Huntington???s disease (HD) is a progressive neurodegenerative disorder caused by the dominant mutation of the huntingtin gene on chromosome 4, first described by G. Huntington in 1872. The mutation results in a progressive atrophy of the basal ganglia. The neurodegenerative process is very selective, affecting the only GABAergic medium-sized spiny neurons, that project from the striatum to the globus pallidus and nucleus subthalamicus. In order to reproduce the striatal neuropathological features of HD, quinolinic acid (QA) represents the most commonly used glutamate agonist in both rodent and primate models of HD, leading to the selective loss of striatal GABAergic neurons. Since it is unable to cross the blood-brain barrier, QA needs to be directly injected into the striatum. Here, we have performed a single unilateral intrastriatal infusion of 240 nmol QA in Sprague-Dawley male rats (weighing 250-300g).This induced an evident rotational behavior of the animal, immediately after injury. Fifteen days after the injection, the animals were sacrificed and the brains dissected. By Nissl staining, we analyzed the brain parenchyma and measured the ipsilateral ventricle volume by Neurolucida software: its size was markedly increased (about 23%) compared to the contralateral one. Additionally we observed a massive astrogliosis (highlighted by GFAP-immunohistochemistry - IHC) and a strong microglial activation (IBA1 IHC). To assess potential restorative therapies, one week after QA injection, a separate group of injured rats received 100,000 EGFP-positive embryonic (E14) neural cells obtained from the murine medial ganglionic eminence. To reduce the immune response induced by graft, animals received an oral treatment with10mg/kg cyclosporine A, the day before transplantation and the following 2 days. After 15 days from transplantation, we found a great number of injected survived cells, in absence of signs of reactions of the host. Grafted cells were differentiated in both glial and neuronal cells, which extended their processes in the host tissue, thus appearing well integrated into the host tissue. Although these are preliminary observations, we are currently extending our studies to improve the lesion model, to better evaluate transplanted cell survival and differentiation, their role within striatum and the possibility to create connections with other host neuronal populations. Supported by MIUR grant to AV.
ALLOGRAFT TRANSPLANTATION OF GANGLIONIC EMINENCE EMBRYONIC STEM CELLS IN A RAT MODEL OF HUNTINGTON???S DISEASE.
BOIDO, Marina Maria;VERCELLI, Alessandro
2013-01-01
Abstract
Huntington???s disease (HD) is a progressive neurodegenerative disorder caused by the dominant mutation of the huntingtin gene on chromosome 4, first described by G. Huntington in 1872. The mutation results in a progressive atrophy of the basal ganglia. The neurodegenerative process is very selective, affecting the only GABAergic medium-sized spiny neurons, that project from the striatum to the globus pallidus and nucleus subthalamicus. In order to reproduce the striatal neuropathological features of HD, quinolinic acid (QA) represents the most commonly used glutamate agonist in both rodent and primate models of HD, leading to the selective loss of striatal GABAergic neurons. Since it is unable to cross the blood-brain barrier, QA needs to be directly injected into the striatum. Here, we have performed a single unilateral intrastriatal infusion of 240 nmol QA in Sprague-Dawley male rats (weighing 250-300g).This induced an evident rotational behavior of the animal, immediately after injury. Fifteen days after the injection, the animals were sacrificed and the brains dissected. By Nissl staining, we analyzed the brain parenchyma and measured the ipsilateral ventricle volume by Neurolucida software: its size was markedly increased (about 23%) compared to the contralateral one. Additionally we observed a massive astrogliosis (highlighted by GFAP-immunohistochemistry - IHC) and a strong microglial activation (IBA1 IHC). To assess potential restorative therapies, one week after QA injection, a separate group of injured rats received 100,000 EGFP-positive embryonic (E14) neural cells obtained from the murine medial ganglionic eminence. To reduce the immune response induced by graft, animals received an oral treatment with10mg/kg cyclosporine A, the day before transplantation and the following 2 days. After 15 days from transplantation, we found a great number of injected survived cells, in absence of signs of reactions of the host. Grafted cells were differentiated in both glial and neuronal cells, which extended their processes in the host tissue, thus appearing well integrated into the host tissue. Although these are preliminary observations, we are currently extending our studies to improve the lesion model, to better evaluate transplanted cell survival and differentiation, their role within striatum and the possibility to create connections with other host neuronal populations. Supported by MIUR grant to AV.
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