)薄的纤维壁厚和宽的纤维腔径有利于水分的贮存,7)具胶质纤维.相似生境条件下生长的海桑属植物次生木质部数量特征的测量结果表明海桑和拟海桑水分输导效率高,但水分输导安全性差,而杯萼海桑水分输导效率低,但其输导安全性高.与高潮位生长的海桑和杯萼海桑相比,低潮位生长的海桑和杯萼海桑次生木质部导管分子更加"小型化"(有更小的导管分子长度、管孔弦向直径、管孔面积),有更高的导管聚合度和管孔密度,数量特征随生境不同的种内变动有利于海桑和杯萼海桑提高水分输导的安全性,但海桑和杯萼海桑输导安全性的获得是以牺牲水分输导的有效性为前提的.与生长在高潮位生境的海桑和杯萼海桑相比,低潮位生境生长的海桑和杯萼海桑的木材结构有更小的管孔面积,更厚的纤维壁,更低的导管比率和射线比率,更高的纤维比率,胶质纤维分布均匀且数量增加,这些特征有利于低潮位生境生长的海桑和杯萼海桑提高木材的韧性和强度,增强抗风浪冲击的能力.     

Formation of steady-state oxygen gradients in vitro: application to liver zonation

We have developed a perfusion bioreactor system that allows the formation of steady state oxygen gradients in cell culture. In this study, gradients were formed in cultures of rat hepatocytes to study the role of oxygen in modulating cellular functions. A model of oxygen transport in our flat-plate reactor was developed to estimate oxygen distribution at the cell surface. Experimental measurements of outlet oxygen concentration from various flow conditions were used to validate model predictions. We showed that cell viability was maintained over a 24-h period when operating with a physiologic oxygen gradient at the cell surface from 76 to 5 mmHg O(2) at the outlet. Oxygen gradients have been implicated in the maintenance of regional compartmentalized metabolic and detoxification functions in the liver, termed zonation. In this system, physiologic oxygen gradients in reactor cultures contributed to a heterogeneous distribution of phosphoenolpyruvate carboxykinase (predominantly localized upstream) and cytochrome p450 2B (predominantly localized downstream) that correlates with the distribution of these enzymes in vivo. The oxygen gradient chamber provides a means of probing the oxygen effects in vitro over a continuous range of O(2) tensions. In addition, this system serves as an in vitro model of zonation that could be further extended to study the role of gradients in ischemia-reperfusion injury, toxicity, and bioartificial liver design.     

Parameters affecting the early seedling development of four neotropical trees under oxygen deprivation stress

Some of the parameters that determine flooding resistance-and consequently habitat zonation-were investigated in four neotropical trees (Schizolobium parahyba, Sebastiania commersoniana, Erythrina speciosa and Sesbania virgata). The constitutive parameters of seeds (size, nature and amount of reserves) only partly influenced resistance to flooding, mainly through a high carbohydrate : size ratio. Parameters describing metabolic efficiency under stress conditions were more important. Among them, fermentation capacity and levels of ATP and of total adenylates played a key role. The highest resistance to anoxia was associated with increased availability of free sugars, elevated alcohol dehydrogenase activity and corresponding mRNA levels, more efficient removal of ethanol and lactate, and higher adenylate levels. Finally, as a lethal consequence of energy shortage, free fatty acids were released on a massive scale in the flooding-sensitive species Schizolobium parahyba, whereas lipid hydrolysis did not occur in the most resistant species Sesbania virgata.     

Hepatic heterogeneity in the response to ATP studied in the bivascularly perfused rat liver

The zonation of the purinergic action of ATP in the hepatic parenchyma was investigated in the bivascularly perfused rat liver by means of anterograde and retrograde perfusion. Livers from fed rats were used, and ATP was infused according to four different experimental protocols: (A) anterograde perfusion and ATP infusion via the portal vein; (B) anterograde perfusion and ATP via the hepatic artery; (C) retrograde perfusion and ATP via the hepatic vein; (D) retrograde perfusion and ATP via the hepatic artery. The following metabolic parameters were measured: glucose release, lactate production and oxygen consumption. The hemodynamic effects were evaluated by measuring the sinusoidal mean transit times by means of the indicator-dilution technique. ATP was infused during 20 min at four different rates (between 0.06-0.77 µmol min_-1 g liver_-1; 20-200 µM) in each of the four experimental protocols.The results that were obtained allow several conclusions with respect to the localization of the effects of ATP along the hepatic acini: (1) In retrograde perfusion the sinusoidal mean transit times were approximately twice those observed in anterograde perfusion. ATP increased the sinusoidal mean transit times only in retrograde perfusion (protocols C and D). The effect was more pronounced with protocol D. These results allow the conclusion that the responsive vasoconstrictive elements are localized in a pre-sinusoidal region; (2) All hepatic cells, periportal as well as perivenous, were able to metabolize ATP, so that concentration gradients were generated with all experimental protocols. Extraction of ATP was more pronounced in retrograde perfusion, an observation that can be attributed, partly at least, to the longer sinusoidal transit times. In anterograde perfusion, the extraction of ATP was time-dependent, a phenomenon that cannot be satisfactorily explained with the available data; (3) ATP produced a transient initial inhibition of oxygen uptake when protocols A and B were employed. These protocols are the only ones in which the cells situated shortly after the intrasinusoidal confluence of the portal vein and the hepatic artery were effectively supplied with ATP. The decrease in oxygen consumption was more pronounced at low ATP infusions when protocol B was employed. These observations allow the conclusion that the former phenomenon is localized mainly in cells situated shortly after the intrasinusoidal confluence of the portal vein and hepatic artery. Oxygen consumption in all other cells, especially the proximal periportal ones, is increased by ATP; (4) In agreement with previous data found in the literature, glycogenolysis stimulation by ATP was more pronounced in the periportal region. The cells that respond more intensively are not the proximal periportal ones, but those situated in the region of the intrasinusoidal confluence of the portal vein and the hepatic artery.     

Influence of puffer predation on vertical distribution of mangrove littorinids in the Gulf of Nicoya, Costa Rica

The eastern Pacific puffer fish Sphoeroides rosenblatti (family Tetraodontidae) preys on the mangrove periwinkles Littorariafasciata and L. varia up to 13 cm above the water surface. Puffers capture snails by jumping partially out of the water and removing them by mouth from Rhizophora spp. prop roots. Field predation trials suggested puffer predation is intense, averaging 40.6% h⁻¹ on L. fasciata tethered at the water surface. L.fasciata <∼8 mm in shell length and L. varia <∼12 mm remained near or below the water surface and were easily crushed by puffers; gut content analyses showed that these snails were commonly eaten by puffers. L.fasciata >∼8 mm and L. varia∼12–16 mm were also crushed by puffers, but were generally located above their reach; these snails were much less abundant in puffer guts. L.varia >∼16 mm were abundant and some were submerged at high tide; however, none were found in puffer guts presumably because puffers encounter difficulty crushing these snails. S.rosenblatti is an important predator of L. fasciata and L. varia and appears to affect their vertical distributions. Received: 21 April 1998 / Accepted: 10 August 1998     

The decision to feed by a scavenger in relation to the risks of predation and starvation

Recent theoretical models have considered how nutritional status and predation risk affect the decision by an individual to forage. We consider the feeding behaviour of a scavenger under risk of predation. The intertidal gastropod Nassarius dorsatus Röding was used to test the following hypotheses: (1) a damaged conspecific should be a less preferred meal than a damaged predator or another species; (2) a scavenger should be able to discriminate between a damaged conspecific in the presence and one in the absence of its predator, preferring the latter; and (3) the decision by a scavenger to feed should reflect a trade-off between nutritional status and the above preferences. Results from the laboratory and field were consistent with predictions. Two hypotheses are suggested from the work: scavengers may generally be less willing to feed on carcases of conspecifics than on those of other species, and intertidal scavengers may commonly face a relative or absolute shortage of food.     

Localization of the fructose 1,6-bisphosphatase at the nuclear periphery

The localization of fructose 1,6-bisphosphatase (D-Fru-1,6-P₂-1-phosphohydrolase, EC 3.1.3.11) in rat kidney and liver was determined immunohistochemically using a polyclonal antibody raised against the enzyme purified from pig kidney. The immunohistochemical analysis revealed that the bisphosphatase was preferentially localized in hepatocytes of the periportal region of the liver and was absent from the perivenous region. Fructose-1,6-bisphosphatase was also preferentially localized in the cortex of the kidney proximal tubules and was absent in the glomeruli, loops of Henle, collecting and distal tubules, and in the renal medulla. As indicated by immunocytochemistry using light microscopy and confirmed with the use of reflection confocal microscopy, the enzyme was preferentially localized in a perinuclear position in the liver and the renal cells. Subcellular fractionation studies followed by enzyme activity assays revealed that a majority of the cellular fructose-1,6-bisphosphatase activity was associated to subcellular particulate structures. Overall, the data support the concept of metabolic zonation in liver as well as in kidney, and establish the concept that the Fructose-1,6-bisphosphatase is a particulate enzyme that can not be considered a soluble enzyme in the classical sense. © 1996 Wiley-Liss, Inc.     

COMPARATIVE PHOTOSYNTHESIS OF TWO SPECIES OF INTERTIDAL EPIPHYTIC MACROALGAE ON MANGROVE ROOTS DURING SUBMERSION AND EMERSION

Photosynthesis and dark respiration rates were measured in water and in air, and the capacity to recover photosynthetic activity from emersion stress was examined for two species of intertidal, epiphytic macroalgae—Bostrychia calliptera (Montagne) Montagne and Caloglossa leprieurii (Montagne) J. Agardh—collected on prop roots of the red mangrove Rhizophora mangle L. in Buenaventura Bay, Pacific coast of Colombia. In both species, net photosynthetic rates were significantly higher under submersed conditions. Maximum photosynthetic rates (P_max) in water and in air were highest in B. calliptera, 126 ± 4 versus 52 ± 9 μmol O₂·mg chl a⁻¹·h⁻¹, respectively. In C. leprieurii, P_max of submerged plants in water and in air were 98 ± 9 versus 30 ± 11 μmol O₂·mg chla⁻¹·h⁻¹. The photoinhibition model of Platt et al. (1980) was used to fit the experimental data in both water and air for both species. Photoinhibition occurred at irradiance as low as 200 μmol·m⁻²·s⁻¹. The photosynthesis–light response curves demonstrated an adaptation to shaded habitats for both species, as light compensation points in water and air for both species were below 17 ± 5 μmol·m⁻²·s⁻¹. The rate of dehydration was significantly lower in thalli of B. calliptera compared to C. leprieurii. An increase of photosynthetic activity in B. calliptera was evident between 5% and 15% water loss, but rates decreased thereafter with declining water content. In C. leprieurii, desiccation negatively influenced photosynthetic rates that significantly decreased linearly with declining water content. In B. calliptera, net photosynthesis reached zero only at a water content between 29% and 35%, whereas in C. leprieurii no net photosynthesis occurred in plants containing less than about 50% of their relative water content. Resubmerged plants ofB. calliptera exhibited 100% photosynthetic recovery after 45 min, whereas C. leprieurii recovered 100% at about 120 min. On the basis of the comparison of rates of light-saturated net photosynthesis for B. calliptera in air versus in water, aerial photosynthetic activity ranged from 35% to 42% of that in water, whereas the emersed photosynthetic capacity of C. leprieurii ranged from 24% to 29% of that in water. Using tidal predictions and the emersed photosynthetic rates, a carbon balance model was constructed for both species over a single daylight period. The calculations indicated that emersed photosynthesis increased average daily carbon production of B. calliptera by 17% and C. leprieuri by 12%. The physiological responses to desiccation stress and the photosynthetic recovery capacities between species correlated with, and may determine, their vertical distribution in the mangrove habitats of Buenaventura Bay.