Experimental Dissection of Metalloproteinase Inhibition-Mediated and Toxic Effects of Phenanthroline on Zebrafish Development

Abstract

Metalloproteinases are zinc-dependent endopeptidases that function as primary effectors of tissue remodeling, cell-signaling, and many other roles. Their regulation is ferociously complex, and is exquisitely sensitive to their molecular milieu, making in vivo studies challenging. Phenanthroline (PhN) is an inexpensive, broad-spectrum inhibitor of metalloproteinases that functions by chelating the catalytic zinc ion, however its use in vivo has been limited due to suspected off-target effects. PhN is very similar in structure to phenanthrene (PhE), a well-studied poly aromatic hydrocarbon (PAH) known to cause toxicity in aquatic animals by activating the aryl hydrocarbon receptor (AhR). We show that zebrafish are more sensitive to PhN than PhE, and that PhN causes a superset of the effects caused by PhE. Morpholino knock-down of the AhR rescues the effects of PhN that are shared with PhE, suggesting these are due to PAH toxicity. The effects of PhN that are not shared with PhE (specifically disruption of neural crest development and angiogenesis) involve processes known to depend on metalloproteinase activity. Furthermore these PhN-specific effects are not rescued by AhR knock-down, suggesting that these are bona fide effects of metalloproteinase inhibition, and that PhN can be used as a broad spectrum metalloproteinase inhibitor for studies with zebrafish in vivo.

Genome editing reveals reproductive and developmental dependencies on specific types of vitellogenin in zebrafish (Danio rerio)

“…1). These abnormailites have been associated with a broad variety of conditions including, as examples, rearing systems for Eurasian perch, Perca fluviatilis (Alix et al, 2017), larval rearing temperatures for Atlantic halibut, Hippoglossus hippoglossus L. (Ottesen and Bolla, 1998), embryo cryopreservation practices for streaked prochilod, Prochilodus lineatus (Costa et al, 2017), and, in zebrafish, phenanthroline toxicity (Ellis and Crawford, 2016), influenza A virus infection (Gabor et al, 2014), knockdown or KO of genes related to kidney function or development, respectively (Hanke et al, 2013; Zhang et al, 2018), knockdown of the wwox tumor suppressor gene (Tsuruwaka et al, 2015), deletion of a gene ( pr130 ) encoding a protein essential for myocardium formation and cardiac contractile function (Yang et al, 2016), and mutagenesis of genes involved in thyroid morphogenesis and function (Trubiroha et al, 2018), among others. The edemas may ultimately result from many different proximal causes such as cardiac, kidney, liver or osmoregulatory failure, and researchers are just beginning to develop screens to differentiate between them (Hanke et al, 2013).…”

Mononuclear gold(III) complexes with phenanthroline ligands as efficient inhibitors of angiogenesis: A comparative study with auranofin and sunitinib

“…However, the previous studies revealed that 1,10-phen and the corresponding gold(III) complex, [Au(1,10phen)Cl 2 ]Cl, had no selective activity against tumor cell lines in comparison to normal human and mammalian cell lines [22,24,26,27], whereas 1,10-phen exerted equal to or even higher cytotoxicity on cancer cell lines than the corresponding platinum(II) and gold(III) complexes [25,26]. Recently, Ellis and Crawford [20] found that 1,10-phen possesses ability to inhibit blood vessels development in zebrafish embryos, eliciting the toxic response in embryos upon an effective anti-angiogenic concentrations. All these biological properties limit the future application of 1,10-phen as a ligand for design and synthesis of metal-based anti-angiogenic drugs.…”

“…An important class of ligands for the synthesis of gold(III) complexes comprises aromatic nitrogen-containing heterocycles (N-heterocycles), since they represent structural moieties of many natural products and biologically active compounds possessing diverse pharmacological properties, such as antitumor, antimicrobial, anticonvulsant, antiinflammatory and anti-angiogenic [17][18][19][20]. Among these compounds, 1,10-phenanthroline…”

“…Based on the discovery of anti-angiogenic property of 1,10-phen [20], and prompted by findings that phenanthrolines with differentially positioned nitrogen atoms, such as 1,7phenanthroline (1,7-phen) and 4,7-phenanthroline (4,7-phen), are significantly less toxic, mutagenic and genotoxic than 1,10-phen [35,36], but nevertheless have not been used for the synthesis of gold(III) complexes, we decided to synthesize two new gold (III) complexes, [AuCl 3 (1,7-phen-κN7)] (1) and [AuCl 3 (4,7-phen-κN4)] (2). These complexes were characterized by NMR ( 1 H and 13 C), IR and UV-vis spectroscopic and single-crystal X-ray diffraction analysis.…”

The developmental and genetic trajectory of coloration in the guppy (Poecilia reticulata)

“…Genes known to be involved in coloration were extracted from the list of differentially expressed genes ( Supplementary Table S2) (Braasch et al, 2009;Ellis & Crawford, 2016Eom et al, 2012Svetic et al, 2007;Walsh, Dale, McGraw, Pointer, & Mundy, 2012). Attention was paid to genes that were involved in production of black melanin or orange/ yellow pteridine pigments.…”