Two-photon laser-scanning microscopy for single and repetitive imaging of dorsal and lateral spinal white matter in vivo.

We developed appropriate surgical procedures for single and repetitive multi-photon imaging of spinal cord in vivo. By intravenous anesthesia, artificial ventilation and laminectomy, acute experiments were performed in the dorsal and lateral white matter. By volatile anesthesia and minimal-invasive surgery, chronic repetitive imaging up to 8 months were performed in the dorsal column through the window between two adjacent spines.

In vivo recording of nerve conduction velocity of spinal CNS fibers in the mouse.

Anesthetic and surgical procedures and an electrophysiological method were developed for recording nerve conduction velocity (NCV) of CNS fibers in the murine spinal cord. Under intravenous anesthesia and artificial ventilation the lumbar spinal cord segments L1 to L4 and dorsal roots L3 to L5 on the left side were exposed by laminectomy. After stimulation of the dorsal root L4, a compound action potential (CAP) was recorded at the ipsilateral left fasciculus gracilis at the spinal cord level L1.

Oligodendroglial NMDA Receptors Regulate Glucose Import and Axonal Energy Metabolism.

Oligodendrocytes make myelin and support axons metabolically with lactate. However, it is unknown how glucose utilization and glycolysis are adapted to the different axonal energy demands. Spiking axons release glutamate and oligodendrocytes express NMDA receptors of unknown function.

Contractile characteristics of gastrocnemius-soleus muscle in the SOD1G93A ALS mouse model.

Objectives: In the SOD1G93A mouse model of amyotrophic lateral sclerosis (ALS), a selective degeneration of fast-fatigable motor units and consequently an early decline of contractile force in individual fast-twitch muscles have been observed in the preclinical stage. However, most human muscles include fast and slow motor units. Gastrocnemius-soleus group (GS) contains such a mixture of units.

What does gamma coherence tell us about inter-regional neural communication?

Neural oscillations have been measured and interpreted in multitudinous ways, with a variety of hypothesized functions in physiology, information processing and cognition. Much attention has been paid in recent years to gamma-band (30-100 Hz) oscillations and synchrony, with an increasing interest in 'high gamma' (>100 Hz) signals as mesoscopic measures of inter-regional communication. The biophysical origins of the measured variables are often difficult to precisely identify, however, making their interpretation fraught with pitfalls.

Long lasting activity of nociceptive muscular afferents facilitates bilateral flexion reflex pattern in the feline spinal cord.

Chronic muscular limb pain requires the adoption of motor patterns distinct from the classic ipsilateral flexion, crossed extension and corresponding reciprocal inhibitions to acute exteroceptive stimulation. Using selective chemical activation of group III/IV afferents in gastrocnemius-soleus (GS) muscles we investigated bilaterally their reflex responses conditioned by (a) acute 'myositis' induced by intramuscular carrageenan; and (b) sub-acute 'myositis' induced by infusion of complete Freund's adjuvant (CFA). Reflex transmission was detected by monosynaptic testing and c-fos staining used to identify increased neuronal activity.

Theta phase segregation of input-specific gamma patterns in entorhinal-hippocampal networks.

Precisely how rhythms support neuronal communication remains obscure. We investigated interregional coordination of gamma oscillations using high-density electrophysiological recordings in the rat hippocampus and entorhinal cortex. We found that 30-80 Hz gamma dominated CA1 local field potentials (LFPs) on the descending phase of CA1 theta waves during navigation, with 60-120 Hz gamma at the theta peak.

Local generation and propagation of ripples along the septotemporal axis of the hippocampus.

A topographical relationship exists between the septotemporal segments of the hippocampus and their entorhinal-neocortical targets, but the physiological organization of activity along the septotemporal axis is poorly understood. We recorded sharp-wave ripple patterns in rats during sleep from the entire septotemporal axis of the CA1 pyramidal layer. Qualitatively similar ripples emerged at all levels.

Reflex transmission to lumbar α-motoneurones in the mouse similar and different to those in the cat.

Investigation and interpretation of defective motor circuitries in transgenic mice required further basic results from wild-type mice. Therefore, we investigated the lumbar motor reflex pattern in anaesthetised mice using intracellular motoneuronal recording and monosynaptic reflex testing. Thresholds and latencies in mice were similar to those in cats: thresholds for monosynaptic (group I) EPSPs were slightly above 1T (T=threshold for the lowest threshold fibres), around 1.

Influence of methylene blue on microglia-induced inflammation and motor neuron degeneration in the SOD1(G93A) model for ALS.

Mutations in SOD1 cause hereditary variants of the fatal motor neuron disease amyotrophic lateral sclerosis (ALS). Pathophysiology of the disease is non-cell-autonomous, with toxicity deriving also from glia. In particular, microglia contribute to disease progression.

The spiking component of oscillatory extracellular potentials in the rat hippocampus.

When monitoring neural activity using intracranial electrical recordings, researchers typically consider the signals to have two primary components: fast action potentials (APs) from neurons near the electrode, and the slower local field potential (LFP), thought to be dominated by postsynaptic currents integrated over a larger volume of tissue. In general, a decrease in signal power with increasing frequency is observed for most brain rhythms. The 100-200 Hz oscillations in the rat hippocampus, including "fast gamma" or "epsilon" oscillations and sharp wave-ripples (SPW-Rs), are one exception, showing an increase in power with frequency within this band.

In vivo measurement of conduction velocities in afferent and efferent nerve fibre groups in mice.

Electrophysiological investigations in mice, particularly with altered myelination, require reference data of the nerve conduction velocity (CV). CVs of different fibre groups were determined in the hindlimb of anaesthetized adult mice. Differentiation between afferent and efferent fibres was performed by recording at dorsal roots and stimulating at ventral roots, respectively.

Major contribution of Aδ-fibres to increased reflex transmission in the feline spinal cord during acute muscle inflammation.

The contribution of activated nociceptive muscle afferents to pathologically increased muscle tone remains obscure. The aim of the present study was to investigate whether an acute myositis of the gastrocnemius-soleus (GS) influences spinal reflex activity and to test whether Aδ-fibre or C-fibre were mainly responsible for any effects. In high spinal cats monosynaptic reflexes (MRs) of flexors and extensors and transmission in reflex pathways from group III and IV muscle afferents (activated by intra-arterial KCl injection) were investigated.

Role of L-DOPA in spinal nociceptive reflex activity: higher sensitivity of Aδ versus C fibre-evoked nociceptive reflexes to L-DOPA.

The role of L-DOPA in spinal nociceptive reflex activity has been re-evaluated. In high spinal cats, with supraspinal loops being excluded, the onset of reflex facilitation induced by noxious radiant heat is delayed after injection of L-DOPA by 4 to 10 s, i.e.

In vivo imaging reveals rapid morphological reactions of astrocytes towards focal lesions in an ALS mouse model.

Pathophysiology of the motoneuron disease amyotrophic lateral sclerosis (ALS) is non-cell-autonomous. In mouse models of familiar ALS, neurotoxicity is derived not only from mutant motor neurons but also from mutant neighbouring glial cells. In vivo imaging by two-photon laser-scanning microscopy was used to study rapid morphological reactions of astroglial cells towards laser-induced axonal transection in ALS-linked transgenic SOD1(G93A) mice.

In Vivo imaging reveals distinct inflammatory activity of CNS microglia versus PNS macrophages in a mouse model for ALS.

Mutations in the enzyme superoxide dismutase-1 (SOD1) cause hereditary variants of the fatal motor neuronal disease Amyotrophic lateral sclerosis (ALS). Pathophysiology of the disease is non-cell-autonomous: neurotoxicity is derived not only from mutant motor neurons but also from mutant neighbouring non-neuronal cells. In vivo imaging by two-photon laser-scanning microscopy was used to compare the role of microglia/macrophage-related neuroinflammation in the CNS and PNS using ALS-linked transgenic SOD1(G93A) mice.

Spinal motor actions of the μ-opioid receptor agonist DAMGO in the cat.

For further evaluation of opioidergic spinal motor functions the action of the μ-opioid receptor agonist DAMGO was tested on transmission in different non-nociceptive and nociceptive spinal reflex pathways from flexor reflex afferents (FRA), and in non-FRA reflex pathways in spinal cats. The action of DAMGO was complex, not following a simple pattern with selective depression of nociceptive pathways compared to non-nociceptive ones. Monosynaptic reflexes of the flexor posterior biceps semitendinosus (PBSt) and transmission in nociceptive as well as non-nociceptive excitatory FRA pathways to PBSt were depressed, while the specific excitatory nociceptive non-FRA pathway from the central foot pad to foot extensors was mainly not depressed but rather facilitated by DAMGO.

Fatigability of spinal reflex transmission in a mouse model (SOD1(G93A) ) of amyotrophic lateral sclerosis.

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease characterized by progressive loss of motor neurons. To analyze the progressive motor deficits during the course of this disease, we investigated fatigability and ability of recovery of spinal motor neurons by testing monosynaptic reflex transmission with increasing stimulus frequencies in the lumbar spinal cord of the SOD1(G93A) mouse model for ALS in a comparison with wild-type (WT) mice. Monosynaptic reflexes in WT and SOD1(G93A) mice without behavioral deficits showed no difference with respect to their resistance to increasing stimulus frequencies.

Differentiation between Aδ and C fibre evoked nociceptive reflexes by TTX resistance and opioid sensitivity in the cat.

The contribution of nociceptive Aδ-fibres and C-fibres of the central pad of the foot to nociceptive spinal flexor reflex pathways (FRA-type) and to nociceptive excitatory reflex pathways to foot extensors (non-FRA type) was investigated in high spinal cats. Persisting effects after complete blocking of A-fibres by tetrodotoxin (TTX) application were thus attributed to nociceptive C-fibres. The results revealed that both Aδ and C-fibre afferents contributed to nociceptive reflexes of a FRA-pattern and of a non-FRA pattern.

Purinergic activation of dorsal root ganglion neurones in vivo.

Functional relevance of non-synaptic purinergic receptors on dorsal root ganglion cells was tested in vivo by the influence of ATP using 2P-LSM and Ca imaging. Within a few seconds after local application of ATP, neurones in dorsal root ganglion were activated indicated by an increase of their calcium signal. The signal reached its maximum within a few seconds and declined to control values after about 30 s.

Long-lasting post-mortem activity of spinal microglia in situ in mice.

As CNS macrophages, microglia show a high spontaneous motility of their processes, continuously surveying their microenvironment. Upon CNS injury, microglia react by immediate cellular polarization and process extension toward the lesion site as well as by subsequent amoeboid lesion-directed migration and phagocytosis. To determine the ability of microglia to fulfill their role within distinctively lesioned tissue in the absence of life support, we investigated microglial activity and responsiveness to laser-induced axonal injuries in the spinal dorsal columns in situ after cardiac and respiratory arrest, i.

NO mediates microglial response to acute spinal cord injury under ATP control in vivo.

To understand the pathomechanisms of spinal cord injuries will be a prerequisite to develop efficient therapies. By investigating acute lesions of spinal cord white matter in anesthetized mice with fluorescently labeled microglia and axons using in vivo two-photon laser-scanning microscopy (2P-LSM), we identified the messenger nitric oxide (NO) as a modulator of injury-activated microglia. Local tissue damages evoked by high-power laser pulses provoked an immediate attraction of microglial processes.

Acute muscle inflammation enhances the monosynaptic reflexes and c-fos expression in the feline spinal cord.

The aim of this research was to study the changes of the motor reflex activity (monosynaptic reflex (MSR) of the flexor and extensor muscles) and Fos immunoreactivity in lumbo-sacral spinal cord after acute induced myositis of m. gastrocnemius-soleus (GS). The experiments were carried out on ischaemic decerebrated, spinalized in C1 cats.