Distribution Of Fos-like Immunoreactivity Research Articles

Distribution of Fos-Like Immunoreactivity, Catecholaminergic and Serotoninergic Neurons Activated by the Laryngeal Chemoreflex in the Medulla Oblongata of Rats.

The laryngeal chemoreflex (LCR) induces apnea, glottis closure, bradycardia and hypertension in young and maturing mammals. We examined the distribution of medullary nuclei that are activated by the LCR and used immunofluorescent detection of Fos protein as a cellular marker for neuronal activation to establish that the medullary catecholaminergic and serotoninergic neurons participate in the modulation of the LCR. The LCR was elicited by the infusion of KCl-HCl solution into the laryngeal lumen of adult rats in the experimental group, whereas the control group received the same surgery but no infusion. In comparison, the number of regions of Fos-like immunoreactivity (FLI) that were activated by the LCR significantly increased in the nucleus of the solitary tract (NTS), the vestibular nuclear complex (VNC), the loose formation of the nucleus ambiguus (AmbL), the rostral ventral respiratory group (RVRG), the ventrolateral reticular complex (VLR), the pre-Bötzinger complex (PrBöt), the Bötzinger complex (Böt), the spinal trigeminal nucleus (SP5), and the raphe obscurus nucleus (ROb) bilaterally from the medulla oblongata. Furthermore, 12.71% of neurons with FLI in the dorsolateral part of the nucleus of the solitary tract (SolDL) showed tyrosine hydroxylase-immunoreactivity (TH-ir, catecholaminergic), and 70.87% of neurons with FLI in the ROb were serotoninergic. Our data demonstrated the distribution of medullary nuclei that were activated by the LCR, and further demonstrated that catecholaminergic neurons of the SolDL and serotoninergic neurons of the ROb were activated by the LCR, indicating the potential central pathway of the LCR.

Robust Up-Regulation of Nuclear Red Fluorescent-Tagged Fos Marks Neuronal Activation in Green Fluorescent Vasopressin Neurons after Osmotic Stimulation in a Double-Transgenic Rat

The up-regulation in the expression of mRNA or protein encoded by the c-fos gene is widely used as a marker of neuronal activation elicited by various stimuli. To facilitate the detection of activated neurons, we generated transgenic rats expressing a fusion gene consisting of c-fos coding sequences in frame with monomeric red fluorescent protein 1 (mRFP1) under the control of c-fos gene regulatory sequences (c-fos-mRFP1 rats). In c-fos-mRFP1 transgenic rats, 90 min after hypertonic saline ip administration, nuclear mRFP1 fluorescence was observed abundantly in brain regions known to be osmosensitive, namely the median preoptic nucleus, organum vasculosum lamina terminalis, supraoptic nucleus, paraventricular nucleus, and subfornical organ. Immunohistochemistry for Fos protein confirmed that the distribution of Fos-like immunoreactivity in nontransgenic rats was similar to those of mRFP1 fluorescence after ip administration of hypertonic saline in the transgenic rats. Several double-transgenic rats were obtained from matings between transgenic rats expressing an arginine vasopressin-enhanced green fluorescent protein fusion gene (AVP-eGFP rats) and c-fos-mRFP1 rats. In these double-transgenic rats, almost all eGFP neurons in the supraoptic nucleus and PVN expressed nuclear mRFP1 fluorescence 90 min after hypertonic saline administration. The c-fos-mRFP1 rats are a powerful tool that enables the facile identification of activated neurons in the nervous system. Furthermore, when combined with transgenes expressing another fluorophore under the control of cell-specific regulatory sequences, activation of specific neuronal cell types in response to physiological cues can be readily detected.

Taste Reactivity and Fos Expression in GAD1-EGFP Transgenic Mice

The central distribution of QHCl-elicited Fos-like immunoreactivity (FLI) suggests the location of a brain stem circuit that controls the oral rejection response. Although many species display an oral rejection response to bitter stimuli, the distribution of FLI associated with this response has been investigated only in rats. Fos data are minimal for the mouse, a species of increasing importance, due to its use in molecular and transgenic studies and taste-evoked oromotor responses are also only incompletely described in these rodents. We investigated these questions in FVB/NJ mice and a related transgenic strain (FVB-Tg(GadGFP)4507) that expresses green fluorescent protein in a subset of GAD1-containing neurons. QHCl, sucrose, or water delivered through intraoral cannulae yielded behavioral profiles that clearly differentiated QHCl from sucrose. Similar to rat, the number of neurons expressing FLI in the medial third of the solitary nucleus was elevated following QHCl compared with the other stimuli. In mice expressing green fluorescent protein, there was a pronounced distribution of GABAergic neurons in the ventral half of the solitary nucleus. Approximately 15% of solitary neurons expressing Fos were GABAergic, but this proportion did not differ according to stimulus.

Effects of cholecystokinin (CCK)-8 on hypothalamic oxytocin-secreting neurons in rats lacking CCK-A receptor

Peripheral administration of cholecystokinin (CCK)-8 selectively activates oxytocin (OXT)-secreting neurons in the supraoptic (SON) and the paraventricular nuclei (PVN) with the elevation of plasma OXT level in rats. We examined the effects of intravenous (iv) administration of CCK-8 on the neuronal activity of hypothalamic OXT-secreting neurons and plasma OXT level in Otsuka Long–Evans Tokushima Fatty (OLETF) rats that have a congenital defect in the expression of the CCK-A receptor gene. In situ hybridization histochemistry (ISH) for c- fos mRNA revealed that the expression of the c- fos gene was not induced in the SON, the PVN, the nucleus of the tractus solitarius (NTS) and the area postrema (AP) 30 min after iv administration of CCK-8 (20 and 40 μg/kg) in OLETF rats. In Long–Evans Tokushima Otsuka (LETO) rats (controls), c- fos mRNA was detected abundantly in those nuclei 30 min after iv administration of CCK-8 (20 μg/kg). Immunohistochemistry for c- fos protein (Fos) showed that the distributions of Fos-like immunoreactivity (LI) were identical to the results obtained from ISH. Dual immunostaining for OXT and Fos revealed that Fos-LI was mainly observed in OXT-secreting neurons in the SON and the PVN of LETO rats 90 min after iv administration of CCK-8 (20 μg/kg). Radioimmunoassay for OXT and arginine vasopressin (AVP) showed that iv administration of CCK-8 did not cause significant change in the plasma OXT and AVP levels in OLETF rats, while iv administration of CCK-8 caused a significant elevation of plasma OXT level without changing the plasma AVP level in LETO rats. These results suggest that peripheral administration of CCK-8 may selectively activate the hypothalamic OXT-secreting neurons and brainstem neurons through CCK-A receptor in rats.

Distribution of fos-like immunoreactivity in the auditory pathway evoked by bipolar electrical brainstem stimulation

Objective—The auditory brainstem implant (ABI) represents a new modality for the treatment of patients deafened as a result of complete excision of a bilateral VIIIth nerve tumor. However, little work has been done on the effect of the ABI on the mammalian auditory pathway. The aim of this study was to demonstrate the effect of the ABI using Fos-like immunoreactivity.Material and Methods—A bipolar electrode was implanted in the dorsal cochlear nucleus of bilaterally deafened Sprague–Dawley rats, and electrical stimulation was presented at an intensity four times that of threshold.Results—Fos-like immunoreactivity was induced in the neurons of various auditory brainstem nuclei and observed in the low-to-middle frequency area. In the ipsilateral dorsal cochlear nucleus, ipsilateral posterior ventral cochlear nucleus and bilateral inferior colliculus, Fos-like immunoreactive neurons were observed as a distinct banding pattern.Conclusions—This study showed that Fos-like immunoreactivity was observed in the restricted area of the primary brainstem auditory pathway with the appropriate tonotopicity. These results indicate that the ABI can provide auditory information suitable for speech recognition.

Isoflurane, but not halothane, depresses c-fos expression in rat spinal cord at concentrations that suppress reflex movement after supramaximal noxious stimulation.

We investigated the effects of isoflurane and halothane on the induction of fos-like immunoreactivity (FLI) in the rat lumbosacral spinal cord after supramaximal noxious mechanical stimulation of the hindpaw. Compared with unstimulated controls (0.9% isoflurane), noxious stimulation at 0.9%-1.5% elicited significant (0.9%-1.5% isoflurane) increases in FLI bilaterally. FLI was distributed mainly in the superficial dorsal horn (laminae I-III) and, to a lesser extent, in the deep dorsal horn (laminae IV-VI) and intermediate zone (lamina VII), with three- to fivefold greater labeling ipsilaterally. At 1.8% isoflurane, mean FLI counts in all laminar regions were significantly smaller (1.7 +/- 1.3 per section) compared with the other concentrations (11.4 +/- 9.5, 7.5 +/- 6.8, and 9.7 +/- 6.6 at 0.9%, 1.2%, and 1.5%, respectively) but were not different from unstimulated controls. At sacral levels, we observed a bilateral distribution of FLI primarily in superficial laminae in unstimulated controls that was not significantly different at any isoflurane concentration. FLI counts were not significantly different across groups receiving halothane (0.9%-1.5%). FLI was reduced only at isoflurane concentrations that depressed both gross, purposeful movement and reflex withdrawal, whereas halothane did not cause depression even at concentrations that depressed withdrawal reflexes. Isoflurane and halothane may have differing effects on neuronal function and responses to noxious stimulation. Isoflurane depressed neuronal activity in the spinal cord as measured with fos-like immunoreactivity (FLI), but this occurred only when reflex withdrawal responses were abolished. Halothane, however, did not depress FLI, even at concentrations sufficient to block reflex withdrawal. These two anesthetics may have differing effects on neuronal function and responses.

Quinine and citric acid elicit distinctive Fos-like immunoreactivity in the rat nucleus of the solitary tract.

The present experiment investigated Fos-like immunoreactivity (FLI) in the nucleus of the solitary tract (NST) after intraoral infusions of 0.1 M citric acid, 0.3 M NaCl, and 0.3-30 mM quinine monohydrochloride (QHCl) in awake, behaving rats. Increases in QHCl concentration produced increases in the numbers of FLI-labeled neurons in the rostral part of the intermediate (i(r)) and rostral (r) NST, but the topographic distribution of FLI was consistent across QHCl concentrations and distinctive compared with effects of citric acid. Quinine elicited FLI concentrated in the medial third of the nucleus; acid elicited more broadly distributed FLI concentrated farther laterally. Surprisingly, in contrast to QHCl and citric acid, NaCl produced FLI that was indistinguishable from that produced by water. Although the functional significance of these patterns is unknown, citric acid and QHCl are nonpreferred stimuli but produced different oromotor behaviors. QHCl (30 mM) elicited approximately 3.2 times as many gapes as citric acid (0.1 M), and acid elicited more ingestive responses. Parallel differences in FLI expression suggest that different NST regions may have distinctive roles in triggering oromotor behaviors.

C-fos induction in rat superficial dorsal horn following cutaneous application of noxious chemical or mechanical stimuli

The method of c-fos immunodetection was used to map the distributions of neurons in the lumbar spinal dorsal horn that were activated following intracutaneous (i.c.) microinjection, or iontophoretic application, of different irritant chemicals to the lateral hindpaw of rats. Microinjections (1 microl) of histamine, serotonin (5-HT), nicotine, capsaicin, or formalin each elicited similar distributions of Fos-like immunoreactivity (FLI) in laminae I-II of the ipsilateral superficial dorsal horn, with little or no FLI in deeper laminae or contralaterally. In laminae I and II, FLI cell counts were significantly higher following i.c. histamine, 5-HT, capsaicin, formalin, and noxious pinch, compared to i.c. saline controls. Capsaicin-evoked FLI was dose-dependent. Multivariate analysis of variance revealed no significant difference in spatial distributions of FLI induced by any of the chemical or pinch stimuli. Iontophoretic application of histamine, 5-HT, or nicotine also elicited similar distributions of FLI in the superficial dorsal horn, and cell counts of FLI were significantly higher compared to controls receiving iontophoretic vehicle (methyl cellulose). These results indicate either that individual laminae I-II neurons are activated by each of the irritant chemicals, or that neurons selectively responsive to a given irritant are comingled without any apparent laminar segregation.

CFos induction during conditioned taste aversion expression varies with aversion strength

Fos-like immunoreactivity (FLI) can indicate the location of neurons activated following expression of conditioned taste aversion (CTA). After one conditioning trial FLI has been identified in the intermediate nucleus of the solitary tract (iNTS) with little expression in other brain regions. The present study assessed the effect of increasing aversion strength on the magnitude and anatomical distribution of FLI during CTA expression. When animals received three rather than one conditioning trial, significant FLI was seen not only in the iNTS but also in the parabrachial nucleus (PBN), and the central nucleus of the amygdala (CNA), regions thought to be important in taste aversion learning.

Distribution of Fos-like immunoreactivity within the rat brain following intraventricular injection of the selective NK(3) receptor agonist senktide.

Neurokinin B (NKB) is one member of an evolutionarily conserved family of neuropeptides, the tachykinins. Preferential binding of NKB to endogenous NK(3) receptors affects a variety of biological and physiological processes, including endocrine secretions, sensory transmission, and fluid and electrolyte homeostasis. In light of its widespread biological actions, immunohistochemical detection of the c-Fos protein product was used to study the distribution of neuronal activation in the rat brain caused by intraventricular (icv) injections of the selective NK(3) receptor agonist (succinyl-[Asp(6), N-Me-Phe(8)] substance P [6-11]), senktide. Quantitative analysis revealed that treatment with isotonic saline or 200 ng senktide resulted in the differential expression of Fos-like immunoreactivity (FLI) throughout the brain. Senktide induced the highest number of FLI neurons in the lateral septum, bed nucleus of the stria terminalis, amygdala, paraventricular nucleus of the hypothalamus, median preoptic nucleus, organum vasculosum of the lamina terminalis, supraoptic nucleus, periaqueductal gray, and medial nucleus of the solitary tract compared to isotonic saline controls. Additional regions that contained elevated FLI following icv injection of senktide, relative to saline injection, included the cerebral cortex, lateral hypothalamic nucleus, suprachiasmatic nucleus, ventral tegmental area, substantia nigra, inferior colliculus, locus coeruleus, zona incerta, and arcuate nucleus. Our data indicate that activation of NK(3) receptors induces the expression of FLI within circumscribed regions of the rat brain. This pattern of neuronal activation overlaps with nuclei known to regulate homeostatic processes, such as endocrine secretion, cardiovascular function, salt intake, and nociception.

Distribution of Fos-like immunoreactivity in guinea-pig brain following administration of the neurokinin-1 receptor agonist, [Sar 9,Met(O 2) 11]substance P

The tachykinins are a family of peptides with putative neurotransmitter roles in the nervous system. They mediate their effects via neurokinin-1, neurokinin-2 and neurokinin-3 receptors. There has been increasing interest in the therapeutic application of the tachykinin neurokinin-1 receptor antagonists in the treatment of pain and emesis, and more recently in depression. However, the central role of neurokinin-1 receptors is not well understood. The aims of the present study were to determine the behavioural responses of guinea-pigs, and the distribution of Fos-like immunoreactivity in the guinea-pig brain, following intracerebroventricular administration of the neurokinin-1 receptor-selective agonist, [Sar 9,Met(O 2) 11]substance P. The effects of pretreatment with the neurokinin-1 receptor antagonist, SR 140333, were also investigated. Administration of [Sar 9,Met(O 2) 11]substance P induced increased locomotor activity, as well as face washing, grooming and wet-dog shake behaviours, all of which were inhibited by the neurokinin-1 receptor antagonist, SR 140333, indicating the involvement of neurokinin-1 receptors. In order to localize the brain regions activated by [Sar 9,Met(O 2) 11]substance P, the distribution of neurons expressing Fos-like immunoreactivity was examined. [Sar 9,Met(O 2) 11]substance P induced increased Fos-like immunoreactivity in widespread areas, including the frontal cortex, hippocampus, amygdala, thalamus, hypothalamus, periaqueductal gray, area postrema and nucleus of the solitary tract. SR 140333 reduced Fos-like immunoreactivity induced by [Sar 9,Met(O 2) 11]substance P in most areas. Thus, brain regions associated with emotion, sensation, learning and memory, autonomic regulation and emesis were activated by stimulation of neurokinin-1 receptors. The present data have added a functional domain to previous neurokinin-1 receptor localization studies by describing the extensive regions of the CNS that may be activated by stimulation of these receptors, and the potential of neurokinin-1 receptor antagonists to inhibit activation of these regions.

Fos-like immunoreactivity in the brain stem following oral quinine stimulation in decerebrate rats.

The present study compared the distribution of Fos-like immunoreactivity (FLI) following intraoral stimulation with quinine monohydrochloride (QHCl) in awake intact rats to the pattern obtained in chronic supracollicular decerebrate (CD) rats. Because the behavioral rejection response to QHCl is evident in the CD rat, it was hypothesized that the pattern of FLI in the lower brain stem should be similar in both groups. Overall, the distribution of FLI in the brain stem was quite similar in both intact and CD groups, and QHCl stimulation increased FLI in the rostral (gustatory) nucleus of the solitary tract, the parabrachial nucleus (PBN), and the lateral reticular formation (RF) compared with an unstimulated control group. The CD group differed from the intact group, however, with a trend toward less FLI in the RF and a shift in the pattern of label away from the external subdivision of the PBN. CD rats also had increased FLI in the caudal nucleus of the solitary tract, with or without intraoral infusions. The distribution of QHCl-induced FLI in the brain stem of intact rats thus indicates both local sensorimotor processing as well as the influence of forebrain structures.

Fos expression in the ferret trigeminal nuclear complex following tooth pulp stimulation

The aim of this study was to establish which regions of the trigeminal nucleus are activated by tooth pulp stimulation in the normal ferret. The distribution of Fos-like immunoreactivity was examined following electrical stimulation of the tooth pulp in the awake and anaesthetized ferret. Stimulus-specific labelling was found in subnuclei caudalis and oralis of the trigeminal spinal nucleus. Three groups of chronically prepared animals; conscious, anaesthetized (alphaxolone/alphadolone) and anaesthetized-paralysed (alphaxolone/alphadolone with gallamine triethiodide), received electrical stimuli to both the upper and lower left canine teeth (1 Hz train of 3 x 0.5 ms at 200 Hz) at an amplitude of 10 times the threshold of the jaw opening reflex. Three control groups were treated identically except no stimulus was given. In stimulated anaesthetized and anaesthetized-paralysed animals, Fos-positive profiles were seen in laminae I and II of subnucleus caudalis and in the medial part of subnucleus oralis. There was no labelling evident in subnucleus interpolaris or the main sensory nucleus, or contralaterally in any of the subnuclei. In all conscious stimulated animals there was additional bilateral Fos-positive labelling, mainly in the deeper laminae of subnucleus caudalis. This bilateral labelling was not stimulus-specific as it was also seen in conscious non-stimulated animals. After correction for this bilateral labelling no significant difference wasfound between conscious, anaesthetized and anaesthetized-paralysed groups of stimulated animals or between the different groups of control animals. These results support the concept that the rostral parts of the trigeminal spinal nucleus are involved in processing of nociceptive input. They also demonstrate that light alphaxolone/alphadolone anaesthesia has no effect on stimulus-specific Fos expression following tooth pulp stimulation. The second aim of this study was to develop a clearly defined model for future studies in which Fos expression is no different to that seen in the conscious state. As in the conscious animal, labelling not associated with the stimulus is difficult to distinguish from stimulus specific labelling, further studies using this model of trigeminal nociceptive pathways would be best carried out in lightly anaesthetized animals.

Localization of Fos-like immunoreactivity induced by the NK3 tachykinin receptor agonist, senktide, in the guinea-pig brain.

1. The effects of intracerebroventricular (i.c.v.) administration of the NK3 tachykinin receptor agonist, senktide (10 nmol each side), in guinea-pigs pretreated with the selective NK3 tachykinin receptor antagonist, SR142801 (3 mg kg(-1) subcutaneous, s.c., 30 min before senktide), or its less active enantiomer, SR142806 (3 mg kg(-1) s.c. 30 min before senktide), on behaviour and on the distribution of Fos-like immunoreactivity (Fos-LI) in central neurones were investigated. Guinea-pigs were chosen for the study since they possess NK3 tachykinin receptors with pharmacological characteristics similar to those in man. 2. Wet-dog shakes, but not locomotor activity, elicited by senktide i.c.v. were significantly reduced by SR142801 but not by SR142806, confirming the involvement of NK3 tachykinin receptors in wet-dog shake behaviour. 3. Senktide induced increased numbers of Fos-LI neurones in the following brain areas: frontal, parietal and piriform cortex, the lateral septum, the CA1, CA2, subiculum and dentate gyrus of the hippocampus, most areas in the amygdala, thalamus and hypothalamus, medial geniculate nucleus and the ventral cochlear nucleus. Pretreatment with SR142801, but not with SR142806, before administration of senktide inhibited Fos-LI expression in the cingulate cortex, dentate gyrus of the hippocampus, some regions of the thalamus, hypothalamus and amygdala and the ventral cochlear nucleus. 4. The present results are the first demonstration that senktide induces Fos-LI in widespread areas of the guinea-pig brain. It is proposed that NK3 tachykinin receptors may play a more extensive role in the control of diverse brain functions, including cortical processing, learning and memory, neuroendocrine and behavioural regulation, than is currently recognized.

Distribution of Fos-Like Immunoreactivity in the Medullary Reticular Formation of the Rat after Gustatory Elicited Ingestion and Rejection Behaviors

The distribution of neurons in the medullary reticular formation (RF) activated by the ingestion of sucrose or rejection of quinine was examined using standard immunohistochemical techniques to detect the expression of the Fos protein product of the immediate-early gene c-fos. Double-labeling techniques were used to gain further insight into the possible functional significance of RF neurons exhibiting Fos-like immunoreactivity (FLI). Compared with sucrose and unstimulated controls, quinine elicited significantly more FLI neurons in three specific RF subdivisions: parvocellular reticular nucleus (PCRt), intermediate reticular nucleus (IRt), and dorsal medullary reticular nucleus (MdD). Moreover, the number of FLI neurons in the RF of quinine-stimulated animals was significantly correlated with the degree of oromotor activity. Thus, the distinct distribution of FLI neurons throughout the RF after quinine may reflect the activation of a specific oral rejection circuit. The double-labeling results indicated a high degree of segregation between FLI neurons and premotor projection neurons to the hypoglossal nucleus (mXII) retrogradely labeled with Fluorogold. Thus, although there were a significant number of double-labeled neurons in the RF, the major concentration of premotor projection neurons to mXII in IRt were medial to the preponderance of FLI neurons in the PCRt. In contrast, there was substantial overlap between FLI neurons in the RF and labeled fibers after injections of the anterograde tracer, biotinylated dextran into the rostral (gustatory) portion of the nucleus of the solitary tract. These results support a medial (premotor)/lateral (sensory) functional topography of the medullary RF.

Analysis of nociceptive neurones in the rat superior colliculus using c-fos immunohistochemistry.

The superior colliculus (SC) has an established role in the sensory guidance of motor commands required to orient an animal towards novel stimuli. In addition to the representations of visual, auditory, and somatosensory stimuli, the SC also contains a large population of nociceptive neurones. The purpose of the present investigation was to see if nociceptive neurones in the SC can be characterised with c-fos immunohistochemistry as a prelude to establishing anatomical connectivity with specific target regions in the brainstem. To ensure comparability with previous electrophysiological investigations, the present study was conducted in animals anaesthetised with urethane. A series of independent issue relating to basic aspects of experimental protocol were investigated. The principal findings were: (i) Despite minimising the exposure of animals to extraneous stimuli, basal levels of immunostaining were observed. (ii) Urethane anaesthesia induced an increase in Fos-like immunoreactivity (FLI) over the basal condition. (iii) No additional labelling was induced by non-noxious tactile stimulation of the hindpaw. (iv) Unilateral noxious mechanical stimulation elicited a reliable increase in FLI over all control conditions. (v) This increase in FLI was expressed bilaterally and restricted largely to the intermediate white layer. (vi) The induction of FLI was related to noxious stimulation intensity. (vii) No reliable differences in the spatial topography of FLI expression were observed when unilateral noxious mechanical stimulation was administered to the face or hind foot. (vii) A higher level of urethane anaesthesia had a generally suppressive effect on FLI expression. (ix) There were no differences in the distribution of FLI induced by noxious mechanical or noxious chemical stimulation. (x) The increase in FLI induced by noxious pinch was abolished by a naloxone reversible pre-treatment with morphine. These data confirm that c-fos immunohistochemistry can be used to characterise nociceptive cells in the rat superior colliculus, and generally complement recent electrophysiological data. The identification of nociceptive cells in the stratum album intermediale, the source of collicular input to regions of the contralateral brainstem involved in orienting, suggests the SC may play a significant role in the localisation of pain.

Distribution of Fos-like immunoreactivity in the caudal medullary reticular formation following noxious facial stimulation in the rat.

To investigate the topographic organization of nociceptive neurons in the caudal medullary reticular formation, the distribution of cells that exhibit c-fos expression was examined following a unilateral noxious facial stimulus: subcutaneous injection of formalin into the vibrissal pad of awake rats. Labelling for Fos-like immunoreactivity was present in a somatotopic distribution in a region of the lateral reticular formation adjacent to trigeminal nucleus caudalis, which corresponds approximately to lamina V of the medullary dorsal horn. Labelling in adjacent regions of the reticular formation showed no somatotopy but was predominantly ipsilateral. Contralateral labelling was concentrated ventrolaterally around the lateral reticular nucleus and dorsally near the nucleus of the solitary tract.

Distribution of Fos-like immunoreactivity in the caudal brainstem of the rat following noxious chemical stimulation of the temporomandibular joint.

Central expression of the protooncogene c-fos was used to examine areas receiving noxious sensory input from the rat temporomandibular joint (TMJ). Fos-like immunoreactivity (Fos-LI) in the caudal brainstem was visualized 2 hours after unilateral injection of the small-fiber-specific excitant/inflammatory irritant mustard oil into the TMJ region. Control animals received injection of either mustard oil into the subcutaneous fascia overlying the masseter muscle or mineral oil vehicle into the TMJ region. In all groups, Fos-LI was consistently observed ipsilaterally in the spinal trigeminal nucleus and cervical dorsal horn and, bilaterally, in the nucleus of the solitary tract and the ventrolateral medulla. The expression of Fos-LI ipsilaterally in the paratrigeminal nucleus was variable. Within the trigeminal sensory complex, Fos-LI was restricted to subnucleus caudalis and the caudal portions of subnucleus interpolaris near the level of the obex. Approximately 12% of Fos-LI cells in subnucleus caudalis and in the cervical dorsal horn were found in laminae III-VI. Compared to TMJ mustard oil injection, mineral oil injection produced less Fos-LI at all rostrocaudal levels, whereas subcutaneous mustard oil injection produced less Fos-LI in caudal subnucleus caudalis but similar amounts in the cervical dorsal horn. Neither of these injections yielded significant ipsilateral responses in subnucleus caudalis, indicating that Fos-LI in this region following TMJ mustard oil injection could be ascribed solely to small-fiber stimulation in the deep TMJ region. The wide rostrocaudal distribution of Fos-LI within the caudal brainstem reflects the distribution of TMJ-responsive nociceptive neurons that may underlie the spread and referral of pain from the TMJ region.

Caudal portions of the spinal trigeminal complex are necessary for autonomic responses and display Fos-like immunoreactivity after corneal stimulation in the cat

Corneal input to the spinal trigeminal nucleus (Vsp) was assessed by examining Fos-like immunoreactivity (Fos-LI) after chemical irritant stimulation by mustard oil in chloralose-anesthetized cats. The distribution of Fos-LI within the ipsilateral Vsp was bimodal: a dominant group of cells within the superficial laminae at caudal levels of subnucleus caudalis and a second group of cells within the ventrolateral pole of Vsp at obex levels and within the adjacent interstitial islands. Few Fos-positive cells were seen within the Vsp rostral to the mid-portion of subnucleus interpolaris or within the contralateral Vsp. To assess the involvement of caudal portions of the Vsp in mediating the adrenal and autonomic responses to corneal stimulation, mustard oil was applied before and after lidocaine blockade of the Vsp at obex levels in a second group of cats. Corneal stimulation alone increased significantly ( P < 0.001) the adrenal secretion of catecholamines, adrenal blood flow, mean arterial pressure and heart rate. With the exception of heart rate, the adrenal and autonomic responses to mustard oil were greatly attenuated or abolished by lidocaine blockade of the ipsilateral Vsp at the level of the obex, a region that displayed a high number of Fos-positive cells after corneal stimulation. These results indicate that neurons within the Vsp at or more caudal than the level of the obex process chemical irritant input from the cornea and are necessary for corneal-evoked changes in adrenal and autonomic function.

Distribution of fos-like immunoreactivity in the medullary and upper cervical dorsal horn produced by stimulation of dural blood vessels in the rat

Neurophysiological studies have generally failed to find evidence of a specific ascending pathway for visceral nociception. However, pain that arises from deep or visceral tissues typically differs from cutaneous pain, particularly in its diffuse, poorly localized quality. In this study, the c-fos mapping technique was used in order to investigate possible differences in the distribution of central neurons activated by afferent pathways from cutaneous and deep tissues that may be related to the differing quality of the sensations they evoke. The distribution of neurons in the upper cervical and medullary dorsal horn that displayed fos-like immunoreactivity (fos-LI) was examined following mechanical stimulation of dural blood vessels (transverse and superior sagittal sinuses), and was compared to that found following mechanical, thermal, and chemical stimulation of facial sites. Dural stimulation was carried out Brevital anesthesia in rats that had received a chronic surgical exposure of the transverse and superior sagittal sinuses 2 d earlier. Localized mechanical stimulation of the dural surface of the transverse sinus produced a predominantly ipsilateral increase in the number of fos-LI neurons in the medullary and upper cervical dorsal horn (primarily laminae I and V), and in the transition region between the trigeminal nucleus caudalis and interpolaris. Stimulation of the superior sagittal sinus produced increases in fos-LI labeling that were generally smaller than those produced by transverse sinus stimulation. The distribution of fos-LI labeling in the dorsal horn induced by dural stimulation differed from that induced by facial stimulation in two ways. (1) Dural stimulation produced a more diffuse distribution of fos-LI than facial stimulation in the dorsal horn. Whereas facial stimulation produced a dense, localized zone of fos-LI labeling in the dorsal horn, dural stimulation produced fos-LI labeling that extended from the midlevel of caudalis to C2/C3, and also extended across a large portion of the ventrolateral-to-dorsomedial axis of the dorsal horn. This distribution roughly corresponds to the representation of most of the dorsal half of the head and face. (2) Dural stimulation produced a more restricted laminar distribution of fos-LI labeling than facial stimulation, in that the dural-induced labeling in the superficial dorsal horn was primarily restricted to lamina I, whereas facial stimulation typically induced substantial labeling in both lamina I and the outer part of lamina II. These differences in the central organization of the afferent pathways from dural and facial sites may contribute to the differences in the quality of sensations evoked by these pathways.