Computational modeling of the effects of auditory nerve dysmyelination

Our previous study showed that exposure to loud sound leading to hearing loss elongated the auditory nerve (AN) nodes of Ranvier and triggered notable morphological changes at paranodes and juxtaparanodes. Here we used computational modeling to examine how theoretical redistribution of voltage gated Na+, Kv3.1, and Kv1.1 channels along the AN may be responsible for the alterations of conduction property following acoustic over-exposure. Our modeling study infers that changes related to Na+ channel density (rather than the redistribution of voltage gated Na+, Kv3.1, and Kv1.1 channels) is the likely cause of the decreased conduction velocity and the conduction block observed after acoustic overexposure (AOE)

A step-by-step guide to non-linear regression analysis of experimental data using a Microsoft Excel spreadsheet,

Abstract The objective of this present study was to introduce a simple, easily understood method for carrying out non-linear regression analysis based on user input functions. While it is relatively straightforward to fit data with simple functions such as linear or logarithmic functions, fitting data with more complicated non-linear functions is more difficult. Commercial specialist programmes are available that will carry out this analysis, but these programmes are expensive and are not intuitive to learn. An alternative method described here is to use the SOLVER function of the ubiquitous spreadsheet programme Microsoft Excel, which employs an iterative least squares fitting routine to produce the optimal goodness of fit between data and function. The intent of this paper is to lead the reader through an easily understood step-by-step guide to implementing this method, which can be applied to any function in the form y=f(x), and is well suited to fast, reliable analysis of data in all fields of biology

Contribution of glycogen in supporting axon conduction in the peripheral and central nervous systems: the role of lactate

The role of glycogen in the central nervous system is intimately linked with the glycolytic pathway. Glycogen is synthesized from glucose, the primary substrate for glycolysis, and degraded to glucose-6-phosphate. The metabolic cost of shunting glucose via glycogen exceeds that of simple phosphorylation of glucose to glucose-6-phosphate by hexokinase; thus, there must be a metabolic advantage in utilizing this shunt pathway. The dogmatic view of glycogen as a storage depot persists, based on initial descriptions of glycogen supporting neural function in the face of aglycemia. The variable latency to conduction failure, dependent upon tissue glycogen levels, provided convincing evidence of the role played by glycogen in supporting neural function. Glycogen is located predominantly in astrocytes in the central nervous system, thus for glycogen to benefit neural elements, intercellular metabolic communication must exist in the form of astrocyte to neuron substrate transfer. Experimental evidence supports a model where glycogen is metabolized to lactate in astrocytes, with cellular expression of monocarboxylate transporters and enzymes appropriately located for lactate shuttling between astrocytes and neural elements, where lactate acts as a substrate for oxidative metabolism. Biosensor recordings have demonstrated a significant steady concentration of lactate present on the periphery of both central white matter and peripheral nerve under unstimulated baseline conditions, indicating continuous cellular efflux of lactate to the interstitium. The existence of this lactate pool argues we must reexamine the “on demand” shuttling of lactate between cellular elements, and suggests continuous lactate efflux surplus to immediate neural requirements

Revision workshops in elementary mathematics enhance student performance in routine laboratory calculations

The ability to understand and implement calculations required for molarity and dilution computations that are routinely undertaken in the laboratory are essential skills that should be possessed by all students entering an undergraduate Life Sciences degree. However, it is increasingly recognized that the majority of these students are ill equipped to reliably carry out such calculations. There are several factors that conspire against students' understanding of this topic, with the alien concept of the mole in relation to the mass of compounds and the engineering notation required when expressing the relatively small quantities typically involved being two key examples. In this report, we highlight teaching methods delivered via revision workshops to undergraduate Life Sciences students at the University of Nottingham. Workshops were designed to 1) expose student deficiencies in basic numeracy skills and remedy these deficiencies, 2) introduce molarity and dilution calculations and illustrate their workings in a step-by-step manner, and 3) allow students to appreciate the magnitude of numbers. Preworkshop to postworkshop comparisons demonstrated a considerable improvement in students' performance, which attenuated with time. The findings of our study suggest that an ability to carry out laboratory calculations cannot be assumed in students entering Life Sciences degrees in the United Kingdom but that explicit instruction in the form of workshops improves proficiency to a level of competence that allows students to prosper in the laboratory environment

Acute effect of pore-forming Clostridium perfringens ε-toxin on compound action potentials of optic nerve of mouse

ε-Toxin is a pore forming toxin produced by Clostridium perfringens types B and D. It is synthesized as a less active prototoxin form that becomes fully active upon proteolytic activation. The toxin produces highly lethal enterotoxaemia in ruminants, has the ability to cross the blood–brain barrier (BBB) and specifically binds to myelinated fibers. We discovered that the toxin induced a release of ATP from isolated mice optic nerves, which are composed of myelinated fibers that are extended from the central nervous system. We also investigated the effect of the toxin on compound action potentials (CAPs) in isolated mice optic nerves. When nerves were stimulated at 100 Hz during 200 ms, the decrease of the amplitude and the area of the CAPs was attenuated in the presence of ε-toxin. The computational modelling of myelinated fibers of mouse optic nerve revealed that the experimental results can be mimicked by an increase of the conductance of myelin and agrees with the pore forming activity of the toxin which binds to myelin and could drill it by making pores. The intimate ultrastructure of myelin was not modified during the periods of time investigated. In summary, the acute action of the toxin produces a subtle functional impact on the propagation of the nerve action potential in myelinated fibers of the central nervous system with an eventual desynchronization of the information. These results may agree with the hypothesis that the toxin could be an environmental trigger of multiple sclerosis (MS)

Demyelination and axonal preservation in a transgenic mouse model of Pelizaeus-Merzbacher disease

It is widely thought that demyelination contributes to the degeneration of axons and, in combination with acute inflammatory injury, is responsible for progressive axonal loss and persistent clinical disability in inflammatory demyelinating disease. In this study we sought to characterize the relationship between demyelination, inflammation and axonal transport changes using a Plp1-transgenic mouse model of Pelizaeus-Merzbacher disease. In the optic pathway of this non-immune mediated model of demyelination, myelin loss progresses from the optic nerve head towards the brain, over a period of months. Axonal transport is functionally perturbed at sites associated with local inflammation and 'damaged' myelin. Surprisingly, where demyelination is complete, naked axons appear well preserved despite a significant reduction of axonal transport. Our results suggest that neuroinflammation and/or oligodendrocyte dysfunction are more deleterious for axonal health than demyelination per se, at least in the short ter

A method for reducing animal use whilst maintaining statistical power in electrophysiological recordings from rodent nerves

The stimulus evoked compound action potential, recorded from ex vivo nerve trunks such as the rodent optic and sciatic nerve, is a popular model system used to study aspects of nervous system metabolism. This includes (1) the role of glycogen in supporting axon conduction, (2) the injury mechanisms resulting from metabolic insults, and (3) to test putative benefits of clinically relevant neuroprotective strategies. We demonstrate the benefit of simultaneously recording from pairs of nerves in the same superfusion chamber compared with conventional recordings from single nerves. Experiments carried out on mouse optic and sciatic nerves demonstrate that our new recording configuration decreased the relative standard deviation from samples when compared with recordings from an equivalent number of individually recorded nerves. The new method reduces the number of animals required to produce equivalent Power compared with the existing method, where single nerves are used. Adopting this method leads to increased experimental efficiency and productivity. We demonstrate that reduced animal use and increased Power can be achieved by recording from pairs of rodent nerve trunks simultaneously

Energy Metabolism in Mouse Sciatic Nerve A Fibres during Increased Energy Demand

The ability of sciatic nerve A fibres to conduct action potentials relies on an adequate supply of energy substrate, usually glucose, to maintain necessary ion gradients. Under our ex vivo experimental conditions, the absence of exogenously applied glucose triggers Schwann cell glycogen metabolism to lactate, which is transported to axons to fuel metabolism, with loss of the compound action potential (CAP) signalling glycogen exhaustion. The CAP failure is accelerated if tissue energy demand is increased by high-frequency stimulation (HFS) or by blocking lactate uptake into axons using cinnemate (CIN). Imposing HFS caused CAP failure in nerves perfused with 10 mM glucose, but increasing glucose to 30 mM fully supported the CAP and promoted glycogen storage. A combination of glucose and lactate supported the CAP more fully than either substrate alone, indicating the nerve is capable of simultaneously metabolising each substrate. CAP loss resulting from exposure to glucose-free artificial cerebrospinal fluid (aCSF) could be fully reversed in the absence of glycogen by addition of glucose or lactate when minimally stimulated, but imposing HFS resulted in only partial CAP recovery. The delayed onset of CAP recovery coincided with the release of lactate by Schwann cells, suggesting that functional Schwann cells are a prerequisite for CAP recovery

Novel hypoglycemic injury mechanism: N-methyl-D-aspartate receptor-mediated white matter damage

Objective: Hypoglycemia is a common adverse event and can injure central nervous system (CNS) white matter (WM). We determined if glutamate receptors were involved in hypoglycemic WM injury. Methods: Mouse optic nerves (MON), CNS WM tracts, were maintained at 37°C with oxygenated artificial cerebrospinal fluid (ACSF) containing 10 mM glucose. Aglycemia was produced by switching to 0 glucose ACSF. Supra-maximal compound action potentials (CAPs) were elicited using suction electrodes and axon function was quantified as the area under the CAP. Amino acid release was measured using HPLC. Extracellular [lactate] was measured using an enzyme electrode. Results: About 50% of MON axons were injured after 60 min of aglycemia (90% after 90 min); injury was not affected by animal age. Blockade of NMDA-type glutamate receptors improved recovery after 90 min of aglycemia by 250%. Aglycemic injury was increased by reducing [Mg2+]o or increasing [glycine]o, and decreased by lowering pHo, expected results for NMDA receptor-mediated injury. Extracellular pH increased during aglycemia, due to a drop in [lactate-]o. Aglycemic injury was dramatically reduced in the absence of [Ca2+]o. Extracellular aspartate, a selective NMDA receptor agonist, increased during aglycemia. Interpretation: Aglycemia injured WM by a unique excitotoxic mechanism involving NMDA receptors (located primarily on oligodendrocytes). During WM aglycemia, the selective NMDA agonist, aspartate, is released, probably from astrocytes. Injury is mediated by Ca2+ influx through aspartate-activated NMDA receptors made permeable by an accompanying alkaline shift in pHo caused by a fall in [lactate-]o. These insights have important clinical implications

Moderators of changes in smoking, drinking and quitting behaviour associated with the first COVID‐19 lockdown in England

Aim To estimate changes in smoking, drinking and quitting behaviour from before to during the first COVID-19 lockdown in England, and whether changes differed by age, sex or social grade. Design Representative cross-sectional surveys of adults, collected monthly between August 2018 and July 2020. Setting England. Participants A total of 36 980 adults (≥ 18 years). Measurements Independent variables were survey month (pre-lockdown: August–February versus lockdown months: April–July) and year (pandemic: 2019/20 versus comparator: 2018/19). Smoking outcomes were smoking prevalence, cessation, quit attempts, quit success and use of evidence-based or remote cessation support. Drinking outcomes were high-risk drinking prevalence, alcohol reduction attempts and use of evidence-based or remote support. Moderators were age, sex and occupational social grade (ABC1 = more advantaged/C2DE = less advantaged). Findings Relative to changes during the same time period in 2018/19, lockdown was associated with significant increases in smoking prevalence [+24.7% in 2019/20 versus 0.0% in 2018/19, adjusted odds ratio (aOR) = 1.35, 95% confidence interval (CI) = 1.12–1.63] and quit attempts (+39.9 versus –22.2%, aOR = 2.48, 95% CI = 1.76–3.50) among 18–34-year-olds, but not older groups. Increases in cessation (+156.4 versus –12.5%, aOR = 3.08, 95% CI = 1.86–5.09) and the success rate of quit attempts (+99.2 versus +0.8%, aOR = 2.29, 95% CI = 1.31–3.98) were also observed, and did not differ significantly by age, sex or social grade. Lockdown was associated with a significant increase in high-risk drinking prevalence among all socio-demographic groups (+39.5 versus –7.8%, aOR = 1.80, 95% CI = 1.64–1.98), with particularly high increases among women (aOR = 2.17, 95% CI = 1.87–2.53) and social grades C2DE (aOR = 2.34, 95% CI = 2.00–2.74). Alcohol reduction attempts increased significantly among high-risk drinkers from social grades ABC1 (aOR = 2.31, 95% CI = 1.78–3.00) but not C2DE (aOR = 1.25, 95% CI = 0.83–1.88). There were few significant changes in use of support for smoking cessation or alcohol reduction, although samples were small. Conclusions In England, the first COVID-19 lockdown was associated with increased smoking prevalence among younger adults and increased high-risk drinking prevalence among all adults. Smoking cessation activity also increased: more younger smokers made quit attempts during lockdown and more smokers quit successfully. Socio-economic disparities in drinking behaviour were evident: high-risk drinking increased by more among women and those from less advantaged social grades (C2DE), but the rate of reduction attempts increased only among the more advantaged social grades (ABC1)