Mihai Moldovan

Intrinsic properties of lumbar motor neurones in the adult G127insTGGG superoxide dismutase-1 mutant mouse in vivo: evidence for increased persistent inward currents

Aim:  Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disease characterized by a preferential loss of motor neurones. Previous publications using in vitro neonatal preparations suggest an increased excitability of motor neurones in various superoxide dismutase‐1 (SOD1) mutant mice models of ALS which may contribute to excitotoxicity of the motor neurones.

Mechanisms of hyperpolarization in regenerated mature motor axons in cat

We found persistent abnormalities in the recovery of membrane excitability in long‐term regenerated motor nerve fibres in the cat as indicated in the companion paper. These abnormalities could partly be explained by membrane hyperpolarization. To further investigate this possibility, we compared the changes in excitability in control nerves and long‐term regenerated cat nerves (3–5 years after tibial nerve crush) during manoeuvres known to alter axonal membrane Na+–K+ pump function: polarization, cooling to 20°C, reperfusion after 10 min ischaemia, and up to 60 s of repetitive stimulation at 200 Hz. The abnormalities in excitability of regenerated nerves were reduced by depolarization and cooling and increased by hyperpolarization and during postischaemia. Moreover, the time course of recovery of excitability from repetitive stimulation and ischaemia was prolonged in regenerated nerves. Our data are consistent with an increased demand for electrogenic Na+–K+ pumping in regenerated nerves leading to membrane hyperpolarization. Such persistent hyperpolarization may influence the ability of the axon to compensate for changes in membrane potential following normal repetitive activity.

Collagen Conduit Versus Microsurgical Neurorrhaphy: 2-Year Follow-Up of a Prospective, Blinded Clinical and Electrophysiological Multicenter Randomized, Controlled Trial

PURPOSE To compare repair of acute lacerations of mixed sensory-motor nerves in humans using a collagen tube versus conventional repair. METHODS In a prospective randomized trial, we repaired the ulnar or the median nerve with a collagen nerve conduit or with conventional microsurgical techniques. We enrolled 43 patients with 44 nerve lacerations. We performed electrophysiological tests and hand function using a standardized clinical evaluation instrument, the Rosen scoring system, after 12 and 24 months. RESULTS Operation time using the collagen conduit was significantly shorter than for conventional neurorrhaphy. There were no complications in terms of infection, extrusion of the conduit, or other local adverse reaction. Thirty-one patients with 32 nerve lesions, repaired with collagen conduits or direct suture, attended the 24-month follow-up. There was no difference between sensory function, discomfort, or total Rosen scores. Motor scores were significantly better for the direct suture group after 12 months, but after 24 months, there were no differences between the treatment groups. There was a general further recovery of both motor and sensory conduction parameters at 24 months compared with 12 months. There were no statistically significant differences in amplitudes, latencies, or conduction velocities between the groups. CONCLUSIONS Use of a collagen conduit produced recovery of sensory and motor functions that were equivalent to direct suture 24 months after repair when the nerve gap inside the tube was 6 mm or less, and the collagen conduit proved to be safe for these nerve lacerations in the forearm. TYPE OF STUDY/LEVEL OF EVIDENCE Therapeutic II.

Acute energy restriction triggers Wallerian degeneration in mouse

Acute exposure of peripheral axons to the free radical Nitric Oxide (NO) may trigger conduction block and, if prolonged, Wallerian degeneration. It was hypothesized that this neurotoxic effect of NO may be due primarily to energy restriction by inhibition of mitochondrial respiration. We compared the neurotoxic effect of NO with the effect of the mitochondrial uncoupler 2,4-dinitrophenol (DNP) on electrically active axons of mouse sciatic nerve. The right tibial nerve was stimulated at the ankle. Muscle responses were recorded from plantar muscles and ascending nerve action potentials were recorded form the exposed sciatic nerve by means of a hook electrode. The sciatic nerve was focally immersed over a length of 1 cm in either phosphate buffered saline (PBS), a solution of approximately 4 microM NO obtained from 10 mM of the NO-donor DETA NONOate, or a solution of up to 1 mM DNP. Following 3 hours of 200 Hz stimulation, the nerves were washed in PBS for 1 hour, the surgical wounds were closed and the mice were left to recover. Following repetitive stimulation in PBS, the nerve responses recovered within 1 hour and the muscle responses within 1 day. The effects of focal acute exposure to NO or DNP were similar: (i) a transient conduction failure that rapidly normalized within one hour of washout and (ii) subsequent Wallerian degeneration of some axons confirmed at morphological studies. Taken together, these data support the hypothesis that neurotoxicity may be caused by energy restriction. Since the pharmacologic effect of NO and DNP was only transient, our data suggest that even a brief period of focal energy restriction can trigger Wallerian degeneration.

Persistent abnormalities of membrane excitability in regenerated mature motor axons in cat

The purpose of our study was to assess by threshold tracking internodal and nodal membrane excitability during the maturation process after tibial nerve crush in cat. Various excitability indices (EI) were computed non‐invasively by comparing the threshold of a submaximal compound motor potential at different stimulation durations (strength–duration relationship), after a conditioning nerve impulse (recovery of excitability), or during the application of a polarizing current (threshold electrotonus). Four months after the lesion, regenerated nerves showed a higher rheobase, shorter chronaxie, shorter refractory period and higher than normal threshold variations during threshold electrotonus (TE). A partial recovery was observed during the first 2 years of maturation. The recovery to depolarizing TE seemed complete but all other EI remained abnormal even after 5 years of regeneration, the most pronounced being the 157 ± 8% (mean ±S.E.M.) increase in threshold during hyperpolarizing TE compared with 94 ± 4% in controls. These EI abnormalities are consistent with increased input impedance. Nevertheless, the time course of maturation and incomplete recovery of EI could only be partially explained by changes in fibre morphology. The highly abnormal response to hyperpolarizing but not to depolarizing TE suggests that voltage‐dependent membrane function also remained abnormal, possibly due to membrane hyperpolarization.

Evaluation of Na+/K+ pump function following repetitive activity in mouse peripheral nerve

After conduction of prolonged trains of impulses the increased Na+/K+ pump activity leads to hyperpolarization. The aim of this study was to develop a mouse model to investigate the Na+/K+ pump function in peripheral nerve by measuring the decrease in excitability during activity-dependent hyperpolarization. Acute electrophysiological investigations were carried out in seven adult mice. Nerve excitability was evaluated by tracking the change in threshold current after 5 min of 100 Hz stimulation of the tibial nerve at ankle. We developed a threshold tracking system that allowed us to follow several excitability measures simultaneously from the evoked plantar compound muscle action potential (CMAP) and sciatic compound nerve action potential (CNAP). Three minutes after repetitive supramaximal stimulation maximal CMAP and CNAP amplitudes recovered but the threshold was increased approximately 40% for motor axons approximately 34% for axons generating CNAP. The threshold recovered with a rate of 3.8%/minute that was similar for nerve and motor responses. By tracking the effect of polarizing currents we found evidence of activity dependent hyperpolarization, and our data suggest that the observed threshold change after repetitive stimulation of the mouse tibial nerve is an indicator of the Na+/K+ pump function in vivo. Evaluation of activity-dependent hyperpolarization may be an important indicator of axonal ability to cope with Na+ load.

Nerve conduction and excitability studies in peripheral nerve disorders.

Purpose of reviewThe review is aimed at providing information about the role of nerve excitability studies in peripheral nerve disorders. It has been known for many years that the insight into peripheral nerve pathophysiology provided by conventional nerve conduction studies is limited. Nerve excitability studies are relatively novel but are acquiring an increasingly important role in the study of peripheral nerves. Recent findingsBy measuring responses in nerve that are related to nodal function (strength–duration time constant, rheobase and recovery cycle) and internodal function (threshold electrotonus, current–threshold (I/V) relationship) it is possible to assess the function of transient and persistent Na+, fast and slow K+ and HCN inward rectifying channels as well as ion pumps. This has allowed insight into normal axon physiology and normal fluctuations of electrolyte concentrations. Studies of different metabolic neuropathies have assessed the influence of uremia, diabetes and ischemia, and the use of these methods in toxic neuropathies has allowed pinpointing damaging factors. Various mutations in ion channels associated with central nervous system disorders have been shown to have counterparts in the peripheral nervous system, in some instances without peripheral nervous system symptoms. Both hereditary and acquired demyelinating neuropathies have been studied and the effects on nerve pathophysiology have been compared with degeneration and regeneration of axons. SummaryExcitability testing holds promise for further understanding of peripheral nerve pathophysiology but is as yet not universally available. Interpretation may be challenging as changes in parameters may have different explanations, and modeling has been helpful in the use of the methods in clinical neurophysiology.

Coffee drinking enhances the analgesic effect of cigarette smoking.

Nicotine (from cigarette smoke) and caffeine (from coffee) have analgesic effects in humans and experimental animals. We investigated the combined effects of coffee drinking and cigarette smoking on pain experience in a group of moderate nicotine-dependent, coffee drinking, young smokers. Pain threshold and pain tolerance were measured during cold pressor test following the habitual nocturnal deprivation of smoking and coffee drinking. Smoking increased pain threshold and pain tolerance in both men and women. Coffee drinking, at a dose that had no independent effect, doubled the increase in pain threshold induced by smoking. The effect could not be explained by a cumulative raise in blood pressure. Our data suggest that caffeine enhances the analgesic effect of nicotine.

Axonal voltage-gated ion channels as pharmacological targets for pain.

Upon peripheral nerve injury (caused by trauma or disease process) axons of the dorsal root ganglion (DRG) somatosensory neurons have the ability to sprout and regrow/remyelinate to reinnervate distant target tissue or form a tangled scar mass called a neuroma. This regenerative response can become maladaptive leading to a persistent and debilitating pain state referred to as chronic pain corresponding to the clinical description of neuropathic/chronic inflammatory pain. There is little agreement to what causes peripheral chronic pain other than hyperactivity of the nociceptive DRG neurons which ultimately depends on the function of voltage-gated ion channels. This review focuses on the pharmacological modulators of voltage-gated ion channels known to be present on axonal membrane which represents by far the largest surface of DRG neurons. Blockers of voltage-gated Na(+) channels, openers of voltage-gated K(+) channels and blockers of hyperpolarization-activated cyclic nucleotide-gated channels that were found to reduce neuronal activity were also found to be effective in neuropathic and inflammatory pain states. The isoforms of these channels present on nociceptive axons have limited specificity. The rationale for considering axonal voltage-gated ion channels as targets for pain treatment comes from the accumulating evidence that chronic pain states are associated with a dysregulation of these channels that could alter their specificity and make them more susceptible to pharmacological modulation. This drives the need for further development of subtype-specific voltage-gated ion channels modulators, as well as clinically available neurophysiological techniques for monitoring axonal ion channel function in peripheral nerves.

Axonal elongation through long acellular nerve segments depends on recruitment of phagocytic cells from the near-nerve environment: Electrophysiological and morphological studies in the cat

The distal nerve stump plays a central role in the regeneration of peripheral nerve but the relative importance of cellular and humoral factors is not clear. We have studied this question by freezing the tibial nerve distal to a crush lesion in cat. The importance of constituents from the near-nerve environment was assessed by modification of the contact between the tibial nerve and the environment. Silicone cuffs, containing electrodes for electrophysiological assessment of nerve regeneration, were placed around the tibial nerve distal to the crush site. The interaction between long acellular frozen nerve segments (ANS) and the near-nerve environment was ascertained by breaching the silicone cuff to allow access of cellular or humoral components. Tibial nerves were crushed and frozen for 40 mm and enclosed in nerve cuffs with 0.45-microm holes or 2.0-mm holes to allow access of humoral factors or tissue ingrowth, respectively. In a second set of experiments, tibial nerves were crushed and either frozen for 20+20 mm, leaving a 10 mm segment with viable cells in the center (stepping-stone segment) or frozen for 50 mm. These nerves were enclosed in cuffs with 2.0 mm holes corresponding to the viable nerve segment. The regeneration was monitored electrophysiologically by implanted electrodes and after 2 months the nerves were investigated by light and electron microscopy. The results indicate that soluble substances in the near-nerve environment, such as nutrients, oxygen or tropic substances did not exert any independent beneficial effect on the outgrowing axons. However, phagocytic cells entering the acellular segment from the near-nerve environment were crucial for axonal outgrowth in long ANS.