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Cited by 32Neuroscience and Neuropharmacology ResearchMotor Control and AdaptationMemory and Neural Mechanisms

In vitro formation of corticospinal synapses in an organotypic slice co-culture

Takuma, H, Sakurai, M, Kanazawa, I·2002·Neuroscience
DOI 10.1016/s0306-4522(01)00472-9
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Abstract

In order to study biological properties of the corticospinal tract, we have reconstructed this system in an in vitro slice culture preparation. Motor cortex and spinal cord slices, prepared from newborn rats, were co-cultured on pored membranes for 16-24 days. Anterograde labeling with biocytin showed that substantial neural connections had formed between the cortex and spinal cord slices. Retrograde labeling with horseradish peroxidase or 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate demonstrated that the parent cells were located primarily in the deeper layer of the cortex, as is found in vivo. Stimulation of the deep layer of the cortex elicited extracellular postsynaptic responses and intracellular excitatory postsynaptic potentials (EPSPs) in the co-cultured spinal cord that were mediated by the 1-amino-3-hydroxy-5-methyl-4-isoxazolepropionate/ kainate-type glutamate receptor. The intracellular injection of biocytin after EPSPs were recorded showed that one-third of these cells were large stellate cells, which are thought to be motoneurons, while a large portion of the remaining labeled cells were bipolar cells of smaller sizes. Using this reconstructed in vitro preparation, we recorded field EPSPs (fEPSPs) along a 100-microm-interval lattice in the spinal gray matter, which allowed the quantitative evaluation of synapse formation. The fEPSP amplitudes were more than two-fold larger when the forelimb cortex was co-cultured with cervical cord rather than lumbar cord. However, hindlimb cortex did not show this preference. The fEPSP amplitudes were more than twice as large when the dorsal side of the spinal cord was adjacent to the cortex than the ventral side. In summary, we have reconstructed the corticospinal projection and synapses in vitro using cortical and spinal explants. This system allows for an efficient quantitative evaluation of synapse formation and for studies of postsynaptic cells. Our results suggest that synapse formation shows preferences along and perpendicular to the neuraxis of the spinal cord.

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  • Long-range neural projections have been observed in cortico-thalamic assembloids, organotypic mouse co-cultures, and sliced cortical organoids with rodent spinal cord explants.

    “Long-range neural projections have been observed in a cortico-thalamic assembloid (Xiang et al., 2019), in organotypic mouse co-cultures (Takuma et al., 2002) and in a preparation that used sliced cortical organoids and rodent spinal cord explants (Giandomenico et al., 2019).”

    From Generation of Functional Human 3D Cortico-Motor Assembloids (2020) · cites this paper as Takuma et al., 2002

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Authors
Takuma, H., Sakurai, M., & Kanazawa, I.
Affiliations
The University of Tokyo (JP)
Corresponding author
Sakurai, M
ORCID
Sakurai, M
Venue
Neuroscience
Publisher
Elsevier BV
Published
2002-01
Volume
109
Issue
2
Pages
359-370
ISSN
0306-4522 (ISSN-L)
DOI
10.1016/s0306-4522(01)00472-9
OpenAlex
W2079045959
Cited by
32
References
63
Citations by year
2020: 1 · 2017: 4 · 2016: 1 · 2015: 3 · 2014: 1 · 2013: 2 · 2012: 3
FWCI
0.58
Type
journal article
Language
en
Open access
closed
Licence
Locations
  • Neuroscience (publishedVersion)
  • PubMed (publishedVersion)
Funders
Ministry of Education, Culture, Sports, Science and Technology
Topics
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Neuroscience › Cognitive Neuroscience › Motor Control and Adaptation
  • Life Sciences › Neuroscience › Cognitive Neuroscience › Memory and Neural Mechanisms
Keywords
corticospinal tract, synaptogenesis, motor cortex, spinal cord, glutamatergic synapses, excitatory postsynaptic potentials, stellate cells, motor neurons, field EPSP
MeSH terms
Animals, Animals, Newborn, Cell Differentiation, Electric Stimulation, Magnesium, Motor Cortex, Motor Neurons, Neurons, Organ Culture Techniques, Pyramidal Tracts, Reaction Time, Spinal Cord, Synapses, Receptors, AMPA, GABA Antagonists, Coculture Techniques, Excitatory Postsynaptic Potentials, Cell Size, Rats

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Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

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