Transcriptome sequencing supports a conservation of macrophage polarization in fish.

Mammalian macrophages can adopt polarization states that, depending on the exact stimuli present in their extracellular environment, can lead to very different functions. Although these different polarization states have been shown primarily for macrophages of humans and mice, it is likely that polarized macrophages with corresponding phenotypes exist across mammals. Evidence of functional conservation in macrophages from teleost fish suggests that the same, or at least comparable polarization states should also be present in teleosts. However, corresponding transcriptional profiles of marker genes have not been reported thus far. In this study we confirm that macrophages from common carp can polarize into M1- and M2 phenotypes with conserved functions and corresponding transcriptional profiles compared to mammalian macrophages. Carp M1 macrophages show increased production of nitric oxide and a transcriptional profile with increased pro-inflammatory cytokines and mediators, including il6, il12 and saa. Carp M2 macrophages show increased arginase activity and a transcriptional profile with increased anti-inflammatory mediators, including cyr61, timp2b and tgm2b. Our RNA sequencing approach allowed us to list, in an unbiased manner, markers discriminating between M1 and M2 macrophages of teleost fish. We discuss the importance of our findings for the evaluation of immunostimulants for aquaculture and for the identification of gene targets to generate transgenic zebrafish for detailed studies on M1 and M2 macrophages. Above all, we discuss the striking degree of evolutionary conservation of macrophage polarization in a lower vertebrate.

Fish Macrophages Show Distinct Metabolic Signatures Upon Polarization.

Macrophages play important roles in conditions ranging from host immune defense to tissue regeneration and polarize their functional phenotype accordingly. Next to differences in the use of L-arginine and the production of different cytokines, inflammatory M1 macrophages and anti-inflammatory M2 macrophages are also metabolically distinct. In mammals, M1 macrophages show metabolic reprogramming toward glycolysis, while M2 macrophages rely on oxidative phosphorylation to generate energy. The presence of polarized functional immune phenotypes conserved from mammals to fish led us to hypothesize that a similar metabolic reprogramming in polarized macrophages exists in carp. We studied mitochondrial function of M1 and M2 carp macrophages under basal and stressed conditions to determine oxidative capacity by real-time measurements of oxygen consumption and glycolytic capacity by measuring lactate-based acidification. In M1 macrophages, we found increased nitric oxide production and irg1 expression in addition to altered oxidative phosphorylation and glycolysis. In M2 macrophages, we found increased arginase activity, and both oxidative phosphorylation and glycolysis were similar to control macrophages. These results indicate that M1 and M2 carp macrophages show distinct metabolic signatures and indicate that metabolic reprogramming may occur in carp M1 macrophages. This immunometabolic reprogramming likely supports the inflammatory phenotype of polarized macrophages in teleost fish such as carp, similar to what has been shown in mammals.

Malaria mosquitoes use leg push-off forces to control body pitch during take-off.

Escaping from a blood host with freshly acquired nutrition for her eggs is one of the most critical actions in the life of a female malaria mosquito. During this take-off, she has to carry a large payload, up to three times her body weight, while avoiding tactile detection by the host. What separates the malaria mosquito from most other insects is that the mosquito pushes off gently with its legs while producing aerodynamic forces with its wings. Apart from generating the required forces, the malaria mosquito has to produce the correct torques to pitch-up during take-off. Furthermore, the fed mosquito has to alter the direction of its aerodynamic force vector to compensate for the higher body pitch angle due to its heavier abdomen. Whether the mosquito generates these torques and redirection of the forces with its wings or legs remains unknown. By combining rigid-body inverse dynamics analyses with computational fluid dynamics simulations, we show that mosquitoes use leg push-off to control pitch torques and that the adaption of the aerodynamic force direction is synchronized with modulations in force magnitude. These results suggest that during the push-off phase of a take-off, mosquitoes use their flight apparatus primarily as a motor system and they use leg push-off forces for control.

A chordwise offset of the wing-pitch axis enhances rotational aerodynamic forces on insect wings: a numerical study.

Most flying animals produce aerodynamic forces by flapping their wings back and forth with a complex wingbeat pattern. The fluid dynamics that underlies this motion has been divided into separate aerodynamic mechanisms of which rotational lift, that results from fast wing pitch rotations, is particularly important for flight control and manoeuvrability. This rotational force mechanism has been modelled using Kutta-Joukowski theory, which combines the forward stroke motion of the wing with the fast pitch motion to compute forces. Recent studies, however, suggest that hovering insects can produce rotational forces at stroke reversal, without a forward motion of the wing. We have conducted a broad numerical parametric study over a range of wing morphologies and wing kinematics to show that rotational force production depends on two mechanisms: (i) conventional Kutta-Joukowski-based rotational forces and (ii) a rotational force mechanism that enables insects with an offset of the pitch axis relative to the wing's chordwise symmetry axis to generate rotational forces in the absence of forward wing motion. Because flying animals produce control actions frequently near stroke reversal, this pitch-axis-offset dependent aerodynamic mechanism may be particularly important for understanding control and manoeuvrability in natural flyers.