FIGURE

Fig. 1

ID

ZDB-FIG-180712-80

Publication

Kelu et al., 2018 - TPC2-mediated Ca²⁺ signaling is required for the establishment of synchronized activity in developing zebrafish primary motor neurons.

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Fig. 1

Effect of TPC2-knockdown, heterozygous-knockout, and mRNA rescue on the spontaneous Ca²⁺ activity of the CaPs in SAIGFF213A;UAS:GCaMP7a embryos at ~24 hpf. SAIGFF213A;UAS:GCaMP7a embryos were injected with: (A) Standard control-MO; (B) p53-MO; (C) TPCN2-MO-T with p53-MO; or (D) TPCN2-MO-T with p53-MO and the mutant tpcn2-mRNA. In addition, (E) untreated SAIGFF213A;UAS:GCaMP7a fish (termed tpcn2^+/+ controls) were imaged and (F) they were crossed with homozygous tpcn2^dhkz1a mutants to generate double-transgenic heterozygous mutants (tpcn2^+/-). (Ai-Fiv) Time-lapse fluorescence images showing the changes in GCaMP7a fluorescence in the CaPs at different time intervals in the various treatment groups. The embryos are in a dorsal orientation and regions of interest (ROIs) on two (of the three) selected CaP cell bodies on the left (L) and right (R) sides of the spinal cord are shown. The arrowheads indicate GCaMP7a signals in the CaPs. Ant. and Pos. are anterior and posterior, respectively. Scale bar, 50 µm. (Av-Fv) Line graphs showing the ΔF/F₀ against time (over a period of ~ 300 s) in the ROIs of the representative embryos shown in (Ai-Fiv), respectively. The time period that corresponds to the Ca²⁺ signaling events shown in the time-lapse fluorescence images (Ai-Fiv) is denoted by a grey vertical bar in (Av-Fv). Also see Movies S1-S3, which illustrate the distinct right/left alternation of Ca²⁺ transients in the controls and how this is disrupted when TPC2-mediated Ca²⁺ release was inhibited.

Expression Data

Gene:	GCaMP
Fish:	nksaigff213aGt; zf415Tg nksaigff213aGt; zf415Tg + MO1-tpcn2 + MO4-tp53 nksaigff213aGt; zf415Tg + MO4-tp53 tpcn2^{dhkz1a/dhkz1a}; nksaigff213aGt; zf415Tg
Knockdown Reagents:	MO1-tpcn2 MO4-tp53
Anatomical Term:	CaP motoneuron
Stage:	Prim-5

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	nksaigff213aGt; zf415Tg + MO1-tpcn2 + MO4-tp53 tpcn2^{dhkz1a/dhkz1a}; nksaigff213aGt; zf415Tg
Knockdown Reagents:	MO1-tpcn2 MO4-tp53
Observed In:	CaP motoneuron release of sequestered calcium ion into cytosol
Stage:	Prim-5

Phenotype Detail

Acknowledgments

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Reprinted from Developmental Biology, 438(1), Kelu, J.J., Webb, S.E., Galione, A., Miller, A.L., TPC2-mediated Ca²⁺ signaling is required for the establishment of synchronized activity in developing zebrafish primary motor neurons., 57-68, Copyright (2018) with permission from Elsevier. Full text @ Dev. Biol.