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Biomedical subjects

P Doherty

Publications and source records attributed to P Doherty.

At least 73 records · Page 4Linked to original sources
PubMed1996

Cell adhesion molecules and neuronal regeneration.

Cell adhesion molecules (CAMs) are multifunctional proteins and are involved in a number of important regulatory processes in the brain, including cell growth, migration and regeneration. Recent studies using model in vitro systems have identified additional binding interactions in which CAMs, particularly those of the Ig superfamily, can participate. Signal transduction pathways are activated following CAM action in the process of neurite outgrowth. Key components in these pathways, such as kinases and phosphatases, are being identified. Receptor phosphatases themselves contain protein motifs characteristic of CAMs and may themselves be involved in adhesion-mediated cell recognition events.

Animals↗
PubMed1996

Review: a role for the FGF receptor in the axonal growth response stimulated by cell adhesion molecules?

Cell adhesion molecules (CAMs) have been shown to stimulate axonal growth. The molecular basis of this response has been extensively studied and a range of agents that promote or inhibit CAM stimulated axonal growth have now been identified. These studies have led to the suggestion that following homophilic and/or heterophilic interactions CAM specific signal transduction pathways are activated which are directly responsible for promotion of axonal growth. In this review we will suggest that the axonal growth response stimulated by three CAMs (NCAM, N-cadherin and L1) can be operationally divided into a number of phases. During the first phase homophilic and/or heterophilic binding between the CAMs expressed on the axonal growth cone and cellular substrate take place. This is followed by an interaction of the neuronal CAMs with the fibroblast growth factor receptor (FGFR), leading to receptor activation by autophosphorylation. This results in the recruitment and activation of additional effector molecules via interactions of their SH2 domains with the activated receptor. In this context the key event in terms of neurite outgrowth appears to be the activation of phospholipase C gamma (PLC gamma) which sets into motion a second messenger cascade that ultimately leads to a modification, most likely by phosphorylation, of cytoskeletal elements that are involved in growth cone motility.

Animals↗
PubMed1996

Shared cell adhesion molecule (CAM) homology domains point to CAMs signalling via FGF receptors.

A number of cell adhesion molecules (CAMs) promote neurite outgrowth following transfection and expression in a variety of monolayer cells. We have shown that N-cadherin, L1 and some isoforms of NCAM can stimulate neurite outgrowth from PC12 cells and primary neurons following transfection and expression at physiologically relevant levels in NIH-3T3 cells. A number of observations suggest that these CAMs stimulate neurite outgrowth by activating a convergent second messenger pathway in neurons rather than by modulating adhesion per se, and that an early or initial step in the pathway involves activation of a tyrosine kinase. The observation that the fibroblast growth factor receptor (FGFR) contains an evolutionarily conserved sequence with homology to the above CAMs (the CAM homology domain-CHD) points to the possibility that CAMs might interact with, and signal via, FGFR tyrosine kinases. This hypothesis has been substantiated by a number of independent experimental tests. We present a speculative model in which the evolutionary conservation of a pair of complementary binding motifs can account for a direct binding interaction between FGFR and the above three CAMs.

Animals↗
PubMed1995

Mouse model for the lysosomal disorder galactosialidosis and correction of the phenotype with overexpressing erythroid precursor cells.

The lysosomal storage disorder galactosialidosis results from a primary deficiency of the protective protein/cathepsin A (PPCA), which in turn affects the activities of beta-galactosidase and neuraminidase. Mice homozygous for a null mutation at the PPCA locus present with signs of the disease shortly after birth and develop a phenotype closely resembling human patients with galactosialidosis. Most of their tissues show characteristic vacuolation of specific cells, attributable to lysosomal storage. Excessive excretion of sialyloligosaccharides in urine is diagnostic of the disease. Affected mice progressively deteriorate as a consequence of severe organ dysfunction, especially of the kidney. The deficient phenotype can be corrected by transplanting null mutants with bone marrow from a transgenic line overexpressing human PPCA in erythroid precursor cells. The transgenic bone marrow gives a more efficient and complete correction of the visceral organs than normal bone marrow. Our data demonstrate the usefulness of this animal model, very similar to the human disease, for experimenting therapeutic strategies aimed to deliver the functional protein or gene to affected organs. Furthermore, they suggest the feasibility of gene therapy for galactosialidosis and other disorders, using bone marrow cells engineered to overexpress and secrete the correcting lysosomal protein.

Abnormalities, Multiple↗
PubMed1995

Eugenics in China.

Explore the source record for details and available documents.

Abortion, Induced↗
PubMed1995

The neural cell adhesion molecule and synaptic plasticity.

Highly stereotyped patterns of neuronal connections are laid down during the development of the nervous system via a range of activity independent and activity dependent mechanisms. Whereas the coarse hard-wiring of the nervous system appears to rely on molecular recognition events between the neuron, its pathway, and its target, the establishment of precisely patterned functional circuits is thought to be driven by neuronal activity. In this review we discuss the role that the neuronal cell adhesion molecule (NCAM) plays in morphological plasticity. Recent studies on NCAM and its probable species homologue in Aplysia (apCAM) suggests that an individual CAM can function to both promote synaptic plasticity and maintain the structure of the synapse. In the adult brain, changes between stability and plasticity are likely to underlie dynamic morphological changes in synaptic structures associated with learning and memory. In this review we use NCAM as an example to illustrate mechanisms that can change the function of an individual CAM from a molecule that promotes plasticity to one that does not. We also discuss evidence that NCAM promotes plasticity by activating a conventional signal transduction cascade, rather than by modulating adhesion per se. Finally, we consider the evidence that supports a role for NCAM in learning and memory.

Animals↗
PubMed1995

NCAM requires a cytoplasmic domain to function as a neurite outgrowth-promoting neuronal receptor.

The neural cell adhesion molecule (NCAM) promotes axonal growth via a homophilic binding mechanism by acting both as a neuronal receptor and a substratum ligand. We have previously shown that the GPI-linked 120-kDa isoform of NCAM, which lacks a cytoplasmic domain, is effective at promoting neurite outgrowth as a cellular ligand. To test its ability to function as a neuronal receptor, we have transfected PC12 cells with a cDNA encoding human GPI-linked NCAM and tested clones displaying stable cell surface expression of this isoform for their ability to respond to NCAM in a cellular substratum. Although they continued to express endogenous transmembrane rat isoforms of NCAM (140 and 180 kDa), PC12 cells expressing the GPI-linked NCAM lost their ability to extend neurites in response to substratum associated NCAM. However, their outgrowth response to N-cadherin and other activators of axonal growth was undiminished. Removal of GPI-linked NCAM from the surface of these clones using phosphatidylinositol-specific phospholipase C (PIPLC) fully restored their responsiveness to NCAM, indicating that the inhibition was a direct consequence of cell surface expression of this "dominant negative" isoform of NCAM. We have previously shown that expression of transfected 140- and 180-kDa isoforms of human NCAM in PC12 cells does not result in a loss of the neurite outgrowth response to NCAM. However, we show that deletion of the cytoplasmic domain of the 140-kDa isoform has the same effect as expression of GPI-linked NCAM. We conclude that the cytoplasmic domain of NCAM is required for an appropriate neurite outgrowth response.

Animals↗
PubMed1995

A Ca2+/calmodulin kinase inhibitor, KN-62, inhibits neurite outgrowth stimulated by CAMs and FGF.

We have used monolayers of parental 3T3 fibroblasts and 3T3 cells expressing transfected cell adhesion molecules (CAMs, NCAM, N-cadherin, or L1) as a culture substrate for cerebellar neurons. Previous studies suggest that the transfected CAMs promote neurite outgrowth by activating a second messenger pathway within the responding neuron that involves influx of calcium into neurons as a consequence of activation of an FGF receptor. The same neurite outgrowth response can be induced by FGF or a number of agents that directly activate defined steps in the CAM signaling pathway. In the present study we show that the neurite outgrowth stimulated by the above three CAMs, FGF, arachidonic acid (AA), and K+ depolarization can be abolished by the Ca2+/calmodulin-dependent (CaM) kinase inhibitor, KN-62. We also demonstrate that neurite outgrowth over astrocytes, which represent a more physiologically relevant cellular substrate, can be substantially inhibited by a number of agents that block the CAM signaling pathway, including KN-62. However, neurite outgrowth induced by activation of protein kinase A is unaffected by inhibition of CaM kinase activity as is basal neurite outgrowth over 3T3 monolayers or a polylysine/laminin substrate. These results suggest that CaM kinase activity is specifically required downstream of calcium influx in the CAM and FGF signaling pathway leading to axonal growth.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1995

The anaesthetic management of the child with Eisenmenger's syndrome.

There is little clinical data in the literature on the anaesthetic management of paediatric patients with Eisenmenger's syndrome undergoing non-cardiac surgery. This paper reviews our experiences with either such patients who underwent a total of 11 surgical procedures. Of the eight children, six had Down's syndrome and an atrio-ventricular septal defect, one had a ventricular septal defect and one an atrial septal defect. Nine of the eleven operations consisted of minor dental, plastic or ENT procedures, while one patient underwent two laparotomies. Premedication (trimeprazine/ meperidine combination or midazolam) was administered on three occasions. Induction of anaesthesia was achieved by either inhalation of halothane (2), or intravenously with thiopentone (6), ketamine (2) or propofol (1). Muscle relaxation and mechanical ventilation were employed only for both intra-abdominal procedures, otherwise patients were allowed to breathe spontaneously with, or without, manual assistance. Halothane (8), isoflurane (2) and enflurane (1) were all used for maintenance of anaesthesia. Non-invasive monitoring was applied intraoperatively for minor procedures, and arterial and central venous catheters inserted for the laparotomies. Postoperative analgesia for both these cases was provided by an epidural infusion of bupivacaine 0.125% and fentanyl 5 micrograms x ml(-1). A single im bolus of morphine was required following a dental clearance, otherwise pain relief for the rest of the cases was achieved by local anaesthetic infiltration and NSAIDS. With the exception of a single episode of bradycardia, induction, maintenance and recovery from anaesthesia were well tolerated in all cases. In conclusion, our experience suggests that despite theoretical risks, children with Eisenmenger's syndrome appear to tolerate a variety of anaesthetic techniques.

Anesthesia↗
PubMed1995

Premedication for ambulatory surgery in preschool children: a comparison of oral midazolam and rectal thiopentone.

Seventy five ASA 1 and 2 children, aged between six months and five years were randomized to receive oral midazolam 0.5 mg.kg-1, rectal thiopentone 35 mg.kg-1 or no premedication to compare the safety and efficacy of, and parental attitudes to, both premedicants. Cardio-respiratory variables were from the time of premedication to awakening from anaesthesia. In addition, anxiety and sedation scores and patients' acceptance of both premedicant and mask at induction, were all recorded using four-point rating scales. Times to recovery and discharge, and parental satisfaction with the premedication their child had received were also recorded. Children receiving rectal thiopentone had higher sedation scores and were more accepting of the mask than were the other two groups (P < 0.001). Their acceptance of the premedication was similar to that of the midazolam group. Times to spontaneous eye opening and discharge were longer in the thiopentone group (P < 0.005). Parental preoperative satisfaction rating was higher for thiopentone, but not midazolam, than no premedication (P < 0.05). When asked their premedication preferences for subsequent general anaesthetics, a higher proportion of parents whose children were not premedicated requested an alternative regimen (P < 0.01). In conclusion the study found that premedication with rectal thiopentone provided superior induction characteristics to oral midazolam, but with a longer recovery period.

Administration, Oral↗
PubMed1995

A soluble chimeric form of the L1 glycoprotein stimulates neurite outgrowth.

Cerebellar neurons, cultured on monolayers of 3T3 fibroblasts or on a polylysine/extracellular matrix-coated substratum, responded to a soluble recombinant L1-Fc chimera by extending longer neurites than controls. The response was inhibited by pretreating neurons with antibodies to L1 or antibodies to the fibroblast growth factor (FGF) receptor. The response could also be inhibited by a range of pharmacological reagents that inhibit various steps in the signal transduction cascade which underlie a neurite outgrowth response to basic FGF. The response was of a similar magnitude and not additive with that induced by L1 expressed in a cellular substrate. These data show that L1 in neurons is capable of directing a neurite outgrowth response to a soluble L1-Fc chimera, and that neuronal FGF receptor function is required for this response. The data also show that the ability of cell adhesion molecules (CAMs) to stimulate neurite outgrowth can be dissociated from their ability to function as substrate-associated adhesion molecules and point to the potential of using CAM-Fc chimeras to promote nerve regeneration.

3T3 Cells↗
PubMed1995

FGF inhibits neurite outgrowth over monolayers of astrocytes and fibroblasts expressing transfected cell adhesion molecules.

We have cultured cerebellar neurons on monolayers of cortical astrocytes in control medium or medium containing recombinant basic fibroblast growth factor (FGF). FGF was found to inhibit neurite outgrowth, with a significant effect seen at 0.5 ng/ml and a maximal effect at 10 ng/ml. FGF increased the production of arachidonic acid (AA) in cerebellar neurons, and when added directly to cultures or generated endogenously via activation of phospholipase A2 using melittin, this second messenger could mimic the inhibitory effect of FGF. FGF and AA could also specifically inhibit neurite outgrowth stimulated by three cell adhesion molecules (NCAM, N-cadherin and L1) expressed in transfected fibroblasts, or in the case of L1 bound to a tissue culture substratum. These data demonstrate that, in certain cellular contexts, FGF can act as an inhibitory cue for axonal growth and that arachidonic acid is the second messenger responsible for this activity. We discuss the possibility that arachidonic acid inhibits neurite outgrowth by desensitising the second messenger pathway underlying neuronal responsiveness to cell adhesion molecules.

3T3 Cells↗
PubMed1994

Activation of the FGF receptor underlies neurite outgrowth stimulated by L1, N-CAM, and N-cadherin.

Cell contact-dependent neurite outgrowth stimulated by CAMs requires activation of a second messenger pathway that requires the function of a tyrosine kinase upstream from calcium influx into neurons. In the present study, we present evidence that implicates activation of the fibroblast growth factor receptor (FGFR) in the pathway underlying neurite outgrowth stimulated by L1, N-CAM, and N-cadherin. We have identified a CAM homology domain in the FGF family of receptors and show that antibodies which bind to this domain specifically inhibit neurite outgrowth stimulated by the above CAMs. We also show that synthetic peptides derived from this domain can differentially and specifically inhibit neurite outgrowth stimulated by L1, N-CAM, and N-cadherin. In addition, a soluble L1-Fc chimera is shown to stimulate an increase in phosphotyrosine on the same set of neuronal proteins that are phosphorylated following activation of the FGFR with basic FGF.

Amino Acid Sequence↗
PubMed1994

A novel role for myelin-associated glycoprotein as an inhibitor of axonal regeneration.

Following nerve injury, axons in the CNS do not normally regenerate. It has been shown that CNS myelin inhibits neurite outgrowth, though the nature of the molecules responsible for this effect are not known. Here, we demonstrate that the myelin-associated glycoprotein (MAG), a transmembrane protein of both CNS and PNS myelin, strongly inhibits neurite outgrowth from both developing cerebellar and adult dorsal root ganglion (DRG) neurons in vitro. This inhibition is reversed by an anti-MAG antibody. In contrast, MAG promotes neurite outgrowth from newborn DRG neurons. These results suggest that MAG may be responsible, in part, for the lack of CNS nerve regeneration in vivo and may influence, both temporally and spatially, regeneration in the PNS.

Animals↗
PubMed1994

Signal transduction events underlying neurite outgrowth stimulated by cell adhesion molecules.

During development of the nervous system, cell adhesion molecules (CAMs) promote cell migration and axonal growth; yet, at other times, CAMs inhibit these events by maintaining stable adhesion between cells. In the present review, we consider recent results that help to explain the paradoxical findings that individual CAMs can both promote and inhibit neuronal plasticity. In particular, we discuss the accumulating evidence that axonal growth stimulated by CAMs depends upon the activation of a second messenger pathway that culminates in calcium entry into neurons rather than on adhesion per se.

Animals↗
PubMed1994

The production of arachidonic acid can account for calcium channel activation in the second messenger pathway underlying neurite outgrowth stimulated by NCAM, N-cadherin, and L1.

We have used monolayers of control 3T3 fibroblasts and 3T3 fibroblasts expressing transfected cell adhesion molecules (CAMs)--NCAM, N-cadherin, and L1--as a culture substrate for cerebellar neurones. The transfected CAMs promote neurite outgrowth by activating a second messenger pathway that culminates in calcium influx into neurones through N- and L-type calcium channels. We show that the same neurite outgrowth response can be directly induced by arachidonic acid (10 microM) and that this response can be inhibited by N- and L-type calcium channel antagonists. In cells, arachidonic acid can be generated by phospholipase A2 or by the sequential activities of a phospholipase C (to generate diacylglycerol) and diacylglycerol lipase. In the present study we show the neurite outgrowth stimulated by CAMs (but not by various other agents) can be abolished by an inhibitor of diacylglycerol lipase acting at a site upstream from calcium channel activation. The results suggest that arachidonic acid and/or one of its metabolites is the second messenger that activates calcium channels in the CAM signalling pathway leading to axonal growth, and this is supported by recent evidence that shows the same concentrations of arachidonic acid can increase voltage-dependent calcium currents in cardiac myocytes.

3T3 Cells↗