Behavioral repertoire of high-shore littorinid snails reveals novel adaptations to an extreme environment.

Species that inhabit high-shore environments on rocky shores survive prolonged periods of emersion and thermal stress. Using two Hong Kong high-shore littorinids (Echinolittorina malaccana and E. radiata) as models, we examined their behavioral repertoire to survive these variable and extreme conditions. Environmental temperatures ranged from 4°C in the cool season to 55.5°C in the hot season, with strong seasonal and daily fluctuations. In the hot season, both species allocated >35% of their activity budgets to stress-mitigating thermoregulatory behaviors (e.g. standing, towering) and relatively small proportions to foraging (<20%) and reproduction (<10%). In the assumedly benign cool season, greater proportions (>70%) of activity budgets were allocated to stress mitigation behaviors (crevice occupation, aggregation formation). Both species exhibited multifunctional behaviors that optimized time use during their tidally-constrained activity window in the hot season. Females mated while foraging when awash by the rising tide, and some males crawled on top of females prior to ceasing movement to form 'towers', which have both thermoregulatory benefits and reduce searching time for mates during subsequent activity. The function of such behaviors varies in a state-dependent manner, for example, the function of trail following changes over an activity cycle from mate searching on rising tides, to stress mitigation on falling tides (aiding aggregation formation), and to both functions through tower formation just before movement stops. Many of these behavioral responses are, therefore, multifunctional and can vary according to local conditions, allowing snails in this family to successfully colonize the extreme high-shore environment.

Tissue distribution of triphenyltin compounds in marine teleost fishes.

Continuous release of the highly toxic triphenyltin compounds (TPT) from antifouling paints and fungicides has caused serious pollution to urbanized coastal marine environments worldwide since the 1960s. Using gas-chromatography mass-spectrometry (GC-MS), this study investigated the distribution profile of TPT in 15 types of tissues of four marine teleost fish species collected from Hong Kong waters. Concentrations of TPT in various tissues had a significant positive correlation with protein contents in the tissues (r = 0.346, p < 0.001) and, to a lesser extent with lipid contents (r = 0.169, p = 0.020). Highest concentrations of TPT were consistently found in liver, ranging from 1074.9 to 3443.7 ng/g wet weight; whereas fish scales always contained the least concentration of TPT in all species, ranging from 10.4 to 48.5 ng/g wet weight. Through mass balance models and regression analyses, muscle tissues were found to contribute most to the total TPT body burden, and the average TPT concentration of both dorsal and ventral muscles was identified as the best predictor for estimating TPT burden in the entire fish. Hence, further investigations of bioaccumulation and biomagnification of TPT in fishes should adopt this modelling approach in estimating its total body burden in individual fish.

High thermal stress responses of Echinolittorina snails at their range edge predict population vulnerability to future warming.

Populations at the edge of their species' distribution ranges are typically living at the physiological extreme of the environmental conditions they can tolerate. As a species' response to global change is likely to be largely determined by its physiological performance, subsequent changes in environmental conditions can profoundly influence populations at range edges, resulting in range extensions or retractions. To understand the differential physiological performance among populations at their distribution range edge and center, we measured levels of mRNA for heat shock protein 70 (hsp70) as an indicator of temperature sensitivity in two high-shore littorinid snails, Echinolittorina malaccana and E. radiata, between 1°N to 36°N along the NW Pacific coast. These Echinolittorina snails are extremely heat-tolerant and frequently experience environmental temperatures in excess of 55 °C when emersed. It was assumed that animals exhibiting high temperature sensitivity will synthesize higher levels of mRNA, which will thus lead to higher energetic costs for thermal defense. Populations showed significant geographic variation in temperature sensitivity along their range. Snails at the northern range edge of E. malaccana and southern range edge of E. radiata exhibited higher levels of hsp70 expression than individuals collected from populations at the center of their respective ranges. The high levels of hsp70 mRNA in populations at the edge of a species' distribution range may serve as an adaptive response to locally stressful thermal environments, suggesting populations at the edge of their distribution range are potentially more sensitive to future global warming.

Modulation of infection-mediated migration of neutrophils and CXCR2 trafficking by osteopontin.

Osteopontin (OPN) is a pro-inflammatory protein that paradoxically protects against inflammation and bone destruction in a mouse model of endodontic infection. Here we have tested the hypothesis that this effect of OPN is mediated by effects on migration of innate immune cells to the site of infection. Using the air pouch as a model of endodontic infection in mice, we showed that neutrophil accumulation at the site of infection with a mixture of endodontic pathogens is significantly reduced in OPN-deficient mice. Reduced neutrophil accumulation in the absence of OPN was accompanied by an increase in bacterial load. OPN-deficiency did not affect neutrophil survival, CXCR2 ligand expression, or the production of inflammatory cytokines in the air pouch. In vitro, OPN enhanced neutrophil migration to CXCL1, whereas in vivo, inhibition of CXCR2 suppressed cellular infiltration in air pouches of infected wild-type mice by > 50%, but had no effect in OPN-deficient mice. OPN increased cell surface expression of CXCR2 on bone marrow neutrophils in an integrin-αv -dependent manner, and suppressed the internalization of CXCR2 in the absence of ligand. Together, these results support a model where the protective effect of OPN results from enhanced initial neutrophil accumulation at sites of infection resulting in optimal bacterial killing. We describe a novel mechanism for this effect of OPN: integrin-αv -dependent suppression of CXCR2 internalization in neutrophils, which increases the ability of these cells to migrate to sites of infection in response to CXCR2 ligands.

Osteopontin binding to the alpha 4 integrin requires highest affinity integrin conformation, but is independent of post-translational modifications of osteopontin.

Osteopontin (OPN) is a ligand for the α4ß1 integrin, but the physiological importance of this binding is not well understood. Here, we have assessed the effect of post-translational modifications on OPN binding to the α4 integrin on cultured human leukocyte cell lines and compared OPN interaction with α4 integrin to that of VCAM and fibronectin. Jurkat cells, whose α4 integrins are inherently activated, adhered to different preparations of OPN in the presence of Mn(2+): the EC50 of adhesion was not affected by phosphorylation or glycosylation status. Thrombin cleavage of OPN at the C-terminus of the α4 integrin-binding site also did not affect binding affinity. THP-1 cells express a low-affinity conformation of the integrin and adhered to OPN only in the presence of Mn(2+) plus PMA or an activating antibody. This was in contrast to VCAM and fibronectin: THP-1 cells adhered to these ligands without integrin activation. Studies with ligand-induced binding site antibodies demonstrated that the SVVYGLR peptide of OPN bound to the α4 integrin with a similar affinity as the LDV peptide of fibronectin, suggesting that a high off-rate is responsible for the reduced binding of OPN to the low-affinity forms of this integrin. Together, the results suggest OPN has very low affinity for the α4 integrin on human leukocytes under physiological conditions.