Figures and data in NG2 glia are required for vessel network formation during embryonic development

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6 figures and 2 tables

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Figure 1

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NG2⁺ glia are in close contact with blood vessels.

(**A–D**) Double immunohistochemistry for NG2 and Isolectin (**A1–A3, B1–B3**) and for NG2 and PECAM (**C1–C3, D1–D2**) on coronal cingulate cortex (CCi) and cingulate bundle (CI) sections of wild-type mice (n=3 each) at E12.5 (**A1–A3**), at E14.5 (**B1–B3** and **C1–C3**), and at E16.5 (**D1–D2). A3, A2, B2, B3, C2, C3**,and D2 are higher power views of the region in **A1, B1, C1,** and D1, respectively (white arrowheads). D3 is an isosurface reconstruction of the labeling seen in D2. The processes of the NG2⁺ glia are in close contact with adjacent blood vessels (open arrowheads in **A3, B3,** and C3). Bar = 675 μm in **A1, B1,** and D1; 50 μm in **A2, B2, C1,** and D2; 40 μm in **A3, B3, C2,** and **C3. CSB**, corticoseptal boundary at the midline where the corpus callosum will form.

https://doi.org/10.7554/eLife.09102.003

Figure 2 with 2 supplements

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NG2⁺ glia of the dorsal telencephalon are derived from Nkx2.1⁺ progenitors of the subpallium.

(**A–E**) Double immunohistochemistry for the YFP and NG2 (**A1–A2**) (n=3), the YFP and Olig2 (B) (n=3), the YFP and S100β (C) (n=3), the YFP and GLAST (D) (n=3), and the YFP and GFAP (E) (n=3) on telencephalic coronal slices of *Cspg4-cre⁺/Rosa-EYFP* mice at E16.5 (B and D) and E18.5 (**A1, A2, C,** and E). (**F–J**) Double immunohistochemistry for the YFP and NG2 (**F1–F2**) (n=5), the YFP and Olig2 (G) (n=5), the YFP and S100β (H) (n=4), the YFP and GLAST (I) (n=4), and the YFP and GFAP (J) (n=3) on telencephalic coronal slices of *Nkx2.1-cre⁺/Rosa-EYFP* mice at E16.5 (**F1, F2,** and H) and E18.5 (**G, I,** and J). A2 and F2 are higher power views of the cingulate region in A1 and F1, respectively. The *NG2*-derived and the *Nkx2.*1-derived YFP⁺ cells co-express polydendroglial markers, NG2 and Olig2, together with S100β (white arrowheads) but lack expression of GLAST and GFAP (open arrowheads in D and I). Bar=675 μm in **A1, F1**; 100 μm in **E, J**; 50 μm in **A2, B, C, D, F2, G, H, I**.

https://doi.org/10.7554/eLife.09102.004

Figure 2—figure supplement 1

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YFP signal in *Nkx2.1-cre⁺/Rosa-EYFP* and *Cspg4-cre⁺/Rosa-EYFP* mice is present in NG2 glia.

(A) Bars (means ± SEM) represent the percentage of YFP-labeled NG2 glia in corpus callosum (CC), cingulate bundle (CI), and cingulate cortex (CCi) sections of E18.5 *Cspg4-cre⁺/Rosa-EYFP* mice (n=3). The YFP signal in *Cspg4-cre⁺/Rosa-EYFP* mice was detected in a majority of NG2⁺ embryonic glia of the dorsal telencephalon (71.7 ± 14.6% in the CC, 57 ± 3.8% in the CI and 69 ± 5.3% in the CCi at E16.5, n=3). (**B–C**) Double immunohistochemistry for NG2 and PECAM in CCi coronal sections of *Cspg4-cre^-/Rosa-DTA* (n=3) (B) and *Cspg4-cre⁺/Rosa-DTA* (n=3) (C) mutant mice at E18.5. Scale bar = 50 µm in B and C. (D) Bars (means ± SEM; unpaired Student’s t-test) represent the percentage of remaining NG2 glia in the telencephalon of E18.5 *Cspg4-cre⁺/Rosa-DTA* mice compared to control mice. A drastic loss of NG2⁺ glia followed by severe vascular defects was observed in *Cspg4-cre⁺/Rosa-DTA* mice compared to *Cspg4-cre^-/Rosa-DTA*. (E) Bars (means) represent the percentage of glial markers expression by YFP-labeled NG2 glia in the CC, Cl, and CCi of E18.5 *Cspg4-cre⁺/Rosa-EYFP* mice. In *Cspg4-cre⁺/Rosa-EYFP* mice, all YFP-labeled NG2 glia of the dorsal telencephalon expressed NG2⁺/Olig2⁺ (n=2), many S100β⁺(n=2) but no GLAST⁺ (n=3) or GFAP⁺ (n=2). (F) Bars (means)represent the percentage of NG2 markers expression by YFP-labeled cells in the telencephalon of E18.5 *Nkx2.1-cre⁺/Rosa-EYFP* mice (n=4) (47.24% in the CC and 27.2% in the Cl). CC, corpus callosum; CCi, cingulate cortex; Cl, cingulate bundle.

https://doi.org/10.7554/eLife.09102.005

Figure 2—figure supplement 2

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*Nkx2.1*-derived NG2 and Olig2 glia are transient and gradually disappear from the dorsal pallium at postnatal ages.

(**A–C**) Double immunohistochemistry for GFP and NG2 on coronal telencephalicsections from *Nkx2.1-cre⁺/Rosa-EYFP* mice at P0 (n=3) (**A1–A2**), P2 (n=3) (**B1–B2**), and P8 (n=2) (**C1–C2**). (D) Double immunohistochemistry for GFP and Olig2 on coronal telencephalicsections from *Nkx2.1-cre⁺/Rosa-EYFP* mice at P8 (n=2) (**D1–D3**). Cell nuclei were counterstained in blue with Hoechst (**A1, B1,** and C1). **A2, B2, C2,** and D3 are high-power views of the cingulate bundle (Cl) seen in **A1, B1, C1**, and D1, respectively. In the Cl of *Nkx2.1-cre⁺/Rosa-EYFP* mice brains at P2, only very few *Nkx2.1*-derived NG2 glia remained (arrowheads B2). At P8, *Nkx2.1*-derived NG2 and Olig2 glia disappeared completely and were replaced by NG2 and Olig2 glia that did not expressed the YFP and were not derived from Nkx2.1 germinal domains (open arrowheads in A2 and C2). At all postnatal ages many *Nkx2.1*-derived GABAergic interneurons found in this region were not labeled for NG2 (arrows in B2 and C2).Scale bar = 675 µm in **A1, B1, C1** and D1; 40 µm in **A2, B2** and **C2;** 100 µm in D2 and D3. CC, corpus callosum; CCi, cingulate cortex; LV, lateral ventricles; SEP, septum.

https://doi.org/10.7554/eLife.09102.006

Figure 3 with 2 supplements

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NG2⁺ glia, but not pericytes, control blood vessels formation.

(**A–C**) Double immunohistochemistry for the YFP and Isolectin (**A1–A2**) or PDGFRβ (**B1–B2, C1–C2**) on coronal cingulate cortex (CCi) and cingulate bundle (CI) sections of *Nkx2.1-cre⁺/Rosa-EYFP* (**A1, C1–C2**) and *Cspg4-cre⁺/Rosa-EYFP* (**A2, B1–B2**) mice (n=3) at E16.5 (**A1–A2**) and E18.5 (**B1–B2** and **C1–C2**). (**A–C**) From E16.5 to E18.5, numerous YFP⁺ NG2 glia are surrounding cortical blood vessels. (**B–C**) The Cre-mediated recombination, visualized by the YFP signal, can be observed in only very few pericytes surrounding blood vessels in *Cspg4-cre⁺/Rosa-EYFP* mice (B2, boxed region showing high magnification of region marked with white arrowhead), but not in *Nkx2.1-cre⁺/Rosa-EYFP* mice (**C2,** boxed region showing high magnification of region marked with arrow). (**D–F**) Double immunohistochemistry for NG2 and PDGFRβ on coronal CI sections in wild-type (**D1–D2**), *Nkx2.1-cre⁺/Rosa-DTA* (**E1–E2**) (n=3) and *Cspg4-cre⁺/Rosa-DTA* (**F1–F2**) (n=3) mice at E18.5. (**D–F**) The NG2⁺ glia form a complex cellular network around the cortico-cerebral blood vessels outlined by NG2 and PDGFRβ staining. The DTA under the control of *Nkx2.1* (E) and *Cspg* or *NG2* (F) promoters selectively depletes NG2⁺ glia but not pericytes. **D2, E2,** and F2 are higher power views of the cingulate region in **D1, E1,** and F1, respectively (white arrowheads). (G) Bars (means ± SEM) represent the percentage of YFP-negative and YFP-positive PDGFRβ labeled pericytes in dorsal telencephalon sections of E18.5 *Cspg4-cre⁺/Rosa-EYFP* mice (n=10). The YFP signal in *Cspg4-cre⁺/Rosa-EYFP* mice was not detected in PDGFRβ⁺ embryonic pericytes of the dorsal telencephalon (95.05 ± 1.54% of pericytes are YFP-negative in the CI at E18.5, n=10). (H) Bars (means ± SEM; unpaired Student’s t-test) represent the percentage of remaining PDGFRβ+ pericytes in cingulate bundle (Cl) sections of E18.5 *Cspg4-cre⁺/Rosa-DTA* mice (n=11) compared to control mice (n=11). No loss of PDGFRβ+ pericytes was observed in *Cspg4-cre⁺/Rosa-DTA* mice compared to *Cspg4-cre^-/Rosa-DTA*. Bar = 100 μm in **B1, C1**; 50 μm in **A1, A2, D1, E1, F1**; 40 μmin **B2, C2, D2, E2, F2**.

https://doi.org/10.7554/eLife.09102.007

Figure 3—figure supplement 1

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Drastic depletion of GAD67-GFP⁺ neurons in *Nkx2.1^-/-, Nkx2.1-cre⁺/Rosa-DTA* and *Nkx2.1-cre⁺/Eno2-DTA* cortices.

(**A–D**) Double immunohistochemistry for the GFP and L1 on coronal CI sections in *Gad1-EGFP* (n=8) (**A1–A2**), *Nkx2.1-cre⁺/Rosa-DTA:Gad1-EGFP* (n=5) (**B1–B2**) and *Nkx2.1-cre⁺/Eno2-DTA:Gad1-EGFP* (n= 6) (**C1–C2**) mice at E18.5. **A2, B2,** and C2 are higher power views showing the quantification of the GAD67-GFP⁺ interneurons (white spots) within the CI region pointed with a white arrowhead in **A1, B1,** and C1, respectively. (D) Bars (means ± SEM; unpaired Student’s t-test) represent the percentage of remaining GAD67-GFP⁺ interneurons in the CI of E18.5 *Nkx2.1-cre⁺/Rosa-DTA:Gad1-EGFP* and *Nkx2.1-cre⁺/Eno2-DTA:Gad1-EGFP* mice compared to *Gad1-EGFP* mice. Both the mutant mice exhibited equivalent loss of neurons when compared to wild-type mice. (E) Table of the corresponding cell density values (cell number*10³/mm³) (unpaired Student’s t-test). Scale bar = 675 µm in **A1, B1,** and C1; 100 µm in **A2, B2,** and C2. CC, corpus callosum; CCi, cingulate cortex; Cl, cingulate bundle; SEP, septum.

https://doi.org/10.7554/eLife.09102.008

Figure 3—figure supplement 2

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GLAST⁺ astrocytes are not affected in *Nkx2.1-cre⁺/Rosa-DTA* cingulate cortex at E18.5.

(**A–B**) Double immunohistochemistry for Olig2 and GLAST in cingulate cortex (CCi) coronal sections of *Nkx2.1-cre^-/Rosa-DTA* mutant mice (n=4) (A) and *Nkx2.1-cre⁺/Rosa-DTA* control mice (n=4) (B) at E18.5. Scale bar = 50 µm in A and B. (C) Bars (means ± SEM; unpaired Student’s t-test) represent the density number of remaining GLAST+ astroglia in the CCi of E18.5 *Nkx2.1-cre⁺/Rosa-DTA* mice compared to control mice. No significant loss of GLAST⁺ astroglia was observed in *Nkx2.1-cre⁺/Rosa-DTA* CCi compared to control mice.

https://doi.org/10.7554/eLife.09102.009

Figure 4 with 1 supplement

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Drastic depletion of embryonic NG2⁺ glia in *Nkx2.1-cre+/Rosa-DTA* and *Cspg4-cre+/Rosa-DTA* midline dorsal telencephalon.

DAB staining for NG2 in wild-type (**A1–A3**) (n=6), *Nkx2.1-Cre⁺/Rosa-DTA* (**B1–B3**) (n=3)*, Cspg4-cre⁺/Rosa-DTA* (**C1–C3**) (n=3), and *Nkx2.1-cre⁺/Eno2-DTA* (**D1–D3**) (n=3) telencephalic coronal slices at E18.5. NG2⁺ glia were completely depleted from the corpus callosum (CC), the cingulate cortex (CCi), and cingulate bundle (Cl) of *Nkx2.1-cre⁺/Rosa-DTA* (**B1–B3**) mutant mice compared to wild-type mice (**A1–A3**). In *Cspg4-cre⁺/Rosa-DTA* mutant mice (**C1–C3**), there was also a drastic loss of NG2⁺ cells in medial cortical areas of the dorsal telencephalon with only few remaining cells. In *Nkx2.1-cre⁺/Eno2-DTA* (**D1–D3**) mutant mice, there was no loss of NG2⁺ glia in all the observed regions. **A2–A3, B2–B3, C2–C3,** and **D2–D3** are higher power views of the Cl regions in **A1, B1, C1,** and **D1,** respectively (white arrowheads). Bar = 500 μm in **A1, B1, C1,** and D1; 250 μm in **A2, B2, C2,** and D2; 125 μm in **A3, B3, C3,** and D3.

https://doi.org/10.7554/eLife.09102.010

Figure 4—figure supplement 1

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Drastic depletion of embryonic NG2⁺ glia in *Nkx2.1-cre⁺/Rosa-DTA* midline dorsal telencephalon starts at E16.5.

(**A–D**) Immunostaining for NG2 in *Nkx2.1-cre^-/Rosa-DTA* (**A1–A2** to **C1–C2**) and *Nkx2.1-cre⁺/Rosa-DTA* (**D1–D2** to **F1–F2**) telencephalic coronal slices at E14.5 (**A1–A2, D1–D2**), E16.5 (**B1–B2, E1–E2**) and E18.5 (**C1–C2, F1–F2). A2, B2, C2, D2, E2,** and D2 are higher power views of the region in **A1, B1, C1, D1, E1,** and F1, respectively (white arrowheads). At E14.5, there were no significant differences in NG2⁺ glia observed in the cingulate regionof *Nkx2.1-cre⁺/Rosa-DTA* mutant mice (n=3) compared to wild-type mice (n=3) (**A1–A2, D1–D2**). By contrast, later at E16.5, there was already a drastic loss of NG2⁺ glia populating the medio-dorsal telencephalon regionsin the *Nkx2.1-cre⁺/Rosa-DTA* (n=3) mutant mice compared to control mice (n=3) (**B1–B2, E1–E2**). This loss was maintained at E18.5 in the *Nkx2.1-cre⁺/Rosa-DTA* (n=3) mutant mice compared to control mice (n=3) (**C1–C2, F1–F2**). Scale bar = 675 µm in **A1, B1, C1, D1, E1,** and F1; 40 μmin **A2, B2, C2, D2, E2,** and F2.

https://doi.org/10.7554/eLife.09102.011

Figure 5 with 4 supplements

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Blood vessel branching is similarly impaired in *Nkx2.1Cre⁺/Rosa-DTA* and *Cspg4-cre⁺/Rosa-DTA* mice.

(**A–C**) DAB staining for PECAM and reconstitution of the vascular network using the Neurolucida tracing tool in wild-type (**A1–A3**) (n=8), *Nkx2.1-cre⁺/Rosa-DTA* (**B1–B3**) (n=4) and *Cspg4-cre⁺/Rosa-DTA* (**C1–C3**) (n=4) cortical coronal sections at E18.5. **A2, B2,** and C2 are higher magnified views of the boxed regions seen in **A1, B1,** and **C1,** respectively. (**D–F**) DAB staining for erythrocytes in wild-type (**D1–D2**) (n=10), *Nkx2.1-cre⁺/Rosa-DTA* (**E1–E2**) (n=5), and *Cspg4-cre⁺/Rosa-DTA* (**F1–F2**) (n=5) cortical coronal sections at E18.5. (G) Bars (means ± SEM) represent the percentages of vessel nodes, vessel branches and volume of the vascular network in mutants compared to wild-type (n=4; unpaired Student’s t-test). The respective absolute values per CCi section are given in Table 1. All the quantified parameters were significantly decreased in mutant mice. (H) Bars (means ± SEM) represent the percentage of erythrocyte clusters and individual erythrocytes in mutants compared to wild-type. The respective absolute values per CCi section are given in Table 2 (n=5; unpaired Student’s t-test).The number of erythrocyte clusters showed a significant increase in all mutants. The total number of individual erythrocytes remained unchanged. Bar = **A1, B1, C1:** 250 μm; **D1, E1, F1:** 125 μm; **A2, A3, B2, B3, C2, C3:** 62.5 μm; **D2, E2, F2:** 50 μm. CCi, cingulate cortex.

https://doi.org/10.7554/eLife.09102.012

Figure 5—figure supplement 1

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Embryonic *Nkx2.1*-derived NG2 glia do not control cortical blood vessels outgrowth before E16.5.

(**A–D**) DAB staining for PECAM in *Nkx2.1-cre^-/Rosa-DTA* (n=4) (**A1-A2, C1–C2**) and *Nkx2.1-cre⁺/Rosa-DTA* (n=4) (**B1–B2, D1–D2**) telencephalic coronal slices at E14.5 (**A1–A2, B1–B2**) and E16.5 (**C1–C2, D1–D2**). At E14.5, there was no significant blood vessel network defect observed in the cingulate region of *Nkx2.1-cre⁺/Rosa-DTA* (**B1–B2**) mutant mice compared to wild-type mice (**A1–A2**). By contrast, later at E16.5, the blood vessel network is less developed in the *Nkx2.1-cre⁺/Rosa-DTA* (**D1–D2**) mutant mice (**D1–D2**) compared to control mice (**C1–C2**). Scale bar = 250 µm in **A1, B1, C1,** and D1; 125 µm in **A2, B2, C2,** and D2.

https://doi.org/10.7554/eLife.09102.013

Figure 5—figure supplement 2

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Blood vessel structure is impaired in the septum of *Nkx2.1-cre⁺/Rosa-DTA* and *Cspg4-cre⁺/Rosa-DTA* mice at E18.5.

(**A–D**) DAB staining for PECAM in wild-type (n=12) (**A1–A2**), *Nkx2.1Cre⁺/Rosa-DTA* (n=4) (**B1–B2**), *Cspg4-cre⁺/Rosa-DTA* (n=4) (**C1–C2**) and *Nkx2.1-cre⁺/Eno2-DTA* (n=4) (**D1–D2**) telencephalic coronal slices at E18.5. The blood vessel network was significantly impaired in the septal region (SEP) of *Nkx2.1-cre⁺/Rosa-DTA* (**B1–B2**) and *Cspg4-cre⁺/Rosa-DTA* (**C1–C2**) mutant mice compared to wild-type mice (**A1–A2**). In *Nkx2.1-cre⁺/Eno2-DTA* (**D1–D2**) mutant mice, there was no significant blood vessel network defect observed in the septal region. Scale bar = 500 µm in **A1, B1, C1,** and D1; 125 µm in **A2, B2, C2,** and D2.

https://doi.org/10.7554/eLife.09102.014

Figure 5—figure supplement 3

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Embryonic *Nkx2.1*-derived GABAergic neurons do not control the development and function of cortical blood vessels.

DAB staining for PECAM (**A1–A2, B1–B2**) and for erythrocytes (**C1–C2, D1–D2**) in coronal slices of the dorsal telencephalon of E18.5 wild-type (n=3) (**A1–A2, C1–C2**) and *Nkx2.1-cre⁺/Eno2-DTA* (n=3) (**B1–B2, D1–D2**) mutant mice. (**A2, B2, C2,** and D2) are higher magnified views of the cingulate bundle (CI) region pointed with a black arrowhead in (**A1, B1, C1,** and D1), respectively. PECAM and erythrocyte staining revealed that the loss of GABAergic neurons in *Nkx2.1-cre⁺/Eno2-DTA* mutant mice did not cause any vascular anomalies (B2 and D2) compared to control mice (A2 and C2). Scale bar = 500 µm in **A1, B1, C1,** and D1; 125 µm in **A2, B2, C2** ,and D2. CC, corpus callosum; CCi, cingulate cortex.

https://doi.org/10.7554/eLife.09102.015

Figure 5—figure supplement 4

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No significant blood vessel network defects are observed in *Cspg4-cre⁺/Rosa-DTA* postnatal brains.

(**A–D**) DAB staining for PECAM in *Cspg4-cre^-/Rosa-DTA* (n=3) (**A1–A2**) and *Cspg4-cre⁺/Rosa-DTA* (n=3) (**B1–B2**) telencephalic coronal slices at P17-P19 (**A1–A2, B1–B2**). At all postnatal ages, there was no significant blood vessel network defect observed in the cingulate region and corpus callosum of *Cspg4-cre⁺/Rosa-DTA* (**B1–B2**) mutant mice compared to control *Cspg4-cre^-/Rosa-DTA* (**A1–A2**) mice. Scale bar = 500 μm in **A1–A2** and **B1–B2**.

https://doi.org/10.7554/eLife.09102.016

Figure 6

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Macrophages and tip cells are not affected in *Nkx2-1Cre⁺/Rosa-DTA* mice.

(**A–B**) Double immunohistochemistry for Isolectin and Ter119, to visualize erythrocytes, on 250-μm-thick coronal of corpus callosum (CC) and cingulate bundle (CI) sections in wild-type (**A1–A4**) (n=5) and *Nkx2.1-cre⁺/Rosa-DTA* (**B1–B4**) (n=5) mice at E18.5. In the CC and the CI (**B1–B2**) of *Nkx2.1-cre⁺/Rosa-DTA* mice, the blood vessels have a twisted shape and the erythrocytes are clustered (white arrowheads in B1 and B2) compared to the wild-type vessels that formed a regular network (white arrowheads in A1 and A2). In the CC and the CI of both wild-type (**A3–A4**) and *Nkx2.1-cre⁺/Rosa-DTA* mice (**B3–B4**), guidepost macrophages labeled by the isolectin (white arrowheads in **A3–A4** and **B3–B4**) are found in the close vicinity of the tip cells (white arrows in **A3–A4** and **B3–B4**). The tip cells exhibit the same morphology and the same number of filopodia with similar length in both circumstances. Bar = 60 μm in **A1, B1**; 40 μm in **A2, B2, A3, A4, B3, B4**.

https://doi.org/10.7554/eLife.09102.017

Tables

Table 1

Nodes, branches and volume of the vascular network visualized per CCi section in control and transgenic mice used to ablate Nkx2.1-derived or only NG2⁺ cells. The values (mean ± SEM) corresponding to the number of nodes, the number of branches and the volume of the vascular network per CCi section are given for: (1) Nkx2.1-cre⁺/Rosa-DTA mice and their corresponding control Nkx2.1-cre^-/Rosa-DTA mice, and (2) Cspg4-cre⁺/Rosa-DTA mice and their corresponding control Cspg4-cre^-/Rosa-DTA mice (n=4 each; unpaired Student’s t-test).

https://doi.org/10.7554/eLife.09102.018

	Values (mean ± SEM) for the vascular network per CCi section
Genotype	Nodes number	Branches number	Vascular volume * 10³ (µm³)
Nkx2.1-cre^-/Rosa-DTA	103.44 ± 7.60	241.77 ± 14.65	20302.06 ± 1290.76
Nkx2.1-cre⁺/Rosa-DTA	84.63 ± 6.65	207.75 ± 11.56	16692.46 ± 879.03
Cspg4-cre^-/Rosa-DTA	135.50 ± 8.60	299.50 ± 16.89	20361.28 ± 1429.69
Cspg4-cre⁺/Rosa-DTA	109.25 ± 6.20	250.87 ± 13	15921.88 ± 403.44

Table 2

Number of erythrocyte clusters and of individual erythrocytes visualized per CCi section in control and transgenic mice used to ablate Nkx2.1-derived or only NG2⁺ cells. The values (mean ± SEM) corresponding to the number of erythrocyte clusters and the number of individual erythrocytes per CCi are given for: (1) Nkx2.1-cre⁺/Rosa-DTA mice and their corresponding control Nkx2.1-cre^-/Rosa-DTA mice, and (2) Cspg4-cre⁺/Rosa-DTA mice and their corresponding control Cspg4-cre^-/Rosa-DTA mice (n=5 each; unpaired Student’s t-test).

https://doi.org/10.7554/eLife.09102.019

	Values (mean ± SEM) per CCi section
Genotype	Number of erythrocyte clusters per CCi section	Number of individual erythrocytes per CCi section
Nkx2.1-cre^-/Rosa-DTA	6.60 ± 0.88	302.33 ± 19.12
Nkx2.1-cre⁺/RosaDTA	13.00 ± 2.62	273.42 ± 41.44
Cspg4-cre^-/Rosa-DTA	5.83 ± 0.62	397.80 ± 18.96
Cspg4-cre⁺/Rosa-DTA	14.95 ± 1.77	447.30 ± 22.91

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Shilpi Minocha
Delphine Valloton
Isabelle Brunet
Anne Eichmann
Jean-Pierre Hornung
Cecile Lebrand

(2015)

NG2 glia are required for vessel network formation during embryonic development

eLife 4:e09102.

https://doi.org/10.7554/eLife.09102

Figures

NG2⁺ glia are in close contact with blood vessels.

NG2⁺ glia of the dorsal telencephalon are derived from Nkx2.1⁺ progenitors of the subpallium.

YFP signal in Nkx2.1-cre⁺/Rosa-EYFP and Cspg4-cre⁺/Rosa-EYFP mice is present in NG2 glia.

Nkx2.1-derived NG2 and Olig2 glia are transient and gradually disappear from the dorsal pallium at postnatal ages.

NG2⁺ glia, but not pericytes, control blood vessels formation.

Drastic depletion of GAD67-GFP⁺ neurons in Nkx2.1^-/-, Nkx2.1-cre⁺/Rosa-DTA and Nkx2.1-cre⁺/Eno2-DTA cortices.

GLAST⁺ astrocytes are not affected in Nkx2.1-cre⁺/Rosa-DTA cingulate cortex at E18.5.

Drastic depletion of embryonic NG2⁺ glia in Nkx2.1-cre+/Rosa-DTA and Cspg4-cre+/Rosa-DTA midline dorsal telencephalon.

Drastic depletion of embryonic NG2⁺ glia in Nkx2.1-cre⁺/Rosa-DTA midline dorsal telencephalon starts at E16.5.

Blood vessel branching is similarly impaired in Nkx2.1Cre⁺/Rosa-DTA and Cspg4-cre⁺/Rosa-DTA mice.

Embryonic Nkx2.1-derived NG2 glia do not control cortical blood vessels outgrowth before E16.5.

Blood vessel structure is impaired in the septum of Nkx2.1-cre⁺/Rosa-DTA and Cspg4-cre⁺/Rosa-DTA mice at E18.5.

Embryonic Nkx2.1-derived GABAergic neurons do not control the development and function of cortical blood vessels.

No significant blood vessel network defects are observed in Cspg4-cre⁺/Rosa-DTA postnatal brains.

Macrophages and tip cells are not affected in Nkx2-1Cre⁺/Rosa-DTA mice.

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NG2+ glia are in close contact with blood vessels.

NG2+ glia of the dorsal telencephalon are derived from Nkx2.1+ progenitors of the subpallium.

YFP signal in Nkx2.1-cre+/Rosa-EYFP and Cspg4-cre+/Rosa-EYFP mice is present in NG2 glia.

Nkx2.1-derived NG2 and Olig2 glia are transient and gradually disappear from the dorsal pallium at postnatal ages.

NG2+ glia, but not pericytes, control blood vessels formation.

Drastic depletion of GAD67-GFP+ neurons in Nkx2.1-/-, Nkx2.1-cre+/Rosa-DTA and Nkx2.1-cre+/Eno2-DTA cortices.

GLAST+ astrocytes are not affected in Nkx2.1-cre+/Rosa-DTA cingulate cortex at E18.5.

Drastic depletion of embryonic NG2+ glia in Nkx2.1-cre+/Rosa-DTA and Cspg4-cre+/Rosa-DTA midline dorsal telencephalon.

Drastic depletion of embryonic NG2+ glia in Nkx2.1-cre+/Rosa-DTA midline dorsal telencephalon starts at E16.5.

Blood vessel branching is similarly impaired in Nkx2.1Cre+/Rosa-DTA and Cspg4-cre+/Rosa-DTA mice.

Embryonic Nkx2.1-derived NG2 glia do not control cortical blood vessels outgrowth before E16.5.

Blood vessel structure is impaired in the septum of Nkx2.1-cre+/Rosa-DTA and Cspg4-cre+/Rosa-DTA mice at E18.5.

Embryonic Nkx2.1-derived GABAergic neurons do not control the development and function of cortical blood vessels.

No significant blood vessel network defects are observed in Cspg4-cre+/Rosa-DTA postnatal brains.

Macrophages and tip cells are not affected in Nkx2-1Cre+/Rosa-DTA mice.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

NG2⁺ glia are in close contact with blood vessels.

NG2⁺ glia of the dorsal telencephalon are derived from Nkx2.1⁺ progenitors of the subpallium.

YFP signal in Nkx2.1-cre⁺/Rosa-EYFP and Cspg4-cre⁺/Rosa-EYFP mice is present in NG2 glia.

NG2⁺ glia, but not pericytes, control blood vessels formation.

Drastic depletion of GAD67-GFP⁺ neurons in Nkx2.1^-/-, Nkx2.1-cre⁺/Rosa-DTA and Nkx2.1-cre⁺/Eno2-DTA cortices.

GLAST⁺ astrocytes are not affected in Nkx2.1-cre⁺/Rosa-DTA cingulate cortex at E18.5.

Drastic depletion of embryonic NG2⁺ glia in Nkx2.1-cre+/Rosa-DTA and Cspg4-cre+/Rosa-DTA midline dorsal telencephalon.

Drastic depletion of embryonic NG2⁺ glia in Nkx2.1-cre⁺/Rosa-DTA midline dorsal telencephalon starts at E16.5.

Blood vessel branching is similarly impaired in Nkx2.1Cre⁺/Rosa-DTA and Cspg4-cre⁺/Rosa-DTA mice.

Blood vessel structure is impaired in the septum of Nkx2.1-cre⁺/Rosa-DTA and Cspg4-cre⁺/Rosa-DTA mice at E18.5.

No significant blood vessel network defects are observed in Cspg4-cre⁺/Rosa-DTA postnatal brains.

Macrophages and tip cells are not affected in Nkx2-1Cre⁺/Rosa-DTA mice.