Dissection and culturing of adult lateral entorhinal cortex layer II neurons from APP/PS1 Alzheimer model mice

Hanssen, Katrine Sjaastad; Sandvig, Axel; Sandvig, Ioanna; Kobro‐Flatmoen, Asgeir

doi:10.1016/j.jneumeth.2023.109840

Cited by 2 publications

(14 citation statements)

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“…To further demonstrate the wider applicability of our model system for reverse engineering anatomically relevant networks for preclinical disease modelling, we used this system to culture neurons extracted from brain regions of interest from adult AD model animals. This method for the dissection and culturing of layer specific entorhinal and hippocampal neurons from AD model rats and mice builds upon our recently published protocol for extraction and culturing of LEC LII neurons from adult AD model APP/PS1 mice (39). We found that adult neurons from both AD model rats-( Figure 5 ) and mice ( Figure S2 ) were able to re-form structural connections in vitro .…”

Section: Resultsmentioning

confidence: 99%

“…Our neural cultures comprised either commercially available rat embryonic cortical and hippocampal neurons (hereafter referred to as cortical-hippocampal networks), or lateral entorhinal cortex layer II neurons (LEC-LII), Dentate Gyrus granular neurons (DG-gl), CA3 pyramidal neurons (CA3-pyr) and CA1 pyramidal neurons (CA1-pyr), freshly dissected from adult McGill-R-Thy1-APP AD-model rats or APP/PS1 AD-model mice (hereafter referred to as adult entorhinal-hippocampal AD networks). The method for the dissection and culturing of adult hippocampal neurons was refined from our previously reported protocol for adult lateral-most LEC LII (lLEC-LII) neurons from APP/PS1 model mice (39). For all cultures, coating and establishment of an astrocytic feeder layer was conducted using the same protocol.…”

Section: Methodsmentioning

confidence: 99%

“…The APP/PS1 mice were genotyped using a KAPA-kit as described in Hanssen et al . (39). The McGill-R-Thy1-APP rat model is a transgenic familial AD model with a Wistar (HsdBrl:WH) genetic background, co-expressing mutated hAPP (APP K670M671delinsNL) and (APP V717F) (67).…”

Section: Methodsmentioning

confidence: 99%

“…Thus, this network is of high interest to study at preclinical stages of AD. We and others have demonstrated that adult entorhinal and hippocampal subregion specific cells from AD model mice are able to reconnect in culture (37–39). We also recently reported a method for long-term (>2 months) culturing and recording of lateral entorhinal cortex layer II (LEC LII) neurons from AD model animals in vitro (39).…”

Section: Introductionmentioning

confidence: 99%

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Reverse Engineering of Feedforward Cortical-Hippocampal Neural Networks Relevant for Preclinical Disease Modelling

Hanssen,

Winter-Hjelm,

Niethammer

et al. 2023

Preprint

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Engineered biological neural networks are indispensable tools for investigating neural function in both healthy and diseased states from the subcellular to the network level. Neurons in vitro self-organize over time into networks of increasing structural and functional complexity, thus maintaining emergent dynamics of neurons in the brain. While in vitro neural network model systems have advanced significantly over the past decade, there is still a need for models able to recapitulate topological and functional organization of brain networks. Especially relevant in this context are interfaces which can support the establishment of multinodal interconnected networks of different neural populations, which at the same time enable control of the direction of connectivity between the nodes. An added required feature of such interfaces is compatibility with electrophysiological techniques and optical imaging tools to facilitate studies of neural network structure-function dynamics. In this study, we applied a custom-designed microfluidic device with Tesla-valve inspired microchannels for structuring multinodal neural networks with controllable feedforward connectivity between rat primary cortical and hippocampal neurons. By interfacing these devices with nanoporous microelectrode arrays, we demonstrate how both spontaneously evoked and stimulation induced activity propagates, as intended, in a feedforward pattern between the different interconnected neural nodes. Moreover, we show that these multinodal networks exhibit functional dynamics suggestive of capacity for both segregated and integrated activity. To advocate the broader applicability of this model system, we also provide proof of concept of long-term culturing of subregion- and layer specific neurons extracted from the entorhinal cortex and hippocampus of adult Alzheimers-model mice and rats. We show that these neurons re-form structural connections and develop spontaneous spiking activity after 15 days in vitro. Our results thus highlight the suitability and potential of our approach for reverse engineering of biologically and anatomically relevant multinodal neural networks supporting the study of dynamic structure-function relationships in both healthy and pathological conditions.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Reverse Engineering of Feedforward Cortical-Hippocampal Neural Networks Relevant for Preclinical Disease Modelling

Hanssen,

Winter-Hjelm,

Niethammer

et al. 2023

Preprint

Self Cite

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“…Many of the limitations of this model hippocampal system are readily evident such as lack of behavioral and other brain modulatory inputs. Our 2D reductionist hippocampus does not model the lamellar organization of the native hippocampus including the layered inputs of the EC into the 3D hippocampus (Marks et al 2020;Hanssen 2023). We have not yet taken advantage of the easy pharmacological access to further define the inhibitory components of the networks.…”

Section: Limitationsmentioning

confidence: 99%

Hippocampal network axons respond to patterned theta burst stimulation with lower activity of initially higher spike train similarity from EC to DG and later similarity of axons from CA1 to EC

Chen,

Vakilna,

Lassers

et al. 2023

J. Neural Eng.

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Objective. Decoding memory functions for each hippocampal subregion involves extensive understanding of how each hippocampal subnetwork processes input stimuli. Theta burst stimulation (TBS) recapitulates natural brain stimuli which potentiates synapses in hippocampal circuits. TBS is typically applied to a bundle of axons to measure the immediate response in a downstream subregion like the cornu ammonis 1 (CA1). Yet little is known about network processing in response to stimulation, especially because individual axonal transmission between subregions is not accessible. Approach. To address these limitations, we reverse engineered the hippocampal network on a micro-electrode array partitioned by a MEMS four-chambered device with interconnecting microfluidic tunnels. The micro tunnels allowed monitoring single axon transmission which is inaccessible in slices or in vivo. The four chambers were plated separately with entorhinal cortex (EC), dentate gyrus (DG), CA1, and CA3 neurons. The patterned TBS was delivered to the EC hippocampal gateway. Evoked spike pattern similarity in each subregions was quantified with Jaccard distance metrics of spike timing. Main results. We found that the network subregion produced unique axonal responses to different stimulation patterns. Single site and multisite stimulations caused distinct information routing of axonal spikes in the network. The most spatially similar output at axons from CA3 to CA1 reflected the auto association within CA3 recurrent networks. Moreover, the spike pattern similarities shifted from high levels for axons to and from DG at 0.2 s repeat stimuli to greater similarity in axons to and from CA1 for repetitions at 10 s intervals. This time-dependent response suggested that CA3 encoded temporal information and axons transmitted the information to CA1. Significance. Our design and interrogation approach provide first insights into differences in information transmission between the four subregions of the structured hippocampal network and the dynamic pattern variations in response to stimulation at the subregional level to achieve probabilistic pattern separation and novelty detection.

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Dissection and culturing of adult lateral entorhinal cortex layer II neurons from APP/PS1 Alzheimer model mice

Cited by 2 publications

References 45 publications

Reverse Engineering of Feedforward Cortical-Hippocampal Neural Networks Relevant for Preclinical Disease Modelling

Reverse Engineering of Feedforward Cortical-Hippocampal Neural Networks Relevant for Preclinical Disease Modelling

Hippocampal network axons respond to patterned theta burst stimulation with lower activity of initially higher spike train similarity from EC to DG and later similarity of axons from CA1 to EC

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