Does blood flow limit acute hypoxia performance in larval zebrafish (Danio rerio)?

Oxygen uptake (ṀO₂) in larval zebrafish prior to maturation of the gill relies on cutaneous O₂ transfer. Under normoxic conditions, rates of cutaneous O₂ transfer are unaffected by haemoglobin availability but are diminished in fish lacking a functional circulatory system, suggesting that internal convection is critically involved in setting the resting ṀO₂ in zebrafish larvae, even when relying on cutaneous O₂ transfer. The reliance of ṀO₂ on blood circulation led to the first objective of the current study, to determine whether loss of internal convection would reduce acute hypoxia performance (as determined by measuring critical PO₂; P_crit) in larval zebrafish under conditions of moderate hypoxia (PO₂ = 55 mmHg) at 28.5 and 34 °C. Internal convection was eliminated by preventing development of blood vessels using morpholino knockdown of vascular endothelial growth factor (VEGF); these fish are termed VEGF morphants. Breathing frequency (f_V) and heart rate (f_H) also were measured (at 28.5 °C) to determine whether any detriment in performance might be linked to cardiorespiratory dysfunction. Although ṀO₂ was reduced in the VEGF morphants, there was no significant effect on P_crit at 28.5 °C. Raising temperature to 34 °C resulted in the VEGF morphants exhibiting a higher P_crit than the shams, suggesting an impairment of hypoxia tolerance in the morphants at the higher temperature. The usual robust increase in f_V during hypoxia was absent or attenuated in VEGF morphants at 4 and 5 days post fertilization (dpf), respectively. Resting f_H was reduced in the VEGF morphants and unlike the sham fish, the morphants did not exhibit hypoxic tachycardia at 4 or 5 dpf. The number of cutaneous neuroepithelial cells (presumptive O₂ chemoreceptors) was significantly higher in the VEGF morphants and thus the cardiorespiratory impairment in the morphants during hypoxia was unlikely related to inadequate peripheral O₂ sensing.