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Biomedical subjects

Stephen B Dunnett

Publications and source records attributed to Stephen B Dunnett.

At least 19 recordsLinked to original sources
PubMed2007

Stem cell transplantation for Huntington's disease.

By way of commentary on a recent report that transplanted adult neural progenitor cells can alleviate functional deficits in a rat lesion model of Huntington's disease [Vazey, E.M., Chen, K., Hughes, S.M., Connor, B., 2006. Transplanted adult neural progenitor cells survive, differentiate and reduce motor function impairment in a rodent model of Huntington's disease. Exp. Neurol. 199, 384-396], we review the current status of the field exploring the use of stem cells, progenitor cells and immortalised cell lines to repair the lesioned striatum in animal models of the human disease. A remarkably rich range of alternative cell types have been used in various animal models, several of which exhibit cell survival and incorporation in the host brain, leading to subsequent functional recovery. In comparing the alternatives with the 'gold standard' currently offered by primary tissue grafts, key issues turn out to be: cell survival, differentiation prior to and following implantation into striatal-like phenotypes, integration and connectivity with the host brain, the nature of the electrophysiological, motor and cognitive tests used to assess functional repair, and the mechanisms by which the grafts exert their function. Although none of the alternatives yet has the capacity to match primary fetal tissues for functional repair, that standard is itself limited, and the long term goal must be not just to match but to surpass present capabilities in order to achieve fully functional reconstruction reliably, flexibly, and on demand.

Animals↗
PubMed2006

Comparison of incremental and accelerating protocols of the rotarod test for the assessment of motor deficits in the 6-OHDA model.

The rotarod test, in which animals must balance on a rotating drum, is widely used to assess motor deficit in neurodegenerative disease models in rodents. Performance is measured by the duration that an animal stays on the rod as a function of drum speed. Two different protocols are widely used, incremental fixed speeds or an accelerating protocol, but there is little information on their equivalence or the relative power, reliability and sensitivity of the two protocols. The present study was undertaken to compare the incremental fixed-speed and accelerating rotarod protocols on two different lesions of the ascending forebrain dopamine pathways. Three groups of rats were used, controls, rats with 6-OHDA lesions of nigrostriatal bundle, and rats with terminal 6-OHDA lesions within the striatum. Rats were tested at different time points after the lesion. We report that whereas the incremental protocol is more sensitive to detect the presence of a lesion, the accelerating protocol provides a more discriminative test to correlate motor deficits against lesion size.

Acceleration↗
PubMed2006

An investigation of the problem of two-layered immunohistochemical staining in paraformaldehyde fixed sections.

In sections of paraformaldehyde fixed brain tissue, stained using immunohistochemical methods, the distribution of staining within the sections is not uniform. Whilst stained cells are seen at the top and bottom surfaces, the central thicknesses of the sections contain little or no immunoreactivity. This presents a major problem for quantification, as each section contains a population of cells that is not visualized by the staining method. Following extensive investigation of this phenomenon, we report that the failure of full thickness, immunohistochemical staining is not a failure of the immunohistochemical methodology per se, nor is it related directly to the thickness of the sections used. Rather, the problem lies in the chemistry of the tissue itself, and originates during fixation of the tissues using paraformaldehyde-based perfusion methods, which render the cell membranes impermeable to one or more components of the staining protocol. We show that this impermeability is affected by addition of membrane-disrupting agents to the fixative, and by a reduction of exposure to paraformaldehyde during fixation. The present investigation contributes to the development of new fixation protocols, optimised for use in both immunohistochemical methods and morphometric analyses.

Adenoviridae↗
PubMed2006

Amphetamine induced rotation in the assessment of lesions and grafts in the unilateral rat model of Parkinson's disease.

In the unilateral rat model of Parkinson's disease (PD), amphetamine induced rotation is widely used as an index of both lesion deficits and of graft-derived recovery. We have analysed the time course of the rotational response in lesioned rats, and in rats with lesions and dopamine grafts. In lesioned rats, the rotation exhibited a typical dose-dependent response, with low rates of rotation in the first 10 min after injection, rising gradually to a maximum after 20-30 min. Grafted rats exhibited a peak of rotation in the first 10 min after injection, which then fell to a minimum after 30 min. We demonstrate that the response seen in grafted rats is both drug and dose-dependent and show that the rotational profile results from interaction of the grafted and intact striata which exhibit differential temporal responses to the amphetamine.

Amphetamine↗
PubMed2006

Hippocampal lesions impair performance on a conditional delayed matching and non-matching to position task in the rat.

The hippocampus is thought to be involved in a range of cognitive processes, from the ability to acquire new memories, to the ability to learn about spatial relationships. Humans and monkeys with damage to the hippocampus are typically impaired on delayed matching to sample tasks, of which the operant delayed matching to position task (DMTP) is a rat analogue. The reported effects of hippocampal damage on DMTP vary, ranging from delay-dependent deficits to no deficit whatsoever. The present study investigates a novel memory task; the conditional delayed matching/non-matching to position task (CDM/NMTP) in the Skinner box. CDM/NMTP uses the presence of specific stimulus cues to signify whether a particular trial is matching or non-matching in nature. Thus, it incorporates both the task contingencies within one session, and supplements the requirement for remembering the side of the lever in the sample phase with attending to the stimulus and remembering the conditional discrimination for the rule. Rats were trained preoperatively and the effects of bilateral excitotoxic lesions of the hippocampus were examined on postoperative retention of the task. Rats with lesions of the hippocampus incurred a significant impairment on the task that was manifest at all delays intervals. Despite a bias towards matching during training, trials of either type were performed with equivalent accuracy and neither rule was affected differentially by the lesion. This task may prove useful in determining the cognitive roles of a range of brain areas.

Animals↗
PubMed2006

Double dissociation between hippocampal and prefrontal lesions on an operant delayed matching task and a water maze reference memory task.

The hippocampus and prefrontal cortex have both been implicated in various aspects of the acquisition, retention and performance of delayed matching to position (DMTP) tasks in the rat, although their precise respective contributions remain unclear. In the present study, rats were trained preoperatively on DMTP before receiving excitotoxic bilateral lesions of either the entire hippocampus or the medial prefrontal cortex. Rats with lesions of the prefrontal cortex exhibited a significant delay-dependent impairment on retention of the DMTP task, whereas hippocampal lesions were without effect. Rats were also exposed to a switch in the contingencies to a 'non-matching' rule, as an analogue of switching between decision rules in the human Wisconsin Card Sorting Test, in which human patients with prefrontal damage are impaired. Both lesion groups acquired the new contingency at control levels, providing no evidence towards a role for either of these areas in this type of rule-switching. The same rats were also assessed in a spatial reference memory task in the water maze, which revealed an impairment in escape latencies and path length that was specific to the hippocampal lesions. The results corroborate previous evidence that the hippocampus is not necessary for at least some aspects of working memory performance in the DMTP task, whereas the delay-dependent deficit in the prefrontal lesion group support this task as a potentially powerful tool for assessing the cognitive changes associated with frontal damage and repair.

Animals↗
PubMed2006

Assessment of the relationship between pre-chip and post-chip quality measures for Affymetrix GeneChip expression data.

BACKGROUND: Gene expression microarray experiments are expensive to conduct and guidelines for acceptable quality control at intermediate steps before and after the samples are hybridised to chips are vague. We conducted an experiment hybridising RNA from human brain to 117 U133A Affymetrix GeneChips and used these data to explore the relationship between 4 pre-chip variables and 22 post-chip outcomes and quality control measures. RESULTS: We found that the pre-chip variables were significantly correlated with each other but that this correlation was strongest between measures of RNA quality and cRNA yield. Post-mortem interval was negatively correlated with these variables. Four principal components, reflecting array outliers, array adjustment, hybridisation noise and RNA integrity, explain about 75% of the total post-chip measure variability. Two significant canonical correlations existed between the pre-chip and post-chip variables, derived from MAS 5.0, dChip and the Bioconductor packages affy and affyPLM. The strongest (CANCOR 0.838, p < 0.0001) correlated RNA integrity and yield with post chip quality control (QC) measures indexing 3'/5' RNA ratios, bias or scaling of the chip and scaling of the variability of the signal across the chip. Post-mortem interval was relatively unimportant. We also found that the RNA integrity number (RIN) could be moderately well predicted by post-chip measures B_ACTIN35, GAPDH35 and SF. CONCLUSION: We have found that the post-chip variables having the strongest association with quantities measurable before hybridisation are those reflecting RNA integrity. Other aspects of quality, such as noise measures (reflecting the execution of the assay) or measures reflecting data quality (outlier status and array adjustment variables) are not well predicted by the variables we were able to determine ahead of time. There could be other variables measurable pre-hybridisation which may be better associated with expression data quality measures. Uncovering such connections could create savings on costly microarray experiments by eliminating poor samples before hybridisation.

Artifacts↗
PubMed2006

Fronto-striatal disconnection disrupts operant delayed alternation performance in the rat.

The hypothesis that an intact corticostriatal system is necessary to mediate accurate performance of an operant version of a classic prefrontal cortical task, delayed alternation, is tested using a crossover lesion paradigm in rats. Following an initial midline transection of the genu of the corpus callosum to separate the hemispheres, crossed lesions of the striatum in one hemisphere and the prefrontal cortex in the other produced a significant and stable impairment in delayed alternation performance whereas similar lesions made on the same side had little detectable effect. Accuracy and signal detection analysis of performance across different intertrial intervals indicated that the crossed lesions induced delay-dependent deficits in working memory aspects of corticostriatal function, without changes in low levels of response bias.

Animals↗
PubMed2006

Impaired bidirectional synaptic plasticity and procedural memory formation in striatum-specific cAMP response element-binding protein-deficient mice.

The striatum has a well documented role in procedural learning and memory. However, the synaptic and molecular mechanisms of acquisition and storage of this form of memory remain poorly understood. We examined procedural memory and plasticity in transgenic mice reversibly expressing a dominant-negative cAMP response element-binding protein (CREB) mutant in the dorsal striatum. In these transgenic mice, corticostriatal long-term potentiation and depression are abolished, indicating that CREB function is essential for bidirectional long-term synaptic plasticity in this structure. Importantly, CREB-deficient animals show reversible alterations in several forms of striatum-dependent memory, including footshock avoidance learning and "response" learning in the cross maze. These findings implicate transcriptional regulation by CREB family transcription factors in striatum-dependent information processing and provide the first clear correlation between procedural learning and memory and synaptic plasticity at the corticostriatal synapse.

Animals↗
PubMed2006

Striatal grafts alleviate bilateral striatal lesion deficits in operant delayed alternation in the rat.

In order to assess the capacity of striatal grafts to alleviate cognitive deficits of the frontal type that arise following bilateral striatal lesions, control, lesion and grafted rats were tested in an operant test of delayed alternation. Bilateral striatal lesions induced a marked impairment in choice accuracy, and signal detection analysis indicated that the lesion animals were reliably impaired on both parametric and non-parametric indices of discriminative sensitivity but not of response bias. The impairment was apparent at all intertrial interval delays, including the very shortest, suggesting the deficit is one of frontal-type executive function rather than of short-term memory. The grafted animals exhibited a significant alleviation of the deficit, again apparent at all delays. Histological analyses indicated good graft survival, and injections of a dextran amine anterograde tracer bilaterally into the host prefrontal cortex indicated reformation of extensive projections into the grafted tissues. Since performance of the operant delayed alternation task is dependent upon the integrity of corticostriatal connections, which is disrupted bilaterally by the lesions and restored to the grafts in the transplanted animals, the results corroborate the hypothesis that striatal grafts can alleviate complex cognitive functions of the frontal type by a mechanism that involves functional integration of the grafted neurons into the neural circuits of the host brain.

Animals↗
PubMed2006

The effects of lateralized training on spontaneous forelimb preference, lesion deficits, and graft-mediated functional recovery after unilateral striatal lesions in rats.

The ability of striatal embryonic grafts to promote functional recovery on complex behavioral tasks depends on various factors, including the amount of striatal-like tissue within the grafts and the duration of post-graft training. However, how the innate paw bias of animals is affected by experience, or influences recovery following injury, is less known. Here, we have examined the effects of intrinsic side bias and lateralized limb use training on spontaneous forelimb preference and graft-mediated functional recovery in a skilled reaching task in a rodent model of Huntington's disease. Naïve rats were assessed on their baseline paw preferences when reaching between the bars of their cage to retrieve sugar pellets from a tray attached outside. Next, rats were lesioned unilaterally in the lateral dorsal striatum with quinolinic acid, and 7-10 days later, half of the animals were given suspension grafts prepared from E15 whole ganglionic eminence implanted into the lesioned striatum. The animals then received extensive unilateral training, either ipsi- or contralateral to the side of the lesion and graft in separate subgroups, on the 'staircase' task until asymptotic performance was obtained. As reported previously, the grafts alleviated lesion-induced deficits in retrieving pellets from the contralateral staircase. Spontaneous biases were then reassessed in the cage-reaching task. Irrespective of whether the animal received ipsilateral or contralateral staircase training, the unilateral lesions induced a significant shift in spontaneous bias towards the ipsilateral paw. Grafted animals showed a similar shift in bias if staircase training was given to the ipsilateral paw but showed no change in spontaneous bias (similar to controls) if they had received contralateral training during the post-transplantation period. The results suggest that striatal grafts can alleviate lesion-induced changes in their spontaneous side preferences, but only if they receive extensive training in the use of the contralateral limb, compatible with the notion that recovery is use-dependent.

Animals↗
PubMed2006

Pharmaceutical, cellular and genetic therapies for Huntington's disease.

HD (Huntington's disease) is a devastating neurodegenerative disorder caused by a polyglutamine expansion in the gene encoding the huntingtin protein. Presently, there is no known cure for HD and existing symptomatic treatments are limited. However, recent advances have identified multiple pathological mechanisms involved in HD, some of which have now become the focus of therapeutic intervention. In this review, we consider progress made towards developing safe and effective pharmaceutical-, cell- and genetic-based therapies, and discuss the extent to which some of these therapies have been successfully translated into clinical trials. These new prospects offer hope for delaying and possibly halting this debilitating disease.

Animals↗
PubMed2006

Re-examining the ontogeny of substantia nigra dopamine neurons.

Recently, the need to detail the precise ontogeny of nigrostriatal dopamine neurons has grown significantly. It is now thought that the gestational day on which the majority of these neurons are born is important not only for maximizing the yield of primary cells for transplantation but also for extracting suitable dopamine neural precursors (as stem cells) for expansion in vitro. Historically, peak ontogeny of substantia nigra pars compacta (SNc) dopamine neurons in the rat has been considered to occur around embryonic day (E)14. However, such a concept is at odds with recent studies that reveal not only that substantial numbers of tyrosine hydroxylase-immunopositive cells reside in the ventral mesencephalic region of rats at E14 but that many of these cells have matured extensive axonal projections to the ventral forebrain. Here, then, the ontogeny of SNc neurons in rats commonly used as a source of donor tissue for experimental cell transplantation in animal models of Parkinson's disease has been re-examined. Using a combination of bromodeoxyuridine (BrdU) administration at E11, E12, E13 or E14 with immunocytochemical stainings for both BrdU and tyrosine hydroxylase after 4 weeks of postnatal development, this characterization reveals that the vast majority (perhaps 80%) of SNc dopamine neurons are probably born on E12 in Sprague-Dawley rats. Such findings are important in refining the use of embryonic tissues for primary cell transplantation and may provide more precise timing for identifying the cellular and molecular events that drive neural stem cells toward a dopaminergic phenotype during development.

Animals↗
PubMed2006

Morphological and cellular changes within embryonic striatal grafts associated with enriched environment and involuntary exercise.

Environmental enrichment (EE) and exercise have been implicated in influencing behaviour and altering neuronal processes associated with cellular morphology in both 'normal' and injured states of the CNS. Using a rodent model of Huntington's disease, we investigated whether prolonged EE or involuntary exercise can induce morphological and cellular changes within embryonic striatal transplants. Adult rats were trained on the Staircase test--requiring fine motor control to reach and collect reward pellets--prior to being lesioned unilaterally in the dorsal neostriatum with quinolinic acid. The lesioned animals received E15 whole ganglionic eminence cell suspension grafts followed by housing in EE or standard cages. Half of the animals in standard cages received daily forced exercise on a treadmill. The grafted animals showed significant functional recovery on both the Staircase test and in drug-induced rotation. Neither the housing conditions nor the training had an impact on the behaviour, with the exception of the treadmill reducing the ipsilateral drug-induced rotation observed amongst the lesioned animals. However, the animals housed in the EE had significantly increased striatal brain-derived neurotrophic factor (BDNF) levels, and graft neurons in these animals exhibited both greater spine densities and larger cell volumes. Animals on forced exercise regime had reduced BDNF levels and grafted cells with sparser spines. The study suggests that the context of the animal can affect the plasticity of transplanted cells. Appropriately exploiting the underlying, and yet unknown, mechanisms could lead the way to improved anatomical and potentially functional integration of the graft.

Animals↗
PubMed2005

EGF and FGF-2 responsiveness of rat and mouse neural precursors derived from the embryonic CNS.

EGF and FGF-2 induce the proliferation of embryonic neural precursors (ENPs) in vitro from a number of different species. In this study, we demonstrate that embryonic age is a crucial determinant of the number and differentiation potential of rat embryonic neural precursor cells responding to either EGF and/or FGF-2, in that (i) there is a differential response to the two growth factors (both alone and in combination) according to the gestational age of isolation and (ii) when allowed to differentiate, there are temporal changes in the ability of these cells to produce neurons. Furthermore, for cultures of all gestational ages, there is a defined pattern of senescence, with cultures expanding longest when cells are isolated earlier in gestation. The suggestion is that rat ENPs in this study consist predominantly of neural progenitor cells with limited division potential rather than self-renewing multipotential neural stem cells. In contrast, mouse ENPs appeared to expand indefinitely and thus allow for longer studies to be carried out looking at the effects of growth factor concentrations. The effect of varying the concentration of EGF was assessed using mouse ENPs.

Animals↗
PubMed2005

The Corridor Task: a simple test of lateralised response selection sensitive to unilateral dopamine deafferentation and graft-derived dopamine replacement in the striatum.

In this experiment, we report a novel drug-free behavioural test of lateralised neglect which is sensitive to unilateral dopamine-denervating lesions and subsequent graft-derived striatal dopamine replacement. For the task, white plastic lids containing sugar pellets were placed along the left and right sides of the floor of a long narrow corridor at regular intervals. Hungry female Sprague-Dawley rats were placed individually into the corridor where they were allowed to make up to 20 pellet retrievals. The number of retrievals each rat made from its left and right sides was counted. Complete mesencephalic or partial nigrostriatal lesions were induced by injection of 6-hydroxydopamine into the medial forebrain bundle or striatum, respectively. Both lesions induced a pronounced ipsilateral retrieval bias in the task. Five weeks after lesion surgery, half of the rats from each lesion group were given E14 ventral mesencephalic cell suspension transplants into the denervated striatum, and were then re-tested in the Corridor Task 5 and 10 weeks later. There was no amelioration of the side bias in rats with medial forebrain bundle lesions. In contrast, in nigrostriatal-lesioned rats, the graft significantly reduced the lesion-induced ipsilateral bias. We conclude that the Corridor Task is a sensitive test of lateralised sensorimotor response selection, and is suitable for assessing deficits and recovery associated with lesions and grafts within the nigrostriatal system.

Animals↗
PubMed2005

Comparison of 6-hydroxydopamine-induced medial forebrain bundle and nigrostriatal terminal lesions in rats using a lateralised nose-poking task with low stimulus-response compatibility.

Experimental therapies for Parkinson's disease are commonly validated in unilateral rat lesion models using simple tests of motor asymmetry such as rotation. However, the human disorder is considerably more complex than this, and alternative tests that yield a more relevant evaluation of the lesion-induced deficit could provide a powerful behavioural tool for analysis of novel therapies. This study evaluated the potential of a lateralised nose-poking task for detailed assessment of the deficit in rats with complete unilateral and partial nigrostriatal terminal lesions induced by unilateral injection of 6-hydroxydopamine (6-OHDA) into the medial forebrain bundle (MFB) or striatum, respectively. To maximise the task's potential for revealing chronic deficits, a version of the task that requires rats to nose-poke away from visual stimuli associated with food rewards was used. Because of its low stimulus-response compatibility, performance of this version is not mediated by the rats innate tendencies, and thus, is less likely to spontaneously recover after partial lesions. Two weeks and 4 months after lesion surgery, both groups of lesioned rats developed an ipsilateral responding bias with a corresponding drop in contralateral accuracy. Rats with complete (but not partial) lesions also developed a deficit in attempting trials, reacting to stimuli requiring a contralateral nose-poke and executing movements bilaterally. This experiment suggests that the version of the lateralised nose-poking task requiring an arbitrary stimulus-response association may be a powerful behavioural tool for assessment of complete or partial dopamine lesions, and any subsequent dopamine replacement strategy.

Animals↗