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Eleanor M Donnelly,
Nicolas N Madigan,
Gemma E Rooney,
Andrew Knight,
Bingkun Chen,
Bret Ball,
Lisa Kinnavane,
Yolanda Garcia,
Peter Dockery, John Fraher,
Padraig M Strappe,
Anthony J Windebank,
Timothy O'Brien,
Siobhan S McMahon
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ABSTRACT: Abstract Background aims. In this study we investigated the effect of neurotrophin-3 (NT-3) and knockdown of NG2, one of the main inhibitory chondroitin sulfate proteoglycans (CSPG), in the glial scar following spinal cord injury (SCI). Methods. Short hairpin (sh) RNA were designed to target NG2 and were cloned into a lentiviral vector (LV). A LV was also constructed containing NT-3. LV expressing NT-3, shRNA to NG2 or combinations of both vectors were injected directly into contused rat spinal cords 1 week post-injury. Six weeks post-injection of LV, spinal cords were examined by histology for changes in scar size and by immunohistochemistry for changes in expression of CSPG, NT-3, astrocytes, neurons and microglia/macrophages. Motor function was assessed using the Basso, Beattie and Bresnahan (BBB) locomotor scale. Results. Animals that received the combination treatment of LV shNG2 and LV NT-3 showed reduced scar size. These animals also showed an increase in levels of neurons and NG2, a decrease in levels of astrocytes and a significant functional recovery as assessed using the BBB locomotor scale at 2 weeks post-treatment. Conclusions. The improvement in locomotor recovery and decrease in scar size shows the potential of this gene therapy approach as a therapeutic treatment for SCI.
Cytotherapy 11/2012; 14(10):1235-44. · 3.63 Impact Factor
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ABSTRACT: A combination of dissection and computed tomography scanning has provided significant novel insights into the structure and function of the Dermochelys coriacea larynx and its associated muscles. Several previously unknown features of the laryngeal aditus (glottis) are described and their functional significance in its opening and closure are considered. The tongue plays an essential part in producing and maintaining closure during dives and feeding bouts. Closure is brought about by compression of the glottis under the action of the two hyoglossus muscles. The tongue thus plays the role of the epiglottis of mammals, sealing the entrance to the larynx. As is already clear, opening is brought about by abduction of the arytenoid cartilages. In addition, there is a powerful mechanism for maintaining the larynx in close apposition to the hyoid plate during feeding and neck flexion, thereby enhancing the efficiency of feeding.
Journal of Experimental Biology 12/2010; 213(Pt 24):4137-45. · 3.00 Impact Factor
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Eleanor M Donnelly,
Padraig M Strappe,
Lisa M McGinley,
Nicolas N Madigan,
Elizabeth Geurts,
Gemma E Rooney,
Anthony J Windebank, John Fraher,
Peter Dockery,
Timothy O'Brien,
Siobhan S McMahon
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ABSTRACT: Following spinal cord injury, a highly inhibitory environment for axonal regeneration develops. One of the main sources of this inhibition is the glial scar that is formed after injury by reactive astrocytes. The inhibitory environment is mainly a result of chondroitin sulphate proteoglycans (CSPGs). NG2, [corrected] one of the main inhibitory CSPGs, is up-regulated following spinal cord injury.
Small interfering RNA (siRNA) was designed to target NG2 and this short hairpin RNA (shRNA) was cloned into a lentiviral vector (LV). The neurotrophic factor neurotrophin-3 (NT-3) promotes the growth and survival of developing neurites and has also been shown to aid regeneration. NT-3 was also cloned into a LV. In vitro assessment of these vectors using a coculture system of dorsal root ganglia (DRG) neurones and Neu7 astrocytes was carried out. The Neu7 cell line is a rat astrocyte cell line that overexpresses NG2, thereby mimicking the inhibitory environment following spinal cord injury.
These experiments show that both the knockdown of NG2 via shRNA and over-expression of NT-3 can significantly increase neurite growth, although a combination of both vectors did not confer any additional benefit over the vectors used individually. These LVs show promising potential for growth and survival of neurites in injured central nervous system tissue (CNS).
The Journal of Gene Medicine 11/2010; 12(11):863-72. · 2.48 Impact Factor
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ABSTRACT: Adult leatherbacks are large animals (300-500 kg), overlapping in size with marine pinniped and cetacean species. Unlike marine mammals, they start their aquatic life as 40-50 g hatchlings, so undergo a 10,000-fold increase in body mass during independent existence. Hatchlings are limited to the tropics and near-surface water. Adults, obligate predators on gelatinous plankton, encounter cold water at depth (<1280 m) or high latitude and are gigantotherms that maintain elevated core body temperatures in cold water. This study shows that there are great ontogenetic changes in tracheal structure related to diving and exposure to cold. Hatchling leatherbacks have a conventional reptilian tracheal structure with circular cartilaginous rings interspersed with extensive connective tissue. The adult trachea is an almost continuous ellipsoidal cartilaginous tube composed of interlocking plates, and will collapse easily in the upper part of the water column during dives, thus avoiding pressure-related structural and physiological problems. It is lined with an extensive, dense erectile vascular plexus that will warm and humidify cold inspired air and possibly retain heat on expiration. A sub-luminal lymphatic plexus is also present. Mammals and birds have independently evolved nasal turbinates to fulfil such a respiratory thermocontrol function; for them, turbinates are regarded as diagnostic of endothermy. This is the first demonstration of a turbinate equivalent in a living reptile.
Journal of Experimental Biology 11/2009; 212(Pt 21):3440-7. · 3.00 Impact Factor
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ABSTRACT: Adult leatherback turtles are gigantothermic/endothermic when foraging in cool temperate waters, maintaining a core body temperature within the main body cavity of ca. 25 degrees C despite encountering surface temperatures of ca. 15 degrees C and temperatures as low as 0.4 degrees C during dives. Leatherbacks also eat very large quantities of cold, gelatinous prey (medusae and pyrosomas). We hypothesised that the head and neck of the leatherback would have structural features to minimise cephalic heat loss and limit cooling of the head and neck during food ingestion. By gross dissection and analytical computed tomography (validated by ground truthing dissection) of an embalmed specimen we confirmed this prediction. 21% of the head and neck was occupied by adipose tissue. This occurred as intracranial blubber, encapsulating the salt glands, medial portions of the eyeballs, plus the neurocranium and brain. The dorsal and lateral surfaces of the neck featured thick blubber pads whereas the carotid arteries and jugular veins were deeply buried in the neck and protected laterally by blubber. The oesophagus was surrounded by a thick sheath of adipose tissue whereas the oropharyngeal cavity had an adipose layer between it and the bony proportion of the palate, providing further ventral insulation for salt glands and neurocranium.
Journal of Experimental Biology 10/2009; 212(17):2753-9. · 3.00 Impact Factor
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ABSTRACT: The floor plate of the neural tube is of major importance in determining axonal behaviour, such that, having crossed, decussating axons do not cross back again. The ventral commissure (VC) of the spinal cord forms immediately ventral to the floor plate shortly after neural tube closure. It is the principal location in which decussating axons cross the midline. It is probably also of major importance in neural tube development, but has received relatively little attention. This study analyses the growth and development of the rat VC and also axon-glial relationships within it throughout the crucial prenatal period of extensive transmedian axon growth, when key biochemical interactions between the two tissues are taking place. The morphometric, stereological and immunohistochemical methods used show that the axonal and glial populations remain in a finely balanced equilibrium throughout a period of almost a hundred-fold growth of both elements. At all stages axons are highly segregated into small bundles of constant size by glial processes, to which they are closely apposed. Thus, glial-axon contact is remarkably precocious, uniquely intimate and persists throughout VC development. This suggests that the relationship between the two tissues is highly controlled through interactions between them. The VC is likely to be the physical basis of a second set of glial-axonal interactions, namely, those which are well known to influence axon crossing behaviour. In mediating these, the extensive axon-glial contact is an ideal arrangement for molecular transfer between them, and is probably the substrate for altering axon responsiveness and ensuring reliable transmedian decussation. The VC is therefore a segregating matrix temporally and spatially specialised for a range of key developmental axon-glial interactions.
Journal of Neurocytology 10/2004; 33(5):489-501. · 1.94 Impact Factor
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John Fraher
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ABSTRACT: At most vertebrate nerve transitional zones (TZs) there is a glial barrier which is pierced by axons passing between the CNS and PNS. Myelinated axons traverse this in individual tunnels. The same is true of larger non-myelinated axons. This holds widely among the vertebrates, for example, the large motor axons of the sea-lamprey Petromyzon (which also possess TZ specializations not found in mammals). Smaller non-myelinated axons traverse the TZ glial tunnels as fascicles and so the barriers are correspondingly less comprehensive for them. Accordingly, in nerves composed of non-myelinated axons, such as the vomeronasal or the olfactory, a TZ barrier stretching across the nerve is effectivelyabsent. The chordateAmphioxus differsfrom the vertebrates in lacking a TZ barrier throughout. Invertebrates also lack glial barriers at the TZs between ganglia and interconnecting nerve trunks. The glial barrier at the dorsal spinal root TZ (DRTZ) has considerable value for analysing protocols aimed at achieving CNS regeneration, because it provides a useful model of the gliotic reaction at sites of CNS injury. Also, it is especially amenable to morphometric analysis, and so enables objective quantification of different protocols. Being adjacent to the subarachnoid space, it is accessible for experimental intervention. The DRTZ was used to investigate the value of neurotrophin 3 (NT3) in promoting axon regeneration across the TZ barrier and into the CNS following dorsal root crush. It promoted extensive regeneration and vigorous non-myelinated axonal ensheathment. On average, around 40% of regenerating axons grew across the interface, compared with virtually none in its absence. These may have traversed the interface through loci occupied by axons prior to degeneration. Many regenerating axons became myelinated, both centrally and peripherally.
Journal of Anatomy 05/2002; 200(4):415-30. · 2.37 Impact Factor
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J Fraher
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ABSTRACT: This Anatomical Society symposium, held at University College, Cork in September 1995 was the first of its kind. The objective was to stimulate and facilitate constructive interaction between experts on topics relating to the various types of glial barrier which form partitions within the nervous system. Some of these barriers are transient and are found only during development, for example, those which define the limits of developing nuclei or fibre bundles. Others are permanent, such as those at the transitional zones which separate the CNS and PNS milieux at nerve root attachments to the neuraxis. Still others, such as glial scars, are seen only following injury when they tend to inhibit neurite regeneration. In experimental circumstances, for example following irradiation or chemical damage, glial barriers may be broken down and even relocated. This may be associated with Schwann cell invasion of the CNS and, experimentally, with Schwann cell and glial cell transplantation into demyelinated areas. Such transplantation studies are in turn related to remyelination and CNS axon regeneration and the factors which facilitate these. Twelve review lectures were given on these topics. Five articles based on these communications are reproduced here. The underlying theme was the relationship between advances in the understanding of fundamental nervous tissue biology, especially as related to glial cells, and potential developments aimed at treating CNS demyelinating diseases and achieving CNS regeneration.
Journal of Anatomy 02/1997; 190(Pt 1):3. · 2.37 Impact Factor
