A mitochondrial nexus in major depressive disorder: Integration with the psycho-immune-neuroendocrine network.

Nervous system processes, including cognition and affective state, fundamentally rely on mitochondria. Impaired mitochondrial function is evident in major depressive disorder (MDD), reflecting cumulative detrimental influences of both extrinsic and intrinsic stressors, genetic predisposition, and mutation. Glucocorticoid 'stress' pathways converge on mitochondria; oxidative and nitrosative stresses in MDD are largely mitochondrial in origin; both initiate cascades promoting mitochondrial DNA (mtDNA) damage with disruptions to mitochondrial biogenesis and tryptophan catabolism. Mitochondrial dysfunction facilitates proinflammatory dysbiosis while directly triggering immuno-inflammatory activation via released mtDNA, mitochondrial lipids and mitochondria associated membranes (MAMs), further disrupting mitochondrial function and mitochondrial quality control, promoting the accumulation of abnormal mitochondria (confirmed in autopsy studies). Established and putative mechanisms highlight a mitochondrial nexus within the psycho-immune neuroendocrine (PINE) network implicated in MDD. Whether lowering neuronal resilience and thresholds for disease, or linking mechanistic nodes within the MDD pathogenic network, impaired mitochondrial function emerges as an important risk, a functional biomarker, providing a therapeutic target in MDD. Several treatment modalities have been demonstrated to reset mitochondrial function, which could benefit those with MDD.

Using biochemical markers to assess the effects of imposed temperature stress on freshwater decapod crustaceans: Cherax quadricarinatus as a test case.

The effects of thermal stress can impact negatively on the abundance and distribution of temperature-sensitive species, particularly freshwater crustaceans. This study investigated the effects of thermal stress on physiological and biochemical parameters at five treatment temperatures resulting in minimal (25 °C), moderate (27, 29 °C) or severe (31, 33 °C) thermal stress in the common tropical freshwater crayfish Cherax quadricarinatus. The aim was to develop a suite of stress-sensitive assays to use on threatened populations of freshwater crustaceans, particularly those restricted to cooler temperatures and only found in high altitude refugia. Significant increases in indicators of oxidative and metabolic stress were observed at 29 °C and were elevated further at 33 °C. After a 50-day acclimation to an imposed temperature stress, significant changes in the level of total glutathione, total lipids, muscular protein, total haemocyte count, lipid peroxidation and protein carbonyls were observed between treatments while superoxide dismutase activity and haemolymph protein concentrations did not change. The data provided proof of concept that measuring key biochemical responses to high temperature can provide a means of contrasting the level of thermal stress experienced between individuals of the same species adapted to different temperatures. The methods developed are expected to be of use in research on wild populations of other freshwater poikilothermic organisms, particularly those susceptible to increased environmental temperatures associated with climate change.

Ecophysiology of neuronal metabolism in transiently oxygen-depleted environments: evidence that GABA is accumulated pre-synaptically in the cerebellum.

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.

The effect of exercise on innate mucosal immunity.

METHODS: The authors conducted a prospective observational study comparing salivary lactoferrin and lysozyme concentration over 5 months (chronic changes) in elite rowers (n=17, mean age 24.3+/-4.0 years) with sedentary individuals (controls) (n=18, mean age=27.2+/-7.1 years) and a graded exercise test to exhaustion (acute changes) with a cohort of elite rowers (n=11, mean age 24.7+/-4.1). RESULTS: Magnitudes of differences and changes were interpreted as a standardised (Cohen's) effect size (ES). Lactoferrin concentration in the observational study was approximately 60% lower in rowers than control subjects at baseline (7.9+/-1.2 microg/ml mean+/-SEM, 19.4+/-5.6 microg/ml, p=0.05, ES=0.68, 'moderate') and at the midpoint of the season (6.4+/-1.4 microg/ml mean +/- SEM, 21.5+/-4.2 microg/ml, p=0.001, ES=0.89, 'moderate'). The concentration of lactoferrin at the end of the study was not statistically significant (p=0.1) between the groups. There was no significant difference between rowers and control subjects in lysozyme concentration during the study. There was a 50% increase in the concentration of lactoferrin (p=0.05, ES=1.04, 'moderate') and a 55% increase in lysozyme (p=0.01, ES=3.0, 'very large') from pre-exercise to exhaustion in the graded exercise session. CONCLUSION: Lower concentrations of these proteins may be indicative of an impairment of innate protection of the upper respiratory tract. Increased salivary lactoferrin and lysozyme concentration following exhaustive exercise may be due to a transient activation response that increases protection in the immediate postexercise period.

Low mass-specific brain Na+/K+-ATPase activity in elasmobranch compared to teleost fishes: implications for the large brain size of elasmobranchs.

Elasmobranch fishes have long been noted for having unusually large brains for ectotherms, and therefore may be exceptions to the rule that vertebrates in general devote less than 8% of their resting metabolic rate to the central nervous system. The brain mass of sharks, skates and rays is often several times larger than that of teleost fishes of the same size. Still, the underlying reasons for this have remained unclear. Ion pumping by the Na+/K+-ATPase is the single most energy consuming process in the brain. In this study, Na+/K+-ATPase activity was measured in the brain of four species of elasmobranchs and 11 species of teleosts. While the average brain mass of the elasmobranchs examined was approximately three times that of the teleosts, the mean specific Na+/K+-ATPase activity was only about one-third of that of the teleosts. Thus, the total brain Na+/K+-ATPase activity was similar in elasmobranchs and teleosts. This suggests that the large brain size of elasmobranchs is at least partly related to a low mass-specific rate of brain energy use.

Neuronal oxidative hypometabolism in the brainstem of the epaulette shark (Hemiscyllium ocellatum) in response to hypoxic pre-conditioning.

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.

Increased nitric oxide synthase in the vasculature of the epaulette shark brain following hypoxia.

Epaulette sharks inhabiting reef platforms are exposed to hypoxic and hyperoxic cycles. The adaptive mechanisms used to prevent neurological damage during these cycles have not been examined. Nitric oxide has a neuroprotective role in some hypoxia-tolerant species. We examined epaulette brains following a severe experimental hypoxic regimen (0.39 mgO2/l for 2 h) and compared nitric oxide synthase (NOS) expression with that in normoxic controls using NADPH-diaphorase staining. Intense NOS activity occurred in microvasculature following exposure to a severely hypoxic environment in contrast to the low levels seen in controls. We established for the first time that the epaulette shark was hypoxia-tolerant because there was no delayed phase of neuronal apoptosis. Enhanced NOS production in response to hypoxia may cause vasodilation, which would maintain the appropriate metabolic environment for continued neuronal survival during exposure to hypoxia.

Hypoxia stimulates cerebral blood flow in the estuarine crocodile (Crocodylus porosus).

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.

Changes in the numbers of neurons and astrocytes during the postnatal development of the rat inferior olive.

In the developing nervous system, cell death is an important component of refining axonal projections. In the developing rat inferior olive, previous studies have demonstrated cell death as temporally incongruent with both initial axon-target interactions and subsequent axon collateral regression. Furthermore, these studies identified a late rise in neuron numbers that is concurrent with climbing fibre regression. As axonal regression has not previously been associated with increasing neuron numbers, and since immature neurons and glia have similar morphological characteristics, it was decided to reassess the timing of cell death within the inferior olive in animals in which neurons and glia had been differentially stained. Glia were identified by the presence of glial cytoskeletal proteins, S100, or glial fibrillary acidic protein, and stereological counts were made of both neurons and glia in the inferior olive from rats of ages 0, 5, 10, 15, and 30 days. The number of inferior olivary neurons was approximately 22,000 between birth and day 10, which decreased to about 17,500 by day 30 (P<0.05). In contrast, the number of glia rose from about 5,000 at birth to approximately 15,000 by day 10 (P<0.001), after which there was no further increase. The changes in neurons and glia caused the neuron-to-glia ratio to fall to approximately 1.5 by the time of functional maturation within the olive. These results confirm that there is neuronal death in the inferior olive but that it is temporally correlated with both climbing fibre regression and functional maturation of the olivocerebellar projection.

Brain blood flow and blood pressure during hypoxia in the epaulette shark Hemiscyllium ocellatum, a hypoxia-tolerant elasmobranch.

The key to surviving hypoxia is to protect the brain from energy depletion. The epaulette shark (Hemiscyllium ocellatum) is an elasmobranch able to resist energy depletion and to survive hypoxia. Using epi-illumination microscopy in vivo to observe cerebral blood flow velocity on the brain surface, we show that cerebral blood flow in the epaulette shark is unaffected by 2 h of severe hypoxia (0.35 mg O2 l-1 in the respiratory water, 24 C). Thus, the epaulette shark differs from other hypoxia- and anoxia-tolerant species studied: there is no adenosine-mediated increase in cerebral blood flow such as that occurring in freshwater turtles and cyprinid fish. However, blood pressure showed a 50 % decrease in the epaulette shark during hypoxia, indicating that a compensatory cerebral vasodilatation occurs to maintain cerebral blood flow. We observed an increase in cerebral blood flow velocity when superfusing the normoxic brain with adenosine (making sharks the oldest vertebrate group in which this mechanism has been found). The adenosine-induced increase in cerebral blood flow velocity was reduced by the adenosine receptor antagonist aminophylline. Aminophylline had no effect upon the maintenance of cerebral blood flow during hypoxia, however, indicating that adenosine is not involved in maintaining cerebral blood flow in the epaulette shark during hypoxic hypotension.

Localization of enkephalin immunoreactivity in the spinal cord of the long-tailed ray Himantura fai.

Enkephalin-like immunoreactivity (ENK-LI) was found throughout the spinal cord of the long-tailed ray Himantura fai. The densest ENK-LI was in the superficial portion of lamina A of the dorsal horn. Lamina B and the deeper parts of lamina A contained radially oriented, labelled fibres. Laminae C, D, and E contained many longitudinally oriented fascicles which were surrounded by a reticulum of transversely oriented, labelled fibres, some of which projected into the ventral and lateral funiculi. Labelled fibres were found in the dorsal commissure and around the central canal, but the later did not cross the midline. One-third of all enkephalinergic cells were found throughout laminae A and B, while two-thirds were located in the medial half of C, D, and E. Occasionally a labelled cell was located in the lateral funiculus. The ventral horn (laminae F and G) contained many enkephalinergic fibres but no labelled nuclei. A few dorsal column axons contained ENK-LI. In the lateral funiculus there were two groups of labelled axons, a superficial, dorsolateral group, and a deeper group, occupying a crescent-shaped region. The ventral funiculus also contained many labelled axons. The central projection of the dorsal root passed through the substantia gelatinosa and divided into rostrally and caudally projecting fascicles within lamina C. The root, and these fascicles, both lacked ENK-LI. In contrast, the fascicles in laminae D and E did contain enkephalinergic fibres. The origin of the various fibre systems and the role of enkephalin in the regulation of sensory processing and motor output are discussed.

Neuronal bungarotoxin displaces (125I) alpha-bungarotoxin binding at the neuromuscular junction as well as to the spinal cord during embryogenesis.

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.

Substance P, bombesin, and leucine-enkephalin immunoreactivities are restored in the frog tectum after optic nerve regeneration.

Extensive regeneration of the optic nerve takes place in adult Amphibia. In this study, we have determined whether one aspect of retinotectal organisation, namely immunoreactive laminae in the retinorecipient layers of the optic tectum, is restored after optic nerve regeneration. To do so, the distributions of substance-P, bombesin, and leucine-enkephalin immunoreactivities were examined in the optic tectum of the frog Litoria (Hyla) moorei. Results of a normal series were compared with those at intervals up to 84 days and at 196 days after either unilateral deafferentation or optic nerve crush. In the normal series, distinct neuropeptide immunoreactive laminae were located within the retinorecipient tectal layers. There were two major laminae with substance-P, two with bombesin, and one with leucine-enkephalin immunoreactivities. Additional faint laminae of both substance-P and bombesin immunoreactivity were present in the tectal region that receives input from the visual streak. In addition, labelling of cell bodies and dendrites was seen elsewhere in the tectum. All except one immunoreactive lamina changed after deafferentation. The deeper of those with substance-P immunoreactivity, along with both bombesin laminae, were eventually lost; the lamina with leucine-enkephalin immunoreactivity was halved in intensity. We assume that these laminae are wholely or, in the case of the leucine-enkephalin lamina, partially associated with primary optic input. By contrast, the more superficial lamina with substance-P immunoreactivity remained unchanged and is presumably not directly related to visual input. During nerve regeneration, the intensity of all laminae associated with optic input initially fell as in the deafferentation series but, in the long term, recovered to approximately 80% of normal intensities. We conclude that ganglion cells associated with each of the immunoreactivities tested had successfully regenerated. The reduced intensity of immunoreactivities after regeneration is due presumably in part to the cell loss from the ganglion cell population. Furthermore, we discuss the findings of similar studies for Rana pipiens (Kuljis and Karten [1983] J. Comp. Neurol. 217:239-251 and [1985] 240:1-15) in light of the present findings. We argue that some of the previous observations can be reinterpreted to indicate that regeneration was not limited to ganglion cells associated with substance-P immunoreactivity as first thought.

Alpha-BTX lowers neuronal metabolism during the arrest of motoneurone apoptosis.

Changes in the metabolic activity of embryonic chick spinal cords were examined following alpha-bungarotoxin (alpha-BTX) administration, in order to investigate a potential mechanism by which this toxin arrests motoneurone apoptosis during neurogenesis. Chick embryos were injected i.p. with alpha-BTX and after 25 h the metabolic markers 2-deoxyglucose and cytochrome oxidase were examined in alternate serial sections of the brachial and lumbar spinal cord. Glucose uptake and cytochrome oxidase activity were reduced throughout the spinal cord and pronounced in the lateral motor columns. Iodinated alpha-BTX reaches and binds to neuronal alpha-BTX-sensitive nicotinic cholinoceptors. Binding of alpha-BTX to these neuronal receptors and to those at the neuromuscular junction has now been shown to have a demonstrable effect on neuronal metabolism. The decreased metabolic activity in spinal cord neurones as a result of toxin treatment may have an important role in the prevention of motoneurone apoptosis at a critical developmental phase.

[125I]-alpha-bungarotoxin binding co-varies with motoneurone density during apoptosis.

During chick embryogenosis, apoptosis removes 50% of the initial population of spinal motoneurones. Administration of alpha-bungarotoxin during this critical phase arrests apoptosis. Intraperitoneally administered [125I]alpha-bungarotoxin reaches and binds to the spinal cord at the onset of motoneurone apoptosis. It is not known whether specific [125I]alpha-bungarotoxin binding is directly associated with motoneurones. This study was carried out to examine the level of toxin binding after the final number of motoneurones in the lateral motor column had been experimentally manipulated during the critical period of apoptosis. The level of specific [125I]alpha-bungarotoxin binding significantly co-varied with the number of motoneurones present, suggesting that the toxin could affect receptor-mediated aspects of motoneurone function which, in turn, could regulate final motoneurone number.