Early Age- and Sex-Dependent Regulation of Astrocyte-Mediated Glutamatergic Synapse Elimination in the Rat Prefrontal Cortex: Setting an Organotypic Brain Slice Culture Investigating Tool

Eugenia Vivi; Lea R Seeholzer; Anastasia Nagumanova; Barbara Di Benedetto

doi:10.20944/preprints202310.1752.v2

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preprints.org > biology and life sciences > neuroscience and neurology > doi: 10.20944/preprints202310.1752.v2

Preprint Article Version 2 Preserved in Portico This version is not peer-reviewed

Early Age- and Sex-Dependent Regulation of Astrocyte-Mediated Glutamatergic Synapse Elimination in the Rat Prefrontal Cortex: Setting an Organotypic Brain Slice Culture Investigating Tool

Version 1 : Received: 26 October 2023 / Approved: 26 October 2023 / Online: 27 October 2023 (07:16:08 CEST)
Version 2 : Received: 28 November 2023 / Approved: 28 November 2023 / Online: 29 November 2023 (09:43:31 CET)

A peer-reviewed article of this Preprint also exists.

Vivi, E.; Seeholzer, L.R.; Nagumanova, A.; Di Benedetto, B. Early Age- and Sex-Dependent Regulation of Astrocyte-Mediated Glutamatergic Synapse Elimination in the Rat Prefrontal Cortex: Establishing an Organotypic Brain Slice Culture Investigating Tool. Cells 2023, 12, 2761. Vivi, E.; Seeholzer, L.R.; Nagumanova, A.; Di Benedetto, B. Early Age- and Sex-Dependent Regulation of Astrocyte-Mediated Glutamatergic Synapse Elimination in the Rat Prefrontal Cortex: Establishing an Organotypic Brain Slice Culture Investigating Tool. Cells 2023, 12, 2761.

Abstract

Clinical and pre-clinical studies of neuropsychiatric (NP) disorders show altered astrocyte properties and synaptic networks. These are refined during early postnatal developmental (PND) stages. Thus, investigating early brain maturational trajectories is essential to understand NP disorders. However, animal experiments are highly time-/resource-consuming, thereby calling for alternative methodological approaches. The function of MEGF10 in astrocyte-mediated synapse elimination (pruning) is crucial to refine neuronal networks during development and adulthood. To investigate the impact of MEGF10 during PND in the rat prefrontal cortex (PFC) and its putative role in brain disorders, we established and validated an Organotypic Brain Slice Culture (OBSC) system.Using Western blot, we characterized the expression of MEGF10 and the synaptic markers synaptophysin and PSD95 in the cortex of developing pups. We then combined immunofluorescent-immunohistochemistry with Imaris-supported 3D-analysis to compare age- and sex-dependent astrocyte-mediated pruning within the PFC in pups and OBSCs. We thereby validated this system to investigate age-dependent astrocyte-mediated changes in pruning during PND. However, further optimizations are required to use OBSCs for revealing sex-dependent differences. In conclusion, OBSCs offer a valid alternative to study physiological astrocyte-mediated synaptic remodelling during PND and might be exploited to investigate pathomechanisms of brain disorders with aberrant synaptic development.

Keywords

organotypic brain slice culture; astrocyte; synaptic phagocytosis; critical period; sex differences

Subject

Biology and Life Sciences, Neuroscience and Neurology

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Comments (1)

Comment 1

Received: 29 November 2023
Commenter: Barbara Di Benedetto
Commenter's Conflict of Interests: Author

Comment: This is an updated version after revisions suggested by peer-reviewers.

Sincerely,
Barbara Di Benedetto

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