Changes in the ultrastructure of synapses and neurons of the ventral horns of the spinal cord in cats with tetany as a result of the removal of the parathyroid glands

An electron microscopic investigation on ultrastructure of synapses and ventral horns of the lumbar thickening in the spinal cord of cats with parathyroprival tetany has been performed. When motor disorders are clearly seen on the 2d - 4th days after removal of the parathyroid glands, together with the changes demonstrating functional activity of the synapses, certain irreversible degenerative changes of the "dark" type are seen in the axodendritic and axosomatic synapses. A complex of structural-metabolic changes has been revealed in the motor neurons, they have both functional-compensatory and degenerative character. It is possible to suggest that the changes revealed at parathyroprival tetany result from a disturbed metabolism and increased afferent influences.     

Monoaminergic Modulation of Spinal Viscero-Sympathetic Function in the Neonatal Mouse Thoracic Spinal Cord

Descending serotonergic, noradrenergic, and dopaminergic systems project diffusely to sensory, motor and autonomic spinal cord regions. Using neonatal mice, this study examined monoaminergic modulation of visceral sensory input and sympathetic preganglionic output. Whole-cell recordings from sympathetic preganglionic neurons (SPNs) in spinal cord slice demonstrated that serotonin, noradrenaline, and dopamine modulated SPN excitability. Serotonin depolarized all, while noradrenaline and dopamine depolarized most SPNs. Serotonin and noradrenaline also increased SPN current-evoked firing frequency, while both increases and decreases were seen with dopamine. In an in vitro thoracolumbar spinal cord/sympathetic chain preparation, stimulation of splanchnic nerve visceral afferents evoked reflexes and subthreshold population synaptic potentials in thoracic ventral roots that were dose-dependently depressed by the monoamines. Visceral afferent stimulation also evoked bicuculline-sensitive dorsal root potentials thought to reflect presynaptic inhibition via primary afferent depolarization. These dorsal root potentials were likewise dose-dependently depressed by the monoamines. Concomitant monoaminergic depression of population afferent synaptic transmission recorded as dorsal horn field potentials was also seen. Collectively, serotonin, norepinephrine and dopamine were shown to exert broad and comparable modulatory regulation of viscero-sympathetic function. The general facilitation of SPN efferent excitability with simultaneous depression of visceral afferent-evoked motor output suggests that descending monoaminergic systems reconfigure spinal cord autonomic function away from visceral sensory influence. Coincident monoaminergic reductions in dorsal horn responses support a multifaceted modulatory shift in the encoding of spinal visceral afferent activity. Similar monoamine-induced changes have been observed for somatic sensorimotor function, suggesting an integrative modulatory response on spinal autonomic and somatic function.     

Time Course of Immediate Early Gene Protein Expression in the Spinal Cord following Conditioning Stimulation of the Sciatic Nerve in Rats

Long-term potentiation induced by conditioning electrical stimulation of afferent fibers is a widely studied form of synaptic plasticity in the brain and the spinal cord. In the spinal cord dorsal horn, long-term potentiation is induced by a series of high-frequency trains applied to primary afferent fibers. Conditioning stimulation (CS) of sciatic nerve primary afferent fibers also induces expression of immediate early gene proteins in the lumbar spinal cord. However, the time course of immediate early gene expression and the rostral-caudal distribution of expression in the spinal cord have not been systematically studied. Here, we examined the effects of sciatic nerve conditioning stimulation (10 stimulus trains, 0.5 ms stimuli, 7.2 mA, 100 Hz, train duration 2 s, 8 s intervals between trains) on cellular expression of immediate early genes, Arc, c-Fos and Zif268, in anesthetized rats. Immunohistochemical analysis was performed on sagittal sections obtained from Th13- L5 segments of the spinal cord at 1, 2, 3, 6 and 12 h post-CS. Strikingly, all immediate early genes exhibited a monophasic increase in expression with peak increases detected in dorsal horn neurons at 2 hours post-CS. Regional analysis showed peak increases at the location between the L3 and L4 spinal segments. Both Arc, c-Fos and Zif268 remained significantly elevated at 2 hours, followed by a sharp decrease in immediate early gene expression between 2 and 3 hours post-CS. Colocalization analysis performed at 2 hours post-CS showed that all c-Fos and Zif268 neurons were positive for Arc, while 30% and 43% of Arc positive neurons were positive for c-Fos and Zif268, respectively. The present study identifies the spinal cord level and time course of immediate early gene (IEGP) expression of relevance for analysis of IEGPs function in neuronal plasticity and nociception.     

The dorsal root reflex in isolated mammalian spinal cord

1. The dorsal root reflex has been investigated in an isolated preparation of adult mammalian spinal cord. 2. Both evoked and spontaneous activity can be recorded from the cord in the dorsal spinal roots. 3. The spontaneous activity has a characteristic pattern of firing in bursts of action potentials. Spontaneous and evoked activity are optimum at temperatures between 25 and 27 degrees C; little activity can be detected above 35 degrees C. 4. The spontaneous dorsal root activity has been shown to be correlated with negative potentials in the dorsal horn of the cord, and intracellular recordings made from primary afferent fibres have shown spontaneous primary afferent depolarizations (PAD) which underlie the generation of the spontaneous dorsal root activity. 5. The evoked dorsal root reflex has been shown to spread up to 16 spinal segments both rostrally and caudally from the stimulated dorsal root, and to the contralateral side of the cord. 6. The spontaneous dorsal root activity in widely separated segments has been shown by cross-correlation analysis to be linked both ipsi- and contra-laterally. 7. The significance of such a widespread system for the generation of PAD is discussed.     

Measuring Spinal Presynaptic Inhibition in Mice By Dorsal Root Potential Recording In Vivo

Presynaptic inhibition is one of the most powerful inhibitory mechanisms in the spinal cord. The underlying physiological mechanism is a depolarization of primary afferent fibers mediated by GABAergic axo-axonal synapses (primary afferent depolarization). The strength of primary afferent depolarization can be measured by recording of volume-conducted potentials at the dorsal root (dorsal root potentials, DRP). Pathological changes of presynaptic inhibition are crucial in the abnormal central processing of certain pain conditions and in some disorders of motor hyperexcitability. Here, we describe a method of recording DRP in vivo in mice. The preparation of spinal cord dorsal roots in the anesthetized animal and the recording procedure using suction electrodes are explained. This method allows measuring GABAergic DRP and thereby estimating spinal presynaptic inhibition in the living mouse. In combination with transgenic mouse models, DRP recording may serve as a powerful tool to investigate disease-associated spinal pathophysiology. In vivo recording has several advantages compared to ex vivo isolated spinal cord preparations, e.g. the possibility of simultaneous recording or manipulation of supraspinal networks and induction of DRP by stimulation of peripheral nerves.     

Role of propriospinal neurons in inhibition of the afferent flow

The effects of stimulation of the dorsal funiculus on dorsal surface potentials (DSPs) of the spinal cord evoked by stimulation of a peripheral nerve and on antidromic action potentials (AAPs) evoked by stimulation of terminal branches of primary afferent fibers and recorded from the afferent nerve or dorsal root, were investigated in acute experiments on spinal cats and on cats anesthetized with pentobarbital and chloralose. Stimulation of the dorsal funiculus led to biphasic inhibition of the N₁-component of the DSP with maxima at the 15th–30th and 60th–80th milliseconds between the conditioning and testing stimuli. Maximal reinforcement of the AAP was found with these intervals. Bilateral division of the dorsal funiculi between the point of application of the conditioning stimuli and the point of recording the DSP abolished the first wave of inhibition of the DSP and the reinforcement of the AAP. After total transection of the cord above the site of conditioning stimulation the picture was unchanged. It is concluded that the initial changes in DSP and AAP are due to activation of the presynaptic inhibition mechanism by antidromic impulses traveling along nerve fibers running in the dorsal funiculus. Repeated inhibition of the DSP, like reinforcement of the AAP, can possibly be attributed to activation of similar inhibitory mechanisms through the propriospinal neurons of the spinal cord.Dnepropetrovsk State University. Translated from Neirofiziologiya, Vol. 5, No. 4, pp. 401–405, July–August, 1973.     

c-Fos expression in the central nervous system elicited by phrenic nerve stimulation.

Phrenic nerve afferents (PNa) have been shown to activate neurons in the spinal cord, brain stem, and forebrain regions. The c-Fos technique has been widely used as a method to identify neuronal regions activated by afferent stimulation. This technique was used to identify central neural areas activated by PNa. The right phrenic nerve of urethane-anesthetized rats was stimulated in the thorax. The spinal cord and brain were sectioned and stained for c-Fos expression. Labeled neurons were found in the dorsal horn laminae I and II of the C3-C5 spinal cord ipsilateral to the site of PNa stimulation. c-Fos-labeled neurons were found bilaterally in the medial subnuclei of the nucleus of the solitary tract, rostral ventral respiratory group, and ventrolateral medullary reticular formation. c-Fos-labeled neurons were found bilaterally in the paraventricular and supraoptic hypothalamic nuclei, in the paraventricular thalamic nucleus, and in the central nucleus of the amygdala. The presence of c-Fos suggests that these neurons are involved in PNa information processing and a component of the central mechanisms regulating respiratory function.     

The role of serotonergic system in body temperature regulation.

Results indicate that vascular responses to temperature stimulation are predominantly impaired in animals with 5-HT deprivation. A hypothesis is therefore raised that the 5-HT system participates in body temperature regulation in such a way as to link the regulatory output with vasomotor pathways. The 5-HT system in the spinal cord has been shown to inhibit the afferent transmission of temperature signals. Therefore, depletion of 5-HT does not prevent sensory transmission, at least at the spinal cord level.     

Fas and FasL Expression in the Spinal Cord Following Cord Hemisection in the Monkey

The changes of endogenous Fas/FasL in injured spinal cord, mostly in primates, are not well known. In this study, we investigated the temporal changes in the expression of Fas and FasL and explored their possible roles in the ventral horn of the spinal cord and associated precentral gyrus following T(11) spinal cord hemisection in the adult rhesus monkey. A significant functional improvement was seen with the time going on in monkeys subjected to cord hemisection. Apoptotic cells were also seen in the ventral horn of injured spinal cord with TUNEL staining, and a marked increase presents at 7 days post operation (dpo). Simultaneously, the number of Fas and FasL immunoreactive neurons in the spinal cords caudal and rostral to injury site and their intracellular optical density (OD) in the ipsilateral side of injury site at 7 dpo increased significantly more than that of control group and contralateral sides. This was followed by a decrease and returned to normal level at 60 dpo. No positive neurons were observed in precentral gyrus. The present results may provide some insights to understand the role of Fas/FasL in the spinal cord but not motor cortex with neuronal apoptosis and neuroplasticity in monkeys subjected to hemisection spinal cord injury.     

脊髓背角痛觉传递和调制的一些化学解剖学观察

本实验研究了脊髓背角内Ｃ纤维末梢的分布和突触学特征及其一些神经递质化学构筑;定量观察了急性痛引起背角的递质变化;显示了初级传入Ｃ纤维,抑制性中间神经元和背角伤害性感受神经元三者之间的突触关系,并探讨它们在痛觉信息传递和调制中的作用。     

Structural-functional connections of the lemniscus system with the caudate nucleus in the cat

Afferent projections were studied of nuclei of the spinal cord dorsal columns to the caudate nuclei. Evoked potentials (EPs) were recorded in the caudate nuclei and the nuclei of dorsal columns of the spinal cord to stimulation of the forelimb against the background and after unilateral elimination of the medial lemniscus. After the section, the EPs in the recorded nuclei sharply intensified in response to stimulation of the forelimb, ipsilateral to the section. Degenerative changes were also shown by electron microscopy in axonal terminals in the caudate nuclei at the operated side. Conclusion is drawn that the caudate nuclei receive direct connections from the contralateral nuclei of the spinal cord dorsal columns, which probably, conduct mainly kinesthetic afferent influences.     

Spinal Astrogliosis in Pain Models: Cause and Effects

Pathological pain has been subjected to intense research to shed light on the underlying mechanisms of key symptoms, such as allodynia and hyperalgesia. The main focus has by and large concerned plasticity of spinal cord neurons and the primary afferent nerves relaying peripheral information to the spinal cord. Animal pain models display an increased presence of reactive astrocytes in the spinal cord, but in contrast to neurons, little is known about how they contribute to abnormal pain sensation. However, astrocytes are now beginning to receive greater attention, and as new information is emerging, it appears that astrocytes undertake critical roles in manifesting pathological pain. Through the secretion of diffusible transmitters, such as interleukins, ATP, and NO, astrocytes may augment primary afferent neuronal signaling or sensitize second order neurons in the spinal cord. In addition, astrocytes might lead to altered pain perception by a direct modulation of synaptic transmission between neurons in the nociceptive pathway or through the creation of astrocytic networks capable of transducing signals for extended distances across and along the spinal cord. Future research in astrocyte activation and signaling may therefore reveal novel drug targets for managing pathological pain.     

Control of locomotion in vitro: I. Deafferentation

We previously described the ability to induce adult-like, coordinated airstepping following electrical stimulation of the brainstem in the hindlimb-attached, in vitro brainstem-spinal cord preparation. These findings suggest the presence at birth of supraspinal systems capable of activating and modulating spinal locomotor mechanisms, which presumably also are present at birth. The current study employed the hindlimb-attached in vitro brainstem-spinal cord preparation from 0- to 4-day-old rats maintained in oxygenated artificial cerebrospinal fluid. After the control threshold-frequency relationship for eliciting airstepping was established, the dorsal roots to the attached limbs were severed and the procedure was repeated. No changes in electrical threshold or major differences in the elicited locomotor pattern were observed after deafferentation, although the amplitude of the electromyograms decreased. The mean frequency of alternation at threshold before deafferentation was similar to that after deafferentation. However, the maximum mean frequency induced by suprathreshold stimulation was significantly higher after deafferentation than that before deafferentation. These results suggest that (1) the supraspinal modulation of spinal locomotor mechanisms is not entirely dependent on afferent input; (2) intrinsic spinal locomotor mechanisms are present in the spinal cord at birth; and (3) afferent input may limit the maximum frequency of alternation of the limbs early in development.     

Electrical excitability of the soma of sensory neurons is required for spike invasion of the soma,but not for through-conduction

The cell soma of primary sensory neurons is electrically excitable, and is invaded by action potentials as they pass from the peripheral nerve, past the dorsal root ganglion (DRG) and toward the spinal cord. However, there are virtually no synapses in the DRG, and no signal processing is known to occur there. Why, then, are DRG cell somata excitable? We have constructed and validated an explicit model of the primary sensory neuron and used it to explore the role of electrical excitability of the cell soma in afferent signaling. Reduction and even elimination of soma excitability proved to have no detectable effect on the reliability of spike conduction past the DRG and into the spinal cord. Through-conduction is affected, however, by major changes in neuronal geometry in the region of the t-junction. In contrast to through-conduction, excitability of the soma and initial segment is essential for the invasion of afferent spikes into the cell soma. This implies that soma invasion has a previously unrecognized role in the physiology of afferent neurons, perhaps in the realm of metabolic coupling of the biosynthesis of signaling molecules required at the axon ends to functional demand, or in cell-cell interaction within sensory ganglia. Spike invasion of the soma in central nervous system neurons may play similar roles.     

Effect of various neurotropic drugs on chronic hyperreflexia in rats after cordotomy]

Substances modulating calcium permeability of cell membrane (verapamil imidazol, 4-aminopyridine), decreasing motor activity (meprobamate) and blocking adrenoreceptors (clophelin, propranolol, droperidol, aminazine++ have been studied for their action on the monosynaptic discharge of the ventral roots (MD VR) reinforced due to chronic cutting of the spinal cord. It is found that verapamil and meprobamate depress more strongly MD VR of rats with chronic cutting of the spinal cord than of those with the acute one. Imidazol and 4-aminopyridine reinforcing MD VR of rats with acute cutting of the spinal cord have no influence on the analogous index of rats with chronic cutting of the spinal cord. Adrenoblockers do not change the amplitude of MD VR in both groups of the animals.     

Changes in the blood vessels, spinal cord neurocytes, pia mater and spinal ganglia in chronic stimulation of the sympathetic trunks

Taking into account the data concerning disturbances in blood supply of the spinal cord as a response to irritation of the sympathetic trunks, the experimental morphological investigation has been performed on rabbits. By means of the injection technique and staining of neurocytes, changes in the spinal cord, in the spinal nodes and in the pia mater have been studied at chronic irritation of the lumbar nodes of the sympathetic trunk. Certain degenerative changes have been revealed in nervous cells and also phenomena of the spinal cord ischemia, decreasing contacts between the nervous cells and the capillaries surrounding them. As the authors believe, these data can be used by clinicians for revealing pathological mechanisms of the spinal cord ischemia as a result of chronic irritation of the sympathetic trunk.     

Normal anatomy and physiology of the spinal cord dorsal horn

The dorsal horn of the spinal cord receives afferent input from innocuous primary afferent neurons via collaterals from the dorsal columns. This input is integrated and relayed primarily by neurons in laminae III-VI. Dorsal horn neurons which encode innocuous inputs project to the medulla and the cervical spinal cord via the dorsal columns and the dorsolateral funiculus. Nociceptive primary afferent neurons enter the spinal dorsal horn via collaterals from Lissauer's tract. Nociceptive input is integrated and relayed by neurons in laminae I, II and V which project to the reticular formation and thalamus via the anterolateral tract.     

Polarization of primary afferent terminals in the lumbar spinal cord during fictitious locomotion

Changes in electrical polarization of primary afferent terminals in the lumbosacral portion of the spinal cord were investigated during fictitious locomotion in immobilized decorticated and spinal cats. Fictitious locomotion was accompanied by stable hyperpolarization of the afferent terminals, against the background of which they were periodically depolarized in rhythm with efferent activity. These tonic and phasic changes were observed in terminals of all groups of afferent fibers tested: cutaneous and muscular (Ia and Ib). Periodic in-phase depolarization was carried out in different ipsilateral segments of the lumbosacral enlargement. During ficitious galloping changes in depolarization of the primary efferents were in phase on different sides; during fictitious walking, these periodic changes were out of phase. On the basis of these results the physiological importance of changes in electrical polarization of primary afferent terminals of the spinal locomotor generator is discussed.A. A. Bogomolets Institute of Physiology, Academy of Sciences of the Ukrainian SSR, Kiev. Translated from Neirofiziologiya, Vol. 12, No. 5, pp. 481–489, September–October, 1980.     

Nerve growth factor in glia and inflammatory cells of the injured rat spinal cord

Nerve growth factor (NGF) is crucial for the development of sympathetic and small-diameter sensory neurons and for maintenance of their mature phenotype. Its role in generating neuronal pathophysiology is less well understood. After spinal cord injury, central processes of primary afferent fibers sprout into the dorsal horn, contributing to the development of autonomic dysfunctions and pain. NGF may promote these states as it stimulates sprouting of small-diameter afferent fibers and its concentration in the spinal cord increases after cord injury. The cells responsible for this increase must be identified to develop a strategy to prevent the afferent sprouting. Using immunocytochemistry, we identified cells containing NGF in spinal cord sections from intact rats and from rats 1 and 2 weeks after high thoracic cord transection. In intact rats, this neurotrophin was present in a few ramified microglia and in putative Schwann cells in the dorsal root. Within and close to the lesion of cord-injured rats, NGF was in many activated, ramified microglia, in a subset of astrocytes, and in small, round cells that were neither glia nor macrophages. NGF-immunoreactive putative Schwann cells were prevalent throughout the thoracolumbar cord in the dorsal roots and the dorsal root entry zones. Oligodendrocytes were never immunoreactive for this protein. Therapeutic strategies targeting spinal cord cells that produce NGF may prevent primary afferent sprouting and resulting clinical disorders after cord injury.