Marco Timmer | University of Cologne (original) (raw)

Papers by Marco Timmer

Research paper thumbnail of Fibroblast growth factor-20 promotes the differentiation of Nurr1-overexpressing neural stem cells into tyrosine hydroxylase-positive neurons

Neurobiology of Disease, 2004

Stem cells are currently considered as alternative cell resources for restorative transplantation... more Stem cells are currently considered as alternative cell resources for restorative transplantation strategies in Parkinson's disease. However, the mechanisms that induce differentiation of a stem cell toward the dopaminergic phenotype are still partly unknown thus hampering the production of dopaminergic neurons from stem cells. In the past, FGF-20 has been found to promote the survival of ventral mesencephalic (VM) dopaminergic (DA) neurons in culture. We hereby provide evidence that FGF-20, a growth factor of the FGF family, is expressed in the adult and 6-OHDA-lesioned striatum and substantia nigra, but is not expressed by VM glia or DA neurons, suggesting that FGF-20 may work on DA neurons in a paracrine-or target-derived manner. We also found that coculture of Nurr1-NSCs with Schwann cells overexpressing FGF-20 induced the acquisition of a neuronal morphology by the NSCs and the expression of tyrosine hydroxylase (TH) as assessed by immunocytochemistry, cell ELISA, and Western blot analysis. RT-PCR showed, that both, Schwann cells and Nurr1-NSCs (differentiated or not), expressed the FGF-1 receptor suggesting that both direct and indirect actions of FGF-20 are possible. We show that differentiated Nurr1 cells retained both neuronal morphology and TH expression after transplantation into the striatum of 6-OHDA-lesioned postnatal or adult rats, but that neuritogenesis was only observed after postnatal grafts. Thus, our results suggest that FGF-20 promotes the differentiation of Nurr1-NSCs into TH-positive neurons and that additional factors are required for the efficient differentiation of DA neurons in the adult brain. D 2004 Elsevier Inc. All rights reserved.

Research paper thumbnail of L-DOPA-induced dyskinesia in the rat Parkinson model: interhemispheric functional coupling of the two nigrostriatal pathways

Aktuelle Neurologie, 2006

Research paper thumbnail of Identification of human cells in brain xenografts and in neural co-cultures of rat by in situ hybridisation with Alu probe

Journal of Neuroscience Methods, 2003

Transplantation of human cells into animal models of neurodegenerative disorders is an important ... more Transplantation of human cells into animal models of neurodegenerative disorders is an important scientific application to analyse the survival and developmental capacity of grafted human cells under in vivo conditions. It is critical, therefore, to have a reliable method to distinguish between human and animal cells. In the present study, we describe a combined in situ hybridisation and immunocytochemistry method for the identification of human cells in cultured rat brain cells and xenografts. The specific Alu probe we utilised, which corresponds to the consensus sequence of human Alu repeats was evaluated by southern blot hybridisation of zoo blot and by in situ hybridisation of primary and neoplastic cells from man, rat, mouse, and hamster. This method allows a definite identification of human cells in neural xenografts and, in combination with additional in situ techniques, a further detection of grafted cells.

Research paper thumbnail of Enhanced survival, reinnervation, and functional recovery of intrastriatal dopamine grafts co-transplanted with Schwann cells overexpressing high molecular weight FGF-2 isoforms

Experimental Neurology, 2004

Dopaminergic (DA) micrografts were co-transplanted with Schwann cells (SC) overexpressing 18 kDa ... more Dopaminergic (DA) micrografts were co-transplanted with Schwann cells (SC) overexpressing 18 kDa and 21/23 kDa FGF-2 into the caudate-putamen unit (CPu) of unilaterally 6-hydroxydopamine-lesioned rats. We report here that SC engineered to overexpress FGF-2 promoted DA-graft-induced restoration, whether co-transplanted at the same site or grafted at a second more distant site within the CPu. In addition, the 21/23 kDa FGF-2 isoforms resulted in a significantly better reinnervation and survival of dopaminergic micrografts when compared to the 18-kDa FGF-2 isoform. However, this effect was not that distinct on functional recovery due to, for example, ceiling effects. One main finding of this study was the influence of the gene promotor on DA survival, respectively, vector-mediated trophism. Therefore, comparisons in terms of survival between 18 kDa and higher molecular weight (HMW) FGF-2 are complicated in the mixed grafted experiments. Furthermore, the first demonstration of the presence of the 21/23 kDa FGF-2 isoforms in the nigrostriatal system and their potent neurotrophic in vivo activities, as shown in the present study, suggest (I) a physiological role of these proteins for dopaminergic neurons and (II) a restorative potential under normal as well as regenerative processes. However, FGF-2-mediated effects are more pronounced after co-transplantation with SC/DA cells mixed in one suspension at the same implantation side than in the side-by-side approach with a spatially and temporally separated transplantation of SC (day 1) and DA-cells (day 3). These findings indicate the necessity of direct contact between FGF-2 and DA-neurons, further elucidate the neurotrophic role of FGF-2 for DA-neurons and highlight the differential restorative potentials of its respective isoforms. We propose that administration of HMW FGF-2 may be used to improve function in the rat Parkinson's disease model. D 2004 Elsevier Inc. All rights reserved.

Research paper thumbnail of Survival and early functional integration of dopaminergic progenitor cells following transplantation in a rat model of Parkinson's disease

Dopaminergic (DA) grafts in rat models of Parkinson's disease (PD) have previously been derived f... more Dopaminergic (DA) grafts in rat models of Parkinson's disease (PD) have previously been derived from embryonic day (E) 14 grafts. Because there is an increasing interest in the restorative capacity of DA stem and progenitor cells, in the present study we examined the survival and early and late functional behavioral effects of DA progenitor cells derived from E12, E13, E14, and E15 grafts transplanted into rats with unilateral 6hydroxydopamin lesions. DA transplant-induced functional recovery was already observed in postural balancing reactions after 10 days and in stepping behavior after 13 days, that is, in spontaneous complex behaviors, and later, after 16 days, in the amphetamine-induced rotation test. Three distinct patterns of functional recovery could be observed at 6-9 weeks posttransplantation. First, behavioral improvements in drug-induced rotational asymmetry, stepping, and skilled forelimb behavior were directly related to DA neuron survival and TH-positive fiber reinnervation. Second, recovery in postural balancing reactions was closely related to a specific developmental time window of donor age, for example, only seen in E13 and E14 grafts. Finally, no functional graft effects were seen in the table lift test. Interestingly, DA neuron graft survival, TH-positive fiber outgrowth, and graft volume were significantly influenced by the developmental time window in which the DA progenitor cells were dissected from the ventral mesencephalon, that is, from E12, E13, E14, or E15 rat embryos. These data highlight the complexity of graft-host interactions and provide novel insights into the dynamics of DA progenitor graft-mediated functional recovery in animal models of Parkinson's disease. V V C 2009 Wiley-Liss, Inc.

Research paper thumbnail of Fibroblast growth factor-20 promotes the differentiation of Nurr1-overexpressing neural stem cells into tyrosine hydroxylase-positive neurons

Neurobiology of Disease, 2004

Stem cells are currently considered as alternative cell resources for restorative transplantation... more Stem cells are currently considered as alternative cell resources for restorative transplantation strategies in Parkinson's disease. However, the mechanisms that induce differentiation of a stem cell toward the dopaminergic phenotype are still partly unknown thus hampering the production of dopaminergic neurons from stem cells. In the past, FGF-20 has been found to promote the survival of ventral mesencephalic (VM) dopaminergic (DA) neurons in culture. We hereby provide evidence that FGF-20, a growth factor of the FGF family, is expressed in the adult and 6-OHDA-lesioned striatum and substantia nigra, but is not expressed by VM glia or DA neurons, suggesting that FGF-20 may work on DA neurons in a paracrine-or target-derived manner. We also found that coculture of Nurr1-NSCs with Schwann cells overexpressing FGF-20 induced the acquisition of a neuronal morphology by the NSCs and the expression of tyrosine hydroxylase (TH) as assessed by immunocytochemistry, cell ELISA, and Western blot analysis. RT-PCR showed, that both, Schwann cells and Nurr1-NSCs (differentiated or not), expressed the FGF-1 receptor suggesting that both direct and indirect actions of FGF-20 are possible. We show that differentiated Nurr1 cells retained both neuronal morphology and TH expression after transplantation into the striatum of 6-OHDA-lesioned postnatal or adult rats, but that neuritogenesis was only observed after postnatal grafts. Thus, our results suggest that FGF-20 promotes the differentiation of Nurr1-NSCs into TH-positive neurons and that additional factors are required for the efficient differentiation of DA neurons in the adult brain. D 2004 Elsevier Inc. All rights reserved.

Research paper thumbnail of L-DOPA-induced dyskinesia in the rat Parkinson model: interhemispheric functional coupling of the two nigrostriatal pathways

Aktuelle Neurologie, 2006

Research paper thumbnail of Identification of human cells in brain xenografts and in neural co-cultures of rat by in situ hybridisation with Alu probe

Journal of Neuroscience Methods, 2003

Transplantation of human cells into animal models of neurodegenerative disorders is an important ... more Transplantation of human cells into animal models of neurodegenerative disorders is an important scientific application to analyse the survival and developmental capacity of grafted human cells under in vivo conditions. It is critical, therefore, to have a reliable method to distinguish between human and animal cells. In the present study, we describe a combined in situ hybridisation and immunocytochemistry method for the identification of human cells in cultured rat brain cells and xenografts. The specific Alu probe we utilised, which corresponds to the consensus sequence of human Alu repeats was evaluated by southern blot hybridisation of zoo blot and by in situ hybridisation of primary and neoplastic cells from man, rat, mouse, and hamster. This method allows a definite identification of human cells in neural xenografts and, in combination with additional in situ techniques, a further detection of grafted cells.

Research paper thumbnail of Enhanced survival, reinnervation, and functional recovery of intrastriatal dopamine grafts co-transplanted with Schwann cells overexpressing high molecular weight FGF-2 isoforms

Experimental Neurology, 2004

Dopaminergic (DA) micrografts were co-transplanted with Schwann cells (SC) overexpressing 18 kDa ... more Dopaminergic (DA) micrografts were co-transplanted with Schwann cells (SC) overexpressing 18 kDa and 21/23 kDa FGF-2 into the caudate-putamen unit (CPu) of unilaterally 6-hydroxydopamine-lesioned rats. We report here that SC engineered to overexpress FGF-2 promoted DA-graft-induced restoration, whether co-transplanted at the same site or grafted at a second more distant site within the CPu. In addition, the 21/23 kDa FGF-2 isoforms resulted in a significantly better reinnervation and survival of dopaminergic micrografts when compared to the 18-kDa FGF-2 isoform. However, this effect was not that distinct on functional recovery due to, for example, ceiling effects. One main finding of this study was the influence of the gene promotor on DA survival, respectively, vector-mediated trophism. Therefore, comparisons in terms of survival between 18 kDa and higher molecular weight (HMW) FGF-2 are complicated in the mixed grafted experiments. Furthermore, the first demonstration of the presence of the 21/23 kDa FGF-2 isoforms in the nigrostriatal system and their potent neurotrophic in vivo activities, as shown in the present study, suggest (I) a physiological role of these proteins for dopaminergic neurons and (II) a restorative potential under normal as well as regenerative processes. However, FGF-2-mediated effects are more pronounced after co-transplantation with SC/DA cells mixed in one suspension at the same implantation side than in the side-by-side approach with a spatially and temporally separated transplantation of SC (day 1) and DA-cells (day 3). These findings indicate the necessity of direct contact between FGF-2 and DA-neurons, further elucidate the neurotrophic role of FGF-2 for DA-neurons and highlight the differential restorative potentials of its respective isoforms. We propose that administration of HMW FGF-2 may be used to improve function in the rat Parkinson's disease model. D 2004 Elsevier Inc. All rights reserved.

Research paper thumbnail of Survival and early functional integration of dopaminergic progenitor cells following transplantation in a rat model of Parkinson's disease

Dopaminergic (DA) grafts in rat models of Parkinson's disease (PD) have previously been derived f... more Dopaminergic (DA) grafts in rat models of Parkinson's disease (PD) have previously been derived from embryonic day (E) 14 grafts. Because there is an increasing interest in the restorative capacity of DA stem and progenitor cells, in the present study we examined the survival and early and late functional behavioral effects of DA progenitor cells derived from E12, E13, E14, and E15 grafts transplanted into rats with unilateral 6hydroxydopamin lesions. DA transplant-induced functional recovery was already observed in postural balancing reactions after 10 days and in stepping behavior after 13 days, that is, in spontaneous complex behaviors, and later, after 16 days, in the amphetamine-induced rotation test. Three distinct patterns of functional recovery could be observed at 6-9 weeks posttransplantation. First, behavioral improvements in drug-induced rotational asymmetry, stepping, and skilled forelimb behavior were directly related to DA neuron survival and TH-positive fiber reinnervation. Second, recovery in postural balancing reactions was closely related to a specific developmental time window of donor age, for example, only seen in E13 and E14 grafts. Finally, no functional graft effects were seen in the table lift test. Interestingly, DA neuron graft survival, TH-positive fiber outgrowth, and graft volume were significantly influenced by the developmental time window in which the DA progenitor cells were dissected from the ventral mesencephalon, that is, from E12, E13, E14, or E15 rat embryos. These data highlight the complexity of graft-host interactions and provide novel insights into the dynamics of DA progenitor graft-mediated functional recovery in animal models of Parkinson's disease. V V C 2009 Wiley-Liss, Inc.