Chapman et al., 2019 - A conserved CCM complex promotes apoptosis non-autonomously by regulating zinc homeostasis. Nature communications   10:1791 Full text @ Nat. Commun.

Fig. 1

KRI-1 regulates MPK-1/ERK1 in the germline. a Di-phosphorylated MPK-1 in the germlines of wild type, kri-1(ok1251), and kri-1(ok1251); mpk-2(ok219) animals is measured along the midline from the distal to proximal end. The red box on each graph corresponds to the pachytene region. Images are representative of at least 30 worms per strain, per condition (×400 magnification). Scale bar is 25 μm. b IR-induced germline apoptosis scored in wild type, lip-1(zh15), kri-1(ok1251), and kri-1(ok1251); lip-1(zh15) animals (n ≥ 60). Red line is mean ± standard deviation. *P < 0.05, two-sided, unpaired t-test. c Schematic of intestinal KRI-1 regulating MPK-1/ERK1 in the germline

Fig. 3

MPK-2/ERK5 and KLF-3 regulate IR-induced apoptosis downstream of KRI-1. a IR-induced germline apoptosis scored after knock-down of the ERK-5/MAPK pathway genes, mekk-3, mek-5, and mpk-2 in wild type, kri-1(ok1251) and daf-2(e1370) animals (n ≥ 60). b IR-induced germline apoptosis scored in wild type, kri-1(ok1251), and kri-1(ok1251); klf-3(on34) animals (n ≥ 45). c Expression of Pmpk-2::gfp. Images are representative of three independent lines (×100 magnification). Scale bar is 50 μm. d IR-induced germline apoptosis scored in wild type, Pmpk-2::mpk-2 “+array”, and Pmpk-2::mpk-2 siblings that have lost the transgene “−array” (n ≥ 30). e IR-induced germline apoptosis scored in wild type, Pelt-2::mpk-2 “+array”, and Pelt-2::mpk-2 siblings that have lost the transgene “−array” (n ≥ 30). ae Red line is mean ± standard deviation. *P < 0.05, two-sided, unpaired t-test

Fig. 5

ZIPT-2.3 promotes zinc storage and IR-induced apoptosis. a, b ZIPT-2.3::GFP expression in wild type animals. Images are representative of three independent lines (a is at ×100 magnification and b is at ×630 magnification). c ZIPT-2.3::GFP in wild type and kri-1(ok1251) animals is measured along the intestinal midline from head to tail. Images are representative of three independent replicates (n ≥ 15). Scale bar is 50 μm. d Stored intestinal zinc measured in the intestines of wild type, kri-1(ok1251), kri-1(ok1251); mpk-2(ok219), kri-1(ok1251); klf-3(on34), zipt-2.3(ok2094), and pgp-2(kx48) animals (n ≥ 7). Scale bar is 25 μm. e Body cavity zinc measured in wild type, kri-1(ok1251), kri-1(ok1251); mpk-2(ok219), kri-1(ok1251); klf-3(on34), ttm-1(ok3505), and cdf-1(n2527) animals (n ≥ 25). Scale bar is 25 μm. f IR-induced germline apoptosis scored in wild type, kri-1(ok1251), kri-1(ok1251); mpk-2(ok219), kri-1(ok1251); let-60(ga89) and kri-1(ok1251); lip-1(zh15) animals after knock-down of zipt-2.3 (n ≥ 55). g IR-induced germline apoptosis scored in wild type, kri-1(ok1251), klf-3(on34), and kri-1(ok1251); klf-3(on34) animals after knock-down of zipt-2.3 (n ≥ 40). h IR-induced germline apoptosis scored in wild type, let-60(ga89), lip-1(zh15), and ced-9(n1653) animals grown on media supplemented with 1 mM zinc (n ≥ 35). dh Red line is mean ± standard deviation. *P < 0.05, two-sided, unpaired t-test

Fig. 6

Zinc localization and SLC39 zinc transporter expression in CCM models. a Zinypr-1 detects zinc in the posterior cardinal vein (PCV) and caudal vein (CV) vasculature of wild type zebrafish 4 days post fertilization (dpf). b Zinc localization in the PCV and CV is lost in the absence of krit1/ccm1 4 dpf. The endothelial cell vasculature is marked in red, and zinc appears green (yellow overlay). cSLC39 zinc transporter expression in CCM lesion tissue from patients (human) and in brain microvasculature endothelial tissue (BMEC) of endothelial-specific knockout mice (BMEC Ccm1)

Acknowledgments:
ZFIN wishes to thank the journal Nature communications for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Nat. Commun.