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Current fishing practices and habitat degradation in most of the world's oceans pose significant threats to marine fish including elasmobranchs. The accurate prediction of survival probability for elasmobranchs subjected to prolonged immobilisation and diminished oxygen availability during capture and a vulnerable state post-release, is reliant on selecting a reliable set of biomarkers to profile as well as using them to design pre-release interventions which minimise elasmobranch death. The purpose of this review is: i) to make a case for the need to develop new biomarkers to use in conjunction with blood chemistry; ii) to briefly present the survival strategies used by other vertebrates subjected to diminished oxygen iii) to discuss new approaches to forecasting the effect that altered physiological and biochemical markers have on long-term survival with a particular emphasis on oxidative stress, the adenylate energy charge, heat shock protein expression and the capacity for repair, so that a more detailed profile of the qualities of elasmobranch survivorship can be constructed. In addition, the review will discuss the relevance of biomarkers to field samples as well as their incorporation into laboratory based research, aimed at providing physiological and biochemical data to inform conservation management.

Interactions between coral reef topography, tide cycles, and photoperiod provided selection pressure for adaptive physiological changes in sheltered hypoxic niches to be exploited by specialized tropical reef fish. The epaulette shark Hemiscyllium ocellatum withstands cyclic hypoxia in its natural environment, many hours of experimental hypoxia, and anoxia for up to 5h. It shows neuronal hypometabolism in response to 5% oxygen saturation. Northern-hemisphere hypoxia- and anoxia-tolerant vertebrates that over-winter under ice alter their inhibitory to excitatory neurotransmitter balance to forestall brain ATP depletion in the absence of oxidative phosphorylation. GABA immunochemistry, HPLC analysis and receptor binding studies in H. ocellatum cerebellum revealed a heterogeneous regional accumulation of neuronal GABA despite no change in its overall concentration, and a significant increase in GABA(A) receptor density without altered binding affinity. Increased GABA(A) receptor density would protect the cerebellum during reoxygenation when transmitter release resumes. While all hypoxia- and anoxia-tolerant teleosts examined to date respond to low oxygen levels by elevating brain GABA, the phylogenetically older epaulette shark did not, suggesting that it uses an alternative neuroprotective mechanism for energy conservation. This may reflect an inherent phylogenetic difference, or represent a novel ecophysiological adaptation to cyclic variations in the availability of oxygen.

To survive anoxia, neural ATP levels have to be defended. Reducing electrical activity, which accounts for 50% or more of neural energy consumption, should be beneficial for anoxic survival. The retina is a hypoxia sensitive part of the central nervous system. Here, we quantify the in vivo retinal light response (electroretinogram; ERG) in three vertebrates that exhibit varying degrees of anoxia tolerance: freshwater turtle (Trachemys scripta), epaulette shark (Hemiscyllium ocellatum) and leopard frog (Rana pipiens). A virtually total suppression of ERG in anoxia, probably resulting in functional blindness, has previously been seen in the extremely anoxia-tolerant crucian carp (Carassius carassius). Surprisingly, the equally anoxia-tolerant turtle, which strongly depresses brain and whole-body metabolism during anoxia, exhibited a relatively modest anoxic reduction in ERG: the combined amplitude of turtle ERG waves was reduced by approximately 50% after 2 h. In contrast, the shark b-wave amplitude practically disappeared after 30 min of severe hypoxia, and the frog b-wave was decreased by approximately 75% after 40 min in anoxia. The specific A(1) adenosine receptor antagonist CPT significantly delayed the suppression of turtle ERG, while the hypoxic shark ERG was unaffected by the non-specific adenosine receptor antagonist aminophylline, suggesting adenosinergic involvement in turtle but not in shark.

The epaulette shark (Hemiscyllium ocellatum) is among the few vertebrates that can tolerate extreme hypoxia for prolonged periods and, as shown here, anoxia. We examined how anoxia affected this shark's level of responsiveness, concentration of brain ATP and adenosine -- an endogenous neuronal depressant. In addition, we investigated how these variables were affected by aminophylline, an adenosine receptor antagonist. Epaulette sharks placed in an anoxic environment (0.02 mg O2 l(-1)) lost their righting reflex after 46.3 +/- 2.8 min, but immediately regained vigilance upon return to normoxia. Then 24 h later, the same sharks were injected with either saline or aminophylline (30 mg kg(-1)) in saline and re-exposed to anoxia. In this second anoxic episode, controls sharks showed a 56% decrease in the time taken to lose their righting reflex but maintained their brain ATP levels; conversely, aminophylline-treated epaulette sharks displayed a 46% increase in the time to loss of righting reflex and had significantly lower brain ATP levels. Since anoxia also caused a 3.5-fold increase in brain adenosine levels, these results suggest that adenosine receptor activation had a pre-emptive role in maintaining brain ATP levels during anoxia. Perhaps because adenosine receptor activation initiates metabolic depression, indicated by the early loss of responsiveness (righting reflex), such a mechanism would serve to reduce ATP consumption and maintain brain ATP levels.

Reduced oxidative demand or neuronal hypometabolism is a neuroprotective strategy used by several anoxia and hypoxia-tolerant species. The epaulette shark, Hemiscyllium ocellatum inhabits shallow reef platforms that can become hypoxic. Hypoxic pre-conditioning (eight cycles of 0.34 mg O(2)/l for 120 min, 12 h apart) was used to determine whether a reduction in oxidative metabolism could be elicited in the epaulette shark brain. Hypoxic pre-conditioning resulted in a significant overall reduction in oxidative activity in coronal sections of the brainstem, but key nuclei displayed heterogeneous levels of oxidative metabolism. Motor nuclei had significantly lower levels of oxidative activity while sensory nuclei did not. The epaulette shark's ability to enter this state of hypometabolism in response to hypoxic pre-conditioning revealed a neuroprotective mechanism, which would not only reduce neuronal damage during hypoxic exposure but also minimise re-oxygenation injury.

The effect of N2 respiration on cerebral blood flow (CBF) velocity on the dorsal surface of cerebellum was examined in the estuarine crocodile, Crocodylus porosus, using epi-illumination microscopy. Twelve minutes of N2 respiration resulted in a 126% increase in CBF velocity. N2 respiration had no effect on blood pressure, indicating an underlying cerebral vasodilation. In addition, heart rate increased significantly. Systemic injections of aminophylline and the NO synthase (NOS) inhibitor nitro-L-arginine (L-NA) did not affect the hypoxia induced increase in CBF. We conclude that C. porosus responds to hypoxia with adenosine and nitric oxide (NO) independent cerebral vasodilation, and that this is likely to be a mechanism protecting the brain from energy deficiency during prolonged dives.

Alpha-Bungarotoxin (alpha BTX) administration in ovo prevents motoneuron apoptosis during development. This process may be mediated by alpha BTX-sensitive nicotinic cholinoceptors in the spinal cord, at the neuromuscular junction or at both sites. In order to differentiate between these possibilities, neuronal bungarotoxin binding (NBTX) binding to embryonic muscle and spinal cord was investigated in the chick.