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S A Shehab

Publications and source records attributed to S A Shehab.

At least 19 recordsLinked to original sources
PubMed2000

Neurokinin 1 receptor expression by neurons in laminae I, III and IV of the rat spinal dorsal horn that project to the brainstem.

Large neurons in laminae III and IV of the spinal cord which express the neurokinin 1 receptor and have dendrites that enter the superficial laminae are a major target for substance P (SP)-containing (nociceptive) primary afferents. Although some of these neurons project to the thalamus, we know little about other possible projection targets. The main aim of this study was to determine whether all cells of this type are projection neurons and to provide information about brainstem sites to which they project. Injections of cholera toxin B subunit were made into four brainstem areas that receive input from the spinal cord, and the proportion of cells of this type in the L4 spinal segment that were retrogradely labelled was determined in each case. The results suggest that most of these cells (>90%) project to the contralateral lateral reticular nucleus (or to a nearby region), while many (>60%) send axons to the lateral parabrachial area and some to the dorsal part of the caudal medulla. However, few of these cells project to the periaqueductal grey matter. As lamina I neurons with the neurokinin 1 receptor appear to be important in the generation of hyperalgesia, we also examined projection neurons in this lamina and found that for each injection site the great majority possessed the receptor. These results demonstrate that dorsal horn neurons which express the neurokinin 1 receptor contribute to several ascending pathways that are thought to be important in pain mechanisms.

Afferent Pathways↗
PubMed1999

GABAergic neurons that contain neuropeptide Y selectively target cells with the neurokinin 1 receptor in laminae III and IV of the rat spinal cord.

Neuropeptide Y (NPY) is contained in a population of GABAergic interneurons in the spinal dorsal horn and, when administered intrathecally, can produce analgesia. We previously identified a strong monosynaptic link between substance P-containing primary afferents and cells in lamina III or IV with the neurokinin 1 (NK1) receptor. Because some of these cells belong to the spinothalamic tract, they are likely to have an important role in pain mechanisms. In this study, we used confocal microscopy to examine the input to lamina III/IV NK1 receptor-immunoreactive neurons from NPY-containing axons. All of the cells studied received a dense innervation from NPY-immunoreactive axons, and electron microscopy revealed that synapses were often present at points of contact. Most NPY-immunoreactive boutons were also GABAergic, which supports the suggestion that they are derived from local neurons. The association between NPY-containing axons and NK1 receptor-immunoreactive neurons was specific, because postsynaptic dorsal column neurons (which were located in laminae III-V but did not possess NK1 receptors) and lamina I neurons with the NK1 receptor received significantly fewer contacts from NPY-immunoreactive axons. In addition, the NK1 receptor-immunoreactive lamina III/IV cells received few contacts from nitric oxide synthase-containing axons (which belong to a different population of GABAergic dorsal horn neurons). The NPY-containing axons appeared to be targeted to the NK1 receptor-immunoreactive neurons themselves rather than to their associated substance P-immunoreactive inputs. The dense innervation of these cells by NPY-containing axons suggests that they may possess receptors for NPY and that activation of these receptors may contribute to NPY-mediated analgesia.

Animals↗
PubMed1998

Cells in laminae III and IV of the rat spinal cord which possess the neurokinin-1 receptor receive monosynaptic input from myelinated primary afferents.

We have previously demonstrated that neurons which have cell bodies in laminae III or IV of the rat spinal cord, dendrites that enter the superficial laminae and which possess the neurokinin-1 receptor receive a major synaptic input from substance P-containing primary afferent axons. In this study we set out to determine whether these cells also receive monosynaptic input from myelinated primary afferents by using transganglionic transport of the B subunit of cholera toxin to identify the central terminals of myelinated afferents from the sciatic nerve. Dual-immunofluorescence and confocal microscopy revealed apparent contacts between labelled primary afferent terminals and all of the neurokinin-1 receptor-immunoreactive cells examined, although these contacts were much less numerous than those which the cells receive from substance P-containing primary afferents. By using a combined confocal and electron microscopic technique we were able to confirm that synapses were present at some of the contacts between primary afferents and neurokinin-1 receptor-immunoreactive neurons. These results suggest that cells of this type will have wide-dynamic range receptive fields, but with a relatively strong input from nociceptors.

Animals↗
PubMed1997

Cells in laminae III and IV of the rat spinal cord that possess the neurokinin-1 receptor and have dorsally directed dendrites receive a major synaptic input from tachykinin-containing primary afferents.

Many neurons with cell bodies in laminae III or IV of the spinal dorsal horn possess the neurokinin 1 receptor and have dorsal dendrites that arborize in the superficial dorsal horn. We have performed a confocal microscopic study to determine whether these cells receive inputs from substance P-containing primary afferents. All neurons of this type received contacts from substance P-immunoreactive axons, and in most cases the contacts onto dorsal dendrites were very numerous. A great majority (90-100%) of substance P-immunoreactive varicosities in contact with these cells were also immunoreactive with antibody to calcitonin gene-related peptide, indicating that they were of primary afferent origin. The density of contacts from substance P-immunoreactive varicosities onto these cells was significantly higher than that seen on cholinergic neurons in lamina III (which do not possess the receptor). Electron microscopy revealed that synapses were present at points of contact between substance P-immunoreactive boutons and dorsal dendrites of cells with the neurokinin 1 receptor. Some cells of this type belong to the spinothalamic tract, and we therefore examined neurons with cell bodies in laminae III or IV that possessed the neurokinin 1 receptor and were labeled retrogradely after thalamic injection of cholera toxin B subunit. These cells also received contacts from substance P-immunoreactive axons on their dorsal dendrites. The results of this study indicate that neurons of this type are a major target for substance P-containing primary afferents.

Afferent Pathways↗
PubMed1996

Evidence for the presence of neurokinin-1 receptors on dorsal horn spinocerebellar tract cells in the rat.

Dorsal horn spinocerebellar tract cells of adult rats were labelled by retrograde axonal transport with the B subunit of cholera toxin. Sections were prepared from lumbar and thoracic spinal segments and incubated with antisera which specifically recognise neurokinin-1 receptor protein and substance P. Labelled cells and immunoreactivity for the receptor and substance P were identified by using three different fluorophores and the relationships between them were assessed in single optical sections with three-colour confocal laser scanning microscopy. Forty-eight cells were examined and 23 of them displayed immunoreactivity for the receptor. Many substance P-immunoreactive profiles were present in lamina V and some formed contacts with spinocerebellar tract cells possessing neurokinin-1 receptor immunoreactivity. The evidence suggests that substance P may influence the activity of a subpopulation of dorsal horn spinocerebellar tract cells by acting through neurokinin-1 receptors.

Animals↗
PubMed1996

Neurokinin-1 receptors on lumbar spinothalamic neurons in the rat.

In order to determine whether spinothalamic neurons in the lumbar spinal cord of the rat process neurokinin-1 (substance P) receptors, we injected cholera toxin B subunit into the thalamus and carried out dual-labelling immunocytochemistry to search for neurons that were immunoreactive with antibodies to cholera toxin and neurokinin-1 receptor. We examined 356 spinothalamic neurons in transverse sections and found that 35% of these were neurokinin-1 receptor-immunoreactive. Double-labelled cells made up the majority of the spinothalamic population in lamina I and the lateral spinal nucleus, and were also present in laminae III-V and the area around the central canal. On the side contralateral to the injection site, 77% of spinothalamic neurons in lamina I also showed neurokinin-1 receptor immunoreactivity, while 33% of those in laminae III-V and 14% of the ventromedial group possessed the receptor. Several of the double-labelled neurons with cell bodies in laminae III and IV had dendrites which could be followed dorsally into the superficial dorsal horn. These results indicate that substance P released from nociceptive primary afferents into the superficial dorsal horn is likely to act on spinothalamic tract neurons in lamina I, and also on those with cells bodies in laminae III-IV and long dorsal dendrites.

Animals↗
PubMed1995

The types of neuron in spinal dorsal horn which possess neurokinin-1 receptors.

In order to provide further information about the types of spinal neuron which possess neurokinin-1 receptors, we have carried out pre-embedding immunocytochemistry on sections of rat lumbar spinal cord with an antiserum raised against a synthetic peptide corresponding to part of the sequence of the receptor, and combined this with post-embedding immunocytochemistry to detect GABA and glycine. Numerous neuronal cell bodies showing neurokinin-1 receptor-immunoreactivity were seen in lamina I, laminae III-VI, the lateral spinal nucleus and the area around the central canal. Most of the cells observed in lamina III were small and had relatively restricted dendritic trees which could often not be followed into lamina II, however some larger cells in laminae III and IV had dendrites which extended through lamina II and into lamina I. Cells of the latter type are likely to represent a major target of substance P released from small-diameter primary afferents in the superficial dorsal horn. The great majority (255 out of 283) of spinal neurons which possessed neurokinin-1 receptor-immunoreactivity, including all of those in lamina I, were not GABA- or glycine-immunoreactive, however a few cells in the deep part of the dorsal horn and the lateral spinal nucleus and several cells near the central canal were GABA-immunoreactive, and some of these were also glycine-immunoreactive. These results suggest that substance P acts through neurokinin-1 receptors mainly on excitatory neurons within the spinal cord.

Afferent Pathways↗
PubMed1994

Some inhibitory neurons in the spinal cord develop c-fos-immunoreactivity after noxious stimulation.

In order to determine which types of spinal neuron produce c-fos in response to noxious stimulation, we have combined pre-embedding detection of c-fos-like immunoreactivity with post-embedding immunocytochemistry using antibodies against GABA and glycine, 2 h after subcutaneous injection of formalin into a hindpaw of anaesthetized rats. Throughout the spinal cord, the majority of c-fos-immunoreactive neurons (72-81%) did not possess GABA- or glycine-like immunoreactivity, while the remaining cells contained one or both types of immunoreactivity. In the superficial dorsal horn (laminae I and II) and dorsal white matter, between 14 and 20% of c-fos-immunoreactive neurons were GABA-immunoreactive, and some of these were also glycine-immunoreactive. A single neuron in lamina I in one animal was glycine- but not GABA-immunoreactive. In the remainder of the spinal cord, between 21 and 35% of the c-fos-immunoreactive cells were GABA- or glycine-immunoreactive, and the majority of these neurons contained both types of immunoreactivity. These results suggest that some inhibitory neurons in both the superficial and deep parts of the dorsal horn are activated by noxious stimuli. It is known that some of the cells which produce c-fos in response to noxious stimulation are projection neurons, with axons ascending to the brainstem or thalamus, however, because of the large number of c-fos-immunoreactive cells in the dorsal horn, it is likely that many are interneurons, and some of these are probably excitatory cells which use glutamate as a transmitter. It therefore appears that after noxious stimulation c-fos is produced in several types of spinal neuron, including projection cells and both excitatory and inhibitory interneurons.

Animals↗
PubMed1991

A quantitative study of neuropeptide immunoreactive cell bodies of primary afferent sensory neurons following rat sciatic nerve peripheral axotomy.

Following peripheral axotomy, fluoride resistant acid phosphatase (FRAP) and most neuropeptides are depleted in the central terminals of axotomised nerves and reduced in their corresponding cell bodies (DRG) but vasoactive intestinal polypeptide (VIP) increases. The increase in VIP probably results from a change in gene expression in other ganglion cells which do not normally express VIP. A quantitative study was performed to investigate the proportion of DRG cells immunoreactive for different peptides at increasing times after sciatic nerve section. Retrograde fluorescent neuronal labelling of sciatic nerve cell bodies by injection of fast blue into the proximal stump was combined with unlabelled antibody immunohistochemistry for CGRP and VIP. The proportion of cells immunoreactive for these peptides was quantified between two and fourteen days post-axotomy. The number of VIP immunoreactive profiles increased significantly in the first 4 days post-axotomy, followed by a slight decrease before rising again. In contrast, the number of and CGRP-immunoreactive cell profiles declined to zero by 14 days post-axotomy. 4 days post-axotomy 50% of VIP positive cells were also immunoreactive for CGRP. There was neither colocalisation between VIP and FRAP nor between CGRP and FRAP. It is concluded that many peptidergic DRG cell bodies switch their expression of peptide to VIP after injury, whereas non-peptide-containing subpopulations do not.

Acid Phosphatase↗
PubMed1990

A polyclonal antibody to goldfish neuronal 145 kDa intermediate filament protein.

A polyclonal antibody raised to a protein from goldfish optic tectum recognises, immunohistochemically, axons throughout normal goldfish visual pathway. In goldfish with injured optic nerve, this antibody recognises degenerating neuronal debris as well as regenerating fibres. On immunoblot, the antibody recognises, primarily, a neuronal intermediate filament protein in the region of 145 kDa. Such an antibody should prove useful in studies pertaining to goldfish visual pathway.

Animals↗
PubMed1990

Preferential histochemical staining of protoplasmic and fibrous astrocytes in rat CNS with GFAP antibodies using different fixatives.

Serial sections of rat brain and spinal cord were fixed in either acid-alcohol or 4% paraformaldehyde, and stained for visualization of astrocytes using GFAP antibodies. With paraformaldehyde, GFAP-positive astrocytes were visualised almost exclusively in the grey matter of all above tissues. In sharp contrast, acid-alcohol treatment gave intensely stained GFAP-containing astrocytes in the white matter. Since fibrous astrocytes are mainly located in the white matter and protoplasmic astrocytes are located in the grey matter, it is concluded that acid-alcohol is a good fixative for fibrous astrocytes while paraformaldehyde is a better fixative for protoplasmic astrocytes.

Animals↗
PubMed1990

Expression of glial fibrillary acidic protein (GFAP) in goldfish optic nerve following injury.

By using an antibody to goldfish glial fibrillary acidic protein (GFAP), the reaction of goldfish optic nerve to injury has been studied by immunoblotting and immunohistochemical methods. Goldfish optic nerve, which normally lacks GFAP immunoreactivity (Nona et al.: Glia, 2:189-200, 1989), expresses GFAP following injury. This immunoreactivity, which is observed as early as 10 days after crush and which is still evident at 30 days after crush, all but disappears by 150 days after crush. Since it is well established that functional restoration of synaptic connections and the recovery of vision takes place in goldfish following optic nerve injury, our results indicate that reactive astrocytes do not represent an impediment to regeneration in goldfish visual system.

Animals↗
PubMed1989

Glial fibrillary acidic protein (GFAP) from goldfish: its localisation in visual pathway.

An intermediate filament fraction, isolated from goldfish brain, contains a prominent protein having a molecular weight of 51 kDa. In normal goldfish visual pathway, this protein is present in tectum and tract, but not in optic nerve. A polyclonal antibody raised to this protein clearly labels ependymal glial profiles in tectum and parallel processes in the tract, whereas optic nerve is unlabelled; Müller fibres in the retina are also labelled. A similar, but less prominent, pattern of staining is observed with antibodies, raised elsewhere, against glial fibrillary acidic protein from human and porcine. These results suggest that the 51 kDa protein is a GFAP, demonstrate the heterogeneity of astrocytes in goldfish visual pathway, and are consistent with the idea that GFAP is well conserved in vertebrate phylogeny.

Animals↗
PubMed1989

Anastomosis of bypass grafts using a low-powered CO2 laser.

The purpose of this study was to evaluate the feasibility of using a low-powered CO2 laser to perform end-to-side anastomoses of bypass grafts. The internal jugular veins of 13 domestic swine were removed and used as grafts to bypass their ligated carotid arteries. Each end of the vein graft was apposed to the artery with four to six temporary stay sutures, and a waveguide delivery CO2 laser with a power density of 900 mW/mm2 was used to perform the anastomoses, after which the sutures were removed. The animals were sacrificed postoperatively at five time intervals: 2 to 4 hours, 5 days, 1 month, 3 months, and 5 months. Of the 10 anastomoses studied at 2 to 4 hours, 70% were fully patent, none thrombosed, and 30% disrupted. Of the 16 anastomoses studied between 5 days and 5 months postoperatively, 88% were patent, 12% thrombosed, and none disrupted. None of the animals developed aneurysms at any stage of this investigation. Histologic analysis of acute studies revealed thermal damage such as charring of tissue, unraveling of the collagen fibrils, and the formation of microvacuoles. Within 3 months, the anastomotic sites showed localized healing with intimal fibromuscular proliferation and dense fibrous tissue. Good apposition of tissue was found to be of great importance in achieving patency of vessels in this procedure.

Animals↗
PubMed1986

Peripheral axotomy of the rat mandibular trigeminal nerve leads to an increase in VIP and decrease of other primary afferent neuropeptides in the spinal trigeminal nucleus.

In the vasoactive intestinal polypeptide (VIP)-rich lumbosacral spinal cord, VIP increases at the expense of other neuropeptides after primary sensory nerve axotomy. This study was undertaken to ascertain whether similar changes occur in peripherally axotomised cranial sensory nerves. VIP immunoreactivity increased in the terminal region of the mandibular nerve in the trigeminal nucleus caudalis following unilateral section of the sensory root of the mandibular trigeminal nerve at the foramen orale. Other primary afferent neuropeptides (substance P, cholecystokinin and somatostatin) were depleted and fluoride-resistant acid phosphatase activity was abolished in the same circumscribed areas of the nucleus caudalis. The rise in VIP and depletion of other markers began 4 days postoperatively and was maximal by 10 days, these levels remaining unchanged up to 1 year postoperatively. VIP-immunoreactive cell bodies were absent from trigeminal ganglia from the unoperated side but small and medium cells stained intensely in the ganglia of the operated side after axotomy. These observations indicate that increase of VIP in sensory nerve terminals is a general phenomenon occurring in both cranial and spinal sensory terminal areas. The intense VIP immunoreactivity in axotomised trigeminal ganglia suggests that the increased levels of VIP in the nucleus caudalis are of peripheral origin, indicating a change in expression of neuropeptides within primary afferent neurons following peripheral axotomy.

Acid Phosphatase↗
PubMed1986

The origins of the sciatic nerve and changes in neuropeptides after axotomy: a double labelling study using retrograde transport of true blue and vasoactive intestinal polypeptide immunohistochemistry.

One month after sciatic nerve section, only dorsal root ganglion cells which take up True blue applied to the cut end of the nerve show vasoactive intestinal polypeptide immunoreactivity. This indicates that VIP expression is only in cells with damaged axons. Motor axons with True blue-positive perikarya in the ventral horn originate from the fourth to sixth lumbar segments whereas unmyelinated and small myelinated sensory nerves terminate in the third to fifth lumbar segments of the spinal cord.

Animals↗
PubMed1986

Vasoactive intestinal polypeptide (VIP) increases in the spinal cord after peripheral axotomy of the sciatic nerve originate from primary afferent neurons.

Following sciatic nerve axotomy, vasoactive intestinal polypeptide (VIP) immunoreactivity increases dramatically in the central terminal areas of the nerve whereas other primary afferent neuropeptides are depleted. The contribution of the peripheral nerve to VIP increases in the spinal cord was investigated by performing sciatic nerve section alone, dorsal rhizotomy of the lumbar roots, axotomy and rhizotomy in combination or section of other peripheral nerves terminating in the same segments as the sciatic nerve. VIP, and for comparison, substance P (SP), cholecystokinin (CCK), somatostatin (SOM), were localized in the lumbar spinal cord and corresponding sensory ganglia using unlabeled antibody immunohistochemistry. After sciatic nerve section, SP, CCK and SOM were depleted in the lumbar dorsal horn whereas VIP increased. After rhizotomy alone all neuropeptide staining including VIP was depleted; axotomy followed by rhizotomy produced the same result. Axotomy of other peripheral nerves terminating in the lumbar cord increased the area of neuropeptide depletion but correspondingly increased the area of VIP staining. A large proportion of small and medium diameter dorsal root ganglion cells were stained for VIP after nerve section or axotomy but not after rhizotomy alone. A radical change in neuropeptide metabolism of dorsal root ganglion cells occurs after peripheral axotomy, in the form of a marked increase in VIP synthesis. An intact dorsal root is necessary for increases in VIP in the spinal cord indicating the primary afferent origin of the response.

Animals↗