scholarly journals Microcontact Printing of Cholinergic Neurons in Organotypic Brain Slices

2021 ◽  
Vol 12 ◽  
Author(s):  
Katharina Steiner ◽  
Christian Humpel
Keyword(s):  
Neurodegenerative Disorder ◽  
Microcontact Printing ◽  
Brain Slices ◽  
Cholinergic Neurons ◽  
Semipermeable Membranes ◽  
Basal Nucleus ◽  
Printing Technique ◽  
The Brain ◽  
Basal Nucleus Of Meynert ◽  
Organotypic Brain Slices

Alzheimer's disease is a severe neurodegenerative disorder of the brain, characterized by beta-amyloid plaques, tau pathology, and cell death of cholinergic neurons, resulting in loss of memory. The reasons for the damage of the cholinergic neurons are not clear, but the nerve growth factor (NGF) is the most potent trophic factor to support the survival of these neurons. In the present study we aim to microprint NGF onto semipermeable 0.4 μm pore membranes and couple them with organotypic brain slices of the basal nucleus of Meynert and to characterize neuronal survival and axonal growth. The brain slices were prepared from postnatal day 10 wildtype mice (C57BL6), cultured on membranes for 2–6 weeks, stained, and characterized for choline acetyltransferase (ChAT). The NGF was microcontact printed in 28 lines, each with 35 μm width, 35 μm space between them, and with a length of 8 mm. As NGF alone could not be printed on the membranes, NGF was embedded into collagen hydrogels and the brain slices were placed at the center of the microprints and the cholinergic neurons that survived. The ChAT+ processes were found to grow along with the NGF microcontact prints, but cells also migrated. Within the brain slices, some form of re-organization along the NGF microcontact prints occurred, especially the glial fibrillary acidic protein (GFAP)+ astrocytes. In conclusion, we provided a novel innovative microcontact printing technique on semipermeable membranes which can be coupled with brain slices. Collagen was used as a loading substance and allowed the microcontact printing of nearly any protein of interest.

scholarly journals S100b Counteracts Neurodegeneration of Rat Cholinergic Neurons in Brain Slices after Oxygen-Glucose Deprivation

2010 ◽  
Vol 2010 ◽  
pp. 1-7 ◽  
Author(s):  
Daniela Serbinek ◽  
Celine Ullrich ◽  
Michael Pirchl ◽  
Tanja Hochstrasser ◽  
Rainald Schmidt-Kastner ◽  
...  
Keyword(s):  
Growth Factor ◽  
Neurodegenerative Disorder ◽  
Brain Slices ◽  
Metabolic Stress ◽  
Cholinergic Neurons ◽  
Glucose Deprivation ◽  
Oxygen Glucose Deprivation ◽  
Basal Nucleus ◽  
Ischemic Events ◽  
Basal Nucleus Of Meynert

Alzheimer's disease is a severe chronic neurodegenerative disorder characterized by beta-amyloid plaques, tau pathology, cerebrovascular damage, inflammation, reactive gliosis, and cell death of cholinergic neurons. The aim of the present study is to test whether the glia-derived molecule S100b can counteract neurodegeneration of cholinergic neurons after oxygen-glucose deprivation (OGD) in organotypic brain slices of basal nucleus of Meynert. Our data showed that 3 days of OGD induced a marked decrease of cholinergic neurons (60% of control), which could be counteracted by 50 μg/mL recombinant S100b. The effect was dose and time dependent. Application of nerve growth factor or fibroblast growth factor-2 was less protective. C-fos-like immunoreactivity was enhanced 3 hours after OGD indicating metabolic stress. We conclude that S100b is a potent neuroprotective factor for cholinergic neurons during ischemic events.

The Organic Cation Transporter 2 Inhibitor Quinidine Modulates the Neuroprotective Effect of Nerve Growth Factor and Memantine on Cholinergic Neurons of the Basal Nucleus of Meynert in Organotypic Brain Slices

Pharmacology ◽  
2021 ◽  
pp. 1-10
Author(s):  
Tugba Gulsun ◽  
Buket Ucar ◽  
Selma Sahin ◽  
Christian Humpel
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Organic Cation ◽  
Brain Slices ◽  
Cholinergic Neurons ◽  
Organic Cation Transporter ◽  
Nerve Growth ◽  
Organic Cation Transporter 2 ◽  
The Brain ◽  
Organotypic Brain Slices

<b><i>Introduction:</i></b> Alzheimer’s disease (AD) is a severe neurodegenerative disorder of the brain characterized by degeneration of cholinergic neurons which is directly linked to cognitive decline. Nerve growth factor (NGF) is the most potent protective factor for cholinergic neurons, additionally the NMDA antagonist memantine blocks glutamate-mediated excitotoxic activity. Quinidine is an inhibitor of organic cation transporter 2 (OCT2). OCT2 is located on cholinergic neurons and plays a role in presynaptic reuptake and recycling of acetylcholine in the brain. We hypothesize that quinidine can modulate the protective effects of NGF and memantine on cholinergic neurons in organotypic brain slices of the nucleus basalis of Meynert (nBM). <b><i>Methods:</i></b> Organotypic brain slices of nBM were incubated with 100 ng/mL NGF, 10 µM memantine, 10 µM quinidine, and combinations of these treatments for 2 weeks. Cholinergic neurons were immunohistochemically stained for choline acetyltransferase (ChAT). <b><i>Results:</i></b> Our data show that NGF as well as memantine counteracted the cell death of cholinergic nBM neurons. Quinidine alone had no toxic effect on cholinergic neurons but inhibited the protective effect of NGF and memantine when applied simultaneously. <b><i>Discussion/Conclusion:</i></b> Our data provide evidence that quinidine modulates the survival of cholinergic nBM neurons via OCT2.

scholarly journals Spreading of Beta-Amyloid in Organotypic Mouse Brain Slices and Microglial Elimination and Effects on Cholinergic Neurons

Biomolecules ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 434
Author(s):  
Kurt Moelgg ◽  
Faryal Jummun ◽  
Christian Humpel
Keyword(s):  
Ventral Striatum ◽  
Brain Slices ◽  
Cholinergic Neurons ◽  
Brain Areas ◽  
Aβ Peptides ◽  
Collagen Hydrogels ◽  
Amyloid Aβ ◽  
Glial Reactions ◽  
Over Time ◽  
Organotypic Brain Slices

The extracellular deposition of b-amyloid (Aβ) is one of the major characteristics in Alzheimer´s disease (AD). The”spreading hypothesis” suggests that a pathological protein (similar to prions) spreads over the entire brain. The aim of the present study was to use organotypic brain slices of postnatal day 8–10 mice. Using collagen hydrogels, we applied different Aβ peptides onto brain slices and analyzed spreading as well as glial reactions after eight weeks of incubation. Our data showed that from all tested Aβ peptides, human Aβ42 had the most potent activity to spread over into adjacent”target“ areas. This effect was potentiated when brain slices from transgenic AD mice (APP_SweDI) were cultured. When different brain areas were connected to the”target slice“ the spreading activity was more intense, originating from ventral striatum and brain stem. Reactive glial-fibrillary acidic protein (GFAP) astrogliosis increased over time, but Aβ depositions co-localized only with Iba1+ microglia but not with astrocytes. Application of human Aβ42 did not cause a degeneration of cholinergic neurons. We concluded that human Aβ42 spreads over into other”target areas“, causing activation of glial cells. Most of the spread Aβ42 was taken up by microglia, and thus toxic free Aβ could not damage cholinergic neurons.

Clinical and neuropathological correlates of depression in Alzheimer's disease

1992 ◽  
Vol 22 (4) ◽  
pp. 877-884 ◽  
Author(s):  
Hans Förstl ◽  
Alistair Burns ◽  
Philip Luthert ◽  
Nigel Cairns ◽  
Peter Lantos ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Cognitive Impairment ◽  
Age Of Onset ◽  
Neuronal Degeneration ◽  
Cholinergic Neurons ◽  
Organic Substrate ◽  
Basal Nucleus ◽  
Alzheimer Pathology ◽  
Basal Nucleus Of Meynert

SynopsisDepressive symptoms have been reported in patients with mild to moderate Alzheimer's disease (AD). Recent evidence suggests that a noradrenergic deficit originating from neuronal degeneration in brainstem nuclei may represent an organic correlate of these disturbances. We examined the neuropathological changes in the locus coeruleus (LC), substantia nigra (SN), basal nucleus of Meynert and cortex of 52 patients (12 male, 40 female, mean age 83·2 ± 6·4 years) with pathologically verified AD. Fourteen patients (1 male, 13 female) showed signs of depression. The majority of these patients suffered from severe physical disability or sensory impairment and developed persistent delusions, but had less cognitive impairment. Neuronal counts in the LC were significantly lower than in the 38 patients without depression (36·9 ± 14 ·0; 51·4 ± 28·0 neuromelaninpigmented cells per section per nucleus;F= 3·4, df = 1, 50,P= 0·04). Neuron counts were higher in the basal nucleus of Meynert in depressed AD patients and there were no differences of the neuron numbers in the SN. Depression (main effect;F= 4·5,P= 0·04) contributed significantly to the variance of neuronal counts in the LC, even when covarying for gender, age of onset, cognitive impairment and cortical Alzheimer pathology. The observed disproportionate loss of noradrenergic and cholinergic neurons in the LC and basal nucleus of Meynert may represent an important organic substrate of depression in AD.

scholarly journals A mode of cell adhesion and migration facilitated by CD44-dependent microtentacles

2020 ◽  
Vol 117 (21) ◽  
pp. 11432-11443 ◽  
Author(s):  
Kayla J. Wolf ◽  
Poojan Shukla ◽  
Kelsey Springer ◽  
Stacey Lee ◽  
Jason D. Coombes ◽  
...  
Keyword(s):  
Tumor Invasion ◽  
Brain Slices ◽  
Connective Tissues ◽  
Superresolution Imaging ◽  
Culture Models ◽  
Cell Adhesion And Migration ◽  
Direct Cell ◽  
And Migration ◽  
The Brain ◽  
Organotypic Brain Slices

The structure and mechanics of many connective tissues are dictated by a collagen-rich extracellular matrix (ECM), where collagen fibers provide topological cues that direct cell migration. However, comparatively little is known about how cells navigate the hyaluronic acid (HA)-rich, nanoporous ECM of the brain, a problem with fundamental implications for development, inflammation, and tumor invasion. Here, we demonstrate that glioblastoma cells adhere to and invade HA-rich matrix using microtentacles (McTNs), which extend tens of micrometers from the cell body and are distinct from filopodia. We observe these structures in continuous culture models and primary patient-derived tumor cells, as well as in synthetic HA matrix and organotypic brain slices. High-magnification and superresolution imaging reveals McTNs are dynamic, CD44-coated tubular protrusions containing microtubules and actin filaments, which respectively drive McTN extension and retraction. Molecular mechanistic studies reveal that McTNs are stabilized by an interplay between microtubule-driven protrusion, actomyosin-driven retraction, and CD44-mediated adhesion, where adhesive and cytoskeletal components are mechanistically coupled by an IQGAP1–CLIP170 complex. McTNs represent a previously unappreciated mechanism through which cells engage nanoporous HA matrix and may represent an important molecular target in physiology and disease.

Brain capillaries and cholinergic neurons persist in organotypic brain slices in the absence of blood flow

2003 ◽  
Vol 18 (1) ◽  
pp. 85-94 ◽  
Author(s):  
Karma V. Moser ◽  
Rainald Schmidt-Kastner ◽  
Hartmann Hinterhuber ◽  
Christian Humpel
Keyword(s):  
Blood Flow ◽  
Brain Slices ◽  
Cholinergic Neurons ◽  
Brain Capillaries ◽  
Organotypic Brain Slices
Sign in / Sign up

Export Citation Format

Share Document