A novel zebrafish model to emulate lung injury by folate deficiency-induced swim bladder defectiveness and protease/antiprotease expression imbalance.

Lung injury is one of the pathological hallmarks of most respiratory tract diseases including asthma, acute respiratory distress syndrome (ARDS) and chronic obstructive pulmonary disease (COPD). It involves progressive pulmonary tissue damages which are usually irreversible and incurable. Therefore, strategies to facilitate drug development against lung injury are needed. Here, we characterized the zebrafish folate-deficiency (FD) transgenic line that lacks a fully-developed swim bladder. Whole-mount in-situ hybridization revealed comparable distribution patterns of swim bladder tissue markers between wild-type and FD larvae, suggesting a proper development of swim bladder in early embryonic stages. Unexpectedly, neutrophils infiltration was not observed in the defective swim bladder. Microarray analysis revealed a significant increase and decrease of the transcripts for cathepsin L and a cystatin B (CSTB)-like (zCSTB-like) proteins, respectively, in FD larvae. The distribution of cathepsin L and the zCSTB-like transcripts was spatio-temporally specific in developing wild-type embryos and, in appropriate measure, correlated with their potential roles in maintaining swim bladder integrity. Supplementing with 5-formyltetrahydrofolate successfully prevented the swim bladder anomaly and the imbalanced expression of cathepsin L and the zCSTB-like protein induced by folate deficiency. Injecting the purified recombinant zebrafish zCSTB-like protein alleviated FD-induced swim bladder anomaly. We concluded that the imbalanced expression of cathepsin L and the zCSTB-like protein contributed to the swim bladder malformation induced by FD and suggested the potential application of this transgenic line to model the lung injury and ECM remodeling associated with protease/protease inhibitor imbalance.

Respiratory responses of domestic fowl to hyperthermia following selective air sac occlusions.

Ventilation together with blood and respiratory gas tensions were measured in adult domestic fowl under normothermic and hyperthermic conditions, following bilateral occlusion of the cranial and caudal thoracic air sacs (series I) or the cranial and caudal thoracic plus the abdominal air sacs (series II). Series I birds showed no significant differences from controls. Both control and experimental animals displayed a typical thermal polypnoea combined with mild hypocapnaemia. A larger drop in PCO2 was demonstrated in the clavicular sac than in the blood, possibly indicating partial failure of inspiratory valving at the ventrobronchi. However, there was no evidence of any effect of thoracic air sac occlusion on inspiratory airflow valving in the palaeopulmo. Series II birds were strongly hypercapnaemic/hypoxaemic in normothermic conditions, with a normal minute volume, but a faster, shallower breathing pattern. During hyperthermia they increased minute ventilation 3-fold, as in control animals, and blood gas tensions were almost restored to normal. Again, there was no evidence that experimental reduction in air sac capacity, in this case up to 70% of the total, had any effect on inspiratory airflow valving in the palaeopulmo, although inevitably in this case airflow in the neopulmo was abolished.