Nerve growth factor and glial cell line-derived neurotrophic factor restore the cholinergic neuronal phenotype in organotypic brain slices of the basal nucleus of Meynert

Weis, Carla; Marksteiner, Josef; Humpel, Christian

doi:10.1016/s0306-4522(00)00452-8

Cited by 66 publications

(88 citation statements)

References 45 publications

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“…Organotypic brain slice cultures were established as described by us in detail [23][24][25][26]. Briefly, for the co-culture system the basal nucleus of Meynert (nBM), ventral mesencephalon (vMes), parietal cortex (Ctx) and dorsal striatum (dStr) of postnatal day 7-9 (P7-9) Sprague Dawley rats was dissected under aseptic conditions, 400 μm slices were cut with a tissue chopper (McIlwain, USA), and the four slices were placed (Fig.…”

Section: Experimental Procedures Organotypic Brain Slice Culturesmentioning

confidence: 99%

Rotenone Induces Cell Death of Cholinergic Neurons in an Organotypic Co-Culture Brain Slice Model

Ullrich¹,

Humpel²

2009

Neurochem Res

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In Alzheimer and Parkinson's disease cell death of cholinergic and dopaminergic neurons are characteristic hallmarks, respectively. It is well established that rotenone, an inhibitor of complex I of the electron transport chain, induces cell death of dopaminergic neurons, however, not much is known on the effects of rotenone on cholinergic neurons. The aim of the present study was to evaluate the effects of rotenone on cholinergic neurons in an organotypic in vitro brain co-slice model. When co-cultures were treated with 10 μM rotenone for 24 h a significantly decreased number of cholinergic neurons was found in the basal nucleus of Meynert but not in the dorsal striatum. This cell death exhibited apoptotic DAPI-positive malformed nuclei and enhanced TUNEL-positive cells. In summary, inhibition of complex I of the electron transport chain may play a role in neurodegeneration of cholinergic neurons.

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Section: Experimental Procedures Organotypic Brain Slice Culturesmentioning

confidence: 99%

Rotenone Induces Cell Death of Cholinergic Neurons in an Organotypic Co-Culture Brain Slice Model

Ullrich¹,

Humpel²

2009

Neurochem Res

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“…The organotypic brain slice model is well established in our research group [5][6][7] and provides an easily accessible in vitro model containing all cells of the brain in close interaction. During the preparation of the slices, the processes of the neurons are cut, but these axotomized neurons had 2 weeks to recover and to seal their cut axons and dendrites.…”

Section: Discussionmentioning

confidence: 99%

“…Organotypic brain slice cultures were established as described by us in detail [5,6]. Briefly, the nBM of postnatal day 7-9 (P7-9) Sprague-Dawley rats was dissected under aseptic conditions, 400 μm slices were cut with a tissue chopper (McIlwain, USA), and the slices were placed on a 30 mm diameter Millicell-CM 0.4 μm membrane insert (Millipore, Austria), where they become attached to the membrane after 2 weeks of incubation.…”

Section: Experimental Procedures Organotypic Brain Slice Culturesmentioning

confidence: 99%

“…Immunohistochemistry was performed similarly as previously described [5]. All incubations were performed free floating for 2 days including 0.1% Triton, such that the antibodies can penetrate from both sides of the slices, which allows good penetration of the antibody into the brain slices.…”

Section: Immunohistochemistrymentioning

confidence: 99%

“…The organotypic brain slice model has been used to study survival of cholinergic neurons of the septum, as well as their nerve fiber innervations to the hippocampus [2]. In our research group we have extensively studied cholinergic neurons of the basal nucleus of Meynert (nBM) in organotypic brain slices when cultured with nerve growth factor [5][6][7].…”

Section: Introductionmentioning

confidence: 99%

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Mini-Ruby is Rapidly Taken up by Neurons and Astrocytes in Organotypic Brain Slices

Ullrich

Humpel

2011

Neurochem Res

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Cholinergic neurons are intensively studied, because they degenerate in Alzheimer's disease. Although neurotracer techniques are widely used to study axonal transport, guidance, regeneration or sprouting it is not clear if cholinergic neurons can be stained by tracer techniques and studied in brain slices. The aim of the present study was to evaluate the characteristics of the neurotracer Miniruby in organotypic brain slices of the basal nucleus of Meynert (nBM), focusing on cholinergic neurons. Miniruby is a biotinylated dextran amine and is taken up very fast by a variety of cells. When 2-week old nerve growth factor-incubated brain slices of the nBM were treated with Mini-ruby crystals for 1 h, only a few (2-3%) cholinergic neurons were clearly labeled as shown by co-localization with choline acetyltransferase. The staining was found in neuN-positive neurons and microtubule associated protein-2 (MAP-2)-positive nerve fibers. A very rapid dynamic change was observed in these labeled varicosities within seconds. However, Mini-ruby was taken up also by many glutamine synthethase-positive astrocytes. At the site of Mini-ruby application an intense CD11b-positive microglial staining was evident. In conclusion, neurons and astrocytes in organotypic brain slices can be labeled very fast with the fluorescent dye Mini-ruby which undergoes dynamic processes.

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Neural cell adhesion molecule ablation in mice causes hippocampal dysplasia and loss of septal cholinergic neurons

Tereshchenko

Morellini

Dityatev

et al. 2011

J of Comparative Neurology

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The neural cell adhesion molecule (NCAM) is implicated in nervous system development and plasticity. In humans, abnormal NCAM expression has been linked to the pathogenesis of neuropsychiatric and neurodegenerative disorders accompanied by cognitive dysfunctions. Impaired cognition is also observed in NCAM-deficient (NCAM(-/-) ) mice. Considering the importance of the septal cholinergic nuclei and the hippocampus for cognition, we performed quantitative morphological analyses of these areas in young and adult (2 and 13 months old, respectively) NCAM(-/-) mice and wild-type (NCAM(+/+) ) littermates. In young mice, we found lower numbers of septal cholinergic neurons in NCAM(-/-) than in NCAM(+/+) mice. Despite deficient numbers of neurons, total lengths of cholinergic axons and choline acetyltransferase protein levels in the hippocampus of NCAM(-/-) mice were normal. The hippocampus of NCAM(-/-) mice was atrophic, notably in the CA3 subfield and the dentate gyrus (DG). The atrophy appeared to have different primary causes in the two subfields: loss of pyramidal cells in CA3 and reduced branching and volume of granule cell dendrites in the DG. The frequency of dendritic spines on dentate granule cells in NCAM(-/-) mice was normal. Numbers of parvalbumin-positive (PV(+) ) interneurons were reduced in NCAM(-/-) mice in proportion to subfield volume loss, and the ratios of principal cells to PV(+) interneurons were similar to those of NCAM(+/+) mice. None of the observed abnormalities was exaggerated or alleviated in adult NCAM(-/-) mice. Our observations indicate that NCAM ablation causes structural abnormalities in the hippocampus and the forebrain cholinergic system in adult mice, which may contribute to impaired cognition in NCAM(-/-) mice.

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Nerve growth factor and glial cell line-derived neurotrophic factor restore the cholinergic neuronal phenotype in organotypic brain slices of the basal nucleus of Meynert

Cited by 66 publications

References 45 publications

Rotenone Induces Cell Death of Cholinergic Neurons in an Organotypic Co-Culture Brain Slice Model

Rotenone Induces Cell Death of Cholinergic Neurons in an Organotypic Co-Culture Brain Slice Model

Mini-Ruby is Rapidly Taken up by Neurons and Astrocytes in Organotypic Brain Slices

Neural cell adhesion molecule ablation in mice causes hippocampal dysplasia and loss of septal cholinergic neurons

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