Fish are exposed to large oxygen fluctuations in their aquatic environment since the inherent properties of water can result in marked spatial and temporal differences in the concentration of oxygen (see oxygenation and underwater). Fish respond to hypoxia with varied behavioral, physiological, and cellular responses to maintain homeostasis and organism function in an oxygen-depleted environment. The biggest challenge fish face when exposed to low oxygen conditions is maintaining metabolic energy balance, as 95% of the oxygen consumed by fish is used for ATP production releasing the chemical energy of nutrients through the mitochondrial electron transport chain. Therefore, hypoxia survival requires a coordinated response to secure more oxygen from the depleted environment and counteract the metabolic consequences of decreased ATP production at the mitochondria.
Hypoxia tolerance A fish's hypoxia tolerance can be represented in different ways. A commonly used representation is the critical O2 tension (Pcrit), which is the lowest water O2 tension (PO2) at which a fish can maintain a stable O2 consumption rate (MO2). A fish with a lower Pcrit is therefore thought to be more hypoxia-tolerant than a fish with a higher Pcrit. But while Pcrit is often used to represent hypoxia tolerance, it more accurately represents the ability to take up environmental O2 at hypoxic PO2s and does not incorporate the significant contributions of anaerobic glycolysis and metabolic suppression to hypoxia tolerance (see below). Pcrit is nevertheless closely tied to a fish's hypoxia tolerance, in part because some fish prioritize their use of aerobic metabolism over anaerobic metabolism and metabolic suppression. It therefore remains a widely used hypoxia tolerance metric.
A fish's hypoxia tolerance can also be represented as the amount of time it can spend at a particular hypoxic PO2 before it loses dorsal-ventral equilibrium (called time-to-LOE), or the PO2 at which it loses equilibrium when PO2 is decreased from normoxia to anoxia at some set rate (called PO2-of-LOE). A higher time-to-LOE value or a lower PO2-of-LOE value therefore imply enhanced hypoxia tolerances. In either case, LOE is a more holistic representation of overall hypoxia tolerance because it incorporates all contributors to hypoxia tolerance, including aerobic metabolism, anaerobic metabolism and metabolic suppression.
Oxygen sensing
Oxygen sensing structures In mammals there are several structures that have been implicated as oxygen sensing structures; however, all of these structures are situated to detect aortic or internal hypoxia since mammals rarely run into environmental hypoxia. These structures include the type I cells of the carotid body, the neuroepithelial bodies of the lungs as well as some central and peripheral neurons and vascular smooth muscle cells. In fish, the neuroepithelial cells (NEC) have been implicated as the major oxygen sensing cells. NEC have been found in all teleost fish studied to date, and are likely a highly conserved structure within many taxa of fish. NEC are also found in all four gill arches within several different structures, such as along the filaments, at the ends of the gill rakers and throughout the lamellae. Two separate neural pathways have been identified within the zebrafish gill arches both the motor and sensory nerve fibre pathways. Since neuroepithelial cells are distributed throughout the gills, they are often ideally situated to detect both arterial as well as environmental oxygen.
Mechanisms of neurotransmitter release in neuroepithelial cells Neuroepithelial cells (NEC) are thought to be neuron-like chemoreceptor cells because they rely on membrane potential changes for the release of neurotransmitters and signal transmission onto nearby cells. Once NEC of the zebrafish gills come in contact with either environmental or aortic hypoxia, an outward K+ "leak" channel is inhibited. It remains unclear how these K+ channels are inhibited by a shortage of oxygen because there are yet to be any known direct binding sites for "a lack of oxygen", only whole cell and ion channel responses to hypoxia. K+ "leak" channels are two-pore-domain ion channels that are open at the resting membrane potential of the cell and play a major role in setting the equilibrium resting membrane potential of the cell. Once this "leak" channel is closed, the K+ is no longer able to freely flow out of the cell, and the membrane potential of the NEC increases; the cell becomes depolarized. This depolarization causes voltage-gated Ca2+ channels to open, and for extracellular Ca2+ to flow down its concentration gradient into the cell causing the intracellular Ca2+ concentration to greatly increase. Once the Ca2+ is inside the cell, it binds to the vesicle release machinery and facilitates binding of the t-snare complex on the vesicle to the s-snare complex on the NEC cell membrane which initiates the release of neurotransmitters into the synaptic cleft.
Signal transduction up to higher brain centres If the post-synaptic cell is a sensory neuron, then an increased firing rate in that neuron will transmit the signal to the central nervous system for integration. Whereas, if the post-synaptic cell is a connective pillar cell or a vascular smooth muscle cell, then the serotonin will cause vasoconstriction and previously unused lamellae will be recruited through recruitment of more capillary beds, and the total surface area for gas exchange per lamella will be increased.
In fish, the hypoxic signal is carried up to the brain for processing by the glossopharyngeal (cranial nerve IX) and vagus (cranial nerve X) nerves. The first branchial arch is innervated by the glossopharyngeal nerve (cranial nerve IX); however all four arches are innervated by the vagus nerve (cranial nerve X). Both the glossopharyngeal and vagus nerves carry sensory nerve fibres into the brain and central nervous system.
Locations of oxygen sensors Through studies using mammalian model organisms, there are two main hypotheses for the location of oxygen sensing in chemoreceptor cells: the membrane hypothesis and the mitochondrial hypothesis. The membrane hypothesis was proposed for the carotid body in mice, and it predicts that oxygen sensing is an ion balance initiated process. The mitochondrial hypothesis was also proposed for the carotid body of mice, but it relies on the levels of oxidative phosphorylation and/or reactive oxygen species (ROS) production as a cue for hypoxia. Specifically, the oxygen sensitive K+ currents are inhibited by H2O2 and NADPH oxidase activation. There is evidence for both of these hypotheses depending on the species used for the study. For the neuroepithelial cells in the zebrafish gills, there is strong evidence supporting the "membrane hypothesis" due to their capacity to respond to hypoxia after removal of the contents of the cell. However, there is no evidence against multiple sites for oxygen sensing in organisms.
Acute responses to hypoxia Many hypoxic environments never reach the level of anoxia and most fish are able to cope with this stress using different physiological and behavioural strategies. Fish that use air breathing organs (ABO) tend to live in environments with highly variable oxygen content and rely on aerial respiration during times when there is not enough oxygen to support water-breathing. Though all teleosts have some form of swim bladder, many of them are not capable of breathing air, and they rely on aquatic surface respiration as a supply of more oxygenated water at the surface of the water. However, many species of teleost fish are obligate water breathers and do not display either of these surface respiratory behaviours.
Typically, acute hypoxia causes hyperventilation, bradycardia and an elevation in gill vascular resistance in teleosts. However, the benefit of these changes in blood pressure to oxygen uptake has not been supported in a recent study of the rainbow trout. It is possible that the acute hypoxia response is simply a stress response, and the advantages found in early studies may only result after acclimatization to the environment.
Behavioral responses Hypoxia can modify normal behavior.Reebs, S.G. (2009) Oxygen and fish behaviour Parental behaviour meant to provide oxygen to the eggs is often affected by hypoxia. For example, fanning behavior (swimming on the spot near the eggs to create a flow of water over them, and thus a constant supply of oxygen) is often increased when oxygen is less available. This has been documented in sticklebacks,Iersel, J.J.A. van. 1953. An analysis of the parental behaviour of the male three-spine stickleback (Gasterosteus aculeatus L.). Behaviour Supplement 3: 1-159.Sevenster, P. 1961. A causal analysis of a displacement activity (fanning in Gasterosteus aculeatus L.). Behaviour Supplement 9: 1-170. gobies, and clownfishes, among others. Gobies may also increase the size of the openings in the nest they build, even though this may increase the risk of predation on the eggs. Rainbow cichlids often move their young fry closer to the water surface, where oxygen is more available, during hypoxic episodes.
Behavioural adaptations meant to survive when oxygen is scarce include reduced activity levels, aquatic surface respiration, and air breathing.
Reduced activity levels
As oxygen levels decrease, fish may at first increase movements in an attempt to escape the hypoxic zone, but eventually they greatly reduce their activity levels, thus reducing their energetic (and therefore oxygen) demands. Atlantic herring show this exact pattern. Other examples of fishes that reduce their activity levels under hypoxia include the common sole, the guppy, the small-spotted catshark, and the viviparous eelpout. Some sharks that ram-ventilate their gills may understandably increase their swimming speeds under hypoxia, to bring more water to the gills.
Aquatic surface respiration In response to decreasing dissolved oxygen level in the environment, fish swim up to the surface of the water column and ventilate at the top layer of the water where it contains relatively higher level of dissolved oxygen, a behavior called aquatic surface respiration (ASR). Oxygen diffuses into water from air and therefore the top layer of water in contact with air contains more oxygen. This is true only in stagnant water; in running water all layers are mixed together and oxygen levels are the same throughout the water column. One environment where ASR often takes place is tidepools, particularly at night. Separation from the sea at low tide means that water is not renewed, fish crowding within the pool means that oxygen is quickly depleted, and absence of light at night means that there is no photosynthesis to replenish the oxygen. Examples of tidepool species that perform ASR include the tidepool sculpin, the three-spined stickleback, and the mummichog.
But ASR is not limited to the intertidal environment. Most tropical and temperate fish species living in stagnant waters engage in ASR during hypoxia. One study looked at 26 species representing eight families of non-air breathing fishes from the North American great plains, and found that all but four of them performed ASR during hypoxia. Another study looked at 24 species of tropical fish common to the pet trade, from tetras to barbs to cichlids, and found that all of them performed ASR. An unusual situation in which ASR is performed is during winter, in lakes covered by ice, at the interface between water and ice or near air bubbles trapped underneath the ice.