Article Figures & Data
Figures
- Figure 1.
Repo-positive glial cells in the adult central brain. A , Glial nuclei labeled by anti-Repo antibody (green), neuronal nuclei labeled by anti-Elav (blue), and neuropil labeled by nc82 (magenta). B , C , Glial membrane labeled with mCD8::GFP driven by repo-GAL4 on brain surface ( B ) and inner brain ( C ). Arrow and arrowhead show glial membrane forming borders among substructures of neuropil and brain cortex, respectively. D–F , Glial nuclei (green) on brain surface ( D ), cortex ( E ), and neuropil ( F ). Glial nuclei (arrows in D and E ), neuronal nuclei, and neuropil were labeled as shown ( A ). G–I , Single cells of small surface glia ( G ), large surface glia ( H ), and cortex glia ( I ) labeled by FLP-out combined with repo-GAL4. J–M , Single cells of neuropil glia labeled by FLP-out combined with repo-GAL4. A dendritic and fibrous lamellar morphology of neuropil glia was labeled around neuropil ( J ). Two different types of neuropil glia arranged nearby (square in J ) are shown at a high magnification ( K ). L , M , High-magnification images of a dendritic ( L ) and fibrous lamellar morphology ( M ) of neuropil glia. Scale bars: A–J , 50 μm; K–M , 25 μm.
- Figure 2.
Surface and cortex glia of adult brain. A–F , Perineurial glia ( A–C , green) and subperineurial glia ( D–F , green) labeled with UAS-GFP ( A , C , D , F ) and UAS-FRT-mCD8::GFP (FLP-out) ( B , E ), driven by NP6293 ( A–C ) and NP2276 ( D–F ), respectively. Glial nuclei and neuronal nuclei were labeled with anti-Repo (magenta) and anti-Elav (blue), respectively. Note that GAL4-negative, Repo-positive surface nuclei (arrowheads, C and F ) were labeled under and over the layer of perineurial glia and subperineurial glia, respectively (arrows in C and F ). G–I , Cortex glial cells (green) labeled with UAS-GFP driven by NP2222 ( G , H ) and with FLP-out combined with NP2222 ( I ). Neuropils were labeled with nc82 (magenta in G ). Glial nuclei (magenta) and neuronal nuclei (blue) were labeled with anti-Repo and anti-Elav, respectively ( H , I ). Arrows show nuclei of cortex glia. Scale bars, 50 μm.
- Figure 3.
Two subtypes of neuropil glia of adult central brain. Ensheathing ( A–D , I , K ) and astrocyte-like ( E–H , J , L ) glia labeled with NP1243 and NP6520, respectively. Entire and single cells of neuropil glial subtype were labeled with UAS-GFP ( A , E , and I–L ), UAS-mCD8::GFP ( B , F ), and UAS-FLP-out ( C , D , G , H ), respectively. Arrows show nuclei of each subtype of glial cells labeled with anti-Repo (magenta in C , G , and I–L ). Neuronal nuclei (blue) were labeled with anti-Elav ( B , F , K , L ) and neuropil (magenta) was labeled with nc82 ( D , H ). Scale bars, 50 μm.
- Figure 4.
Systematic MARCM analysis of glial proliferation during postembryonic development. A–L , Perineurial ( A–F ) and neuropil ( G–L ) glial cells labeled by heat-shock-induced mitotic recombination in mid-first-instar larvae ( A , B ), newly hatched larvae ( G , H ), early third-instar larvae ( C , D , I , J ) and early pupae ( E , F , K , L ). Scale bar, 50 μm. M–R , Quantification of glial cells that were labeled by MARCM system with heat-shock-induced mitotic recombination at different developmental stages (x-axis). Average number ( M , O ) and size ( N , P ) of perineurial glia ( M , N ) and neuropil glia ( O , P ) clusters that were induced at different developmental stage (y-axis). Q , Average number of large perineurial glia (>10 cells) and neuropil glia (>8) clusters per brain. R , Average number of single cell clones of perineurial and neuropil glia. Same samples were used for analysis in M – R . Numbers of examined brains are shown in M . Genotypes: hs-FLP/repo-GAL4; FRTG13, UAS-mCD8::GFP/FRTG13, tubP-GAL80, repo-GAL80 ( A–L ) and hs-FLP/repo-GAL4; FRTG13, UAS-nlsGFP/FRTG13, tubP-GAL80, repo-GAL80 ( M–R ).
- Figure 5.
Precursors of perineurial and neuropil glia. Perineurial glia cluster in a wandering larva ( A , B ) and adult ( C ), which were labeled by MARCM system with heat-shock-induced mitotic recombination in mid-first-instar larvae. Arrows show filopodial processes extending from larval perineurial glia. Multiple neuropil glia clusters in adults ( D–F ) and wandering larvae ( G–L ), which were labeled by MARCM with long heat-shock (40 min) induced mitotic recombination in NHL. Note that labeled cells were localized in posterior-medial ( G , J ), lateral ( H , K ), and dorsal ( I , L ) interface between brain cortex and neuropil. J–L , High-magnification images of G–I , respectively. Glial nuclei were labeled with anti-Repo (magenta, A and G–I ). Scale bars, 50 μm.
- Figure 6.
Function of gcm on postembryonic development of perineurial and neuropil glia. A , B , Cells labeled with GFP driven by gcm-GAL4 in a wandering larva. C , D , Cells labeled by MARCM system with gcm-GAL4. Mitotic recombination was induced in NHL. Glial nuclei were labeled with anti-Repo antibody (magenta, A–D ). High-magnification images of A and C are shown in B and D , respectively. Note that gcm-positive cells were labeled with anti-Repo antibody ( B , D ). E–H , Wild-type ( E , G ) and gcm ΔP1 ( F , H ) MARCM clones of neuropil glia ( E , F ) induced at NHL and perineurial glia ( G , H ) induced at mid-first-instar larvae, respectively. I , J , Quantification of neuropil glia ( I ) and perineurial glia ( J ) that were labeled by MARCM system. Average number of neuropil glia per brain ( I ) and average number of large perineurial glia cluster (>10) per brain were examined in wt, gcm ΔP1, and Df(2L)132 clones. Scale bars, 50 μm. Genotypes: gcm-GAL4/UAS-GFP ( A , B ); hs-FLP/+; gcm-GAL4, FRTG13, UAS-nlsGFP/FRTG13, tubP-GAL80, repo-GAL80 ( C , D ); hs-FLP, UAS-mCD8::GFP/repo-GAL4; FRT40A, tubP-GAL80/FRT40A or FRT40A, gcm ΔP1 ( E–H ); and hs-FLP/repo-GAL4; FRT40A, tubP-GAL80/FRT40A or FRT40A, gcm ΔP1 or FRT40A, Df(2L)132; UAS-nlsGFP/+ ( I , J ).
- Figure 7.
Neuropil glial clones. Ensheathing glial clones ( A , C , E , F ) and astrocyte-like glial clones ( B , D , G ) labeled by standard MARCM ( A–D ) and dual-expression control MARCM ( E–H ). Note that C and D show glial clones located in the antennal lobe. Whereas the ensheathing glial clone was labeled by both rCD2::GFP driven by repo-LexA::GAD (green, E ) and mCD8::RFP driven by NP6520 (magenta, F ), the astrocyte-like glial cone was labeled by only rCD2::GFP driven by repo-LexA::GAD (green, G ), but not by mCD8::RFP driven by NP6520 (magenta, H ). Scale bars, 50 μm. Genotypes: hs-FLP/repo-GAL4; FRTG13, UAS-mCD8::GFP/FRTG13, tubP-GAL80, repo-GAL80 ( A–D ) and hs-FLP/repo-LexA::GAD; FRTG13/FRTG13, tubP-GAL80, repo-GAL80; NP6520/UAS-mCD8::RFP, LexA-operator-rCD2::RFP ( E–H ).
- Figure 8.
Schematic model of postembryonic glial proliferation. A , B , Perineurial glia ( A ) and two subtypes of neuropil glia ( B ).
