Organ Length Control by an ADAMTS Extracellular Protease inCaenorhabditis elegans

Abstract

MIG-17, a secreted protease of the ADAMTS family, acts in the directed migration of gonadal distal tip cells (DTCs) through regulation of the gonadal basement membrane in Caenorhabditis elegans. Here, we show that MIG-17 is also required for the control of pharynx elongation during animal growth. We found that the pharynx was elongated in mig-17 mutants compared with wild type. MIG-17 localized to the pharyngeal basement membrane as well as to the gonadal basement membrane. The number of nuclei in the pharynx, and the pumping rate of the pharynx, were not affected in mig-17 mutants, suggesting that cells constituting the pharynx are elongated, although the pharynx functions normally in these mutants. In contrast to the control of DTC migration, MIG-18, a secreted cofactor of MIG-17, was not essential for pharynx length regulation. In addition, the downstream pathways of MIG-17 involving LET-2/type IV collagen, FBL-1/fibulin-1, and NID-1/nidogen, partly diverged from those in gonad development. These results indicate that basement membrane remodeling is important for organ length regulation, and suggest that MIG-17/ADAMTS functions in similar but distinct molecular machineries in pharyngeal and gonadal basement membranes.

The Role of Tissue Inhibitors of Metalloproteinases in Organ Development and Regulation of ADAMTS Family Metalloproteinases in Caenorhabditis elegans

“…(Guo et al 1991;Sibley et al 1993). Consistent with previous studies, the type IV collagen a1 chain construct EMB-9::mCherry, when expressed from the integrated transgene tkTi1[emb-9::mCherry], localized to the gonadal basement membrane in WT C. elegans (Figure 4, A and B) (Ihara et al 2011;Shibata et al 2016). Conversely, localization of EMB-9::mCherry to the gonadal basement membrane was disrupted in timp-1(tk71) (Figure 4, C and D).…”

“…The following alleles were used for mutant construction: mig-17(k174), mig-17(k113) (Nishiwaki 1999), gon-1(q518), gon-1(e1254) , unc-42(e270), unc-119(ed3), unc-119(e2498), cri-2(gk314) (obtained from the C. elegans Gene Knockout Consortium), timp-1(tk71) (this work), nT1[qIs51], tkTi1[emb-9p::emb-9::mCherry + Cbrunc-119(+)] (Shibata et al 2016), oxIs322[myo-2p::-mCherry::H2B + myo-3p::mCherry::H2B + Cbr-unc-119(+)] (Frøkjaer-Jensen et al 2014), Ex[gon-1p::gon-1 + sur-5;;GFP + punc-119(+)], Ex[mig-17p::mig-17::Venus + punc-119(+)], bkcEx1[timp-1p::timp-1::Venus + punc-119(+)] (this work), and bkcEx2[cri-2p::cri-2::Venus + punc-119(+)] (this work). k174 and q518 are the null alleles of mig-17 and gon-1, respectively.…”

“…Gene inactivation experiments have indicated that ADAMTS family members are required for development of the ovaries, palate, and limbs in mice (Shindo et al 2000;Shozu et al 2005;Brown et al 2006;McCulloch et al 2009;Enomoto et al 2010). These proteases are also required for cell migration, gonadal morphogenesis, proper gonad function, regulation of pharynx length, and synapse formation in Caenorhabditis elegans and detachment of cells from the apical ECM of salivary glands in Drosophila melanogaster Blelloch and Kimble 1999;Nishiwaki et al 2000;Ismat et al 2013;Kurshan et al 2014;Qin et al 2014;Shibata et al 2016). Despite the apparent importance of ADAMTSs in the regulation of ECM remodeling, how their activities are regulated in vivo is not well understood.…”

Toolkits for detailed and high-throughput interrogation of synapses inC. elegans

“… (A) Overview of pharynx and total body length elongation between L1 and adult stages based on previous findings (Witvliet et al, 2021; Shibata et al, 2016). (B-F) Representative images and quantification of number of presynaptic specializations at early larval and adult stages visualized using (B) GFP::RAB-3 in MC ( otEx7505 ), (C) GFP::CLA-1 in I2 ( otEx7497) , (D) GFP::RAB-3 in M3 ( otIs602 ), (E) GFP::RAB-3 in M4 ( otIs597) , and (F) GFP::CLA-1 in NSM ( otEx7499 ) neurons.…”

Mitochondrial dysfunction causes Ca2+overload and ECM degradation–mediated muscle damage inC. elegans

“…Because of the wavy myofilament phenotype and previous reports highlighting the significance of muscle ECM (6), we considered whether a change in muscle attachment with the ECM or the composition of the ECM itself might be responsible for loss of myofilament anchoring and, therefore, wavy sarcomere structure. Collagen is the major ECM component, and we first analyzed changes in muscular collagen IV-α-1 (EMB-9 in C. elegans ) after Antimycin A treatment using the NF3620 strain ( emb-9::mCherry ) (16). A progressive decrease in EMB-9::mCherry fusion protein was observed with increasing concentration of Antimycin A ( Fig.…”