Individual Sensory Neurons Research Articles

Transplantation of dorsal root ganglion into the olfactory bulb of neonatal rats: a histochemical study

We have transplanted encapsulated dorsal root ganglia (DRG) from adult Wistar albino rats unilaterally into partially bulbectomized (n = 20) neonatal (P3-5) rats of the same strain. Three months postoperatively the animals were perfused and their brains processed by direct thiocholine method for cholinesterases (Ch), specific acetylcholinesterase (AChE) and nonspecific butyrylcholinesterase (BuChE) or stained by Cresyl violet. Selected sections were immunohistochemically stained for olfactory marker protein (OMP). In 17 cases we found surviving transplanted DRG. Fifteen transplants were well integrated with the spared portion of the olfactory bulb (OB) as clearly demonstrated by AChE and BuChE histochemistry, while two did not integrate. Regenerated OMP positive olfactory axons originating from neuroepithelium and AChE positive fibres from OB remnant penetrated into the transplants. In one case, fibers connected with BuChE positive Schwann cells grew from the transplanted DRG into the host OB. Individual sensory neurons of the transplants revealed variable intensity of the AChE staining, thus resembling the pattern of AChE activity in normal DRG. BuChE activity was mostly localized on the surface of sensory neurons in the ring of satellite cells. Some BuChE positive blood vessels penetrated into the DRG, and were observed around sensory neurons. The results showed a considerable viability and adaptability of the sensory neurons in the new environment after a long-term transplantation.

Activity-dependent induction of facilitation, depression, and post-tetanic potentiation at an insect central synapse

In Manduca sexta larvae, sensory neurons innervating planta hairs on the tips of the prolegs make monosynaptic excitatory connections with motoneurons innervating proleg retractor muscles. Tactile stimulation of the hairs evokes reflex retraction of the proleg. In this study we examined activity-dependent changes in the amplitude of the excitatory postsynaptic potentials (EPSPs) evoked in a proleg motoneuron by stimulation of individual planta hair sensory neurons. Deflection of a planta hair caused a phasic-tonic response in the sensory neuron, with a mean peak instantaneous firing frequency of greater than 300 Hz, and a tonic firing rate of 10-20 Hz. Direct electrical stimulation was used to activate individual sensory neurons to fire at a range of frequencies including those observed during natural stimulation of the hair. At relatively low firing rates (e.g., 1 Hz), EPSP amplitude was stable indefinitely. At higher instantaneous firing frequencies (greater than 10 Hz), EPSPs were initially facilitated, but continuous stimulation led rapidly to synaptic depression. High-frequency activation of a sensory neuron could also produce post-tetanic potentiation, in which EPSP amplitude remained elevated for several min following a stimulus train. Facilitation, depression, and post-tetanic potentiation all appeared to be presynaptic phenomena. These activity-dependent changes in sensory transmission may contribute to the behavioral plasticity of the proleg withdrawal reflex observed in intact insects.

Sensory neurons and motoneurons of the jaw-closing reflex pathway in rats: a combined morphological and physiological study using the intracellular horseradish peroxidase technique

Motoneurons and muscle spindle afferents of the rat masseter muscle were physiologically and morphologically characterized. Their soma-dendritic morphology and axonal course were investigated using the intracellular horseradish peroxidase method. Following electrical stimulation of the masseter nerve, individual motoneurons were identified by antidromic all-or-none action potentials and individual sensory neurons by orthodromic action potentials. Using threshold separation an excitatory input from muscle spindles to a masseter motoneuron was demonstrated. The short latency difference of 0.34 ms between the mean orthodromic response in the sensory neurons and the beginning of the synaptic potential in the masseter motoneuron suggests a monosynaptic connection between the spindle afferents and the motoneurons. Following intrasomatic horseradish peroxidase injection large multipolar cell bodies of masseter motoneurons were found within the motor nucleus. Their positions corresponded to the topographic organization of the motor trigeminal nucleus as described in retrograde tracing studies. Dendrites of masseter motoneurons were complex and could be found far beyond the nuclear borders. Distal dendrites extended to the mesencephalic trigeminal nucleus, the supratrigeminal nucleus, the lateral lemniscus and the reticular formation. Within the reticular formation dendrites were seen in the intertrigeminal nucleus and the peritrigeminal zone. Unipolar cell bodies of muscle spindle afferents were found in the mesencephalic trigeminal nucleus after intra-axonal injection of horseradish peroxidase. For all reconstructed sensory neurons a similar axonal course was found. Axonal terminals were found ipsilateral in the motor trigeminal nucleus, indicating a direct connection between sensory neurons and motoneurons. Further collaterals were found ipsilateral in the supratrigeminal nucleus and caudal to the motor trigeminal nucleus in the parvocellular reticular nucleus alpha. Since the latter termination areas are important for bilateral control of jaw-movements, the muscle spindle afferents are likely to participate not only in a monosynaptic motor reflex, but also in more complex neuronal circuits involved in jaw-movements.

Chapter 27 Second messenger-mediated actions of norepinephrine on target neurons in central circuits: a new perspective on intracellular mechanisms and functional consequences

Ever since the initial demonstration of a widespread distribution of noradrenergic fibers to functionally diverse regions of the mammalian forebrain, there has been considerable interest in determining the electrophysiological effects of norepinephrine (NE) on individual neurons within these target areas. While early studies showed that NE could directly inhibit cell firing via increased intracellular levels of cyclic AMP, more recent work has revealed a spectrum of noradrenergic actions, which are more accurately characterized as neuromodulatory. More specifically, numerous experimental conditions have been described where NE at levels subthreshold for producing direct depressant effects on spontaneous firing can facilitate neuronal responses to both excitatory and inhibitory synaptic stimuli. The goal of this report is to review recent evidence which suggests that the various modulatory actions of NE on central neurons result from the activation of different adrenoceptor-linked second messenger systems. In particular, we have focused on the candidate signal transduction mechanisms that may underlie NE's ability to augment cerebellar and cortical neuronal responsiveness to GABAergic synaptic inputs. The consequences of such NE-induced changes in synaptic efficacy are considered not only with respect to their influences on feature extraction properties of individual sensory cortical neurons but also with regard to the potential impact such actions would have on the signal processing capabilities of a network of noradrenergically innervated cortical cells.

Ataxic sensory neuropathy and dorsal root ganglionitis associated with Sjögren's syndrome

Thirteen patients, 11 women and 2 men, developed sensory and autonomic neuronopathies in association with features of primary Sjögren's syndrome. In 11, Sjögren's syndrome had not been previously diagnosed at the time of neurological presentation. All had prominent loss of kinesthesia and proprioception. Pain and thermal sensibility were less severely affected. Most had evidence of autonomic insufficiency. In some this was severe, with Adie's pupils, fixed tachycardia, and orthostatic hypotension. The course ranged from an abrupt, devastating onset to indolent progression over years. Stabilization or functional improvement occurred in 6 patients, 2 of whom received no drug therapy. Sensory nerve conduction studies and examination of nerve biopsy specimens demonstrated a wide spectrum in the severity of loss of large myelinated fibers. The cutaneous nerves of 6 patients had perivascular mononuclear infiltrates without necrotizing arteritis. Examination of biopsy specimens of dorsal root ganglia in 3 patients revealed lymphocytic (T-cell) infiltration in the dorsal roots and ganglia, with focal clusters around neurons. In the more mildly affected ganglia, individual sensory neurons were undergoing degeneration. In the most advanced case, very few neurons remained. The possibility of Sjögren's syndrome should be considered in patients, especially women, who develop acute, subacute, or chronic sensory and autonomic neuropathies, with ataxia and kinesthetic loss.

Role of competition among sensory neurons in regulation of pattern of innervation at their central and peripheral targets.

1. The importance of competitive interactions among muscle sensory afferents on their projections to central and peripheral targets was studied by producing large reductions in the number of afferents during development. Removal of the brachial dorsal root ganglion (DRG2), which normally supplies the entire sensory innervation of the forelimb, in bullfrog (Rana catesbeiana) tadpoles caused a smaller number of neurons in the adjacent thoracic ganglion (DRG3) to sprout into the forelimb and into the brachial spinal cord. 2. Horseradish peroxidase labeling in postmetamorphic animals showed that DRG3 neurons innervating the triceps muscle arborize in a novel but now appropriate area of the spinal cord, the region containing motoneuronal dendrites. These foreign afferents do not arborize in inappropriate regions of the spinal gray matter, and their collaterals have the same rostrocaudal distribution as those of normal DRG2 muscle afferents. 3. After metamorphosis, the number of DRG3 sensory axons in individual triceps muscle nerves was determined. Normally, two-thirds of all triceps afferents project to the medial head alone, even though each of the three heads is of similar size and is contacted by similar numbers of motoneurons. After DRG2 removal, although the total number of DRG3 afferents projecting to the triceps muscle was smaller than normal, the medial head still received approximately two-thirds of the axons, just as in normal frogs. These results suggest that the proportional sensory innervation of the triceps muscle-heads is not dependent on competitive interactions among afferents. 4. DRG3 afferents projecting to the forelimb also sprouted to innervate appropriate brachial motoneurons. The average strength of connection between individual sensory and motor neurons was found to be the same as in normal animals, even though there was presumably more central target space available for each afferent axon. This suggests that the number and/or strength of central connections made by individual fibers may be an intrinsic property of muscle sensory neurons.

Intracellular injection of cAMP induces a long-term reduction of neuronal K+ currents.

Intracellular signals that trigger long-term (24-hour) changes in membrane currents in identified neurons of Aplysia have been examined in order to understand the cellular mechanisms underlying long-term sensitization. Adenosine 3',5'-monophosphate (cAMP) was directly injected into individual sensory neurons to mimic the effects of sensitization training at the single cell level. Potassium currents of these cells were reduced 24 hours after injection of cAMP; these currents were similar to those reduced 24 hours after behavioral sensitization. These results suggest that cAMP is part of the intracellular signal that induces long-term sensitization in Aplysia.

The spinal projection of individual identified A-delta- and C-fibers.

1. Recordings were made from individual sensory neurons with an A-delta peripheral conduction velocity, either intrasomally in the L7 dorsal root ganglion, or extracellularly in Lissauer's Tract or in the dorsal root close to the root entry zone. The spinal projection of these afferents was assessed by their antidromic response to stimulation of the dorsal columns (DC) or Lissauer's Tract (LT) at the L5/L6 border. The adequate stimulus was also ascertained. 2. A-delta-fibers could be divided into two groups: high-threshold mechanoreceptors from either skin or muscle (HTMRs) and low-threshold mechanoreceptors (LTMs), primarily Down Hairs. A third group of cells recorded intrasomally had broad spikes with shoulders on the downstroke characteristic of A-delta-nociceptors and were so classified provisionally, although no adequate stimulus could be identified. HTMRs and broad spike cells projected either in DC or LT, but LTMs projected only in DC, never in LT. About one-quarter of both groups failed to project rostrally as far as L5/L6. 3. Cells with unmyelinated axons recorded intrasomally were found to supply either low-threshold or high-threshold mechanoreceptors. Unlike A-delta-cells, all these cells had broad spikes with shoulders on the downstroke. Proportionally fewer C-fibers than A-delta-fibers projected as far as one segment rostral from their root entry zone. Of those that did, axons supplying low-threshold mechanoreceptors projected only in DC, whereas those innervating high-threshold mechanoreceptors could project either through LT or DC. 4. A-delta-fibers supplying LTMs and HTMRs exhibited a similar reduced conduction velocity was reduced even further in the spinal cord but much more for HTMRs than for LTMs. For C-fibers the conduction velocity decrease was more substantial in the dorsal root for HTMRs than for LTMs. 5. These findings suggest that axons innervating different peripheral receptors exhibit characteristic cellular properties. They confirm that the primary afferent component of Lissauer's Tract is specialized as a "pain pathway" but also indicate that the dorsal columns may play some role in the transmission of nociceptive information.

Molecular convergence of presynaptic inhibition and presynaptic facilitation on common substrate proteins of individual sensory neurons of Aplysia.

The behavioral stimuli that serve as reinforcers for learning often have two components: a facilitatory component, important for sensitization and classical conditioning, and an inhibitory component, important for conditioned inhibition or as a break on the facilitation (Konorski 1967). We have encountered both of these components in Aplysia in the course of studying the effects of tail-reinforcing stimuli on the gill-withdrawal reflex of this animal (Carew et al. 1983; Frost et al. 1985; Mackey et al. 1987). These two opposing behavioral components are mediated, at the cellular level, by two opposing modulatory systems that act to regulate the strength of the connections between the sensory and the motor neurons of this reflex. Each of these two modulatory systems utilizes different transmitters, and these modulatory transmitters in turn activate distinct second-messenger systems. As a result, the anatomical pathways of the neuronal modulatory circuits in Aplysia are re-represented at the molecular pathway....

Sensory neurons supplying touch domes near the body midlines project bilaterally in the thoracic spinal cord of rats.

Sensory neurons that project bilaterally to laminae III-V of the dorsal horns in the thoracic spinal cord of rats were described in a previous anatomical study. The electrophysiological experiments reported here show that these neurons innervate skin near the dorsal and ventral midlines. Three series of experiments were undertaken: the regions of skin supplied by subdivisions of thoracic nerves were mapped by recording from the subdivisions while mechanically stimulating the skin; subdivisions containing the axons of contralaterally projecting sensory neurons were identified by antidromic activation from the contralateral dorsal horn; the receptive fields of individual sensory neurons and dorsal horn neurons were characterized by single-unit recording methods. Contralaterally projecting sensory neurons had axons in subdivisions of the thoracic nerves that supply skin adjacent to the body midlines. Their receptive fields were located near the dorsal or ventral midline but did not extend onto the contralateral side of the body. Most of the neurons were slowly adapting "type I" units supplying touch domes. Dorsal horn neurons receiving input from contralateral sensory neurons had receptive fields that overlapped the dorsal or ventral midline. This bilateral representation of skin near the midlines may be important for the fusion of the sensory maps of the right and left sides of the body.

Physiological evidence for specificity of synaptic connections between individual sensory and motor neurons in the brachial spinal cord of the bullfrog

We have studied synaptic connections between individual stretch-sensitive muscle afferents and motoneurons in the brachial spinal cord of bullfrogs. Sensory afferents from a given head of the triceps brachii muscle preferentially innervate motoneurons that project to the same muscle head. This preference is characterized in two ways: each class of sensory axon innervates a greater proportion of corresponding motoneurons than of motoneurons projecting to synergistic or unrelated muscles, and the synaptic potentials in these corresponding motoneurons are motoneurons that project to the same muscle head. This preference is characterized in two ways: each class of sensory axon innervates a greater proportion of corresponding motoneurons than of motoneurons projecting to synergistic or unrelated muscles, and the synaptic potentials in these corresponding motoneurons are of larger amplitude. A novel feature of these experiments is that even the smallest averaged synaptic potential is several times larger than the noise level. These smallest synaptic potentials thus represent the smallest synaptic interaction between a sensory and motor cell, and they could be the physiological correlate of a single sensory bouton on a motoneuronal dendrite.

Mechanoafferent neurons innervating tail of Aplysia. II. Modulation by sensitizing stimulation.

The tail-withdrawal reflex of Aplysia can be sensitized by weak stimulation of a site outside the site used to test the reflex or by repeatedly stimulating the test site itself. The sensitization of tail-withdrawal responses is associated with enhanced activation of the tail motor neurons and heterosynaptic facilitation of the monosynaptic connections between the tail sensory neurons and tail motor neurons. This synaptic facilitation can occur under conditions in which neither posttetanic potentiation nor generalized changes in postsynaptic input resistance contribute to the facilitation. In addition to producing monosynaptic excitatory postsynaptic potentials (EPSPs), action potentials in tail sensory neurons often recruit longer latency polysynaptic input to the tail motor neurons during sensitization. Strong, noxious tail shock similar in intensity to that used previously for sensitization and aversive classical conditioning of other responses in Aplysia produces more heterosynaptic facilitation than does weak sensitizing stimulation. Heterosynaptic facilitation builds up progressively with multiple trials and lasts for hours. Very strong shocks to the tail can change the response characteristics of tail sensory neurons so that a prolonged, regenerative burst of spikes is elicited by a brief intracellular depolarizing pulse. This bursting response produced by sensitizing stimulation has not been described previously in Aplysia sensory neurons and can greatly amplify the synaptic input to tail motor neurons from the sensory neurons. In addition, strong shocks to the tail increase the duration and magnitude of individual sensory neuron action potentials. Sensitizing tail stimulation usually produces long-lasting depolarization of the tail motor neurons and often long-lasting hyperpolarization of the tail sensory neurons. The tail motor and sensory neurons show both increases and decreases of input resistance following sensitizing stimulation. However, the small, occasional increases in input resistance of the motor neuron are insufficient to explain the heterosynaptic facilitation produced by sensitizing stimulation. Serotonin (5-HT) application can mimic many of the effects of sensitizing tail shock, including facilitation of both tail withdrawal and the monosynaptic connections between tail sensory and motor neurons, hyperpolarizing and depolarizing responses in the tail sensory neurons, and an increase in the duration and magnitude of the sensory neuron action potential. In the nearly isolated sensory neuron soma, 5-HT usually produces a slow, decreased conductance depolarizing response, suggesting that the 5-HT-induced hyperpolarizing response see

Associative conditioning of single sensory neurons suggests a cellular mechanism for learning.

A cellular analog of associative learning has been demonstrated in individual sensory neurons of the tail withdrawal reflex of Aplysia. Sensory cells activated by intracellular current injection shortly before a sensitizing shock to the animal's tail display significantly more facilitation of their monosynaptic connections to a tail motor neuron than cells trained either with intracellular stimulation unpaired to tail shock or with tail shock alone. This associative effect is acquired rapidly and is expressed as a temporally specific amplification of heterosynaptic facilitation. The results suggest that activity-dependent neuromodulation may be a mechanism underlying associative information storage and point to aspects of subcellular processes that might be involved in the formation of neural associations.

Regeneration of synaptic connections by sensory neurons in leech ganglia maintained in culture.

Leech ganglia maintained in organ culture were used to follow regeneration and the formation of synaptic connections by individual neurons. From earlier physiological studies on operated animals it is known that the c. n. s. of the leech is able to regenerate and that specific connections can be reformed. The present experiments show that specific regeneration also occurs in vitro but that axons do not simply grow directly back to their targets. (1) Axons were severed by crushing the connectives linking pairs of ganglia at the time of removal from the animal. Light and electron microscopy indicated that the procedure of crushing severed all the axons within the connectives. For several days after the lesion had been made, conduction of impulses from one ganglion to the next was abolished. (2) After 5–10 days in culture, stimulation of the connectives with external electrodes gave rise to impulses that were once again conducted beyond the site of the lesion. Characteristic excitatory and inhibitory synaptic potentials were evoked in identified sensory and motor cells in both ganglia by this indiscriminate stimulation of axons. Electron micrographs of the crushed region showed not only regenerated axons traversing the site of the lesion but also synaptic profiles similar to those seen in the neuropile of normal ganglia. Thus, pre- and post-synaptic specializations had been formed during regeneration in a part of the c. n. s. where they are not normally present. (3) Individual sensory neurons were injected with horseradish peroxid­ase to reveal the course taken by their regenerating axons. At the site of the crush profuse branching occurred by 7 days. The arborization of a single axon was highly complex, with many varicosities present on fine branches. After two weeks in culture, one or more of the processes had usually grown beyond the crush and in certain instances had reached the next ganglion. Other branches ran back towards the ganglion in which the cell body was situated. During the period of the experiments (up to 45 days) no retraction of the sprouted fibres or of the arborization at the crush was observed. In addition to sprouting at the site of the lesion considerable sprouting also occurred within the ganglion, close to the cell body. (4) Individual mechanosensory neurons regenerated and would once again evoke synaptic potentials in their original targets after two weeks in culture. Thus, intracellular stimulation of single sensory cells in one ganglion gave rise to synaptic potentials in the appropriate motor neuron of the neighbouring ganglion. Injection of such sensory cells with horse­radish peroxidase showed that their axons had extended beyond the lesion and ramified in the neuropile of the next ganglion. (5) It is concluded that neurons in leech ganglia are able to regenerate and reform appropriate synaptic connections in culture. The degree of precision is hard to assess because of novel synaptic interactions and numerous additional sprouts that develop during regeneration.

Stimulus coding in topographic and nontopographic afferent modalities: On the significance of the activity of individual sensory neurons.

Stimulus coding in topographic and nontopographic afferent modalities: On the significance of the activity of individual sensory neurons.