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Cited by 250Developmental Biology and Gene RegulationPlant Molecular Biology ResearchNeurobiology and Insect Physiology Research

Dorsoventral patterning in the Drosophila central nervous system: the intermediate neuroblasts defective homeobox gene specifies intermediate column identity

Open access
Weiss, Joseph B., Von Ohlen, Tonia, Mellerick, Dervla M., Dressler, Gregory, Doe, Chris Q., Scott, Matthew P.·1998·Genes & Development
DOI 10.1101/gad.12.22.3591PMC PMC317240
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Abstract

One of the first steps in neurogenesis is the diversification of cells along the dorsoventral axis. In Drosophila the central nervous system develops from three longitudinal columns of cells: ventral cells that express the vnd/nk2 homeobox gene, intermediate cells, and dorsal cells that express the msh homeobox gene. Here we describe a new Drosophila homeobox gene, intermediate neuroblasts defective (ind), which is expressed specifically in the intermediate column cells. ind is essential for intermediate column development: Null mutants have a transformation of intermediate to dorsal column neuroectoderm fate, and only 10% of the intermediate column neuroblasts develop. The establishment of dorsoventral column identity involves negative regulation: Vnd represses ind in the ventral column, whereas ind represses msh in the intermediate column. Vertebrate genes closely related to vnd (Nkx2.1 and Nkx2.2), ind (Gsh1 and Gsh2), and msh (Msx1 and Msx3) are expressed in corresponding ventral, intermediate, and dorsal domains during vertebrate neurogenesis, raising the possibility that dorsoventral patterning within the central nervous system is evolutionarily conserved.

Integrity signals

Registry

Crossref · 10.1101/gad.12.22.3591

Cites

58 of 58 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

8 of 16 cited sources checked · 4 supported · 0 flagged · accuracy not yet measured

  • Premisecitation

    msh expression in the dorsal neuroectoderm is mirrored by Msx1 transcription in the dorsal neural tube.

    “Similarly msh expression in the dorsal neuroectoderm is mirrored by Msx1 transcription in the dorsal neural tube (D' Alessio and Frasch 1996;Isshiki et al. 1997).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites D' Alessio and Frasch 1996 source
    • cites Isshiki et al. 1997 source
  • Premisecitation

    Vertebrate genes closely related to vnd (Nkx2.1 and Nkx2.2), ind (Gsh1 and Gsh2), and msh (Msx1 and Msx3) are expressed in corresponding ventral, intermediate, and dorsal domains during vertebrate neurogenesis.

    “Vertebrate genes closely related to vnd (Nkx2.1 and Nkx2.2), ind (Gsh1 and Gsh2), and msh (Msx1 and Msx3) are expressed in corresponding ventral, intermediate, and dorsal domains during vertebrate neurogenesis”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites Valerius et al. 1995 source

      The In Situ Hybridizations section reports Gsh-1 expression in a restricted mid-ventricular region of the developing mouse neural tube, supporting the intermediate-domain portion of the claim. The Gsh-1 Target DNA Binding Sequence section also reports strong Gsh-1/Gsh-2 homeodomain similarity. Our copy does not establish the proposed relationships to ind, vnd, or msh, or the claimed ventral and dorsal expression patterns of the other listed genes; the passages provide no page numbers. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
    • cites Szucsik et al. 1997 source
  • Findingcitation

    Ind protein-containing cells are immediately adjacent but nonoverlapping with Vnd-containing cells of the ventral column neuroectoderm.

    “The Ind proteincontaining cells are immediately adjacent but nonoverlapping with Vnd-containing cells of the ventral column neuroectoderm (see McDonald et al. 1998).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites McDonald et al. 1998 source
  • Interpretationreasoning

    The similar spatial distribution of Nkx2.2 and Msx1 and Msx3 expression to that of vnd and msh suggests a conserved mechanism of DV patterning across species.

    “The similar spatial distribution of Nkx2.2 and Msx1 and Msx3 expression to that of vnd and msh, respectively suggests a conserved mechanism of DV patterning across species (D' Alessio and Frasch 1996;Wang et al. 1996).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites D' Alessio and Frasch 1996 source
    • cites Wang et al. 1996 source
  • Findingown data

    The three Vnd binding sites each contain one copy of the sequence GTGAACT.

    “The three sites protected in the footprinting assay each contain one copy of the sequence GTGAACT, which has been found to be a recognition sequence for both Vnd and the Tinman-related Nkx2.5 vertebrate protein (Chen and Schwartz 1995;Gruschus et al. 1997).”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites Chen and Schwartz 1995 source

      The DNA-binding results and Discussion (page unavailable) identify nkx-2.5 binding sequences, including the preferred consensus 5′-CTNNAGTG-3′. Our copy does not report three Vnd sites, a footprinting assay of those sites, or the sequence GTGAACT; searches for GTGAACT and Vnd found no passage addressing the claim. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites Gruschus et al. 1997 source
  • Classified as probably not a substantial claim. 4
  • May combine two claims. 22
  • The quote only partly states this claim. 2

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    vnd is thought to promote neuroblast formation by proneural-dependent and proneural-independent pathways.

    “Similarly, vnd is thought to promote neuroblast formation by proneural-dependent and proneural-independent pathways (Jimenez et. al. 1995;McDonald et al. 1998).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites Jimenez et. al. 1995 source
    • cites McDonald et al. 1998 source

      In “Vnd expression during neurogenesis” (page unavailable), the paper proposes that vnd acts with the proneural genes achaete and scute and says it has an additional role beyond activating their expression. It does not establish that this additional role is independent of proneural genes altogether; the passage also mentions lethal of scute in other ventral neuroblasts. (LLM, conf=0.82)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Findingcitation

    vnd function is required to establish the ventral border of ind: in the absence of vnd, expression of ind is observed in the ventral-column neuroectoderm and neuroblasts.

    “In this paper and in McDonald et al. (1998) we show that vnd function is required to establish the ventral border of ind: In the absence of vnd, expression of ind is observed in the ventral-column neuroectoderm and neuroblasts.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites McDonald et al. 1998 source

  • May combine two claims.

    Findingown data

    ind expression is first detected in two parallel lateral columns in stage 5 cellular blastoderm embryos, each about five cells wide, and narrows to two cells wide during gastrulation.

    “Whole-mount in situ hybridization first detects ind expression in two parallel, lateral columns in stage 5 cel-lular blastoderm embryos (Fig. 2).”
    • Also stated in: § A screen for genes regulated by Tinman class homeodomain proteins · plain
      “During gastrulation, the ind expression domain narrows until each column is two cells wide.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Embryos lacking ind function have profound defects in the development of the intermediate column of the CNS, including derepression of dorsal column markers within the intermediate-column neuroectoderm.

    “Embryos lacking ind function have profound defects in the development of the intermediate column of the CNS.”
    • Also stated in: § Discussion · plain
      “The defects include a derepression of dorsal column markers within the intermediate-column neuroectoderm.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    By stage 9, ind mRNA is absent from the neuroectoderm but detectable in all neuroblasts in the intermediate column (Nbs 3-2, 4-2, 5-3, 6-2, and 7-2).

    “By stage 9, ind mRNA is absent from the neuroectoderm, but is detectable in all of the neuroblasts in the intermediate column (Nbs 3-2, 4-2, 5-3, 6-2, and 7-2).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    ind is a Drosophila homeobox gene expressed specifically in the intermediate column cells.

    “Here we describe a new Drosophila homeobox gene, intermediate neuroblasts defective (ind), which is expressed specifically in the intermediate column cells.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In stage 9 wild-type embryos, five neuroblasts constitute the intermediate column in each hemisegment, whereas in ind mutant embryos at most one intermediate-column neuroblast is observed in each hemisegment and the normal number of ventral and dorsal column neuroblasts form.

    “In stage 9 wild-type embryos, five neuroblasts constitute the intermediate column in each hemisegment.”
    • Also stated in: § ind promotes intermediate-column neuroblast formation · plain
      “In ind mutant embryos, at most one intermediate-column neuroblast is observed in each hemisegment, whereas the normal number of ventral and dorsal column neuroblasts form.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In ind mutant embryos, 32 intermediate neuroblasts were observed out of 80 hemisegments, where 400 would be expected in wild-type; 11 expressed Vnd, 12 expressed Msh, and 9 expressed neither marker.

    “A total of 80 hemisegments were scored.In a wild-type embryo 400 intermediate neuroblasts would be expected.In the ind mutant embryo 32 were observed.Eleven expressed Vnd, 12 expressed Msh, and 9 expressed neither marker.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In wild-type embryos, an Eve-protein-containing RP2 is found in every hemisegment (100/100), whereas in ind mutant embryos the Eve-containing RP2 is never detected (0/96).

    “In wild-type embryos, an Eve-protein-containing RP2 is found in every hemisegment (100/100), whereas in ind mutant embryos the Eve-containing RP2 is never detected (0/96) (Fig. 6M,N).”
    Quote in the paperSubstantial claim

  • May combine two claims. The quote only partly states this claim.

    Findingown data

    Ind protein is detected in the nucleus at all stages assayed and is restricted to the intermediate column neuroectoderm and neuroblasts.

    “The Ind protein is detected in the nucleus at all stages assayed, as expected for a homeodomain protein (Fig. 3).”
    • Also stated in: § A screen for genes regulated by Tinman class homeodomain proteins · boosted
      “The antibody staining confirms that Ind is restricted to the intermediate column neuroectoderm and neuroblasts.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Vnd specifically binds the ind genomic DNA fragment isolated in the screen, with three specific binding sites of roughly equal affinity.

    “Vnd specifically binds the fragment of ind genomic DNA isolated in the screen (Fig. 5A).”
    • Also stated in: § A screen for genes regulated by Tinman class homeodomain proteins · plain
      “Three specific binding sites of roughly equal affinity can be identified using footprinting assays (Fig. 5B).”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Findingreasoning

    In Figure 7 the DV order of the Drosophila and mouse genes are clearly the same.

    “In Figure 7 the DV order of the Drosophila and mouse genes are clearly the same.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    At early stage 9, ind is expressed in the intermediate neurectoderm and in S1 intermediate-column neuroblasts.

    “(C,G) Early stage 9 embryos in which ind is expressed in the intermediate neurectoderm and S1, intermediate column neuroblasts.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In ind16.2 mutant embryos, Msh is expanded in the neurectoderm at stage 7, and this expansion persists until stage 9 when the neuroblasts delaminate.

    “(B,D,F).ind 16.2 mutant embryos show Msh is expanded in the neurectoderm at stage 7 (B).This persists until stage 9 when the neuroblasts delaminate (D,F).”
    Quote in the paperSubstantial claim

  • The quote only partly states this claim.

    Findingown data

    A P-element insertion (R0220) obtained from the Drosophila Genome Project was mapped near the coding sequence of ind.

    “A P-element insertion obtained from the Drosophila Genome Project was mapped near the coding sequence of ind (Fig. 1).”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Findingown data

    The ind coding sequence is contained in a single exon.

    “The coding sequence for ind is contained in a single exon, shown as a large box.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Vnd forms a DNA/protein complex with ind DNA nucleotides 5-135 that is competed out by the Vnd-I oligonucleotide but not by the irrelevant H2A oligonucleotide.

    “(B) Electrophoresis mobilityshift assay using ind DNA nucleotides 5-135, which includes all three Vnd-binding regions.A DNA/protein complex is observed upon addition of Vnd.This complex can be competed out with increasing amounts of the Vnd-I oligonucleotide but not with an irrelevant oligonucleotide (H2A).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Wild-type late stage 9 embryos have three columns of neuroblasts, whereas ind16.2 embryos have only two columns of S1 neuroblasts.

    “Pan neural expression of Hunchback in wild-type late stage 9 embryos shows there are three columns of neuroblasts (K).In ind 16.2 embryos only two columns of S1 neuroblasts are observed.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The deletion in the ind79.3 allele is 7 kb long and removes the ind coding sequence.

    “The deletion is 7 kb long and removes the coding sequence for ind.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    ind appears to be the missing regulator, a gene that precisely marks and controls the development of the intermediate column of neuroectoderm and neuroblasts.

    “ind appears to be the missing regulator, a gene that precisely marks and controls the development of the intermediate column of neuroectoderm and neuroblasts.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: No genes were known to be expressed specifically in the intermediate column of the Drosophila CNS before this study.

  • May combine two claims.

    Premisecitation

    Vertebrate orthologs of vnd and msh have been identified: Nkx2.1 and Nkx2.2 and Msx1 and Msx3, respectively.

    “In addition vertebrate orthologs of vnd and msh have been identified: Nkx2.1 and Nkx2.2 and Msx1 and Msx3, respectively (Holland 1991;Barth and Wilson 1995).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Holland 1991 source
    • cites Barth and Wilson 1995 source

  • May combine two claims.

    Interpretationown data

    ind represses msh and achaete gene expression directly or indirectly, and ind is necessary for establishing proper intermediate-column identity within the neuroectoderm.

    “We conclude that ind represses msh and achaete gene expression directly or indirectly, and that ind is necessary for establishing proper intermediate-column identity within the neuroectoderm.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    The ind phenotype is caused specifically by loss of ind function because the ind 79.3 mutation deletes the ind-coding region and 6 kb of flanking DNA but not any other detectable gene.

    “We believe that the ind phenotype described below is caused specifically by the loss of ind function because the ind 79.3 mutation is a small deficiency that deletes the ind-coding region and 6 kb of flanking DNA, but not any other detectable gene (Fig. 1).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    ind is essential for the formation of intermediate column neuroblasts and for the repression of ventral- and dorsal-specific genes within these neuroblasts.

    “Thus, ind is essential for the formation of intermediate column neuroblasts, and for the repression of ventral-and dorsal-specific genes within these neuroblasts.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationnone given

    The three longitudinal columns along the DV axis are defined by the ventral expression and function of vnd, the intermediate expression and function of ind, and the dorsal expression and function of msh.

    “The three longitudinal columns of cells along the DV axis are defined by the ventral expression and function of vnd; the intermediate expression and function of ind; and the dorsal expression and function of msh.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    The sequence of both strands of the ind cDNA was determined by cycle sequencing with an automated ABI sequencer.

    “The sequence of both strands was determined by the cycle sequencing reaction and an automated ABI sequencer (ABI PRISM Dye Terminator Cycle Sequencing Ready Reaction Kit, Perkin Elmer).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Loss of ind function results in loss of intermediate-column neuroblasts and of intermediate-column-derived neurons.

    “Loss of ind function results in loss of intermediate-column neuroblasts and intermediate-column-derived neurons.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The ind mutant alleles used (including the ind79.3 deletion) disrupt only the ind gene, so the observed CNS phenotypes are due to loss of ind function.
27 of 27

  • The paper relies on this without support.

    No genes were known to be expressed specifically in the intermediate column of the Drosophila CNS before this study.

    “Although vnd and msh are candidate genes for establishing ventral and dorsal column fates within the CNS, no genes are known currently to be expressed specifically in the intermediate column of the CNS.”

  • The paper relies on this without support.

    The anti-Ind antibodies used specifically detect Ind protein, so the observed protein distribution reflects Ind.

    “Rat serum and monoclonal antibodies were raised against an Ind peptide conjugated to bovine serum albumin (BSA).After the monoclonal fusion, hybridoma supernatants were screened by ELISA against a peptide-KLH conjugate.Positive hybridoma lines were tested by immunohistochemistry.”

  • The paper relies on this without support.

    The ind mutant alleles used (including the ind79.3 deletion) disrupt only the ind gene, so the observed CNS phenotypes are due to loss of ind function.

    “The deletion is 7 kb long and removes the coding sequence for ind.”
3 of 3

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • The third P-element mutant has two allele numbers

    Minor

    The Results identify the third P-element-derived allele as ind 4.1, while Materials and methods identify it as ind 4.2.

  • Figure 5 assay panels are reversed in the text

    Minor

    The Results assign binding and footprinting assays to panels A and B, respectively. The figure caption labels panel A as footprinting and panel B as the mobility-shift binding assay.

  • Ind protein is both present and absent after stage 11

    Minor

    The text says Ind protein is detectable in all intermediate-column neuroblasts at stage 13, but the next sentence says it is not detectable after stage 11.

3 of 3

Cites

Flagged first, then by citations

Research works printed but not linked: 2

  1. Angerer L. Angerer R. (1992) In situ hybridization to cellular RNA with radiolabelled RNA probes. in In situ hybridization: A practical approach, ed Wilkinson D.G. (Oxford University Press, Oxford, UK), pp 15–32.
  2. Ausubel F.M. Brent R. Kingston R.E. Moore D.D. Seidman J.G. Smith J.A. Struhl K. (1994) Current protocols in molecular biology (John Wiley & Sons, New York, NY).

Cited by

Sorted by citations

Related

Authors
Weiss, J. B., Von Ohlen, T., Mellerick, D. M., Dressler, G., Doe, C. Q., & Scott, M. P.
Affiliations
Howard Hughes Medical Institute (US)
ORCID
Venue
Genes & Development
Publisher
Cold Spring Harbor Laboratory
Published
1998-11-15
Volume
12
Issue
22
Pages
3591-3602
ISSN
  • 0890-9369 (ISSN-L)
  • 1549-5477
DOI
10.1101/gad.12.22.3591
PMC
PMC317240
OpenAlex
W2123933036
Semantic Scholar
d5b2bd0f775838548cd175b48517cdd9cce81c8a
Cited by
250
References
57
Citations by year
2026: 7 · 2025: 6 · 2024: 14 · 2023: 1 · 2022: 7 · 2021: 1 · 2020: 2 · 2019: 7 · 2018: 2 · 2017: 5 · 2016: 8 · 2015: 10 · 2014: 1 · 2013: 8 · 2012: 4
Type
posted content
Language
en
Open access
Locations
  • Genes & Development (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
  • Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant Molecular Biology Research
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neurobiology and Insect Physiology Research
Keywords
dorsoventral patterning, Drosophila central nervous system, homeobox genes, neuroblast differentiation, negative regulation, evolutionary conservation
MeSH terms
Amino Acid Sequence, Animals, Base Sequence, Central Nervous System, Drosophila

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