ZFIN ID: ZDB-FIG-160901-4
Casano et al., 2016 - Developmental Apoptosis Mediates Entry and Positioning of Microglia in the Zebrafish Brain. Cell Reports   16(4):897-906 Full text @ Cell Rep.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Conditions:
Knockdown Reagent:
Anatomical Terms:
Stage Range: Protruding-mouth to Day 4
PHENOTYPE:
Fish:
Conditions:
Knockdown Reagent:
Observed In:
Stage Range: Protruding-mouth to Day 4

Fig. 4

Nucleotide Release Downstream of Neuronal Cell Death Recruits Microglial Precursors into the Brain

(A-E) AO staining (A-D) and quantification (E) of apoptotic nuclei in 4-dpf pU.1-morphant embryos, 24 hr after treatment with DMSO (A), suramin (B), CBX (C), and pannexin1a-MO (D), are shown.

(F-J) Microglial distribution (F-I) and quantification (J) in pU.1::GAL4-UAS::TagRFP embryos, 24 hr after treatment with DMSO (F), suramin (G), CBX (H), and pannexin1a-MO (I), are shown.

(K-N) Representative hindbrain (K-M) and quantification (N) of an mpeg1::GAL4-UAS::Kaede;HB::NTR-mCherry embryo at 4 dpf, 24 hr after treatment with MTZ + suramin (K), MTZ + CBX (L), and MTZ + pannexin1a-MO (M). Dotted lines mark the hindbrain.

(O-Q) Dorsal views of mpeg1::GAL4-UAS::Kaede embryos 24 hr after photo-conversion of microglia and treatment with DMSO (O), suramin (P), and CBX (Q). Newly entered microglia are in green. Microglia derived from converted cells are in red/yellow.

(R) Quantification of microglia deriving from cell invasion and in loco proliferation 24 hr after photo-conversion and drug treatment is shown.

(S) Quantification of mitotic events in the optic tectum of 2.5-dpf mpeg1::GAL4-UAS::Kaede embryos during <15 hr of DMSO, suramin, or CBX treatment is shown.

(T) Number of slc7a7+ microglial precursors accumulating in the anterior head region and in the optic tectum of slc7a7::Kaede embryos, 24 hr after drug treatment (graph on the left) and drug washout (graph on the right). Blue-colored areas in the schematic indicate the regions of interest.

For all quantifications, data are pooled from three independent experiments. Dotted line delimits the areas of interest. n, number of analyzed embryos; NS, non-significant; *p < 0.05, **p < 0.01, and ***p < 0.001. Scale bar represents 30 ┬Ám. See also Figures S3 and S4.

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
Kaede gl24Tg; hdb7Tg; s1999tTg chemical ablation: neuron, chemical treatment by environment: metronidazole, chemical treatment: CARBENOXOLONE Day 4 hindbrain microglial cell IFL
gl24Tg; hdb7Tg; s1999tTg chemical ablation: neuron, chemical treatment by environment: metronidazole, chemical treatment: suramin Day 4 hindbrain microglial cell IFL
gl24Tg; hdb7Tg; s1999tTg + MO2-panx1a chemical ablation: neuron, chemical treatment by environment: metronidazole Day 4 hindbrain microglial cell IFL
gl24Tg; s1999tTg control Protruding-mouth optic tectum microglial cell IFL
gl24Tg; s1999tTg chemical treatment: CARBENOXOLONE Protruding-mouth optic tectum microglial cell IFL
gl24Tg; s1999tTg chemical treatment: suramin Protruding-mouth optic tectum microglial cell IFL
mCherry gl24Tg; hdb7Tg; s1999tTg chemical ablation: neuron, chemical treatment by environment: metronidazole, chemical treatment: CARBENOXOLONE Day 4 hindbrain neuron IFL
gl24Tg; hdb7Tg; s1999tTg chemical ablation: neuron, chemical treatment by environment: metronidazole, chemical treatment: suramin Day 4 hindbrain neuron IFL
gl24Tg; hdb7Tg; s1999tTg + MO2-panx1a chemical ablation: neuron, chemical treatment by environment: metronidazole Day 4 hindbrain neuron IFL
TagRFP hdb2Tg control Protruding-mouth microglial cell IFL
hdb2Tg chemical treatment: CARBENOXOLONE Protruding-mouth microglial cell IFL
hdb2Tg chemical treatment: suramin Protruding-mouth microglial cell IFL
hdb2Tg + MO2-panx1a control Protruding-mouth microglial cell IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
gl24Tg; hdb7Tg; s1999tTg chemical ablation: neuron, chemical treatment by environment: metronidazole Day 4 hindbrain microglial cell increased amount, abnormal
Day 4 hindbrain neuron decreased amount, abnormal
gl24Tg; hdb7Tg; s1999tTg chemical ablation: neuron, chemical treatment by environment: metronidazole, chemical treatment: CARBENOXOLONE Day 4 hindbrain microglial cell amount, ameliorated
Day 4 hindbrain neuron decreased amount, abnormal
gl24Tg; hdb7Tg; s1999tTg chemical ablation: neuron, chemical treatment by environment: metronidazole, chemical treatment: suramin Day 4 hindbrain microglial cell amount, ameliorated
Day 4 hindbrain neuron decreased amount, abnormal
gl24Tg; hdb7Tg; s1999tTg + MO2-panx1a chemical ablation: neuron, chemical treatment by environment: metronidazole Day 4 hindbrain microglial cell amount, ameliorated
Day 4 hindbrain neuron decreased amount, abnormal
gl24Tg; s1999tTg chemical treatment: CARBENOXOLONE Protruding-mouth brain macrophage chemotaxis decreased occurrence, abnormal
gl24Tg; s1999tTg chemical treatment: suramin Protruding-mouth brain macrophage chemotaxis decreased occurrence, abnormal
hdb2Tg chemical treatment: CARBENOXOLONE Protruding-mouth brain microglial cell decreased amount, abnormal
hdb2Tg chemical treatment: suramin Protruding-mouth brain microglial cell decreased amount, abnormal
hdb2Tg + MO2-panx1a control Protruding-mouth brain microglial cell decreased amount, abnormal
hdb4Tg chemical treatment: CARBENOXOLONE Protruding-mouth microglial cell accumulation optic tectum anterior region, abnormal
Protruding-mouth optic tectum microglial cell decreased amount, abnormal
hdb4Tg chemical treatment: suramin Protruding-mouth microglial cell accumulation optic tectum anterior region, abnormal
Protruding-mouth optic tectum microglial cell decreased amount, abnormal
Acknowledgments:
This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Cell Rep.