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S S Cheema

Publications and source records attributed to S S Cheema.

At least 37 records · Page 2Linked to original sources
PubMed1998

Neural precursor differentiation into astrocytes requires signaling through the leukemia inhibitory factor receptor.

The differentiation of precursor cells into neurons or astrocytes in the developing brain has been thought to be regulated in part by growth factors. We show here that neural precursors isolated from the developing forebrain of mice that are deficient in the gene for the low-affinity leukemia inhibitory factor receptor (LIFR-/-) fail to generate astrocytes expressing glial fibrillary acidic protein (GFAP) when cultured in vitro. Precursors from mice heterozygous for the null allele show normal levels of GFAP expression. These findings support the in vivo findings that show extremely low levels of GFAP mRNA in brains of embryonic day 19 LIFR-/- mice. In addition, monolayers of neural cells from LIFR-/- mice are far less able to support the neuronal differentiation of normal neural precursors than are monolayers from heterozygous or wild-type animals, indicating that endogenous signaling through the LIFR is required for the expression of both functional and phenotypic markers of astrocyte differentiation. LIFR-/- precursors are not irreversibly blocked from differentiating into astrocytes: they express GFAP after long-term passaging or stimulation with bone morphogenetic protein-2. These findings strongly implicate the LIF family of cytokines in the regulation of astrocyte differentiation and indeed the LIF-deficient animals show a significant reduction in the number of GFAP cells in the hippocampus. However, because this reduction is only partial it suggests that LIF may not be the predominant endogenous ligand signaling through the LIFR.

Animals↗
PubMed1998

Leukemia inhibitory factor maintains choline acetyltransferase expression in vivo.

Following axotomy most medial septal neurons in the adult rat brain have dramatically reduced numbers of choline acetyltransferase (ChAT) positive neurons. Since leukemia inhibitory factor (LIF) promotes cholinergic expression in several neuronal populations, the aim of this study was to determine if LIF would continue to support cholinergic expression in axotomized medial septal neurons. Mini-osmotic pumps were used to infuse saline or LIF into the lateral cerebral ventricle. Counts of ChAT and low-affinity nerve growth factor (p75NGFR) immunostained neurons indicated that LIF-treated animals retained ChAT expression in > 90% of axotomized neurons whereas in saline-infused animals this was < 30%. Also, LIF was equally effective in maintaining p75NGFR expression levels in axotomized medial septal neurons.

Analysis of Variance↗
PubMed1997

Leukemia inhibitory factor enhances the regeneration of transected rat sciatic nerve and the function of reinnervated muscle.

The cytokine leukemia inhibitory factor (LIF) favors the survival and growth of axons in vitro and in vivo. Fibronectin has been shown to enhance nerve regeneration when added in combination with various growth factors including LIF. The goal of this study was to evaluate the effect of LIF plus fibronectin on the regeneration of transected nerve and functional recovery of reinnervated skeletal muscle, in one experimental model of peripheral nerve repair, at two recovery times. The rat sciatic nerve was cut at mid-thigh level and a silicone cuff containing either saline (control), LIF, or LIF plus fibronectin (L+F) was used to bridge the proximal and distal nerve stumps leaving a 1 cm gap between them. Rats were then explored at 6 or 12 weeks following the initial surgery. Regenerating nerves were assessed by measuring the diameter of myelinated axons, conduction velocity, and number of myelinated fibers. Muscle reinnervation was assessed by measuring muscle mass, force of contraction, and histologically for changes in muscle fiber type (type I and type II). In this report we demonstrate that at 6 weeks there were significant increases in 1) nerve conduction velocity, 2) myelinated axon diameter, and 3) number of myelinated axons over that of control (saline-treated) animals. Both LIF groups demonstrated a shift in type II muscle fiber area compared to saline-treated controls, with the L+F group having a significant increase in muscle mass. At 12 weeks there was an improved recovery over and above that demonstrated at 6 weeks. Muscle mass was 65% and 42% greater than control for LIF and L+F, respectively. Force of contraction, conduction velocity, myelinated fiber number, and diameter were also significantly greater for both LIF- and L+F-treated rats than saline-treated rats. These results demonstrate that LIF significantly improves the regeneration of damaged peripheral nerves and the preservation of muscle viability, resulting in greatly enhanced recovery of skeletal muscle function.

Analysis of Variance↗
PubMed1997

Lhx2, a LIM homeobox gene, is required for eye, forebrain, and definitive erythrocyte development.

We investigated the function of Lhx2, a LIM homeobox gene expressed in developing B-cells, forebrain and neural retina, by analyzing embryos deficient in functional Lhx2 protein. Lhx2 mutant embryos are anophthalmic, have malformations of the cerebral cortex, and die in utero due to severe anemia. In Lhx2-/- embryos specification of the optic vesicle occurs; however, development of the eye arrests prior to formation of an optic cup. Deficient cellular proliferation in the forebrain results in hypoplasia of the neocortex and aplasia of the hippocampal anlagen. In addition to the central nervous system malformations, a cell non-autonomous defect of definitive erythropoiesis causes severe anemia in Lhx2-/- embryos. Thus Lhx2 is necessary for normal development of the eye, cerebral cortex, and efficient definitive erythropoiesis.

Anemia↗
PubMed1996

Reducing p75 nerve growth factor receptor levels using antisense oligonucleotides prevents the loss of axotomized sensory neurons in the dorsal root ganglia of newborn rats.

The low-affinity p75 receptor for nerve growth factor (p75NGFR) has been implicated in mediating neuronal cell death in vitro. A recent in vitro study from our laboratory showed that the death of sensory neurons can be prevented by reducing the levels of p75NGFR with antisense oligonucleotides. To determine if p75NGFR also functions as a death signal in vivo, we have attempted to reduce its expression in peripheral sensory neurons by applying antisense oligonucleotides to the proximal end of the transected sciatic or median and ulnar nerves. We report here that antisense oligonucleotides, when applied to the proximal stump of a transected peripheral nerve, are retrogradely transported and effectively reduce p75NGFR protein levels in sensory neurons located in the dorsal root ganglia. Furthermore, treatment of the proximal nerve stump with antisense p75NGFR oligonucleotides significantly reduced the loss of these axotomized sensory neurons. These findings further support the view that p75NGFR is a death signaling molecule and that it signals death in axotomized neurons in the neonatal sensory nervous system.

Animals↗
PubMed1996

Leukemia inhibitory factor is a myotrophic and neurotrophic agent that enhances the reinnervation of muscle in the rat.

The effects of leukemia inhibitory factor (LIF) on muscle atrophy and the reinnervation of muscle were investigated. The rat medial gastrocnemius (MG) nerve was either cut (denervation groups), crushed (reinnervation group) or left intact (normal group). Muscles were injected with LIF in phosphate buffered saline (PBS) containing pluronic gel, the contralateral control muscles were injected with the vehicle alone. The muscles from the LIF-injected denervation groups were analysed for muscle fibre area; this was found to be significantly larger than controls. The greatest change was observed in the reinnervation group, where the muscle fibre area following LIF treatment was 53% (Type 1) and 84% (Type 2) greater. In addition, nerve fibre diameters were analysed in the reinnervation-treated group and these were also significantly larger. However, LIF injected into normally innervated muscle resulted in a decrease in muscle fibre area. These results show that LIF ameliorates denervation-induced muscle atrophy and improves regeneration of muscle and nerve.

Animals↗
PubMed1996

Up-regulation of leukaemia inhibitory factor and interleukin-6 in transected sciatic nerve and muscle following denervation.

Leukaemia inhibitory factor (LIF) and Interleukin-6 (IL-6) are multifunctional cytokines that are related on the basis of their predicted structural similarities and shared signal transducing receptor components. Both these factors stimulate myoblast proliferation, and whereas LIF is neurotrophic for sensory neurons, and for the motor neurons which innervate muscle, IL-6 has only been reported to act on a population of septal neurons in the brain. We have looked at the effect of peripheral nerve trauma on the expression of these factors. We show here that whereas LIF and IL-6 mRNAs are expressed in low levels in normal sciatic nerve and skeletal muscle, there is significant up-regulation in the nerve segments after injury, with proximally and distally. There is also an increase in LIF and IL-6 expression in the denervated muscle located largely in the muscle cells. In addition, while there is retrograde axonal transport of LIF by the sciatic nerve, IL-6 is not retrogradely transported, and as a result, IL-6 does not stimulate the survival of sensory neurons in vitro. Both growth factors are produced by Schwann cells. These results show a rapid response in the expression of these genes after injury and suggest that LIF and IL-6 act as trauma factors but with different roles in injured peripheral nerve.

Animals↗
PubMed1994

The engraftment of transplanted primary neuroepithelial cells within the postnatal mouse brain.

Primary neuroepithelial precursor cells carrying the reporter gene lacZ were transplanted into postnatal murine brain and assessed for their engraftment capacity. Freshly dissected precursors, derived from lacZ transgenic embryonic day 10 mouse brain, predominantly engrafted as discrete clusters, whereas the same precursors cultured in vitro with fibroblast growth factor-2, engrafted as single cells within the parenchyma of the hippocampus. Approximately 0.5% of the transplanted cells survived in the host brain for up to 3 months. Many of these cells displayed neuronal and astrocyte morphologies. These observations suggest that transplanted primary precursors derived from the embryonic brain can engraft and commit in situ to a variety of developmental fates.

Animals↗
PubMed1994

Leukaemia inhibitory factor rescues motoneurones from axotomy-induced cell death.

The death of spinal motoneurones after axotomy provides a useful model for studying novel factors which prevent motoneurone loss in vivo. Peripheral nerves of newborn rats were unilaterally transected and treated with either a vehicle solution or leukaemia inhibitory factor (LIF). Compared with the vehicle controls, treatment with a gelfoam containing LIF significantly reduced motoneurone loss: from 38% to 22% after 3 days and from 55% to 38% after 7 days. The loss of motoneurones was further reduced by placing the LIF-containing gelfoam inside a silicone chamber: from 39% to 15% after 7 days, which represented a 62% rescue. Thus, LIF is a potential therapeutic agent for preventing the loss of injured or diseased motoneurones.

Animals↗
PubMed1994

Leukemia inhibitory factor prevents the death of axotomised sensory neurons in the dorsal root ganglia of the neonatal rat.

Leukemia inhibitory factor (LIF) has several characteristics of a neurotrophic factor for sensory neurons. Here we have investigated whether LIF also supports the survival of axotomised sensory neurons in vivo. Newborn rat pups received a unilateral sciatic nerve transection and the injury site was treated with gelfoam soaked in phosphate buffered saline (PBS), nerve growth factor (NGF), or LIF. Neuronal nucleoli in the L5 dorsal root ganglia were counted, appropriate corrections applied, and the resultant neuronal loss expressed as a percentage of the contralateral intact side. In animals where LIF was administered neuronal loss was significantly reduced: 2 days after LIF treatment neuronal loss was 19.5% compared to 43% in PBS-treated animals; 3 days after LIF treatment neuronal loss was 20.4% compared to 40.2% in PBS-treated animals; however, 7 days after LIF treatment there was no significant reduction in the number of neurons lost. The degree of rescue of sensory neurons in vivo by LIF was found to be similar to NGF, which was not surprising as both factors supported the survival of a similar population of sensory neurons in vitro. Rescue was not observed when LIF-containing gelfoam was placed away from the axotomised nerve, suggesting that LIF's action may be associated with its retrograde transport or direct signalling at the site of nerve injury.

Animals↗
PubMed1993

Clusterin levels increase during neuronal development.

The expression of clusterin has been shown to be elevated in several models of experimentally induced programmed cell death and in association with a number of neurodegenerative conditions. In order to test whether this protein is expressed in neurons during development, the expression of clusterin was examined in the developing nervous system, using immunohistochemistry and mRNA analysis. Clusterin expression was observed in the earliest neurons of the cortical plate on embryonic day (E) 12. Thereafter, the intensity of clusterin staining continued to increase in an age-dependent manner, with the greatest intensity of staining being found in the postnatal mature brain. Virtually all neurons were clusterin-positive and there was no evidence for the appearance of clusterin-positive cells specifically during epochs of programmed neuronal death in the embryo. This study suggests that clusterin has a role in neuronal maturation and it is unlikely to be associated exclusively with neuronal cell death.

Animals↗
PubMed1990

Thalamic projections to sensorimotor cortex in the macaque monkey: use of multiple retrograde fluorescent tracers.

We used several fluorescent dyes (Fast Blue, Diamidino Yellow, Rhodamine Latex Microspheres, Evans Blue, and Fluoro-Gold) in each of eight macaques, to examine the patterns of thalamic input to the sensorimotor cortex of macaques 12 months or older. Inputs to different zones of motor, premotor, and postarcuate cortex, supplementary motor area, and areas 3b/1 and 2/5 in the postcentral cortex, were examined. Coincident labeling of thalamocortical neuron populations with different dyes (1) increased the precision with which their soma distributions could be related within thalamic space, and (2) enabled the detection by double labeling, of individual thalamic neurons that were common to the thalamic soma distributions projecting to separate, dye-injected cortical zones. Double-labeled thalamic neurons projecting to sensorimotor cortex were rarely seen in mature macaques, even when the injection sites were only 1-1.5 mm apart, implying that their terminal arborizations were quite restricted horizontally. By contrast, separate neuron populations in each thalamic nucleus with input to sensorimotor cortex projected to more than one cytoarchitecturally distinct cortical area. In ventral posterior lateral (oral) (VPLo), for example, separate populations of cells sent axons to precentral medial, and lateral area 4, medial premotor, and postarcuate cortex, as well as to supplementary motor area. Extensive convergence of thalamic input even to the smallest zones of dye uptake in the cortex (approximately 0.5 mm3) characterized the sensorimotor cortex. The complex forms of these projection territories were explored using 3-dimensional reconstructions from coronal maps. These projection territories, while highly ordered, were not contained by the cytoarchitectonic boundaries of individual thalamic nuclei. Their organization suggests that the integration of the diverse information from spinal cord, cerebellum, and basal ganglia that is needed in the execution of complex sensorimotor tasks begins in the thalamus.

Afferent Pathways↗
PubMed1990

Thalamic projections to sensorimotor cortex in the newborn macaque.

In the present experiments thalamocortical projections to different functional areas of the newborn (or prematurely delivered) macaque's sensorimotor cortex were labeled using retrogradely transported fluorescent dyes. Several dyes were used in each animal to (1) enable the direct comparison of the soma distributions of different thalamocortical projections within thalamic space, and (2) identify by double labeling neurons shared between these distributions. The projection patterns in the newborn macaque were compared with those of the mature animal reported by Darian-Smith et al. (J. Comp. Neurol. 1990;298:000-000). The main observations were (1) all thalamocortical projections to the sensorimotor cortex of the mature macaque are well established by embryonic days 146-150, as was shown by labeling these pathways in infants delivered by cesarean section, (2) a significant number of thalamocortical neurons in the newborn were double-labeled following dye injections into different pre- or postcentral areas, and where the margins of the dye uptake zones were separated by 3-8 mm, and (3) extensive projections from the anterior pulvinar nucleus to the motor and premotor cortex, and to the supplementary motor cortex were labeled in the newborn macaque. Both the exuberant terminal arborizations, and the precentral pulvinar projections were diminished by the 6th postnatal month, and absent in the mature macaque. The role of epigenetic determinants of these postnatal events is briefly considered.

Afferent Pathways↗
PubMed1989

Determinants of patchy metabolic labeling in the somatosensory cortex of cats: a possible role for intrinsic inhibitory circuitry.

Despite repeated experimental demonstration that somatic stimulation leads to an intermittent, "column-like" pattern of 2-deoxyglucose (2DG) label in the somatosensory cortex, the functional significance of this pattern remains uncertain. A number of recent studies have suggested that the putative inhibitory neurotransmitter GABA may play an influential role in the cortical processing of sensory information. To test the possibility that GABA-mediated inhibitory processes might participate in the formation of the 2DG patches, the 2DG pattern obtained under "normal" experimental conditions was compared with the pattern observed when cortical inhibition was modified by topical application of the GABA antagonist, bicuculline methiodide (BIC). Under "normal" experimental conditions, we found that somatic stimulation led to an intermittent, patch like distribution of 2DG uptake in cat somatosensory cortex, which exhibited consistent features in animals studied using the same stimulus and experimental condition. Reconstructions of the stimulus-evoked activity patterns revealed that the label was confined to territories known to receive input from the stimulated body region and was organized into elongated strips. Topical application of BIC to the somatosensory cortex dramatically altered the dimension of the metabolic patches, which were often embedded in a field of elevated 2DG uptake. In BIC-treated hemispheres the average width of 2DG patches was 1266 microns, whereas the average width of patches in the opposite untreated hemisphere (elicited by identical stimuli) was 713 microns. Unfolded maps of the labeling pattern revealed that in the BIC-treated hemispheres adjacent "strips" of 2DG label tended to fuse, leading to a less intermittent distribution than that observed in the untreated hemispheres. An important role for GABA in the formation of the normal cortical response to somatic stimulation is suggested.

Animals↗
PubMed1986

Intracellular staining study of the feline cuneate nucleus. I. Terminal patterns of primary afferent fibers.

The terminal arborizations of single identified cutaneous hair follicle and slowly adapting type I receptors and muscle (Ia) afferents have been studied in the cuneate nucleus of cats after intra-axonal injection of horseradish peroxidase. Penetrations were mainly at the middle and caudal levels of the nucleus--i.e., from obex to approximately 7 mm caudal to it. Following histochemical processing, the injected axons, along with their collateral branches and synaptic terminals, were visualized and examined with light and electron microscopy. Cutaneous afferents in middle cuneate (from obex to approximately 4 mm caudal to it) issued collateral branches, along the rostrocaudal axis of the nucleus, at intervals between 100 and 1,000 microns. The terminal field of each collateral's branches encompassed an area elongated largely rostrocaudally and virtually confined to the dorsal part of the middle cuneate. Although adjacent collaterals had nonoverlapping terminal arborizations, each one could give rise to separate foci of terminations. Muscle afferents differed, on the whole, from cutaneous afferents in the location and extent of collateral branching and terminal arborizations. However, because muscle fibers terminated primarily in the ventral region of the cuneate, but nevertheless exhibited sparser terminations in the dorsal part of the middle cuneate, there was some spatial overlap between zones of muscle and cutaneous projection. Synaptic boutons of cutaneous afferent fibers contained round clear vesicles, contacted dendritic profiles (sometimes more than one), and were postsynaptic to small boutons containing polymorphic vesicles. In contrast, boutons of muscle afferent fibers contacted somatic and dendritic profiles and were not postsynaptic to other boutons. The results are in general agreement with previous anatomical and electrophysiological work; however, the extent of the terminal field of single collateral branches may provide for a greater convergence of different receptor classes and of receptive fields on neurons in the middle cuneate than estimated by previous electrophysiological investigations.

Afferent Pathways↗
PubMed1986

Intracellular staining study of the feline cuneate nucleus. II. Thalamic projecting neurons.

Morphological and physiological features of thalamic projecting neurons in the middle region of the cuneate nucleus of cats (from obex to 4 mm below it) have been studied, using intracellular recording and iontophoresis of horseradish peroxidase. All cuneothalamic neurons in the present sample) responded to movement of hairs on wrist, paw, or digits. However, approximately 50% of the neurons could be activated by other types of stimulation (e.g., light or maintained pressure on the skin, movement of claws, etc.). No clear differences were apparent in the physiological responses correlated with the varied dendritic pattern of stained neurons. Dendritic arborizations of most cuneothalamic neurons were more extensive than assumed previously, from Golgi impregnated material. As a consequence, only a few neurons have dendrites ramifying within a restricted region--i.e., corresponding to a typical cluster of the middle cuneate nucleus. Dendrites extending in various directions and spanning a distance up to 500 microns provide cuneothalamic neurons with the ability to receive input from relatively widespread areas. Collateral branches of axons of cuneothalamic neurons were observed in 50% of the stained neurons. Most of these collaterals terminated ventrally within the cuneate nucleus. Extensive collateral arborizations were observed in the dorsal as well as the ventral cuneate. These results, together with those reported in the previous paper, suggest complex interactions of afferent inputs on cuneothalamic neurons. In particular, such neurons are likely to be influenced by convergent input from different receptor classes and, because of their axonal collaterals, probably affect the excitability of other neurons, projecting or intrinsic, in their immediate vicinity or in other nuclear regions.

Afferent Pathways↗
PubMed1984

Light and electron microscopic evidence for a direct corticospinal projection to superficial laminae of the dorsal horn in cats and monkeys.

The anterograde transport of horseradish peroxidase (HRP) and wheat germ agglutin conjugated to horseradish peroxidase (WGA-HRP) was employed in cats and monkeys to investigate, at both the light and electron microscopical levels, the contribution of the corticospinal tract (CST) to the superficial laminae of the dorsal horn. At the light microscopic level, this approach not only confirmed the previously documented pattern of CST termination, but also revealed a sparse projection to laminae VIII and IX of the cat and a prominent projection to the most superficial parts of the brachial dorsal horn, i.e., laminae I and II. Discrete injections involving particular cytoarchitectonic areas (4, 3a, 3b, and 1-2) of monkeys showed that the superficial laminae receive their corticofugal inputs primarily from areas 3b, 1, and 2. Electron microscopic observations were made on CST fibers and boutons which were labelled, after histochemical processing, with the reaction product of anterogradely transported WGA-HRP. The labelled fibers in the superficial laminae were small (+/- 0.5 micron), and boutons established mainly axodendritic contacts, contained mostly clear, spherical, or pleomorphic vesicles, but sometimes also displayed dense core vesicles. These boutons were primarily in lamina I and outer lamina II, but not in inner lamina II. The possible role of a direct monosynaptic pathway from the cerebral cortex to the superficial laminae of the dorsal horn is discussed in relation to the previous reports that laminae I and II play a significant role in nociception.

Animals↗