Microcontact Printing of Cholinergic Neurons in 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 (NG...

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Autores principales:	Katharina Steiner, Christian Humpel
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	cholinergic neurons microcontact printing nerve growth factor organotypic brain slices brain-on-a-chip Neurology. Diseases of the nervous system RC346-429
Acceso en línea:	https://doaj.org/article/41fd6f53194c461fa6766ae0ce9d9c56
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id	oai:doaj.org-article:41fd6f53194c461fa6766ae0ce9d9c56
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spelling	oai:doaj.org-article:41fd6f53194c461fa6766ae0ce9d9c562021-11-17T13:31:13ZMicrocontact Printing of Cholinergic Neurons in Organotypic Brain Slices1664-229510.3389/fneur.2021.775621https://doaj.org/article/41fd6f53194c461fa6766ae0ce9d9c562021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fneur.2021.775621/fullhttps://doaj.org/toc/1664-2295Alzheimer'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.Katharina SteinerChristian HumpelFrontiers Media S.A.articlecholinergic neuronsmicrocontact printingnerve growth factororganotypic brain slicesbrain-on-a-chipNeurology. Diseases of the nervous systemRC346-429ENFrontiers in Neurology, Vol 12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	cholinergic neurons microcontact printing nerve growth factor organotypic brain slices brain-on-a-chip Neurology. Diseases of the nervous system RC346-429
spellingShingle	cholinergic neurons microcontact printing nerve growth factor organotypic brain slices brain-on-a-chip Neurology. Diseases of the nervous system RC346-429 Katharina Steiner Christian Humpel Microcontact Printing of Cholinergic Neurons in Organotypic Brain Slices
description	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.
format	article
author	Katharina Steiner Christian Humpel
author_facet	Katharina Steiner Christian Humpel
author_sort	Katharina Steiner
title	Microcontact Printing of Cholinergic Neurons in Organotypic Brain Slices
title_short	Microcontact Printing of Cholinergic Neurons in Organotypic Brain Slices
title_full	Microcontact Printing of Cholinergic Neurons in Organotypic Brain Slices
title_fullStr	Microcontact Printing of Cholinergic Neurons in Organotypic Brain Slices
title_full_unstemmed	Microcontact Printing of Cholinergic Neurons in Organotypic Brain Slices
title_sort	microcontact printing of cholinergic neurons in organotypic brain slices
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/41fd6f53194c461fa6766ae0ce9d9c56
work_keys_str_mv	AT katharinasteiner microcontactprintingofcholinergicneuronsinorganotypicbrainslices AT christianhumpel microcontactprintingofcholinergicneuronsinorganotypicbrainslices
_version_	1718425540867653632

Microcontact Printing of Cholinergic Neurons in Organotypic Brain Slices

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