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S B Dunnett

Publications and source records attributed to S B Dunnett.

At least 19 recordsLinked to original sources
PubMed1996

GDNF enhances dopaminergic cell survival and fibre outgrowth in embryonic nigral grafts.

Two groups of rats with unilateral 6OHDA lesions received either intrastriatal suspension grafts of embryonic ventral mesencephalon or sham grafts. Three subgroups of each of these received intrastriatal infusions of 1000 ng or 500 ng glial cell-line derived trophic factor (GDNF) or vehicle alone for 10 consecutive days. There was a highly significant dose-dependent effect of GDNF both on the number of TH-positive cells surviving in the grafts and on the density of fibre outgrowth. All grafted rats showed rapid compensation of amphetamine-induced rotation compared with rats with sham grafts. GDNF may provide a powerful tool to enhance the survival and maturation of dopaminergic neurones within mesencephalic transplants.

Animals↗
PubMed1996

Expression of c-fos, jun D and pp60c-src+ mRNAs in the developing and grafted rat striatum.

Expression of the mRNAs of the proto-oncogenes pp60c-src+, c-fos and jun D were studied using in-situ hybridisation histochemistry in the developing striatum and in striatal grafts. The temporal patterns of mRNA expression were monitored in the striatum of the normal developing rat from the 12th day of gestation (E12) to 10 days postnatally, and were compared to the changes in gene expression observed in E13-E14 primordial striatal tissue grafts 7, 15 and 30 days after implantation in the ibotenic acid-lesioned striatum of adult rats. During development, all three proto-oncogenes were most highly expressed just before birth, at E19. Striatal expression of all three proto-oncogenes was markedly reduced after birth and remained at a low level through to adulthood. A different mode of expression was observed in the transplanted striatum which was unique to each particular gene. jun D and pp60c-src+ were expressed for a longer time period in the grafted primordial cells than in normal development, whereas no c-fos expression could be detected in the grafts. These results suggest that transplantation of embryonic neural cells into the host brain may affect the normal developmental regulation of such cells and their expression of some proto-oncogenes.

Animals↗
PubMed1996

The neurotrophin NT4/5, but not NT3, enhances the efficacy of nigral grafts in a rat model of Parkinson's disease.

The neurotrophins NT4/5 and NT3 have previously been shown to improve the survival and fibre outgrowth of embryonic dopaminergic neurons in vitro. In the present study we attempted to augment the efficacy of embryonic nigral grafts in vivo. This was done by directly infusing the neurotrophins intraparenchymally in close proximity to transplanted nigral tissue placed in the dopamine depleted striatum of 6-hydroxydopamine lesioned rats. Our results indicated that NT4/5, but not NT3, stimulated fibre growth from embryonic nigral cells and enhanced functional efficacy of the grafts as assessed by metamphetamine-induced rotation.

Animals↗
PubMed1996

Immortalized neural cells from trisomy 16 mice as models for Alzheimer's disease.

The trisomy 16 mouse (Ts16) is a general accepted animal model for both Downs syndrome (DS) and Alzheimer's Disease (AD). However, the efficacy of this model is severely hampered by the fact that Ts16 is lethal after about 18-20 days of gestation. Chimeras, long-term tissue culture and neural transplantation of Ts16 material have previously been used to overcome this limitation presented by death in utero of the Ts16. In this paper we describe a new strategy to overcome this limitation, i.e. immortalization of primary cells from Ts16 mice with retrovirus-mediated gene transfer of a temperature sensitive immortalizing oncogene. By this method we have obtained a total of 21 stable cell lines from Ts16 hippocampus, Ts16 cortex, normal hippocampus, and normal cortex. So far, two of the cell lines have been karyotyped and as expected, the cell line immortalized from Ts16 embryos has retained three copies of chromosome 16. We are currently characterizing these cell lines with respect to expression of APP, T-antigen, Nestin, GFAP, NF and Map-2. Moreover, the processing and secretion of APP fragments are being investigated by immunoblotting. In summary, we have immortalized CNS cells from Ts16 mice and we expect that these cell lines will be useful as in vitro and in vivo models for studying various aspects of the pathology of Alzheimer's disease.

Alzheimer Disease↗
PubMed1996

Survival and differentiation of rat and human epidermal growth factor-responsive precursor cells following grafting into the lesioned adult central nervous system.

Epidermal Growth Factor (EGF)-responsive stem cells isolated from the developing central nervous system (CNS) can be expanded exponentially in culture while retaining the ability to differentiate into neurons and glia. As such, they represent a possible source of tissue for neural transplantation, providing they can survive and mature following grafting into the adult brain. In this study we have shown that purified rat stem cells generated from either the embryonic mesencephalon or the striatum can survive grafting into the striatum of rats with either ibotenic acid or nigrostriatal dopamine lesions. However, transplanted stem cells do not survive as a large mass typical of primary embryonic CNS tissue grafts, but in contrast form thin grafts containing only a small number of surviving cells. There was no extensive migration of transplanted stem cells labeled with either the lac-z gene or bromodeoxyuridine into the host region surrounding the graft, although a small number of labeled cells were seen in the ventral striatum some distance from the site of implantation. Some of these appeared to differentiate into dopamine neurons, particularly when the developing mesencephalon was used as the starting material for generating the stem cells. EGF-responsive stem cells could also be isolated from the mesencephalon of developing human embryos and expanded in culture, but only grew in large numbers when the gestational age of the embryo was greater than 11 weeks. Purified human CNS stem cells were also transplanted into immunosuppressed rats with nigrostriatal lesions and formed thin grafts similar to those seen when using rat stem cells. However, when primary cultures of human mesencephalon were grown with EGF for only 10 days and this mixture of stem cells and primary neural tissue was transplanted into the dopamine-depleted striatum, large well-formed grafts developed. These contained mostly small undifferentiated cells intermixed with a number of well-differentiated TH-positive neurons. These results show that purified populations of rat or human EGF-responsive CNS stem cells do not form large graft masses or migrate extensively into the surrounding host tissues when transplanted into the adult striatum. However, modifications of the growth conditions in vitro may lead to an improvement of their survival in vivo.

Animals↗
PubMed1996

The time course of loss of dopaminergic neurons and the gliotic reaction surrounding grafts of embryonic mesencephalon to the striatum.

Grafts of embryonic ventral mesencephalic tissue placed in the striatum of 6-hydroxydopamine-lesioned rats survive, and make and receive connections to and from the host brain. The dopaminergic neurons of the graft can grow processes into the host brain, and thereby alleviate many of the behavioral deficits of this form of experimental Parkinson's disease. However, when examined some weeks after implantation, grafted substantia nigra only contains about 5% of the expected complement of dopaminergic neurons. We have examined the time course of loss of grafted neurons. We find that the majority die during the first 7 days after transplantation. However, we have shown previously that three-dimensional cultures with the same dimensions as a graft, made of identical cell suspensions, have much better dopaminergic neuronal survival. There must, therefore, be features in the environment surrounding a graft that are toxic to dopaminergic neurons. A limiting factor in the efficacy of dopaminergic grafts is the small distance over which the neurons are able to grow neurites and form connections in the host brain. We find that the growth of neurites from dopaminergic neurons into the host striatum occurs in two phases. Neurites reach their maximum length within 7 days of transplantation, and this is followed by a much slower process of branch and terminal formation. Since axon growth in the adult brain may be inhibited by a number of factors associated with reactive gliosis, we have immunostained various ages of graft for vimentin, tenascin, chondroitin sulfate proteoglycan (CS-PG) using the CS56 antibody, the DSD-1 proteoglycan, and microglia using the OX-42 antibody. We have compared this staining with that surrounding a simple stab wound. Vimentin staining was initially seen in the graft and in astrocytes immediately surrounding it. By 7 weeks staining was restricted to a ring of astrocytes surrounding the graft. Tenascin, DSD-1, and CS-PG were initially seen in and around the grafts. By 7 weeks they had disappeared from grafts, but CS-PG and tenascin persisted in small amounts around stab wounds. In general, immunostaining of these molecules persisted longer around a stab lesion than around a graft. There was also an intense local microglial reaction surrounding both grafts and stab wounds which had largely resolved by 7 weeks.

Animals↗
PubMed1996

Fetal ventral mesencephalon of human and rat origin maintained in vitro and transplanted to 6-hydroxydopamine-lesioned rats gives rise to grafts rich in dopaminergic neurons.

Free-floating roller tube cultures of human fetal (embryonic age 6-10 weeks post-conception) and rat fetal (embryonic day 13) ventral mesencephalon were prepared. After 7-15 days in vitro, the mesencephalic tissue cultures were transplanted into the striatum of adult rats that had received unilateral injections of 6-hydroxydopamine into the nigrostriatal bundle 3-5 weeks prior to transplantation. Graft survival was assessed in tyrosine hydroxylase (TH)-immunostained serial sections of the grafted brains up to post-transplantation week 4 for the human fetal xenografts and post-transplantation week 11 for the rat fetal allografts. D-amphetamine-induced rotation was monitored up to 10 weeks after transplantation in the allografted animals and compared with that of lesioned-only control animals. All transplanted animals showed large, viable grafts containing TH-immunoreactive (ir) neurons. The density of TH-ir neurons in the human fetal xenografts and in rat fetal allografts was similar. A significant amelioration of the amphetamine-induced rotation was observed in the animals that received cultured tissue allografts. These results promote the feasibility of in vitro maintenance of fetal human and rat nigral tissue prior to transplantation using the free-floating roller tube technique.

Animals↗
PubMed1996

Functional and anatomical reconstruction of the 6-hydroxydopamine lesioned nigrostriatal system of the adult rat.

In an attempt to reconstruct the 6-hydroxydopamine lesioned nigrostriatal system of the adult rat we have combined homotopic grafting of embryonic ventral mesencephalon suspensions with the implantation of long oblique "bridge" grafts of fibroblast growth factor-4-transfected RN-22 schwannoma cells stretching from the site of the neuronal grafts to the striatum. At seven weeks after receiving both grafts, animals were killed and processed for immunohistochemistry against tyrosine hydroxylase. Tyrosine hydroxylase-immunoreactive axons were seen to extend from the nigral grafts, along the bridge graft to the striatum where terminal arborizations could be seen. The retrograde tracer Fluoro-gold was injected intrastriatally in some of the experimental animals and was taken up by grafted neurons confirming their projection to the striatum. In parallel to the anatomical reconstruction of the system, a decrease in amphetamine-induced rotation was demonstrated in those animals receiving both grafts which had received > 98% complete lesions. This decrease was greatest in those animals with the most tyrosine hydroxylase-immunoreactive axons in their bridge grafts. The presence of the bridge graft also led to an increase in neuronal graft survival with twice as many tyrosine hydroxylase-immunoreactive neurons being found in the grafts of those animals that had received both grafts compared to those that had received a neuronal graft but no bridge graft.

Amphetamine↗
PubMed1996

Bridge grafts of fibroblast growth factor-4-secreting schwannoma cells promote functional axonal regeneration in the nigrostriatal pathway of the adult rat.

Axons damaged in the adult mammalian central nervous system are able to regenerate when their inhibitory glial environment is replaced with a more permissive substrate. Here, we have used long oblique "bridge" grafts of fibroblast growth factor-4-transfected RN-22 schwannoma cells to allow mechanically lesioned nigrostriatal axons to regenerate back to their original target in the adult rat brain. Regenerated axons were able to leave the bridge graft to form terminal arborizations and increase the density of tyrosine hydroxylase-immunoreactive fibres within the striatum. Bridge grafting also resulted in an increase in the number of neurons within the substantia nigra pars compacta taking up the fluorescent retrograde tracer Fluoro-Gold from the striatum. Animals which had received RN-22 bridge grafts showed lower rates of amphetamine-induced rotation 10 weeks after a mechanical lesion of the nigrostriatal tract compared to lesioned controls, the magnitude of the behavioural effect being related to the number of regenerated axons, and this comparative reduction was reversed by mechanical section of the bridge graft. It is concluded that our bridge grafting strategy allowed the partial anatomical and functional regeneration of the mechanically lesioned nigrostriatal tract, an unmyelinated central axon bundle, and that bridge grafting therefore represents a realistic approach to the repair of central nervous system lesions involving axon tract damage.

Amphetamine↗
PubMed1996

Reduced retrograde labelling with fluorescent tracer accompanies neuronal atrophy of basal forebrain cholinergic neurons in aged rats.

During ageing, basal forebrain cholinergic neurons are prone to degeneration for unknown reasons. In this study we morphometrically evaluated the retrograde labelling of basal forebrain neurons obtained after injection of FluoroGold into multiple sites in the cerebral neocortex in aged (24-33 months) as compared with young adult (four to six months) male Sprague-Dawley rats. In addition, we looked for differences in the distribution of degenerative changes in topographic subdivisions of the basal forebrain cholinergic complex of neurons identified by immunohistochemical detection of the cholinergic markers choline acetyltransferase or low-affinity neurotrophin receptor. After injection of FluoroGold into the cerebral neocortex, the number of retrogradely labelled neurons in the horizontal diagonal band/ substantia innominata and basal nucleus was significantly lower in aged rats, by 41% and 48%, respectively. In aged rats injected with FluoroGold as well as in non-injected aged rats, the numbers of neurons immunoreactive for choline acetyltransferase and low-affinity neurotrophin receptor were significantly lower, by 23-27% in the basal forebrain system as a whole, with no significant difference in the degree of decline amongst different subdivisions (i.e. medial septum, diagonal band, substantia innominata and basal nucleus). The ratios of the number of neurons labelled with FluoroGold as compared with the number of neurons immunoreactive for either cholinergic marker were significantly lower in aged rats, by 32-37%, indicating that the decline in the number of neurons retrogradely transporting tracer was greater than the decline in the number of immunoreactive neurons in aged animals. Immunoreactive as well as retrogradely labelled neurons showed a significant shrinkage of cell surface area of 6-13% in different subdivisions of the basal forebrain cholinergic system in aged rats. These findings confirm significant loss and atrophy of basal forebrain cholinergic neurons in aged rats, and demonstrate significantly reduced retrograde labelling of these neurons with fluorescent tracer applied to their target cortex. This reduced retrograde labelling suggests an impairment of either uptake or retrograde transport mechanisms in these neurons in aged rats. Such an impairment may contribute to the degenerative changes of basal forebrain cholinergic neurons observed in ageing and age-related degenerative conditions such as Alzheimer's disease.

Acetylcholine↗
PubMed1996

The placement of a striatal ibotenic acid lesion affects skilled forelimb use and the direction of drug-induced rotation.

The motor consequences of excitotoxic striatal damage have been evaluated extensively in the rat, using tests of whole body motor asymmetry and of deficits in skilled paw and limb movements. However conflicting results of both the type and extent of behavioural deficits have been reported, particularly in the direction of rotation observed in response to the dopamine receptor agonist, apomorphine. The present study investigated the effect of unilateral ibotenic acid lesions in the dorsal striatum of the adult rat, placed at either anterior, posterior, medial, or lateral loci, on rotation in response to both amphetamine and apomorphine, and in the "staircase test" of skilled forelimb use. In a 2 x 2 matrix design experiment, adult female albino rats received a double unilateral lesion of 0.5 microliter 0.06 M ibotenic acid injected at each of two sites either anterior (medial and lateral), posterior (medial and lateral), medial (anterior and posterior), or lateral (anterior and posterior). Rats that received posterior lesions showed a marked ipsilateral rotation in response to both amphetamine and apomorphine, while animals receiving anterior lesions showed little ipsilateral or a slight contralateral bias. Rats receiving lateral lesions showed a marked impairment of contralateral paw use on the "staircase test," while animals with medial lesions showed no significant difference to control unoperated animals. These results confirm the somatotopic organisation of the dorsal striatum in its control of motor functions, and indicate the need to take into account the locus of an excitotoxic lesion in the design of lesion and transplantation studies if we are to achieve reliable tests of the behavioural deficits and recovery.

Animals↗
PubMed1996

Selective immunolesioning of the basal forebrain cholinergic system disrupts short-term memory in rats.

Selective depletion of nerve growth factor receptor-bearing neurons in the basal forebrain cholinergic system nuclei by the immunotoxin 192 IgG-saporin offers a new and highly useful tool for the study of the role of the forebrain cholinergic system in cognitive functions. In the present study, we have tested the effects of 192 IpG-saporin in an operant delayed matching-to-position task which has previously been used to discriminate between delay-dependent learning impairments and delay-independent disturbances of non-mnemonic processes. Rats were first trained to criterion performance and then received intraventricular injections of 5 microg of 192 IgG-saporin 4 weeks prior to a second testing session. Rats with 192 IgG-saporin lesions displayed a significant delay-dependent decline in performance compared to normal controls, indicating a deficit in short-term memory. Administration of the muscarinic blocker scopolamine (0.5 mg/kg, i.p.) produced more pronounced impairment in the performance of the normal control rats across all delays, and induced further impairment also in animals with 192 IgG-saporin lesions. These effects were not observed following control injections of methyl scopolamine, suggesting that the impairment induced by scopolamine was due to the blockade of central muscarinic receptors. No improvement in performance was observed in either group following systemic treatment with the muscarinic cholinergic agonist arecoline (1.00 mg/kg). Biochemical and morphological analyses confirmed the selective and severe (>90-95%) depletion of cholinergic neurons throughout the septal-diagonal band area and the nucleus basalis region by the intraventricular 192 IgG-saporin treatment. Although the immunotoxin was observed to produce additional damage to the cerebellar Purkinje cells, no gross motor abnormalities were observed that could contribute to the effects on accuracy in the task used here. In conclusion, the results show that selective combined lesions of the basal forebrain cholinergic neurons in the septal-diagonal band area and nucleus basalis produce long-lasting impairments in short-term memory, thus providing further support for a role of this system in cognitive functions.

Animals↗
PubMed1996

Bilateral striatal lesions impair retention of an operant test of short-term memory.

It has been previously shown that lesions of the dorsal striatum can disrupt performance on a variety of cognitive tasks related to prefrontal cortex function. In order to extend these studies, we have compared the effects of bilateral striatal lesions on retention of an operant test of short-term memory using a delayed matching to position task. Rats were initially pretrained on the matching task. Then, one group of animals received ibotenic acid lesions in the dorsal striatum and a second group received sham lesions. The striatal lesions induced marked deficits on delayed matching performance when the rats were retested one week following surgery. The delay-independent pattern of deficit observed does not suggest a primary impairment in short-term memory function. The lesion induced deficits were temporary, with recovery seen in the lesion group back to a control level of performance in 2 additional test sessions 5 and 10 weeks post-lesion. A follow up study investigating the time course of the deficit suggests that the recovery seen in performance measures cannot be attributed to the elapse of time per se, but rather is due to re-learning. In addition to the performance deficits in the operant task, the lesioned rats exhibited marked nocturnal locomotor hyperactivity. The results indicate that bilateral striatal lesions induce marked deficits in retention of the delayed matching to position task. They suggest a key role for the neostriatum in the execution of pre-learned responses, but do not exclude the possibility of additional involvement in short-term memory functions under certain testing conditions.

Animals↗
PubMed1995

In vivo effects of kFGF on embryonic nigral grafts in a rat model of Parkinson's disease.

Central injections of FGF have been reported to promote the survival of dopamine neurones in nigral grafts. With the goal of developing an improved delivery of trophic molecules, an immortalized RN22 Schwann cell line transfected with a secretory form of FGF, kFGF, was irradiated and co-transplanted with embryonic nigral grafts in the 6-OHDA lesioned rat striatum. Amphetamine-induced turning was alleviated by nigral grafts, but was not further improved by co-grafts, whether or not transfected to secrete kFGF. Histological analysis showed similar numbers of surviving transplanted cells and a similar extent of fibre growth from the nigral grafts whether implanted alone or co-grafted with the Schwann cells. These results suggest that kFGF does not have any clear in vivo effect on embryonic nigral grafts in this model.

Animals↗
PubMed1995

Assessment of striatal graft viability in the rat in vivo using a small diameter PET scanner.

A small diameter positron emission tomography (PET) scanner has been used to monitor [11C]raclopride (D2 receptor) binding in vivo in either intact striatum, denervated striatum following an excitotoxic lesion with ibotenic acid, or lesioned and grafted striatum following implantation of cortical or striatal tissue grafts in rats. Binding of [11C]raclopride was localized in the intact striatum within 20 min of injection of the radioligand, and was much reduced within the lesioned striatum. Cortical grafts exhibited a similar low level of binding to the lesioned striatum, whereas striatal grafts showed specific binding at an intermediate level. The [11C]raclopride binding signal in vivo correlated well with the extent of surviving or grafted striatal tissue observed post morten by Nissl staining and acetylcholinesterase histochemistry. Thus, the distribution of dopamine receptors as seen in the PET scanner are consistent with post mortem anatomical observations of striatal, lesion and graft sizes, and suggest that PET can provide a useful tool for monitoring the viability of implanted striatal graft tissues in vivo.

Animals↗
PubMed1995

The effects of bilateral striatal lesions on the acquisition of an operant test of short term memory.

It has been previously shown that lesions of the dorsal striatum can disrupt performance on a variety of cognitive tasks related to frontal cortex function. In order to extend these studies, we have investigated the effects of bilateral striatal lesions on the acquisition of an operant test of short term memory in the delayed non-matching to position paradigm. The animals received either ibotenic acid or saline control injections into the dorsal striatum prior to training on the non-matching task. Striatal lesions retarded acquisition of the task, although with further training the lesioned rats achieved a similar level of asymptotic performance to the control animals. The lesioned rats also exhibited marked nocturnal locomotor hyperactivity when tested under conditions of food deprivation, but not when tested satiated. The results indicate that bilateral striatal lesions induce mild deficits in the acquisition of the discrimination rules involved in performance of the delayed non-matching to position task. The present study does not support a role for the neostriatum in the specific mediation of short term memory in a operant DNMTP test.

Animals↗
PubMed1995

Functional repair of striatal systems by neural transplants: evidence for circuit reconstruction.

Intrastriatal grafts of nigral and adrenal tissues have been found to be effective in alleviating many of the simple motor and sensorimotor deficits associated with lesions of the nigrostriatal dopamine system. However, the mechanisms by which such grafts exert their effects may be less specific than originally conceived, and both pharmacological and trophic actions play an essential role. Damage to intrinsic cortico-striatal circuits are unlikely to prove similarly amenable to such diffuse mechanisms of repair. Nevertheless, striatal grafts have been found to alleviate cognitive and motor deficits after excitotoxic lesions of the neostriatum. Accumulating evidence suggests that in this particular case many aspects of functional recovery may indeed be attributable to the striatal grafts providing an effective functional reconstruction of damaged neuronal circuits within the host brain.

Adrenal Glands↗