Jehan-Hervé Lignot

Osmoregulation, bioenergetics and oxidative stress in coastal marine invertebrates: raising the questions for future research

ABSTRACT Osmoregulation is by no means an energetically cheap process, and its costs have been extensively quantified in terms of respiration and aerobic metabolism. Common products of mitochondrial activity are reactive oxygen and nitrogen species, which may cause oxidative stress by degrading key cell components, while playing essential roles in cell homeostasis. Given the delicate equilibrium between pro- and antioxidants in fueling acclimation responses, the need for a thorough understanding of the relationship between salinity-induced oxidative stress and osmoregulation arises as an important issue, especially in the context of global changes and anthropogenic impacts on coastal habitats. This is especially urgent for intertidal/estuarine organisms, which may be subject to drastic salinity and habitat changes, leading to redox imbalance. How do osmoregulation strategies determine energy expenditure, and how do these processes affect organisms in terms of oxidative stress? What mechanisms are used to cope with salinity-induced oxidative stress? This Commentary aims to highlight the main gaps in our knowledge, covering all levels of organization. From an energy-redox perspective, we discuss the link between environmental salinity changes and physiological responses at different levels of biological organization. Future studies should seek to provide a detailed understanding of the relationship between osmoregulatory strategies and redox metabolism, thereby informing conservation physiologists and allowing them to tackle the new challenges imposed by global climate change. Summary: From an energy-redox perspective, changes in environmental salinity induce a wide diversity of responses in marine invertebrates. Characterization of mitochondrial function may allow disentangling evolutionary adaptations to different marine environments

Osmoregulation and salinity-induced oxidative stress: is oxidative adaptation determined by gill function?

ABSTRACT Osmoregulating decapods such as the Mediterranean green crab Carcinus aestuarii possess two groups of spatially segregated gills: anterior gills serve mainly respiratory purposes, while posterior gills contain osmoregulatory structures. The co-existence of similar tissues serving different functions allows the study of differential adaptation, in terms of free radical metabolism, upon salinity change. Crabs were immersed for 2 weeks in seawater (SW, 37 ppt), diluted SW (dSW, 10 ppt) and concentrated SW (cSW, 45 ppt). Exposure to dSW was the most challenging condition, elevating respiration rates of whole animals and free radical formation in hemolymph (assessed fluorometrically using C-H2DFFDA). Further analyses considered anterior and posterior gills separately, and the results showed that posterior gills are the main tissues fueling osmoregulatory-related processes because their respiration rates in dSW were 3.2-fold higher than those of anterior gills, and this was accompanied by an increase in mitochondrial density (citrate synthase activity) and increased levels of reactive oxygen species (ROS) formation (1.4-fold greater, measured through electron paramagnetic resonance). Paradoxically, these posterior gills showed undisturbed caspase 3/7 activity, used here as a marker for apoptosis. This may only be due to the high antioxidant protection that posterior gills benefit from [superoxide dismutase (SOD) in posterior gills was over 6 times higher than in anterior gills]. In conclusion, osmoregulating posterior gills are better adapted to dSW exposure than respiratory anterior gills because they are capable of controlling the deleterious effects of the ROS production resulting from this salinity-induced stress. Summary: Gill tissues with different functions (respiratory/osmoregulatory) differentially produce reactive oxygen species and are differently adapted to deal with oxidative stress.

Salinity stress from the perspective of the energy-redox axis: Lessons from a marine intertidal flatworm

In the context of global change, there is an urgent need for researchers in conservation physiology to understand the physiological mechanisms leading to the acquisition of stress acclimation phenotypes. Intertidal organisms continuously cope with drastic changes in their environmental conditions, making them outstanding models for the study of physiological acclimation. As the implementation of such processes usually comes at a high bioenergetic cost, a mitochondrial/oxidative stress approach emerges as the most relevant approach when seeking to analyze whole-animal responses. Here we use the intertidal flatworm Macrostomum lignano to analyze the bioenergetics of salinity acclimation and its consequences in terms of reactive oxygen/nitrogen species formation and physiological response to counteract redox imbalance. Measures of water fluxes and body volume suggest that M. lignano is a hyper-/iso-regulator. Higher salinities were revealed to be the most energetically expensive conditions, with an increase in mitochondrial density accompanied by increased respiration rates. Such modifications came at the price of enhanced superoxide anion production, likely associated with a high caspase 3 upregulation. These animals nevertheless managed to live at high levels of environmental salinity through the upregulation of several mitochondrial antioxidant enzymes such as superoxide dismutase. Contrarily, animals at low salinities decreased their respiration rates, reduced their activity and increased nitric oxide formation, suggesting a certain degree of metabolic arrest. A contradictory increase in dichlorofluorescein fluorescence and an upregulation of gluthathione-S-transferase pi 1 (GSTP1) expression were observed in these individuals. If animals at low salinity are indeed facing metabolic depression, the return to seawater may result in an oxidative burst. We hypothesize that this increase in GSTP1 could be a “preparation for oxidative stress”, i.e. a mechanism to counteract the production of free radicals upon returning to seawater. The results of the present study shed new light on how tolerant organisms carry out subcellular adaptations to withstand environmental change.

Changes in Form and Function of the Gastrointestinal Tract During Starvation: From Pythons to Rats

During episodes of food deprivation, animals enhance survival by minimising energy costs incurred by the metabolically demanding maintenance and activity of their digestive system. The response occurs at physiological, biochemical and molecular levels and mainly concerns the intestinal wall. After a feeding episode, this response leads to a rapid decrease in gut length and mass, as well as enzyme activities, protein synthesis and the expression of many proteolytic-related genes. Intestinal atrophy affects the mucosa, and induces a general decrease in its surface area through the diminishing of the mucosal area (shortening of the intestinal folds), changes in the microvilli surface area and by reducing both the size (hypotrophy) and number of cells along the intestinal barrier (hypoplasia). In most of the species studied, these morphological responses are time dependent and do not alter nutrient transport capacity, at least at the beginning of the fasting period. Fasting is usually anticipated in hibernating mammals, migratory vertebrates and infrequent feeders such as pythons. When preparing itself for fasting, the digestive system mainly uses tools such as hyperphagia, intracellular recycling and the production of new cells that are downregulated at the end of the postprandial period. As food deprivation continues starving animals may respond differently. This has been illustrated in rats that had reached a proteolytic phase during which energy requirements were mostly derived from increased protein utilisation. In these animals, cell proliferation and cell migration were seen to increase while apoptosis at the tip of the intestinal villi ceased. This has been considered as an optimising process that may also exist in other species.

Effects of domestic effluent discharges on mangrove crab physiology: Integrated energetic, osmoregulatory and redox balances of a key engineer species

Mangroves are increasingly used as biofiltering systems of (pre-treated) domestic effluents. However, these wastewater discharges may affect local macrofauna. This laboratory study investigates the effects of wastewater exposure on the mangrove spider crab Neosarmatium meinerti, a key engineering species which is known to be affected by waste waters in effluent-impacted areas. These effects were quantified by monitoring biological markers of physiological state, namely oxygen consumption, the branchial cavity ventilation rate, gill physiology and morphology, and osmoregulatory and redox balance. Adults acclimated to clean seawater (SW, 32 ppt) and freshwater (FW, ∼0 ppt) were compared to crabs exposed to wastewater for 5 h (WW, ∼0 ppt). Spider crabs exposed to WW increased their ventilation and whole-animal respiration rates by 2- and 3-fold respectively, while isolated gill respiration increased in the animals exposed to FW (from 0.5 to 2.3 and 1.1 nmol O2 min-1 mg DW-1 for anterior and posterior gills, respectively) but was not modified in WW-exposed individuals. WW exposure also impaired crab osmoregulatory capacity; an 80 mOsm kg-1 decrease was observed compared to FW, likely due to decreased branchial NKA activity. ROS production (DCF fluorescence in hemolymph), antioxidant defenses (superoxide dismutase and catalase activities) and oxidative damage (malondialdehyde concentration) responses varied according to animal gender. Overall, this study demonstrates that specific physiological parameters must be considered when focusing on crabs with bimodal breathing capacities. We conclude that spider crabs exposed to WW face osmoregulatory imbalances due to functional and morphological gill remodeling, which must rapidly exhaust energy reserves. These physiological disruptions could explain the ecological changes observed in the field.