Selective Glial Cell Line-Derived Neurotrophic Factor Production in Adult Dopaminergic Carotid Body Cells In Situ and after Intrastriatal Transplantation

Laboratorio de Investigaciones Biomédicas, Departamento de Fisiología and Hospital Universitario Virgen del Rocío, Universidad de Sevilla, 41013 Sevilla, Spain.
The Journal of Neuroscience : The Official Journal of the Society for Neuroscience (Impact Factor: 6.34). 05/2005; 25(16):4091-8. DOI: 10.1523/JNEUROSCI.4312-04.2005
Source: PubMed


Glial cell line-derived neurotrophic factor (GDNF) exerts a notable protective effect on dopaminergic neurons in rodent and primate models of Parkinson's disease (PD). The clinical applicability of this therapy is, however, hampered by the need of a durable and stable GDNF source allowing the safe and continuous delivery of the trophic factor into the brain parenchyma. Intrastriatal carotid body (CB) autografting is a neuroprotective therapy potentially useful in PD. It induces long-term recovery of parkinsonian animals through a trophic effect on nigrostriatal neurons and causes amelioration of symptoms in some PD patients. Moreover, the adult rodent CB has been shown to express GDNF. Here we show, using heterozygous GDNF/lacZ knock-out mice, that unexpectedly CB dopaminergic glomus, or type I, cells are the source of CB GDNF. Among the neural or paraneural cells tested, glomus cells are those that synthesize and release the highest amount of GDNF in the adult rodent (as measured by standard and in situ ELISA). Furthermore, GDNF expression by glomus cells is maintained after intrastriatal grafting and in CB of aged and parkinsonian 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated animals. Thus, glomus cells appear to be prototypical abundant sources of GDNF, ideally suited to be used as biological pumps for the endogenous delivery of trophic factors in PD and other neurodegenerative diseases.

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Available from: Simón Méndez-Ferrer
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    • "Accordingly, we went on to ask, can the nano-stromal approach protect and improve the yield of healthy human DA cells under conditions used for transplantation? Here, in addition to LIF-nano, we included nano-stromal cargos to target delivery of BDNF, GDNF and XAV, reasoning firstly that BDNF and GDNF are already known to be pro-survival for DA neurons (Hyman et al., 1991; Villadiego et al., 2005), and secondly that XAV might provide in the future a means to modulate neuronal fate. Fig. 4A outlines the process of hfVM harvest prior to cell preparation and nano-stromal treatment. "
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    ABSTRACT: The endogenous reparative capacity of the adult human brain is low and chronic neurodegenerative disorders of the central nervous system represent one of the greatest areas of unmet clinical need in the developing world. Novel therapeutic strategies to treat them include (i) growth factor delivery to boost endogenous repair and (ii) replacement cell therapy, including dopaminergic neurons for Parkinson's Disease (PD). However these approaches are limited not only by rapid degradation of growth factors, but also by the poor availability and survival of implanted cells that lack the necessary stromal support. We therefore hypothesised that provision of a transient artificial stroma for paracrine delivery of pro-survival factors may overcome both of these issues. Using leukaemia inhibitory factor (LIF) - a proneural, reparative cytokine - formulated as target-specific PLGA nano-particles (LIF-nano) - we discovered that attachment of LIF-nano to freshly isolated fetal dopaminergic cells improved their survival fourfold: furthermore, in vivo, the numbers of surviving human fetal dopaminergic cells tended to be higher at 3 months after grafting into the striatum of nude rats. In addition we also analysed the effect of a novel nano-stroma incorporating XAV939, a potent inhibitor of the developmentally important wnt/β-catenin signalling pathway, to investigate whether it could also promote the survival and differentiation of human fetal dopaminergic precursors and found both TH+ neurons and total neurons were increased in number. This is the first demonstration that LIF-nano-stroma, and XAV-nano-stroma, each have pro-survival effects on human dopaminergic neurons, with potential value for target-specific modulation of neurogenic fate in cell based therapies for PD.
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    • "GDNF protein content was estimated in CB and SCG as previously described (Villadiego et al. 2005). Briefly, tissues were removed and immediately frozen in liquid nitrogen, and then homogenised in lysis buffer ( "
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    ABSTRACT: The carotid body (CB) is the major peripheral arterial chemoreceptor in mammals that mediates the acute hyperventilatory response to hypoxia. The CB grows in response to sustained hypoxia and also participates in acclimatisation to chronic hypoxemia. Knowledge of CB physiology at the cellular level has increased considerably in recent times thanks to studies performed on lower mammals, and rodents in particular. However, the functional characteristics of human CB cells remain practically unknown. Herein, we use tissue slices or enzymatically dispersed cells to determine characteristics of human CB cells. The adult human CB parenchyma contains clusters of chemosensitive glomus (type I) and sustentacular (type II) cells as well as nestin-positive progenitor cells. This organ also expresses high levels of the dopaminotrophic glial cell line-derived neurotrophic factor (GDNF). We found that GDNF production and the number of progenitor and glomus cells were preserved in the CBs of human subjects of advanced age. Moreover, glomus cells exhibited voltage-dependent Na+, Ca2+ and K+ currents that were qualitatively similar to those reported in lower mammals. These cells responded to hypoxia with an external Ca2+-dependent increase of cytosolic Ca2+ and quantal catecholamine secretion, as reported for other mammalian species. Interestingly, human glomus cells are also responsive to hypoglycaemia and together these two stimuli can potentiate each other's effects. The chemosensory responses of glomus cells are also preserved at an advance age. These new data on the cellular and molecular physiology of the CB pave the way for future pathophysiological studies involving this organ in humans.
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    • "Images shown in previous figures were obtained from the GDNF/X-gal mouse line, in which the expression of the β-gal enzyme is under the control of the endogenous GDNF promoter. This reporter mouse line shows a characteristic X-gal staining on neural tissue, presenting discrete X-gal deposits due to the low level of expression of the β-gal enzyme791011. We also tested the validity of our method in a high-expression LacZ reporter line using a TH-ires-Cre/R26R mouse (carrying the Rosa26R-LacZ reporter construct). "
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