Cited by 367Neurogenesis and neuroplasticity mechanismsAxon Guidance and Neuronal SignalingMicrotubule and mitosis dynamics

The DOI matches its Crossref record; 49 of the 49 references it cites were found in a registry or an index; 12 of the 78 sources its claims cite were checked for support.

Immunocytochemical localization of tubulin and microtubule-associated protein 2 during the development of hippocampal neurons in culture

Open access
Caceres, A, Banker, GA, Binder, L·1986·The Journal of Neuroscience

Abstract

n dissociated-cell cultures prepared from the embryonic rat hippocampus, neurons establish both axons and dendrites, which differ in geometry, in ultrastructure, and in synaptic polarity. We have used immunocytochemistry with monoclonal antibodies to study the regional distribution of beta-tubulin and micro-tubule-associated protein 2 (MAP2) in hippocampal cultures and their localization during early stages of axonal and dendritic development. After development for a week or more in culture, when axons and dendrites were well- differentiated, the distribution of these two proteins was quite different. Beta-tubulin was present throughout the nerve cell, in soma, dendrites, and axon. It was also present in all classes of non-neuronal cells, astrocytes, fibroblasts, and a presumptive glial progenitor cell. In contrast, MAP2 was preferentially localized to nerve cells; within neurons, MAP2 was present in soma and dendrites, but little or no immunostaining was detectable in axons. Both beta-tubulin and MAP2 were present in nerve cells at the time of plating. From the earliest stages of process extension, beta-tubulin was present in all neuronal processes, both axons and dendrites. Surprisingly, MAP2 was also initially present in both axons and dendrites, extending as far as the axonal growth cone. With subsequent development, MAP2 staining was selectively lost from the axon so that after 1 week in vitro little or no axonal staining remained. Taken together with earlier results (Caceres et al., 1984a), these data indicate that the establishment of neuronal polarity, as manifested by the molecular differentiation of the axonal and dendritic cytoskeleton, occurs largely under endogenous control, even under culture conditions in which cell interactions are greatly restricted.(ABSTRACT TRUNCATED AT 250 WORDS)

Overview

AI snapshot

Written from the paper’s full text.

TL;DR

In dissociated embryonic rat hippocampal cultures, beta-tubulin was present in axons, dendrites, soma, and non-neuronal cells, whereas MAP2 was preferentially localized to soma and dendrites in mature neurons. Unexpectedly, MAP2 was initially present in both axons and dendrites, including axonal growth cones, and was selectively lost from axons over the first week in culture.

Key findings

  • In mature cultures (7–35 d in vitro), beta-tubulin immunoreactivity was present throughout neurons—soma, dendrites, and even the thinnest axons—and in astrocytes, fibroblasts, and presumptive glial progenitor cells; MAP2 was intensely stained in perikarya and dendrites but showed little or no axonal staining.
  • At plating, both beta-tubulin and MAP2 were present in nerve cells; within 1–2 d, MAP2 immunoreactivity was present in all developing processes, including axons, and could be followed to axonal tips as far as 150 µm from cell bodies.
  • By 3–4 d in culture, MAP2 staining in axons appeared preferentially in distal axonal regions, sometimes restricted to the growth cone and immediately proximal axon; by about the end of the first week, axonal MAP2 staining had largely disappeared.
  • After 1 d in culture, when MAP2 was still uniformly distributed in axons and dendrites, dendrites were already elongating about 5× more slowly than axons, indicating that MAP2 presence does not determine dendritic identity.

Methods

  • Hippocampal neurons were prepared from 17–20-d-old rat fetuses, dissociated after trypsin treatment, and plated onto polylysine-treated coverslips at 5,000–15,000 cells/cm²; coverslips were co-cultured facing confluent astroglial monolayers in MEM with N2 supplement and 0.1% ovalbumin.
  • Immunocytochemistry used monoclonal antibodies against beta-tubulin (clones Tu9b, Tu12, Tu27) and MAP2 (clones AP9, AP13), with several fixation/permeabilization procedures including paraformaldehyde/glutaraldehyde, cold methanol, and Triton X-100 extraction; staining was visualized by peroxidase-antiperoxidase and diaminobenzidine.
  • Analysis was qualitative light-microscopic immunocytochemistry, with controls omitting primary antibody or preabsorbing with purified antigen.

Limitations

  • The paper states that MAP2 distribution in situ has not been studied at comparably early stages, so it is unknown whether axons transiently contain MAP2 during normal development in situ.
  • The paper states that axonal MAP2 staining could be an artifact of dissociation and culture, and that the functional role of MAP2 in developing axons is not known.
  • Evident from the text: the study is qualitative immunocytochemistry without quantification, and the culture system uses glial co-culture and trophic support, so it may not fully reproduce in vivo cell interactions.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1523/jneurosci.06-03-00714.1986

Cites

49 of 49 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Verification

Claims the paper makes

32 Own result · 30 Sources not found · 18 Own interpretation · 8 Not checked yet · 5 Partly supported · 4 Supported · 4 Own method · 3 Supported so far · 3 No full text of the sources · 2 No basis given · accuracy not yet measured

  • Probably not a substantial claim. 28
  • It may state two separate claims as one. 35
  • The quote only partly states this claim. 1
54 of 54
  • The paper relies on this without support.

    Cell interactions are greatly restricted under the culture conditions used.

    “even under culture conditions in which cell interactions are greatly restricted.”
  • The paper relies on this without support.

    Proteins detected in neurons 30 min after plating were present in situ rather than synthesized during the 30 min after plating.

    “The presence of these proteins so soon after plating almost certainly indicates that they were present in situ”
  • The paper relies on this without support.

    The distribution of MAP2 in situ has not yet been studied at comparably early stages.

    “The distribution of MAP2 in situ has not yet been studied at comparably early stages.”
  • The paper relies on this without support.

    MAP2 is bound to the cytoskeleton in the developing dendrites of hippocampal neurons.

    “the MAP2, which is bound to the cytoskeleton in the developing dendrites of hippocampal neurons, is not associated with microtubules.”
4 of 4

Citations

Cites

Show all 39 references

Research works printed but not linked: 3

  1. Hillman, D. E. (1979) Neuronal shape parameters and substructures as a basis of neuronal form. In The Neurosciences: Fourth Study Program, F. 0. Schmidt and F. W. Worden, eds., pp. 477-498, MIT Press, Cambridge, MA.
  2. Imhof. B. A.. H. P. Vollmers. S. L. Goodman. and W. Birchmeier teins: A monoclonal antibody to MAP2 binds to differentiated neu- rons. Proc. Natl. Acad. Sci. USA 77: 4747-4754.
  3. Matus, A., R. Bemhardt, and T. Hugh-Jones (198 1) High molecular weight microtubule associated proteins are preferentially associated with dendritic microtubules in brain. Proc. Natl. Acad. Sci. USA 78: 3010-3014.

Cited by

Sorted by citations

See all 367

Metadata

Authors
Caceres, A., Banker, G., & Binder, L.
Affiliations
Albany Medical College (US)
Venue
The Journal of Neuroscience
Published
1986-03-01
Volume
6
Issue
3
Pages
714-722
ISSN
  • 0270-6474 (ISSN-L)
  • 1529-2401
OpenAlex
W1604664180
Cited by
367
References
39
Citations by year
2026: 2 · 2025: 1 · 2024: 4 · 2022: 8 · 2021: 8 · 2020: 9 · 2019: 4 · 2018: 3 · 2017: 7 · 2016: 5 · 2015: 14 · 2014: 6 · 2013: 9 · 2012: 3
FWCI
7.34
Type
journal article
Language
en
Open access
Locations
  • Journal of Neuroscience (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
  • Europe PMC (PubMed Central) (submittedVersion)
  • PubMed Central (submittedVersion)
Funders
  • National Institutes of Health (NS12333)
  • Fogarty International Center
  • National Institute of Neurological Disorders and Stroke (NS-12333, NS 17112)
Topics
  • Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Axon Guidance and Neuronal Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Microtubule and mitosis dynamics
Keywords
beta-tubulin, MAP2, hippocampal neurons, neuronal polarity, axon development, dendrite formation, axonal growth cones, cytoskeleton, neuronal differentiation, glial progenitor cells, astrocytes, fibroblasts
MeSH terms
Animals, Axons, Dendrites, Female, Hippocampus, Histocytochemistry, Immunologic Techniques, Microtubule-Associated Proteins, Neurons, Pregnancy, Tissue Distribution, Tubulin, Rats

About these checks

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.

A language model drew the claims from the paper on 2026-10-24, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).

A second model labelled each claim on 2026-10-24 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).

Cited sources are checked with a language model’s help (gpt-6-sol).

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review