Dopamine-dependent biphasic behaviour under 'deep diving' conditions in Caenorhabditis elegans.

Underwater divers are susceptible to neurological risks due to their exposure to increased pressure. Absorption of elevated partial pressure of inert gases such as helium and nitrogen may lead to nitrogen narcosis. Although the symptoms of nitrogen narcosis are known, the molecular mechanisms underlying these symptoms have not been elucidated. Here, we examined the behaviour of the soil nematode Caenorhabditis elegans under scuba diving conditions. We analysed wild-type animals and mutants in the dopamine pathway under hyperbaric conditions, using several gas compositions and under varying pressure levels. We found that the animals changed their speed on a flat bacterial surface in response to pressure in a biphasic mode that depended on dopamine. Dopamine-deficient cat-2 mutant animals did not exhibit a biphasic response in high pressure, while the extracellular accumulation of dopamine in dat-1 mutant animals mildly influenced this response. Our data demonstrate that in C. elegans, similarly to mammalian systems, dopamine signalling is involved in the response to high pressure. This study establishes C. elegans as a powerful system to elucidate the molecular mechanisms that underly nitrogen toxicity in response to high pressure.

Power Equation for Predicting the Risk of Central Nervous System Oxygen Toxicity at Rest.

Patients undergoing hyperbaric oxygen therapy and divers engaged in underwater activity are at risk of central nervous system oxygen toxicity. An algorithm for predicting CNS oxygen toxicity in active underwater diving has been published previously, but not for humans at rest. Using a procedure similar to that employed for the derivation of our active diving algorithm, we collected data for exposures at rest, in which subjects breathed hyperbaric oxygen while immersed in thermoneutral water at 33°C (n = 219) or in dry conditions (n = 507). The maximal likelihood method was employed to solve for the parameters of the power equation. For immersion, the CNS oxygen toxicity index is K I = t2 × PO2 10.93, where the calculated risk from the Standard Normal distribution is Z I = [ln(K I 0.5) - 8.99)]/0.81. For dry exposures this is K D = t2 × PO2 12.99, with risk Z D = [ln(K D 0.5) - 11.34)]/0.65. We propose a method for interpolating the parameters at metabolic rates between 1 and 4.4 MET. The risk of CNS oxygen toxicity at rest was found to be greater during immersion than in dry conditions. We discuss the prediction properties of the new algorithm in the clinical hyperbaric environment, and suggest it may be adopted for use in planning procedures for hyperbaric oxygen therapy and for rest periods during saturation diving.

The Effect of Aging on the Ventilatory Response to Wearing a Chemical, Biological, Radiological, and Nuclear Hood Respirator at Rest and During Mild Exercise.

BACKGROUND: Structural changes in the human body resulting from aging may affect the response to altered levels of O2 and CO2. An abnormal ventilatory response to a buildup of CO2 in the inspired air due to rebreathing may result in adverse effects, which will impair the individual's ability to function under stress. The purpose of this study was to evaluate the effect of age on the respiratory response to wearing an escape hood at rest and during mild exercise. METHODS: Subjects were seven healthy, young adult males (20-30 years) and seven healthy, middle-aged males (45-65 years). Inspired CO2 and O2, breathing pattern (tidal volume [VT] and breathing frequency [F]), and mouth inspiratory and expiratory pressures, were measured at rest and during mild exercise (50 w) while wearing the CAPS 2000 escape hood (Shalon Chemical Industries and Supergum-Rubber and Plastic Technology, Tel Aviv, Israel). FINDINGS: Resting inspired CO2 was higher in the middle-aged group compared with the young group (2.25% ± 0.42% and 1.80% ± 0.34%, respectively; p < 0.05). Breathing pattern in the middle-aged group tended to be shallower and faster compared with the young group (VT: 0.69 ± 0.27 L and 0.79 ± 0.32 L, respectively; F: 14.7 ± 4.0 breaths/min and 12.4 ± 2.8 breaths/min, respectively). During exercise, there was a trend toward a high inspired CO2 in the middle-aged group compared with the young group (2.18% ± 0.40% CO2 and 1.94% ± 0.70% CO2, respectively). A correlation was found between age and inspired CO2 when wearing the escape hood (r2 = 0.375; p < 0.05). DISCUSSION: The age-related decrease in pulmonary function, together with the finding in this study of a higher inspired CO2 in middle-aged subjects wearing the CAPS 2000, may represent a greater risk for persons of middle age wearing an escape hood. RECOMMENDATIONS: On the basis of this study, it would appear reasonable to recommend that new respirators be evaluated on subjects from different age groups, to ensure the safety of both young and old.

Evidence for the infiltration of gas bubbles into the arterial circulation and neuronal injury following "yo-yo" dives in pigs.

"Yo-yo" diving may place divers at a greater risk of neurologic decompression illness (DCI). Using a rat model, we previously demonstrated that "yo-yo" diving has a protective effect against DCI. In the current study, we evaluated the risk of neurologic DCI following "yo-yo" dives in a pig model. Pigs were divided into four groups. The Control group (group A) made a square dive, without excursions to the surface ("peeps"). Group B performed two "peeps," group C performed four "peeps," and group D did not dive at all. All dives were conducted on air to 5 atm absolute, for 30-min bottom time. Echocardiography was performed to detect cardiac gas bubbles before the dive, immediately after, and at 90-min postdive. Motor performance was observed during the 5-h postdive period. Symptoms increased dramatically following a dive with four "peeps." Gas bubbles were detected in the right ventricle of all animals except for the sham group and in the left ventricle only after the four-peep dive. Neuronal cell injury was found in the spinal cord in each of the three experimental groups, tending to decrease with an increase in the number of "peeps." A four-peep "yo-yo" dive significantly increased the risk of neurologic DCI in pigs. Following a four-peep dive, we detected a higher incidence of bubbles in the left ventricle, supporting the common concern regarding an increased risk of neurologic DCI, albeit there was no direct correlation with the frequency of "red neurons" in the spinal cord.

Selective pressure modulation of synaptic voltage-dependent calcium channels-involvement in HPNS mechanism.

Exposure to hyperbaric pressure (HP) exceeding 100 msw (1.1 MPa) is known to cause a constellation of motor and cognitive impairments named high-pressure neurological syndrome (HPNS), considered to be the result of synaptic transmission alteration. Long periods of repetitive HP exposure could be an occupational risk for professional deep-sea divers. Previous studies have indicated the modulation of presynaptic Ca(2+) currents based on synaptic activity modified by HP. We have recently demonstrated that currents in genetically identified cellular voltage-dependent Ca(2+) channels (VDCCs), CaV 1.2 and CaV 3.2 are selectively affected by HP. This work further elucidates the HPNS mechanism by examining HP effect on Ca(2+) currents in neuronal VDCCs, CaV 2.2 and CaV 2.1, which are prevalent in presynaptic terminals, expressed in Xenopus oocytes. HP augmented the CaV 2.2 current amplitude, much less so in a channel variation containing an additional modulatory subunit, and had almost no effect on the CaV 2.1 currents. HP differentially affected the channels' kinetics. It is, therefore, suggested that HPNS signs and symptoms arise, at least in part, from pressure modulation of various VDCCs.

The N-methyl-D-aspartate receptor's neglected subunit - GluN1 matters under normal and hyperbaric conditions.

Professional deep-water divers exposed to hyperbaric pressure (HP) above 1.1 MPa develop high-pressure neurological syndrome, which is associated with central nervous system hyperexcitability. It was previously reported that HP augments N-methyl-D-aspartate receptor (NMDAR) synaptic responses, increases neuronal excitability, and potentially causes irreversible neuronal damage. In addition, we have reported that HP (10.1 MPa) differentially affects ionic currents, measured by the two-electrode voltage-clamp technique, of eight specific NMDAR subtypes generated by the co-expression of GluN1-1a or GluN1-1b with one of the four GluN2(A-D) subunits in Xenopus laevis oocytes. We now report that eight GluN1 splice variants, when co-expressed with GluN2A, mediate different ionic currents at normal and HP (5.1 MPa). These data, in conjunction with our previous results, indicate that both GluN1 and GluN2 subunits play a critical role in determining NMDAR currents under normal and HP conditions. These data, given the differential spatial distribution of the different NMDAR subtypes in the central nervous system, may offer a partial explanation for the mechanism governing the complex signs and symptoms of high-pressure neurological syndrome, and an explanation for the suspected long-term HP health decrement due to repetitive deep dives by professional divers.

Selective modulation of cellular voltage-dependent calcium channels by hyperbaric pressure-a suggested HPNS partial mechanism.

Professional deep sea divers experience motor and cognitive impairment, known as High Pressure Neurological Syndrome (HPNS), when exposed to pressures of 100 msw (1.1 MPa) and above, considered to be the result of synaptic transmission alteration. Previous studies have indicated modulation of presynaptic Ca(2+) currents at high pressure. We directly measured for the first time pressure effects on the currents of voltage dependent Ca(2+) channels (VDCCs) expressed in Xenopus oocytes. Pressure selectivity augmented the current in CaV1.2 and depressed it in CaV3.2 channels. Pressure application also affected the channels' kinetics, such as ƮRise, ƮDecay. Pressure modulation of VDCCs seems to play an important role in generation of HPNS signs and symptoms.

Modulation of presynaptic Ca(2+) currents in frog motor nerve terminals by high pressure.

Presynaptic Ca(2+) -dependent mechanisms have already been implicated in depression of evoked synaptic transmission by high pressure (HP). Therefore, pressure effects on terminal Ca(2+) currents were studied in Rana pipiens peripheral motor nerves. The terminal currents, evoked by nerve or direct stimulation, were recorded under the nerve perineurial sheath with a loose macropatch clamp technique. The combined use of Na(+) and K(+) channel blockers, [Ca(2+) ]o changes, voltage-dependent Ca(2+) channel (VDCC) blocker treatments and HP perturbations revealed two components of presynaptic Ca(2+) currents: an early fast Ca(2+) current (ICaF ), possibly carried by N-type (CaV 2.2) Ca(2+) channels, and a late slow Ca(2+) current (ICaS ), possibly mediated by L-type (CaV 1) Ca(2+) channels. HP reduced the amplitude and decreased the maximum (saturation level) of the Ca(2+) currents, ICaF being more sensitive to pressure, and may have slightly shifted the voltage dependence. HP also moderately diminished the Na(+) action current, which contributed to the depression of VDCC currents. Computer-based modeling was used to verify the interpretation of the currents and investigate the influence of HP on the presynaptic currents. The direct HP reduction of the VDCC currents and the indirect effect of the action potential decrease are probably the major cause of pressure depression of synaptic release.

Hyperbaric pressure effects on voltage-dependent Ca+2 channels: relevance to HPNS.

Known and unpublished data regarding hyperbaric pressure (HP) effects on voltage dependent-Ca2+ channels (VDCCs) were reviewed in an attempt to elucidate their role in the development of high-pressure neurological syndrome (HPNS). Most postulated effects from studies performed in the last two decades (e.g., depressed maximal current) rely on indirect findings, derived from extracellular [Ca2+] manipulation or by observing Ca(2+)-dependent processes. More recent experiments have tried to directly measure Ca2+ currents under high pressure conditions, some of which are potentially challenging previous indirect findings on one hand, but support findings from work done on neuronal behavior on the other. Additional support for some of the recent findings is provided by computer simulation of pressure effects on a spinal motor neuron activity. HP effect on different types of VDCCs seems to be selective - i.e., HP may suppress, facilitate or not change their activity. Thus, the specific distribution of the various types of the channels in each synaptic terminal or throughout the neuron will determine their function and will influence the neuronal network behavior under HP. Further research is needed in order to fully understand the HPNS etiology.