ZFIN Figure: Bae et al., 2009, Fig. 9

Fig. 9

Presynaptic terminals of mossy and climbing fibers. (A) Immunoblotting of brain lysates with an anti-Vglut2a antibody. Rabbit anti-Vglut2a (lane 1) and pre-immune sera (lane 2) were used to detect Vglut2a (asterisk). Several bands other than 64-kD Vglut2a were detected in lane 1, but they are likely non-specific as they were also detected in lane 2. (B, C) Immunostaining of Vglut2a. Larva at 3 dpf that received 4 pg of antisense vglut2a MO (C) and control larva (B) stained with anti-Vglut2a (green) and anti-HuC/D (magenta) antibodies. Dorsal views of the hindbrain regions. Anti-Vglut2a but not anti-HuC/D staining was abolished in the vglut2a morphant hindbrain. (D–F) Co-staining of a Tg(vglut2a:EGFP) brain with anti-Vglut2a and anti-EGFP antibodies. EGFP (green, D), Vglut2a (magenta, E) and merged images (F) are shown. Sagittal section. Note that Vglut2a⁺ dots are detected in the distal tips (presynaptic terminals) of vglut2a:EGFP⁺ axons (mossy fibers) in the GCL. (G, H) Co-staining with anti-Vglut2a (magenta) and anti-Pvalb7 (green) antibodies. Low (G) and high (H) magnification views. Vglut2a⁺ synaptic boutons were detected on the soma (arrowheads) of Purkinje cells (asterisks) or abutting the proximal dendrites of Purkinje cells (arrow). (I) Co-staining of a Tg(pou4f1-hsp70l:EGFP) brain with anti-Vglut2a (magenta) and anti-EGFP (green) antibodies. Vglut2a⁺ dots were also pou4f1:EGFP⁺ in the ML (arrowheads), indicating they were presynaptic terminals of climbing fibers. The position of the PCL is indicated by a dotted line. (J) Immunoblotting of brain lysates with anti-Gad1 antibodies. Rabbit anti-Gad1 generated in this study (lane 1), control sera (pre-immune sera, lane 2), and commercially available anti-human GAD1/2 (lane 3) were used to detect zebrafish Gad1 (asterisk). (K, L) Co-staining of the Tg(vglut2a:EGFP) brain with anti-Gad1(magenta) and anti-EGFP (green) antibodies. Low (K) and high (L, in GCL) magnification views. Vglut2a⁺ and Gad1⁺ dots were adjacent in the GCL and indicated the cerebellar glomeruli. The abbreviations are as described in Fig. 1. Scale bars: 100 μm (C, F, G, I, K), 50 μm (L), and 20 μm (H).

Expression Data

Genes:	EGFP gad1b pvalb7 slc17a6b
Antibodies:	Ab1-elavl Ab1-gad1 Ab1-slc17a6
Fish:	WT WT + MO1-slc17a6b rw0110bTg zf139Tg
Knockdown Reagent:	MO1-slc17a6b
Anatomical Terms:	brain corpus cerebelli presynaptic active zone dorsal tegmental nucleus axon granular layer corpus cerebelli Golgi cell granular layer corpus cerebelli presynaptic active zone hindbrain inferior olive axon molecular layer corpus cerebelli molecular layer corpus cerebelli presynaptic active zone molecular layer corpus cerebelli stellate interneuron molecular layer valvula cerebelli optic tectum Purkinje cell dendrite Purkinje cell neuronal cell body
Stage Range:	Protruding-mouth to Adult

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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Reprinted from Developmental Biology, 330(2), Bae, Y.K., Kani, S., Shimizu, T., Tanabe, K., Nojima, H., Kimura, Y., Higashijima, S.I., and Hibi, M., Anatomy of zebrafish cerebellum and screen for mutations affecting its development, 406-426, Copyright (2009) with permission from Elsevier. Full text @ Dev. Biol.