Origin of the rubrospinal tract in neonatal, developing, and mature rats.

This investigation describes the origin of the rubrospinal tract in neonatal (1-10 days old), developing (15-20 days old), and mature (2-4 months old) rats studied by using the horseradish peroxidase (HRP) method of tracing neuronal connections. HRP was administered in the cervical or lumbosacral segments of the spinal cord either in the crystal or solution form. The results showed that the rubrospinal tract extended to the lumbosacral part of the spinal cord at birth. There appeared to be no difference in the pattern of labelled rubrospinal (RS) neurons following the administration of HRP in the cervical or the lumbosacral cord segment of the neonatal, developing, and mature rats. In rats of these three age groups, labelled neurons were found bilaterally in the red nucleus, with a contralateral predominance, and they were found in both the parvicellular and magnocellular portions of the red nucleus. There was a somatotopic arrangement in the labelled RS neurons: Those projecting to the cervical cord segments were located in the dorsal and dorsomedial regions of the red nucleus and those projecting to the lumbosacral cord segments were located in the ventral and ventrolateral regions of the nucleus.

Localizing spinal-cord-projecting neurons in adult albino rats.

Following horseradish peroxidase injection into the cervical and lumbosacral cords of adult albino rats, labeled neurons were seen in the first cervical segment, brain stem, and cerebellar and diencephalic nuclei. A new pathway, the faciospinal projection, originating in the medial portion of the rostral part of the facial nucleus, was traced. Another new pathway, the olivospinal pathway, is probably also present. Our results for neurons projecting to the spinal cord (spinal-projecting neurons) from the nucleus ambiuus, dorsal motor nucleus of the vagus, superior vestibular nucleus and nucleus f, nucleus Darshevch, nucleus Rolleri, nucleus prepositus hypoglossi, and nucleus of the posterior commissure have been reported before in other mammals but not in rats. Projections from the following regions are in general agreement with previous results in rats, but show significant topographical differences: the first cervical segment; nuclei gracilis, cuneatus, and cuneatus lateralis; the midline and lateral reticular nuclear complex; the trigeminal nuclear complex (spinal, principal, and mesencephalic); nucleus of the tractus solitarius; the medial, lateral, and descending vestibular nuclei, nuclei coeruleus and subcoeruleus; superior colliculus; interstitial nucleus of Cajal, and the deep cerebellar nuclei. The distribution of labeled neurons in the nucleus parabrachialis, nucleus tegmentolaterodorsalis, nucleus Kölliker-Fuse, nucleus Edinger-Westphal, and the hypothalamic nuclear complex confirmed that of previous reports in rats. With the exception of a few nuclear groups which project primarily to either lumbosacral (e.g., the paraventricular nucleus of the hypothalamus) or cervical segments (e.g., the facial motor nucleus and the superior colliculus) most of the other nuclear groups project to both the lumbar and cervical levels. There is no distinct somatotopy in the neuronal groups projecting to both cervical and lumbosacral levels. With only a few exceptions (e.g., the superior vestibular nucleus) most of the spinal-projecting neurons are bilaterally distributed, some with contralateral and others with ipsilateral predominance.

Localizing spinal-cord-projecting neurons in neonatal and immature albino rats.

According to results from the horseradish peroxidase (HRP) method of tracing neuronal connections, the spinal cords of neonatal and immature rats receive a large number of descending projections from the first cervical cord segment, various brain-stem nuclei, and deep cerebellar and diencephalic nuclei. All these projections are present at birth, though at this age some of them are not fully established. Thus, only a few cells in the trigeminospinal, solitariospinal, tectospinal, and cerebellospinal groups were labeled after HRP injection in the lumbosacral or cervical cord segments in neonatal animals. They were clearly labeled in older, immature animals. The labeled neurons in other descending pathways appeared to be equal in density in neonatal, immature, and adult rats. This visual impression was stregthened by the counts of neurons in the interstitial nucleus of Cajal, which showed no significant difference in the number of labeled neurons in the three age groups. However, counts of labeled cells in the lateral vestibular nucleus and nucleus of the posterior commissure showed that there is a steady rise in the number of labeled neurons as the animals increase in age.

Ultrastructural study of external cuneothalamic neurons and their synaptic relationships with primary afferents in the gerbil.

The present study examined the synaptic organization of external cuneothalamic neurons and their relationships with primary afferents in the gerbil external cuneate nucleus (ECN) following an injection of horseradish peroxidase (HRP) into the anterodorsal cap of the ventrobasal thalamus in conjunction with a simultaneous injection of HRP into the contralateral brachial and cervical nerve plexuses. The thalamus-projecting neurons have been shown to be confined to the intermediate portion of the caudal half of the ECN at the light microscopic level (Lan et al., 1994c). In this study, HRP-labelled external cuneothalamic neurons were ultrastructurally characterized by their relatively small-sized soma bearing a variable number of somal spines. Their nucleus had a slightly indented contour with an eccentric nucleolus. The HRP-labelled somata were postsynaptic to many axon terminals, which were classified into round (Rs type; 53.0%), pleomorphic (Ps type; 32.7%), and flattened (Fs type; 14.3%) vesicle-containing boutons. The HRP-labelled dendritic elements were postsynaptic to a greater number of axon terminals, which were also classified into the round (Rd; 64.7%), pleomorphic (Pd; 25.2%), and flattened (Fd; 10.1%) type boutons. These presynaptic axonal boutons tended to synapse on distal and secondary dendrites of external cuneothalamic neurons. In the present simultaneous HRP labelling study, some of the primary afferent terminals made direct synaptic contacts with the dendrites of the external cuneothalamic neurons. In view of the multiple inputs onto the external cuneothalamic neurons, impinging particularly on their somata and secondary dendrites, it is suggested that the proprioceptive information reaching these neurons is intensively modulated and integrated before transmission ultimately to the cerebral sensorimotor cortex.

Synaptic relationships between GABA-immunoreactive boutons and primary afferent terminals in the rat cuneate nucleus.

The present study examined the synaptic relation between the primary afferent terminals and intrinsic neuronal elements in the rat cuneate nucleus. For this purpose, experimental degeneration after multiple cervicothoracic dorsal rhizotomies or anterograde transport of wheatgerm agglutinin conjugated to horseradish peroxidase were used to identify the primary afferent terminals, while immunogold postembedding staining was employed to identify the GABA-immunoreactive boutons. The combined procedure allowed us to demonstrate a direct synaptic relationship between the primary afferent terminals and GABA-immunoreactive boutons. At least two types of synaptic relation were observed between the primary afferent terminals, identified by their degenerating features or labeled by wheatgerm agglutinin conjugated to horseradish peroxidase, and the immunogold-labeled GABA-immunoreactive boutons (i) a GABA-immunoreactive bouton making a simple presynaptic contact with the primary afferent terminal; and (ii) a synaptic glomerular complex in which the centrally located primary afferent terminal was postsynaptic to a GABA-immunoreactive bouton and presynaptic to dendrites closely associated with it; both terminals were sometimes presynaptic to a common dendrite. It is speculated from this study that the incoming impulses from the forelimb area are modulated by the GABA-immunoreactive boutons in the cuneate nucleus of the rat.

The synaptic interrelationships between primary afferent terminals, cuneothalamic relay neurons and GABA-immunoreactive boutons in the rat cuneate nucleus.

The present study described an ultrastructural synaptic configurations between primary afferent terminals (PATs), cuneothalamic relay neurons (CTNs) and GABA-immunoreactive (GABA-IR) boutons in the cuneate nucleus of rats using cervicothoracic dorsal rhizotomies, retrograde transport of wheat germ agglutinin conjugated with horseradish peroxidase complex (WGA-HRP) and anti-GABA immunogold labelling methods. With this procedure, direct synaptic relationships between the PATs, CTNs and GABA-IR boutons have been demonstrated. The most remarkable feature was the observation whereby an immunogold-labelled GABA-IR bouton was presynaptic to a WGA-HRP labelled dendrite of CTN and a degenerating PAT; the same PAT was in turn presynaptic to the HRP-labelled dendrite. This was evident in ten out of a total of 133 synaptic configurations that were closely scrutinized. Results of this study support the concept that GABA-IR boutons are not only involved in presynaptic inhibition on the primary afferent input to the cuneothalamic relay neurons, but also exert a simultaneous postsynaptic inhibition on these cells.

Somatic noxious mechanical stimulation induces Fos expression in the postsynaptic dorsal column neurons in laminae III and IV of the rat spinal dorsal horn.

This study was conducted to ascertain the possible expression of Fos-like immunoreactivity (Fos-LI) in the postsynaptic dorsal column (PSDC) neurons in response to noxious mechanical stimulation of the forepaw glabrous area of normal rats. For this purpose, Fos immunohistochemistry along with Fluoro-Gold (FG) retrograde tracing was utilized. After repeated noxious pinching of the forepaw glabrous area, there was a marked increase in number of Fos-LI neurons in the dorsal horn, including Rexed's laminae III and IV, at C5-T1 spinal cord segments ipsilateral to the stimulation. Between segments C5 and T1, about 40% of the Fos-LI neurons in laminae III and IV were distributed at segment C7. In the rats subjected to the noxious pinch coupled with FG injection into the right cuneate nucleus, PSDC neurons double labeled with Fos and FG were localized in the ipsilateral laminae III and IV extending from segment C5 to T1, with about 70% of them distributed at segments C6 and C7. At segment C6 or C7, double-labeled neurons made up about 10% of the PSDC neurons that projected their axons to the cuneate nucleus. Most of the double-labeled neurons appeared fusiform with their primary dendrites projected dorso-ventrally. The present results suggest that the morphologically distinct, subclasses of PSDC neurons in spinal laminae III and IV may contribute to the central transmission of mechanical nociceptive information through the dorsal column into the cuneate nucleus.

Neuronal connections between the auricular skin and the sympathetic pre- and postganglionic neurons of the dog as studied by using pseudorabies virus.

Pseudorabies virus (PrV) as a neuronal tracer was microinjected into the concave surface of the puppy's left pinna to establish the morphological basis of somato-visceral linkage. The virus infected neurons were detected by FITC conjugated with polyclonal swine anti-PrV serum. Labelled neurons were localized in: (1) the trigeminal, geniculate and superior vagal ganglia; (2) the subnucleus caudalis of the spinal trigeminal nucleus; (3) the intermediolateral column (IML) of the thoracolumbar segments and (4) the sympathetic chain ganglia. Present results suggest that when injected into the peripheral nerves, PrV was retrogradely transported to the nerve cell bodies located in the respective sensory ganglia. From the first order sensory neurons, the virus would self-replicate and was transported trans-synaptically via the brainstem nuclei and IML to reach the neurons in the sympathetic ganglia.

Multiple inputs of GABA-immunoreactive neurons in the cuneate nucleus of the rat.

Using anterograde transport of WGA-HRP and the experimental degeneration method for identification of corticocuneate (CCT) and primary afferent (PAT) terminals in conjunction with gamma-amino butyric acid (GABA) and glutamate immunocytochemistry, this study has demonstrated that the GABA-immunoreactive (GABA-IR) neurons in the rat cuneate nucleus were post-synaptic to PATs (some of them being glutamate-IR), GABA-IR and GABA-negative terminals. The HRP-labelled CCTs did not make any synaptic contacts with GABA-IR neurons but with some GABA-negative dendrites. PATs labelled by HRP or showing degenerating features made direct synaptic contacts with the dendrites of GABA-IR neurons. Beside the above GABA-IR boutons also showed axosomatic and axodendritic synapses with the GABA-IR neurons. In 'triple labeling' method for GABA, PAT and glutamate, it was found that the PATs which were usually glutamate-positive were presynaptic to the dendrites of GABA-IR neurons. Furthermore, some glutamate-IR terminals which were of non-PAT's origin also synapsed with the dendrites and somata of GABA-IR neurons. It is concluded from this study that the major inputs of GABA-IR neurons were from glutamate immunopositive PATs and glutamate terminals of non-PATs origin; other GABA-IR terminals either intrinsic or extrinsic also contributed to the afferent sources of GABA-IR neurons. The CCTs contributed very little, if any, to this input. It is suggested that the PATs and glutamate-IR terminals on GABA-IR neurons may be involved in lateral inhibition for increase of spatial precision. The synaptic contacts between GABA-IR boutons and dendrites or somata of GABA-IR neurons may provide a possible means for disinhibition.

Somatostatin-IR neurons are a major subpopulation of the cuneothalamic neurons in the rat cuneate nucleus.

This study was aimed to localize and characterize the somatostatin-immunoreactive (SOM-IR) neurons in the rat cuneate nucleus (CN). By immuno-histochemistry, the SOM-IR neurons, which were widely distributed in the nucleus, were round, spindle or multiangular in shape (mean area = 226.1 +/ -3.1 microm(2), n = 1016). By electron microscopy, the neurons shared all the ultrastructural features of the cuneothalamic neurons (CTNs) which showed a slightly indented nucleus and a fairly rich cytoplasm containing well-developed Golgi apparatuses and rough endoplasmic reticulum (rER). The SOM immunoreaction product filled the cytoplasm of the neurons extending from the soma to the proximal and distal dendrites, which were postsynaptic to unlabeled boutons. In addition to soma and dendrites, SOM-IR boutons were also identified which made axodendritic synaptic contacts with SOM-IR dendrites. The SOM-IR neurons were characterized by using anti-SOM pre-embedding immunolabeling coupled with horseradish peroxidase (HRP) retrograde method, or SOM immunolabeling along with anti-glutamate, gamma-aminobutyric acid (GABA) or glycine post-embedding immunolabeling for identification of CTNs, glutamate-IR, GABA-IR and glycine-IR neurons, respectively. It was shown that more then 80% of the CTNs contained SOM and, furthermore, they contained glutamate but not GABA or glycine. On the basis of present findings, it is suggested the majority of the SOM-IR neurons in the rat CN are CTNs and that they may be involved in modulation of somatosensory synaptic transmission.

An electron microscopic study of the corticorubral fibers after neonatal deep cerebellar nuclear lesions in albino rats.

After a lesion in the sensorimotor and adjacent cortex in normal adult rats, degenerating terminals showing the dense reaction form asymmetrical contacts with spines, dendrites of various sizes, soma and other axonal terminals. Filamentous degeneration is also present. After neonatal deep cerebellar nuclear lesions involving the dentate nucleus and the adjacent interposed nucleus, the cerebrocorticorubral fibers form similar synaptic contacts with somatic, dendritic and axonal profiles. The incidence of axo-dendritic contacts on spine is reduced, while that of axo-dendritic contacts on small, medium-sized and large dendrites and axo-somatic contacts is increased.

An electron microscopic study of the cerebellorubral connections after neonatal lesions in the sensorimotor and adjacent cortex in the albino rat.

After a single right deep cerebellar nuclear (DCN) lesion in the dentate and adjacent interposed nuclei in adult rats, most degenerating cerebellorubral terminals in the rostral parvicellular portion of the contralateral red nucleus make contact with medium-sized and large dendrites. A few of them make contact with small dendrites, somata and other axonal terminals. The degenerating axons and terminals show the dense type of reaction. After left neonatal sensorimotor and adjacent cortical lesions followed by a right DCN lesion 2-10 months later, the pattern of most synaptic contacts resembles that in normal adult rats receiving only DCN lesions. There are, however, noteworthy differences. The incidence of axodendritic contact on dendritic spines is greatly increased. There is also an increase in the incidence of two terminals in close proximity and in contact with other neuronal profiles, and of one terminal in contact with more than one dendritic profile. The incidence of axodendritic contact on medium-sized and large dendrites is slightly reduced. The possibility of a taking over of cerebrocorticorubral projections by cerebellorubral fibers and of axonal and dendritic sprouting is suggested. In animals receiving double lesions, the filamentous type of degeneration could be seen. Degenerating dendrites occur in animals receiving single or double lesions.

The distribution and characterization of NADPH-d/NOS-IR neurons in the rat cuneate nucleus.

Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and nitric oxide synthase (NOS) immunohistochemistry have been used to characterize the nitric oxide (NO)-containing neurons in the rat cuneate nucleus. The present results showed that NADPH-d-positive/NOS-immunoreactive (-IR) neurons were distributed in the entire rostrocaudal extent of the nucleus. In the caudal region (approximately 1-2 mm caudal to the obex), NADPH-d/NOS-IR neurons were aggregated along the dorsal area of the nucleus notably in the lateral aspect. When traced rostrally, labeled neurons were progressively reduced and the cells were randomly distributed. The labeled neurons varied from round, ovoid to spindle-shaped with a mean profile area of about 140.1+/-1.7 microm(2) (n=720). They made up 7-10% of the neuronal population in the cuneate nucleus. By immunoelectron microscopy, the immunoreaction product was deposited throughout the cytoplasm extending from the soma to the proximal and distal dendrites. Results of NADPH-d staining paralleled that of NOS immunohistochemistry. Furthermore, NADPH-d reactivity and NOS-IR were colocalized in the same neurons following double labeling. Using NADPH-d histochemistry along with anti-gamma-aminobutyric acid (GABA) and -glycine postembedding immunolabeling for identification of GABA- and glycine-IR neurons, respectively, about 33% of the NADPH-d-positive neurons contained both GABA and glycine, 26% of them contained only glycine, while 41% of them showed neither GABA nor glycine labeling. Cuneothalamic neurons (CTNs) were identified by injecting the retrograde tracer Fluorogold (FG) into the ventrobasal complex of the thalamus. Numerous FG-labeled neurons were present in the contralateral cuneate nucleus, but none were reactive for NADPH-d. The present results suggest that approximately 60% of the NADPH-d/NOS-IR neurons in the cuneate nucleus are interneurons containing GABA and/or glycine.

A beta-fiber intensity stimulation of chronically constricted median nerve induces c-fos expression in thalamic projection neurons of the cuneate nucleus in rats with behavioral signs of neuropathic pain.

This study was aimed to investigate the possible involvement of neurons in the cuneate nucleus (CN) in the processing of A beta afferent inputs evoked by electrical stimulation of constricted median nerve in rats with behavioral signs of neuropathic pain. Immunohistochemical localization of Fos protein was used to examine the neuronal activation, and the combination of Fos immunohistochemistry with the retrograde labeling of Fluoro-Gold (FG) injected into the ventrobasal complex of the thalamus was used to characterize the activated neurons. Two weeks after unilateral median nerve constriction injury, the rats exhibited behavioral signs of neuropathic pain in the affected forepaws. In rats after nerve injury but without electrical stimulation, some Fos-like immunoreactive (Fos-LI) neurons were detected in the dorsal horn of the seventh cervical segment (C7) but none was found in the CN. Similar features were also noted when the stimulation of the intact median nerve served as an additional control. After A beta-fiber intensity stimulation of the previously constricted median nerve, an increase in number of Fos-LI neurons occurred in the medial half of the ipsilateral C7 dorsal horn as well as in the ipsilateral CN. In the latter, the Fos-LI neurons were located in the median nerve projection territory throughout the nucleus. Most of the Fos-LI neurons were distributed in the middle region of the CN, with about 78% of them emitting FG fluorescence indicating that they were cuneothalamic projection neurons. The results of this study suggest that the dorsal column-medial lemniscal system may contribute to the transmission and modulation of A beta-fiber mediated neuropathic pain signals.

Ultrastructural study of phenylethanolamine-N-methyltransferase, corticotropin-releasing factor and neurotensin immunoreactive neurons in the external cuneate nucleus of the gerbil.

The present study examined the existence of catecholamine-, corticotropin-releasing factor (CRF)- and neurotensin (NT)-containing neurons in the external cuneate nucleus (ECN) of the gerbil using single label pre-embedding immunocytochemistry in an attempt to shed light on the increasing evidence for autonomic involvement of the ECN. Peroxidase immunoreactivity of phenylethanolamine-N-methyl-transferase (PNMT), CRF or NT was identified in the heterogeneous population of the ECN neurons characterized by a deeply infolded nucleus. The label was localized in their somata, dendrites, myelinated axons and axon terminals. The immunolabelled dendrites were contacted by spherical (S) and flattened (F) types of presynaptic boutons containing spherical and flattened synaptic vesicles, respectively. The PNMT-labelled dendrites, however, were postsynaptic to an additional type of axon terminals containing pleomorphic (P) synaptic vesicles. Among the immunoreactive axon terminals, the PNMT-labelled boutons consisted of two types: S and F; in the CRF- and NT-labelled axon terminals, only the S type was observed. The catecholamine-containing ECN neurons differed from the CRF- and NT-immunoreactive neurons in their synaptic organization. The latter two were considered to be of the same cell population because of their similarities in ultrastructural features and synaptic relations. In view of a high frequency (48% for PNMT, 50% for CRF and 46% for NT) of the F-typed boutons associated with the three categories of immunolabelled neurons in the ECN, it is possible that they are under considerable inhibitory control. The presence of catecholamine, CRF and NT in the ECN suggests that the nucleus may be involved in the integration of proprioception-, exercise- or stress-evoked autonomic responses.

Synaptic relationships between corticocuneate terminals and glycine-immunoreactive neurons in the rat cuneate nucleus.

The present study describes the ultrastructural synaptic relationships between corticocuneate terminals (CCTs) and glycine-immunoreactive (glycine-IR) neurons in the cuneate nucleus of rats using anterograde tract-tracing of wheatgerm agglutinin conjugated with horseradish peroxidase (WGA-HRP) and anti-glycine immunoperoxidase labeling methods. The HRP-labeled CCTs made axodendritic synapses preferentially in the ventral part of the cuneate nucleus near the obex. In a total of 182 CCTs surveyed, 14 of them made direct synaptic contacts with immunoperoxidase-labeled glycine-IR dendrites. The present results suggest that cortical modulation on the sensory transmission of cuneate nucleus may be mediated through glycine-IR neurons.

Melatonin attenuates neuronal NADPH-d/NOS expression in the hypoglossal nucleus of adult rats following peripheral nerve injury.

Oxidative stress and massive production of nitric oxide (NO) have been implicated in the neuropathogenesis following peripheral nerve injury. This study was aimed to ascertain whether melatonin would exert its neuroprotective effect on the lesioned hypoglossal neurons after peripheral axotomy, since it is known to reduce the oxidative damage in a variety of experimental neuropathologies in which NO is involved. Right-sided hypoglossal nerve transection was performed in adult rats following which the animals were given two different doses of melatonin administered intraperitoneally for 3, 7, 14, 21 and 30 successive days. Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and neuronal nitric oxide synthase (nNOS) immunohistochemistry were carried out to detect the neuronal NADPH-d/NOS expression in the hypoglossal nucleus (HN). At various time intervals following axotomy, the neurons in the affected HN were induced to express NADPH-d/NOS reactivity on the lesioned side peaking at 14 days. However, the enzyme expression was markedly depressed by melatonin treatment in a dose-dependent manner in terms of frequency of labelled neurons and staining intensity. It is suggested that the suppressive effect of melatonin on NADPH-d/NOS expression may be attributed to its antioxidant properties. Hence, in consideration of therapeutic strategies for reducing the oxidative stress following peripheral nerve injury, melatonin may prove to be beneficial.

Efferent connections from the external cuneate nucleus to the medulla oblongata in the gerbil.

The present study revealed the efferent projections from the external cuneate nucleus (ECN) to various medullary nuclei in the gerbil as demonstrated in fresh living brainstem slices by using in vitro anterogradely tracing with the dextran-tetramethyl-rhodamine-biotin. The tracer-labelled ECN axon terminals were observed (1) in most of the vital autonomic-related nuclei: the nucleus solitary tractus, nucleus ambiguus, rostroventrolateral reticular nucleus and C2 adrenergic area, (2) in the reticular formation: the medullary, parvocellular, intermediate, gigantocellular, dorsal paragigantocellular and lateral paragigantocellular reticular nuclei and medullary linear nucleus, and (3) in sensory nuclei: the cuneate nucleus, spinal trigeminal nuclei caudalis and interpolaris, paratrigeminal nucleus, medial and spinal vestibular nuclei, inferior olive and prepositus hypoglossal nucleus. These new findings are discussed in relation to possible roles of the ECN in cardiovascular, respiratory and sensorimotor controls.

Intrathecally administered c-fos antisense oligodeoxynucleotide decreases formalin-induced nociceptive behavior in adult rats.

c-fos antisense strategy was applied as a pharmacological approach to characterize its dose-dependent role and reversibility in the reduction of formalin-induced hyperalgesia. Nociceptive behavioral responses (weighted score, flinching response, licking/biting) following formalin (50 microl 5%) injection were assessed in adult Wistar rats receiving different doses (50 nM, 250 nM) of intrathecally administered c-fos antisense oligodeoxynucleotides at different times prior to formalin injections. The treatments dose dependently decreased both Fos immunoreactivity expression in dorsal horn of rat lumbar spinal cord and all nociceptive measures in the tonic phase of the formalin test. c-Fos correlated well with weighted pain score and/or flinching responses, but not with licking/biting behavior. With the exception of a 48-120 h period required for licking/biting behavior to be restored to its normal status, the suppressive effect on c-fos expression and other nociceptive behaviors disappeared 48 h following c-fos antisense oligodeoxynucleotide treatment. The results suggest a pharmacological potential of c-fos antisense oligodeoxynucleotides in the central nervous system to block immediate-early genes and their resulting physiological consequence following noxious stimulus.

Protein synthesis inhibitor cycloheximide dose-dependently decreases formalin-induced c-Fos protein and behavioral hyperalgesia in rats.

We had previously demonstrated that c-fos antisense oligodeoxynucleotides dose-dependently suppressed formalin-induced c-Fos protein and behavioral hyperalgesia. To test whether de novo protein synthesis is required for the development of persistent pain after peripheral inflammation, we observed formalin-induced spinal c-Fos protein and nociceptive behaviors following pretreatment with cycloheximide, a protein synthesis inhibitor. Cycloheximide dose-dependently inhibited formalin-induced spinal c-Fos protein and tonic nociceptive responses. The possible non-specific effects other than protein synthesis inhibition on nociceptive behavior were carefully discussed and excluded. These results provide further support to the hypothesis that de novo protein synthesis is essential for the development of behavioral hyperalgesia.