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Early B-cell factors 2 and 3 (EBF2/3) regulate early migration of Cajal-Retzius cells from the cortical hem.

Chiara F, Badaloni A, Croci L, Yeh ML, Cariboni A, Hoerder-Suabedissen A, Consalez GG, Eickholt B, Shimogori T, Parnavelas JG, Rakić S - Dev. Biol. (2012)

Bottom Line: Here, we show that Ebf transcription factors are expressed in forebrain signalling centres-the septum, cortical hem and the pallial-subpallial boundary-known to generate CR cells.Accordingly, using in vitro preparations, we demonstrated that both Ebf2 and Ebf3, singly or together, control the migration of CR cells arising in the cortical hem.These findings provide evidence that Ebfs directly regulate CR cell development.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell and Developmental Biology, University College London, UK.

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Ebfs overexpression and downregulation affect the migration of GN11 cells in vitro. Ebf1 and Ebf3 are expressed in GN11 cells (A). Fewer cells are visible in the Mock sample (B) compared to the Ebf-flag (C). Bar graph shows the increased migration of GN11 cells towards FBS when treated with Ebfs overexpression plasmids (D). GN11 cells do not migrate towards DMEM (D). Migration of GN11 cells when treated with sh-Ebf1 and Ebf3 is impaired compared to Mock- and Ebf2-treated (E–H). Migration is significantly rescued after transfecting with Ebf-flag plasmids sh-Ebf treated cells (G compared to F; H). Scale bar: 25 μm.
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f0035: Ebfs overexpression and downregulation affect the migration of GN11 cells in vitro. Ebf1 and Ebf3 are expressed in GN11 cells (A). Fewer cells are visible in the Mock sample (B) compared to the Ebf-flag (C). Bar graph shows the increased migration of GN11 cells towards FBS when treated with Ebfs overexpression plasmids (D). GN11 cells do not migrate towards DMEM (D). Migration of GN11 cells when treated with sh-Ebf1 and Ebf3 is impaired compared to Mock- and Ebf2-treated (E–H). Migration is significantly rescued after transfecting with Ebf-flag plasmids sh-Ebf treated cells (G compared to F; H). Scale bar: 25 μm.

Mentions: In line with this, we first used the GN11 cell line as a model to study migratory cell behaviour (Maggi et al., 2000) in response to the overexpression or downregulation of Ebf factors. GN11 cells naturally express Ebf1 and Ebf3, but not Ebf2 (Fig. 7A), thus not requiring further manipulation prior our experiments. The plasmids used were previously tested for specificity in COS cells (Figs. S3). GN11 cells, transfected either with Ebf1, Ebf2 or Ebf3-flag, and exposed to a general chemoattractant (1% FBS) in a Boyden chamber assay, displayed a significant increase in chemomigration compared to mock-transfected cells (Mock) (Fig. 7B compared to C; D: Mock 609 ± 36, Ebf1-flag 1522 ± 30, Ebf2-flag 1585 ± 24, Ebf3-flag 1747 ± 48; n = 3; p < 0.001). Interestingly, Ebf2, even if not expressed by GN11 cells, was able, when overexpressed, to increase the rate of migration.


Early B-cell factors 2 and 3 (EBF2/3) regulate early migration of Cajal-Retzius cells from the cortical hem.

Chiara F, Badaloni A, Croci L, Yeh ML, Cariboni A, Hoerder-Suabedissen A, Consalez GG, Eickholt B, Shimogori T, Parnavelas JG, Rakić S - Dev. Biol. (2012)

Ebfs overexpression and downregulation affect the migration of GN11 cells in vitro. Ebf1 and Ebf3 are expressed in GN11 cells (A). Fewer cells are visible in the Mock sample (B) compared to the Ebf-flag (C). Bar graph shows the increased migration of GN11 cells towards FBS when treated with Ebfs overexpression plasmids (D). GN11 cells do not migrate towards DMEM (D). Migration of GN11 cells when treated with sh-Ebf1 and Ebf3 is impaired compared to Mock- and Ebf2-treated (E–H). Migration is significantly rescued after transfecting with Ebf-flag plasmids sh-Ebf treated cells (G compared to F; H). Scale bar: 25 μm.
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f0035: Ebfs overexpression and downregulation affect the migration of GN11 cells in vitro. Ebf1 and Ebf3 are expressed in GN11 cells (A). Fewer cells are visible in the Mock sample (B) compared to the Ebf-flag (C). Bar graph shows the increased migration of GN11 cells towards FBS when treated with Ebfs overexpression plasmids (D). GN11 cells do not migrate towards DMEM (D). Migration of GN11 cells when treated with sh-Ebf1 and Ebf3 is impaired compared to Mock- and Ebf2-treated (E–H). Migration is significantly rescued after transfecting with Ebf-flag plasmids sh-Ebf treated cells (G compared to F; H). Scale bar: 25 μm.
Mentions: In line with this, we first used the GN11 cell line as a model to study migratory cell behaviour (Maggi et al., 2000) in response to the overexpression or downregulation of Ebf factors. GN11 cells naturally express Ebf1 and Ebf3, but not Ebf2 (Fig. 7A), thus not requiring further manipulation prior our experiments. The plasmids used were previously tested for specificity in COS cells (Figs. S3). GN11 cells, transfected either with Ebf1, Ebf2 or Ebf3-flag, and exposed to a general chemoattractant (1% FBS) in a Boyden chamber assay, displayed a significant increase in chemomigration compared to mock-transfected cells (Mock) (Fig. 7B compared to C; D: Mock 609 ± 36, Ebf1-flag 1522 ± 30, Ebf2-flag 1585 ± 24, Ebf3-flag 1747 ± 48; n = 3; p < 0.001). Interestingly, Ebf2, even if not expressed by GN11 cells, was able, when overexpressed, to increase the rate of migration.

Bottom Line: Here, we show that Ebf transcription factors are expressed in forebrain signalling centres-the septum, cortical hem and the pallial-subpallial boundary-known to generate CR cells.Accordingly, using in vitro preparations, we demonstrated that both Ebf2 and Ebf3, singly or together, control the migration of CR cells arising in the cortical hem.These findings provide evidence that Ebfs directly regulate CR cell development.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell and Developmental Biology, University College London, UK.

Show MeSH