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D Lima

Publications and source records attributed to D Lima.

At least 37 records · Page 2Linked to original sources

Activation by cutaneous or visceral noxious stimulation of spinal neurons projecting to the medullary dorsal reticular nucleus in the rat: a c-fos study.

The involvement of spinal neurons in the transmission of cutaneous and visceral nociceptive input to the medullary dorsal reticular nucleus was studied. Rats were injected with cholera toxin subunit B in the left dorsal reticular nucleus and subjected 4 days later to noxious mechanical, thermal or chemical stimulation of the proximal internal aspect of the left thigh, or to chemical stimulation of the urinary bladder. Sections of spinal segments T13-L3 were processed immunocytochemically for cholera toxin subunit B and Fos protein. The percentage of double-labelled cells in the population of Fos-positive cells was higher in lamina I (1-4%) than in deeper laminae (0-0.7%) following all stimuli. The percentage of double-labelled cells in the population of retrogradely labelled cells was 30-53% in lamina I and 0-5% in laminae III-X. Visceral stimulation activated more retrogradely labelled lamina I cells than any kind of cutaneous stimulation. Pyramidal cells were activated in higher numbers than multipolar and flattened cells after thermal cutaneous or visceral stimulation, and in lower numbers than multipolar cells after mechanical stimulation. These results suggest that, in the experimental conditions used, spinal cord cells conveying noxious input to the dorsal reticular nucleus are concentrated in lamina I. They further indicate that the spinal-dorsal reticular nucleus pathway plays a major role in the transmission of nociceptive visceral input, and point to the preferential involvement of pyramidal cells in cutaneous thermal and visceral processing.

Animals↗

The pontine A5 noradrenergic cells which project to the spinal cord dorsal horn are reciprocally connected with the caudal ventrolateral medulla in the rat.

A disynaptic pathway linking the caudal ventrolateral medulla (VLM) to the spinal cord via the A5 noradrenergic cell group of the pons has recently been described in the rat. In the present work, the projections of the A5 to the VLM and to the spinal dorsal horn were studied with double-tracing techniques combined with immunostaining of the noradrenaline-synthesizing enzyme dopamine-beta-hydroxylase. Cholera toxin subunit B (CTb) injected into the VLM and fluoro-gold injected into the spinal dorsal horn produced double retrograde labelling of A5 neurons immunoreactive for dopamine-beta-hydroxylase, which received appositions of fibre varicosities labelled anterogradely with CTb injected into the VLM. After injecting CTb into the A5, retrogradely labelled neurons occurred in the VLM. These neurons were contacted by anterogradely labelled fibres from the A5 group. These observations indicate that the VLM cells acting upon the A5 spinally projecting neurons, which are likely to exert an alpha2-adrenoreceptor-mediated inhibition on the spinal cord, are targeted by collaterals of the A5 spinal cord-bound axons. The A5-VLM pathway may be the anatomical substrate of a negative feedback circuit whereby the modulatory action of the VLM on the spinal cord is self-inhibited through activation of the A5.

Animals↗

The ventrolateral medulla of the rat is connected with the spinal cord dorsal horn by an indirect descending pathway relayed in the A5 noradrenergic cell group.

The pathway conveying the descending inhibitory noradrenergic input elicited from the caudal ventrolateral medulla (VLM) onto the spinal cord dorsal horn was studied in the rat. Retrograde labeling with cholera toxin subunit B (CTb) injected into the dorsal horn was combined with immunostaining for dopamine-beta-hydroxylase (DBH) in the VLM and other brainstem nuclei containing noradrenergic cells. CTb-labeled neurons occurred in the lateral part of the VLM (VLMlat), located ventrolaterally to the DBH-immunoreactive cells of the A1 noradrenergic cell group. Neuronal profiles stained for CTb and DBH (double labeled) occurred in the A5 (31%), A6 (57%), and A7 (12%) noradrenergic cell groups. To ascertain whether noradrenergic cells targeting the spinal cord in those groups received projections from the VLMlat, this area was injected with the anterograde tracer biotinylated dextran amine (BDA). Labeled terminal fibers with boutons en passant were apposed to numerous double-stained neurons in the A5 cell group. Similar appositions occurred in small amounts in the ventral subcoerulear component of the A6. Correlated light and electron microscopic analyses of the labeled appositions revealed that the BDA-labeled axonal boutons contained spherical vesicles and were presynaptic at asymmetrical contacts to somata and dendritic profiles of the double-stained A5 neurons. These data indicate the occurrence of an indirect dysynaptic pathway connecting the VLM to the spinal cord, with a relay in the A5 cells. This pathway may convey the antinociceptive effects mediated by alpha 2-adrenoreceptors, which have been previously observed in the spinal cord following VLM stimulation.

Animals↗

The medullary dorsal reticular nucleus facilitates acute nociception in the rat.

The influence on pain processing caused by destruction or stimulation of the dorsal reticular nucleus (DRt) was studied using the tail-flick and the increasing temperature hot-plate tests. Lesions of the DRt were obtained by injecting quinolinic acid (180 nmol/microliters) unilaterally or bilaterally, and nociceptive responses were evaluated by both tests. Following unilateral lesions, the tail-flick latencies and the hot-plate response temperatures were increased, values differing statistically from controls in the latter test. Bilateral lesions resulted in statistically significant increases of both tail-flick latency and hot-plate response temperature. Stimulation of the DRt was performed by injecting glutamate (100 nmol/microliters) unilaterally, which was followed 1 min later by a significant decrease in the tail-flick latency compared to saline injected controls. These results suggest that the DRt is involved in the facilitation of nociception after acute thermal noxious stimulation. This effect may be mediated through a spino-DRt-spinal loop causing a rebound of excitation in lamina I cells receiving noxious input from their own receptive field.

Animals↗

Hypertension inhibits noxious-evoked c-fos expression in the rat spinal cord.

The effect of hypertension on spinal induction of the c-fos proto-oncogene following noxious mechanical stimulation of the skin was studied in the rat. The occlusion of renal artery raised blood pressure steeply, reaching 52% over initial values. Oral administration of NG-nitro-L-arginine methyl ester provoked a gradual increase in blood pressure which reached up to 62%. The numbers of spinal dorsal horn Fos-immunoreactive nuclei were reduced to 66% and 38% of controls in animals with renal- and pharmacologically-induced hypertension, respectively. These data indicate that hypertension is accompanied by an inhibition of spinal nociceptive neurones which probably accounts for the hypoalgesia observed in hypertensive subjects. They further suggest an influence by the rate of increase of blood pressure on the level of spinal inhibition.

Animals↗

Projection sites of superficial or deep dorsal horn in the dorsal reticular nucleus.

The termination sites in the medullary dorsal reticular nucleus (DRt) of axons arising from superficial or deep dorsal horn cells were analysed by anterograde tracing with biotinylated dextran (BD). Following BD injections in spinal laminae I-III or in these laminae plus the dorsal funiculus, labelled fibres and boutons were located in the dorsal most ipsilateral portion of the DRt. Injections covering laminae III-V or laminae I-V resulted in bilateral labelling of the ventral DRt and ipsilateral labelling of the dorsal DRt. These findings, together with previous data concerning the origin of DRt projections to the superficial or deep dorsal horn, suggest the occurrence of reciprocal anatomical loops connecting the dorsal or the ventral areas of both regions.

Animals↗

Spinal c-fos expression is differentially induced by brief or persistent noxious stimulation.

The influence of stimulus duration on spinal induction of the c-fos proto-oncogene was analysed in the rat by pinching or heating the skin for periods varying from 20 s to 2 h. At stimulation periods shorter than 20 min, c-fos activation occurred in laminae I-IIi following mechanical stimulation and I-IIo following thermal stimulation. Mechanical stimulation produced delayed activation in laminae III-IV, V and VII at 30 min, 60 min and 2 h, respectively, and thermal stimulation in lamina IIi at 50 min. It is suggested that late c-fos activation signals inflammatory pain and is due to sensitization of primary afferent neurones.

Animals↗

Descending projections from the caudal medulla oblongata to the superficial or deep dorsal horn of the rat spinal cord.

The location of neurons in the caudal medulla oblongata that project to the superficial or deep dorsal horn was studied in the rat, by means of retrograde labelling from confined spinal injection sites. The tracer cholera toxin subunit B was injected into laminae I-III (fuve rats) or I-V (three rats) at C4-7 spinal segments. Neurons projecting to the superficial dorsal horn were located in the dorsomedial part of the dorsal reticular nucleus ipsilaterally, the subnucleus commissuralis of the nucleus tractus solitarius bilaterally, and a region occupying the lateralmost part of the ventrolateral reticular formation between the lateral reticular nucleus and the caudal pole of the spinal trigeminal nucleus, pars caudalis, bilaterally. Neurons projecting to the deep dorsal horn, which were only labelled when laminae I-V were filled by the tracer, occurred in the dorsomedial and ventrolateral parts of the dorsal reticular nucleus and in the ventral reticular nucleus bilaterally. A few cells were located in the above described lateralmost portion of the ventrolateral reticular formation bilaterally and in the ventral portion of the ipsilateral cuneate nucleus. In the light of previous data demonstrating that dorsal horn neurons project to the dorsal reticular nucleus, the ventrolateral reticular formation, and the nucleus tractus solitarius, and that neurons in these three medullary regions are involved in pain inhibition at the spinal level, the descending projections demonstrated here suggest the occurrence of spino-medullary-spinal loops mediating the analgesic actions elicited in each nucleus upon the arrival of nociceptive input from the dorsal horn.

Animals↗

Influence of electrical stimulation of locus coeruleus on the rat blood-brain barrier permeability to sodium fluorescein.

The role of central adrenergic innervation of the brain capillaries is still a matter of discussion. The hypothesis that these nerves control the blood-brain barrier permeability was tested by electrically stimulating the locus coeruleus, the major central adrenergic nucleus, in the anaesthetized rat. Frequencies of 5, 15, and 30 Hz were used. A frequency dependent increase in blood-brain barrier permeability to sodium fluorescein was verified. Prior administration of the alpha-adrenoceptor antagonist phenoxybenzamine (10 mg/kg i.p., 24 h before electrical stimulation) totally blocked the effect of 15 Hz stimulation. The same dose of pindolol (a beta-adrenoceptor antagonist) given 1 h before electrical stimulation potentiated the effect of 5 Hz stimulation. Thus, blood-brain barrier permeability is increased, in a frequency dependent manner, by electrical stimulation of the locus coeruleus. The results obtained with phenoxybenzamine and pindolol suggest an opposite effect of alpha and beta-adrenoceptors on the control of sodium fluorescein transport through the blood-brain barrier.

Adrenergic Fibers↗

Activation of the c-fos proto-oncogene in the spinal cord following noxious stimulation of the urinary bladder.

Activation of the c-fos proto-oncogene following mechanical or chemical noxious stimulation of the urinary bladder was studied at T12-L2 and L5-S1, the spinal cord segments of projection of the hypogastric nerve (HGN) and pelvic nerve (PN) fibers, respectively. In intact adult rats, c-fos expression was found at T12-L2 only in lamina I. At L5-S1, Fos cells occurred in lamina I, the intermediolateral gray matter (ILG), and the dorsal commissure (DCM). These two areas contained the highest number of immunoreactive cells. Although more Fos cells were induced by mechanical than by chemical stimulation, the distribution of the reactive neurons was similar after both types of stimuli. In adult rats that had been treated neonatally with capsaicin, there was a marked fall in c-fos activation by mechanical or chemical noxious stimuli in all immunoreactive areas. The loss of Fos cells was more pronounced in ILG and DCM at L5-S1 (95%) than in lamina I at the two spinal domains (70%). The confinement of c-fos activation to lamina I at T12-L2, the spinal cord domain of the HGN, suggests that the input carried from the bladder by this nerve is preferentially used for pain perception. The same function is expected for noxious input reaching lamina I at L5-S1, the spinal cord territory of termination of the PN. However, the striking number of Fos cells in ILG and DCM supports the important role played by this nerve in the control of the micturition reflex.(ABSTRACT TRUNCATED AT 250 WORDS)

Afferent Pathways↗

Neurons in the superficial dorsal horn of the rat spinal cord projecting to the medullary ventrolateral reticular formation express c-fos after noxious stimulation of the skin.

The nociceptive nature of the neurons of the superficial dorsal horn (laminae I-III) which project to the medullary ventrolateral reticular formation is studied in the rat. Medullary injections of Fluoro-Gold showed exclusive retrograde labeling of laminae I-III cells when the tracer filled a zone intermediate between the lateral tip of the lateral reticular nucleus and the spinal trigeminal nucleus, pars caudalis. This zone is here called VLMlat. Following noxious mechanical or thermal stimulation of the skin, double-labeled neurons, which stained retrogradely and were Fos-immunoreactive, prevailed in laminae I and IIo. Double-labeled neurons were few in lamina IIi after thermal stimulation and entirely lacking in lamina III after the two kinds of stimulation. Findings in lamina I confirm previous electrophysiological data (see Menétrey et al., J. Neurophysiol., 52 (1984) 595-611) showing that lamina I cells projecting to the ventrolateral reticular medulla convey noxious messages. The occurrence of numerous double-labeled cells in lamina IIo suggests that this lamina is also involved in nociceptive transmission to the VLMlat.

Animals↗

Periterminal synaptic organization of primary afferents in laminae I and IIo of the rat spinal cord, as shown after anterograde HRP labelling.

The fine structure and periterminal synaptology of the primary afferent terminations in laminae I and IIo are examined in the rat, following anterograde labelling with horseradish peroxidase applied to the right C5-dorsal root. Labelled varicosities observed along the terminal arbors in parasagittal thick sections were relocated in ultrathin sections by electron microscopy. The labelled terminal profiles generated by the three primary afferent plexuses which can be identified by light microscopy in laminae I-IIo had similar fine structural features, except that axo-axonal contacts, although rare, were more frequent in the medial network plexus. Primary boutons were packed with agranular spherical vesicles and some large granular vesicles, and were mostly presynaptic to profiles of dendritic trunks of marginal cells. Unlabelled axonal profiles, either light with some flattened vesicles, or dense with round vesicles, were also presynaptic at symmetrical or asymmetrical contacts, respectively, to those dendritic profiles. It is suggested that such knobs of intrinsic origin are responsible for postsynaptic modulation of the primary noxious input. Although the 20 microns wide lamina IIo belongs cytoarchitectonically to lamina II and can be distinguished from lamina I by a decreased amount of myelinated fibres and large dendritic profiles, the periterminal synaptology was here found to be the same as in lamina I.

Afferent Pathways↗

Descending projections from the medullary dorsal reticular nucleus make synaptic contacts with spinal cord lamina I cells projecting to that nucleus: an electron microscopic tracer study in the rat.

An ultrastructural study is made of the synaptic contacts occurring between structures labelled anterogradely and retrogradely in the superficial dorsal horn following injections of cholera toxin subunit B or horseradish peroxidase in the dorsal reticular nucleus of the medulla oblongata of the rat. Both tracers revealed labelled axonal boutons in lamina I with round synaptic vesicles and a few large granular vesicles making asymmetrical synaptic contacts upon labelled somata and dendrites. After injections of Phaseolus vulgaris leucoagglutinin in the dorsal reticular nucleus, labelled boutons identical to those revealed by the two other tracers were presynaptic to unlabelled somata and dendrites. In addition, dorsoreticular neurons were labelled retrogradely following injections of cholera toxin subunit B into the superficial dorsal horn of the cervical enlargement. These observations show the occurrence of a reciprocal connection between dorsal reticular and lamina I neurons. Considering the putative excitatory nature of the axodendritic contacts in lamina I, a positive feedback circuit is suggested, whereby the nociceptive signals transmitted to the dorsal medullary reticular formation by marginal neurons are intensified.

Afferent Pathways↗

Differential activation of c-fos in spinal neurones by distinct classes of noxious stimuli.

The laminar distribution of spinal cord neurones expressing immunoreactivity to the Fos protein was evaluated in the rat following chemical, thermal or mechanical noxious stimulation of the skin for 2 h. After stimulation by 20% or 5% formalin, Fos-immunoreactive neurones prevailed in lamina I where they accounted for 64% and 59%, respectively, of the entire population of Fos-immunoreactive spinal cells. Values in the remaining laminae were low (2-10%). Following thermal stimulation by radiant heat at 65 degrees C or 58 degrees C, Fos cells were concentrated in laminae I and IIo, amounting to 57% and 62%, respectively, in lamina I, and to 26% and 29% in lamina IIo. Values were lower than 10% in the remaining laminae. Following mechanical stimulation by pinching or needle prick, Fos-positive cells were regularly distributed throughout laminae I-V amounting to 25-26% in lamina I, and 10-20% in each of the remaining laminae. These findings suggest that the spinal neuronal groups upon by prolonged noxious stimulation differ according to the nature of the stimulus.

Animals↗

Structural types of spinal cord marginal (lamina I) neurons projecting to the nucleus of the tractus solitarius in the rat.

The structural types of spinal cord marginal (lamina I) neurons projecting to the nucleus of the tractus solitarius (NTS) were studied. Upon injections of cholera toxin subunit B (CTb) into the caudal part of the NTS, including its lateral and medial portions, labeled cells occurred bilaterally in laminae I, IV-VII, and X, and the lateral spinal nucleus (LSN). After injections into the lateral portion alone, only a few cells were labeled in laminae V, VII, and X, and the LSN, and none in the superficial dorsal horn. Of 1882 labeled marginal cells, 38% belonged to the flattened type, 37% to the pyramidal type, and 25% to the fusiform type. Flattened and pyramidal cells were labeled in considerably greater numbers than those reported when other supraspinal targets of these cells were injected with CTb. Since cells in the NTS are known to be under marked gamma-aminobutyric acidergic (GABA-ergic) inhibition, it is possible that only strong input conveyed by great numbers of flattened and pyramidal cells is capable of overcoming that barrier. Fusiform cells were labeled in numbers similar to those observed previously after tracer injections into the two other targets of this neuronal type, the parabrachial nuclei and the lateral reticular nucleus. Considering that these regions, as well as the NTS, control cardiovascular and respiratory functions, it is suggested that fusiform cells transmit noxious input that will influence autonomic reflexes processed in the three nuclei.

Afferent Pathways↗

Neurons in the substantia gelatinosa rolandi (lamina II) project to the caudal ventrolateral reticular formation of the medulla oblongata in the rat.

Following injections of cholera toxin subunit B in the caudal ventrolateral reticular formation, in an area between the lateral reticular nucleus and the ventrocaudal tip of the spinal trigeminal nucleus, pars caudalis, large numbers of retrogradely labelled cells occurred in lamina II, amounting to 28% and 21% of all labelled neurons in cervical and lumbar enlargements, respectively. Most lamina II cells presented dendritic arbors distributed as narrow radial sheets, resembling the central cells of Ramòn y Cajal. It is suggested that the ventrolateral bulbar reticular formation is the only significant supraspinal target of lamina II tract neurons.

Afferent Pathways↗