Cited by 23Neurogenesis and neuroplasticity mechanismsDevelopmental Biology and Gene RegulationNeuroinflammation and Neurodegeneration Mechanisms

The DOI matches its Crossref record; 64 of the 64 references it cites were found in a registry or an index; 5 of the 47 sources its claims cite were checked for support.

Dynamic expression of Dab2 in the mouse embryonic central nervous system

Open access
Cheung, Kwok-Kuen, Mok, Samuel C, Rezaie, Payam, Chan, Wood Yee·2008·BMC Developmental Biology

Abstract

Background: Dab2, one of two mammalian orthologs of Drosophila Disabled, has been shown to be involved in cell positioning and formation of visceral endoderm during mouse embryogenesis, but its role in neuronal development is not yet fully understood. In this report, we have examined the localization of the Dab2 protein in the mouse embryonic central nervous system (CNS) at different developmental stages. Results: Dab2 protein was transiently expressed in rhombomeres 5 and 6 of the developing hindbrain between E8.5 and E11.5, and in the floor plate of the neural tube from E9.5 to E12.5, following which it was no longer detectable within these regions. Dab2 protein was also identified within circumventricular organs including the choroid plexus, subcommissural organ and pineal gland during their early development. While Dab2 was still strongly expressed in the adult choroid plexus, immunoreactivity within the subcommissural organ and pineal gland was lost after birth. In addition, Dab2 was transiently expressed within a subpopulation of Iba1-positive mononuclear phagocytes (including presumed microglial progenitors) within the neural tube from E10.0 and was lost by E14.5. Dab2 was separately localized to Iba1 positive cells from E9.5 and subsequently to F4/80 positive cells (mature macrophage/myeloid-derived dendritic cells) positioned outside the neural tube from E12.5 onwards, implicating Dab2 expression in early cells of the mononuclear phagocyte lineage. Dab2 did not co-localize with the pan-neuronal marker PGP9.5 at any developmental stage, suggesting that Dab2 positive cells in the developing CNS are unlikely to be differentiating neurons. Conclusion: This is the first study to demonstrate the dynamic spatiotemporal expression of Dab2 protein within the CNS during development.

Overview

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TL;DR

This study maps Dab2 protein expression in the developing mouse central nervous system and shows that it is dynamically and transiently expressed in non-neuronal cells, including hindbrain rhombomeres, floor plate, circumventricular organs, and mononuclear phagocytes. It is the first detailed report of spatiotemporal Dab2 expression during mammalian CNS development.

Key findings

  • Dab2 was transiently expressed in rhombomeres 5 and 6 of the developing hindbrain between E8.5 and E11.5, and in the neural tube floor plate from E9.5 to E12.5; expression was no longer detectable in these regions afterward.
  • Dab2 was detected in developing circumventricular organs—choroid plexus, subcommissural organ, and pineal gland—but while it remained strongly expressed in the adult choroid plexus, immunoreactivity was lost in the subcommissural organ and pineal gland after birth.
  • Dab2 was transiently expressed in a subpopulation of Iba1-positive mononuclear phagocytes within the neural tube from E10.0 and was lost by E14.5; outside the neural tube, Dab2 localized to Iba1-positive cells from E9.5 and to F4/80-positive cells from E12.5 onward.
  • Dab2 did not co-localize with the pan-neuronal marker PGP9.5 at any developmental stage, suggesting Dab2-positive cells in the developing CNS are unlikely to be differentiating neurons.

Methods

  • Random-bred ICR mice were used; embryos from E7.5 to E18.0 were collected, with at least 5–10 embryos analyzed per developmental stage.
  • Dab2 protein localization was examined by immunohistochemistry on 7-μm wax sections using a monoclonal anti-p96/Dab2 antibody, with standard ABC/DAB detection and pre-adsorption or primary-antibody replacement controls; whole-mount immunohistochemistry was also performed.
  • Double immunofluorescence staining used antibodies against Dab2, Iba1, F4/80, and PGP9.5, with confocal microscopy for co-localization analysis.

Limitations

  • Stated by the authors: in the developing pineal gland, they could not rule out that interstitial cells, in addition to pinealocytes, express Dab2; the functional significance of transient Dab2 expression in the pineal gland remains to be investigated.
  • Evident from the text: the study is descriptive localization without functional perturbation or lineage tracing, so roles in CNS and myeloid development are largely inferred.
  • Evident from the text: the anti-p96/Dab2 antibody recognizes an N-terminal epitope shared by the three known Dab2 isoforms p96, p93, and p67, so the study cannot distinguish which isoform(s) are expressed in the reported sites.

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Integrity signals

Registry

Crossref · 10.1186/1471-213x-8-76

Cites

64 of 64 references verified0 citation errors · 0 not checkable

Statistics

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Figures

Figure 1 Dab2 immunoreactivity in the neural tube. In whole mount preparations (A-D), tissues surrounding the neural tube, including the otic vesicle (asterisks in C) had been removed by microdissection. At E8.5, the neural tube (A: dorsal view; A': lateral view) already shows Dab2 expression in rhombomeres 5 and 6 (arrowheads). At E9.0 (B: lateral view) and E9.5 (C: dorsal view; C': lateral view), expression within rhombomeres 5 and 6 (r5 and r6, arrows in C') remains strong, and spreads along the floor plate of the neural tube (arrowheads in B). The lack of immunoreactivity in the pre-adsorption control (D) confirms the specificity of the Dab2 immunoreaction. An oblique section (E) and longitudinal sections (F, G, G") through E9.5 embryos demonstrate strong Dab2 expression in the whole span of r6 and weak immunoreactivity in the ventral part of r5 (G" shows the high magnification of the immunoreactive r5 and r6 in G). The pre-adsorption control section (G') adjacent to the section shown in G does not exhibit any immunoreactivity. The ventral expression of Dab2 in the floor plate of the neural tube extends rostrocaudally from the mesencephalic flexure (arrow, F) to the caudal spinal cord (arrow, G). At E10.5, Dab2 immunoreactivity becomes weak in r6 (arrow, H) (Hb; hindbrain; 4V: fourth ventricle) and absent in r5. Dab2 immunoreactivity along the floor plate of the spinal cord (arrowheads, I to M) is also observed in transverse sections through the rostral (I, J, K, L, M) and caudal spinal cord (I', J', K') at different developmental stage. Also note the weak Dab2 staining in the notochord (arrow, I'). Insets in I to M represent the high magnification of the floor plate shown in corresponding figures. The schematic diagram (N) summarizes changes in Dab2 expression along the floor plate of the neural tube from E9.5 to E12.5. The dotted line indicates the axial level of the mesencephalic flexure and the shading represents Dab2 immunoreactivity observed in the floor plate of the neural tube. Arrowheads in N indicate regions of the neural tube from which sections shown in I, I', K, K', L, M were taken. Note that the onset and down-regulation of Dab2 expression follow a rostral-to-caudal sequence. Scale bar: 100 μm for I; 50 μm for G' and I', 25 μm for insets of I-L, and 200 μm for other panels.
Figure 1 Dab2 immunoreactivity in the neural tube. In whole mount preparations (A-D), tissues surrounding the neural tube, including the otic vesicle (asterisks in C) had been removed by microdissection. At E8.5, the neural tube (A: dorsal view; A': lateral view) already shows Dab2 expression in rhombomeres 5 and 6 (arrowheads). At E9.0 (B: lateral view) and E9.5 (C: dorsal view; C': lateral view), expression within rhombomeres 5 and 6 (r5 and r6, arrows in C') remains strong, and spreads along the floor plate of the neural tube (arrowheads in B). The lack of immunoreactivity in the pre-adsorption control (D) confirms the specificity of the Dab2 immunoreaction. An oblique section (E) and longitudinal sections (F, G, G") through E9.5 embryos demonstrate strong Dab2 expression in the whole span of r6 and weak immunoreactivity in the ventral part of r5 (G" shows the high magnification of the immunoreactive r5 and r6 in G). The pre-adsorption control section (G') adjacent to the section shown in G does not exhibit any immunoreactivity. The ventral expression of Dab2 in the floor plate of the neural tube extends rostrocaudally from the mesencephalic flexure (arrow, F) to the caudal spinal cord (arrow, G). At E10.5, Dab2 immunoreactivity becomes weak in r6 (arrow, H) (Hb; hindbrain; 4V: fourth ventricle) and absent in r5. Dab2 immunoreactivity along the floor plate of the spinal cord (arrowheads, I to M) is also observed in transverse sections through the rostral (I, J, K, L, M) and caudal spinal cord (I', J', K') at different developmental stage. Also note the weak Dab2 staining in the notochord (arrow, I'). Insets in I to M represent the high magnification of the floor plate shown in corresponding figures. The schematic diagram (N) summarizes changes in Dab2 expression along the floor plate of the neural tube from E9.5 to E12.5. The dotted line indicates the axial level of the mesencephalic flexure and the shading represents Dab2 immunoreactivity observed in the floor plate of the neural tube. Arrowheads in N indicate regions of the neural tube from which sections shown in I, I', K, K', L, M were taken. Note that the onset and down-regulation of Dab2 expression follow a rostral-to-caudal sequence. Scale bar: 100 μm for I; 50 μm for G' and I', 25 μm for insets of I-L, and 200 μm for other panels.CC BY · PubMed Central
Figure 2 Dab2 immunoreactivity in the developing choroid plexus, subcommissural organ and pineal gland. Sections through the hindbrain (Hb) at stages from E9.5 to E11.5 (A to C; B' and C' represent the high magnification of the boxes in B and C, respectively) show Dab2 expression in the roof plate of the hindbrain (arrows) and on scattered round cells (mononuclear phagocytes). Note that the adjacent hindbrain neuroepithelium is negative for Dab2 staining (arrowheads in B, C). Sections through the hindbrain (Hb) at stages from E12.5 to adulthood (D to G, D' is the higher magnification of the choroid plexus shown in D) show Dab2 staining in the choroid plexus (e.g. arrow in D) of the fourth ventricle (4V). Note the stronger Dab2 staining in the luminal (apical) surface of the epithelial cells as compared to the corresponding basal surface in embryos (arrowheads in D), but however, in the adult choroid plexus, the basal surface shows stronger Dab2 staining than the luminal (apical) surface of the epithelial cells (arrowheads in G). Also note the scattered and strongly Dab2 positive macrophages (arrows in D'). Sagittal sections show Dab2 staining in the subcommissural organ along the dorsal midline at different stages (H to K; H' and I' represent the higher magnification of the boxes in H and I, respectively). Note that Dab2 protein is expressed in the neuroepithelium of the subcommissural organ (sco) located in the roof plate of the diencephalon (Di). It lies underneath the posterior commissure (pc), anterior to the midbrain (Mb) (I', J, K). Also note the thin processes (arrows in I', J, K), which are also Dab2 positive, extending from the subcommissural organ into the posterior commissure. Coronal section (L) shows absence of Dab2 staining in the adult subcommissural organ. Double labeling of Dab2 (green, M) with PGP9.5 (red, M') shows that they are not co-localized (M"). Arrowheads in M indicate autofluorescence in blood cells. The sagittal section (N) shows Dab2 staining in the developing pineal gland (pg) along the dorsal midline at E16.5. Fb: forebrain; Mb: midbrain; Hb: hindbrain. Di: diencephalon. 4V: 4th ventricle; pc: posterior commissure; sco: subcommissural organ; pg: pineal gland. Scale bar: 50 μm.
Figure 2 Dab2 immunoreactivity in the developing choroid plexus, subcommissural organ and pineal gland. Sections through the hindbrain (Hb) at stages from E9.5 to E11.5 (A to C; B' and C' represent the high magnification of the boxes in B and C, respectively) show Dab2 expression in the roof plate of the hindbrain (arrows) and on scattered round cells (mononuclear phagocytes). Note that the adjacent hindbrain neuroepithelium is negative for Dab2 staining (arrowheads in B, C). Sections through the hindbrain (Hb) at stages from E12.5 to adulthood (D to G, D' is the higher magnification of the choroid plexus shown in D) show Dab2 staining in the choroid plexus (e.g. arrow in D) of the fourth ventricle (4V). Note the stronger Dab2 staining in the luminal (apical) surface of the epithelial cells as compared to the corresponding basal surface in embryos (arrowheads in D), but however, in the adult choroid plexus, the basal surface shows stronger Dab2 staining than the luminal (apical) surface of the epithelial cells (arrowheads in G). Also note the scattered and strongly Dab2 positive macrophages (arrows in D'). Sagittal sections show Dab2 staining in the subcommissural organ along the dorsal midline at different stages (H to K; H' and I' represent the higher magnification of the boxes in H and I, respectively). Note that Dab2 protein is expressed in the neuroepithelium of the subcommissural organ (sco) located in the roof plate of the diencephalon (Di). It lies underneath the posterior commissure (pc), anterior to the midbrain (Mb) (I', J, K). Also note the thin processes (arrows in I', J, K), which are also Dab2 positive, extending from the subcommissural organ into the posterior commissure. Coronal section (L) shows absence of Dab2 staining in the adult subcommissural organ. Double labeling of Dab2 (green, M) with PGP9.5 (red, M') shows that they are not co-localized (M"). Arrowheads in M indicate autofluorescence in blood cells. The sagittal section (N) shows Dab2 staining in the developing pineal gland (pg) along the dorsal midline at E16.5. Fb: forebrain; Mb: midbrain; Hb: hindbrain. Di: diencephalon. 4V: 4th ventricle; pc: posterior commissure; sco: subcommissural organ; pg: pineal gland. Scale bar: 50 μm.CC BY · PubMed Central
Figure 3 Dab2 immunoreactivity in mononuclear phagocytes. A-A": Double labeling (immunofluorescence staining) on a sagittal section through an E10.5 embryos shows co-localization of Dab2 (green, arrows in A) and Iba1 (red, arrows in A') in mononuclear phagocytes (yellow, arrows in A") within the neural tube (Ne, white outline), and in fetal macrophages outside the neural tube. B: Double labeling shows that Dab2 immunoreactivity (green, indicated by arrows) does not overlap with PGP9.5 immunoreactivity (red) within the neural tube (white outline) in a sagittal section through an E11.5 embryo. C: Double labeling at E12.5 shows Iba1+/Dab2- microglial progenitors (arrows, red) within the neural tube (Ne) and Iba1+/Dab2+ fetal macrophages (arrowhead) in the adjacent mesenchyme (mes).D-F: Sagittal sections show that Dab2+ cells (arrows) are often closely apposed to blood vessels (arrowheads) within the neural tube at E10.5 (D) and E11.5 (E), but more scattered by E12.5 (F). G: Some Dab2+ cells (arrowheads) are also observed closely associated with the roof of the fourth ventricle (4V). H-H": Double labeling for Dab2 (green, H) and Iba1 (red, H') at E14.5 shows that Dab2 is only expressed in Iba1+ fetal macrophages (arrows) in the mesenchyme (mf: mesencephalic flexure, white outline) outside the neural tube (Ne) but absent in Iba1+ microglial progenitors within the neural tube. I: At E16.5, Dab2 expression is detected in the wall of blood microvessels (arrowheads), but Dab2+ microglial progenitors are no longer detectable at this stage of development. J, K: Sections of an adult brain show Dab2+ perivascular cells (arrowheads in J) and Iba1+ microglia (arrows in K). L-L": A merged image (L") shows co-localization of F4/80 protein (red, L') in some (arrows in L") but not all of the Dab2+ (green, L) cells lying on the wall of cerebral microvessels (mv, white outline). M: A sagittal section of an E9.5 embryo shows Dab2+ cells in the cranial mensenchyme (mes) outside the neural tube (Ne). N-N": A merged image at E9.5 shows complete co-localization (yellow, brightfield, arrows in N") of Dab2 (green, arrows in N) and Iba1 (red, arrows in N') immunoreactivities in fetal macrophages outside the neural tube (Ne, white outline). O-O": A section through an E14.5 choroid plexus shows complete co-localization (yellow, O") of Dab2 (green, O) and F4/80 (red, O') immunoreactivities. Arrowheads in O' and O" indicate autofluorescence from red blood cells. DAPI counterstained in B, C, H", L". Scale bar: 50 μm.
Figure 3 Dab2 immunoreactivity in mononuclear phagocytes. A-A": Double labeling (immunofluorescence staining) on a sagittal section through an E10.5 embryos shows co-localization of Dab2 (green, arrows in A) and Iba1 (red, arrows in A') in mononuclear phagocytes (yellow, arrows in A") within the neural tube (Ne, white outline), and in fetal macrophages outside the neural tube. B: Double labeling shows that Dab2 immunoreactivity (green, indicated by arrows) does not overlap with PGP9.5 immunoreactivity (red) within the neural tube (white outline) in a sagittal section through an E11.5 embryo. C: Double labeling at E12.5 shows Iba1+/Dab2- microglial progenitors (arrows, red) within the neural tube (Ne) and Iba1+/Dab2+ fetal macrophages (arrowhead) in the adjacent mesenchyme (mes).D-F: Sagittal sections show that Dab2+ cells (arrows) are often closely apposed to blood vessels (arrowheads) within the neural tube at E10.5 (D) and E11.5 (E), but more scattered by E12.5 (F). G: Some Dab2+ cells (arrowheads) are also observed closely associated with the roof of the fourth ventricle (4V). H-H": Double labeling for Dab2 (green, H) and Iba1 (red, H') at E14.5 shows that Dab2 is only expressed in Iba1+ fetal macrophages (arrows) in the mesenchyme (mf: mesencephalic flexure, white outline) outside the neural tube (Ne) but absent in Iba1+ microglial progenitors within the neural tube. I: At E16.5, Dab2 expression is detected in the wall of blood microvessels (arrowheads), but Dab2+ microglial progenitors are no longer detectable at this stage of development. J, K: Sections of an adult brain show Dab2+ perivascular cells (arrowheads in J) and Iba1+ microglia (arrows in K). L-L": A merged image (L") shows co-localization of F4/80 protein (red, L') in some (arrows in L") but not all of the Dab2+ (green, L) cells lying on the wall of cerebral microvessels (mv, white outline). M: A sagittal section of an E9.5 embryo shows Dab2+ cells in the cranial mensenchyme (mes) outside the neural tube (Ne). N-N": A merged image at E9.5 shows complete co-localization (yellow, brightfield, arrows in N") of Dab2 (green, arrows in N) and Iba1 (red, arrows in N') immunoreactivities in fetal macrophages outside the neural tube (Ne, white outline). O-O": A section through an E14.5 choroid plexus shows complete co-localization (yellow, O") of Dab2 (green, O) and F4/80 (red, O') immunoreactivities. Arrowheads in O' and O" indicate autofluorescence from red blood cells. DAPI counterstained in B, C, H", L". Scale bar: 50 μm.CC BY · PubMed Central

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Claims the paper makes

60 Own result · 20 Own interpretation · 19 No full text of the sources · 6 Not checked yet · 2 Partly supported · 2 Own method · 2 No basis given · 1 Supported so far · 1 Sources not found · accuracy not yet measured

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Show all 69 claims

  • Probably not a substantial claim. 15
  • It may state two separate claims as one. 35
44 of 44
  • The paper relies on this without support.

    The presumptive subcommissural organ develops from the region of the roof plate of the diencephalon just anterior to the mesencephalon.

    “The presumptive subcommissural organ (SCO) develops from the region of the roof plate of the diencephalon just anterior to the mesencephalon (Fig 2H, H').”
  • The paper relies on this without support.

    PGP9.5 is a pan-neuronal marker.

    “Double immunofluorescent labeling of Dab2 and PGP9.5, a pan-neuronal marker, was performed in order to determine whether Dab2 was expressed in neurons or nerve fibers that were closely associated with the SCO.”
  • The paper relies on this without support.

    F4/80 antigen is a macrophage differentiation marker.

    “The macrophage differentiation marker, F4/80 antigen, began to be expressed weakly in these fetal macrophages located outside the neural tube at E12.5 and became prominent from E14.5 onwards.”
  • The paper relies on this without support.

    A protein's expression pattern in specific cell types and developmental stages indicates its functional importance.

    “Dynamic expression of Dab2 protein was detected in the neuroepithelium, blood vessels, circumventricular organs and mononuclear phagocytes (fetal macrophages, microglial progenitors), strongly suggesting that Dab2 is an important molecule during mammalian CNS and myeloid cell development.”
  • The paper relies on this without support.

    The very early ontogenetic differentiation of SCO secretory cells is crucial for the smooth circulation of CSF secreted from the choroid plexus during early development.

    “The very early ontogenetic differentiation of SCO secretory cells is crucial for the smooth circulation of CSF secreted from the choroid plexus during early development.”
  • The authors state this as an assumption.

    Copulation occurred around midnight, so that noon of the plug day can be designated as embryonic day 0.5 (E0.5).

    “The noon of the plug day was designated as embryonic day 0.5 (E0.5) assuming that copulation occurred around midnight.”
6 of 6

Citations

Cites

Show all 62 references

Research works printed but not linked: 1

  1. Rezaie P: Microglia, from origins to differentiation, distribution and dynamics within the mammalian central nervous system. Trends in Glial Research - Basic and Applied. Edited by: Dheen ST, Ling EA. 2007, India , Research Signpost, 1-41.

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Metadata

Authors
Cheung, K.-K., Mok, S. C., Rezaie, P., & Chan, W. Y.
Affiliations
  • Hong Kong Polytechnic University (HK)
  • The University of Texas MD Anderson Cancer Center (US)
  • King's College London (GB)
  • Chinese University of Hong Kong (HK)
Corresponding author
Chan, Wood Yee
Venue
BMC Developmental Biology
Published
2008-08-04
Volume
8
Issue
1
Pages
76-76
ISSN
1471-213X (ISSN-L)
OpenAlex
W2075358654
Cited by
23
References
62
Citations by year
2025: 1 · 2024: 3 · 2021: 1 · 2020: 2 · 2018: 1 · 2016: 1 · 2015: 1 · 2014: 3 · 2013: 3
FWCI
0.66
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • BMC Developmental Biology (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Open Research Online - ORO (The Open University) (acceptedVersion)
  • PolyU Institutional Research Archive (Hong Kong Polytechnic University) (submittedVersion, cc-by)
Funders
  • U.S. Department of Health and Human Services
  • Chinese University of Hong Kong
  • National Cancer Institute (R33CA103595, P50CA165009)
Topics
  • Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
Keywords
Dab2, floor plate, circumventricular organs, choroid plexus, subcommissural organ, pineal gland, mononuclear phagocytes, macrophages, myeloid dendritic cells, spatiotemporal gene expression, neuronal development
MeSH terms
Animals, Brain, Embryo, Mammalian, Mice, Inbred ICR, Mononuclear Phagocyte System, Gene Expression Regulation, Developmental, Adaptor Proteins, Vesicular Transport, Adaptor Proteins, Signal Transducing, Apoptosis Regulatory Proteins, Mice, Neural Tube

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