Coagulation and survival in Drosophila melanogaster fondue mutants

A tissue communication network coordinating innate immune response during muscle stress

“…Further supporting a shared connection between muscle maintenance and innate immunity, we recently identified fondue (fon) as essential for maintaining the extracellular matrix (ECM) of the larval muscle attachment site (MAS) (Green et al, 2016). The common suite of secreted hemolymph proteins that accumulate and function at the MAS were also found to act with Fon in its previously described role in coagulation (Bajzek et al, 2012;Lindgren et al, 2008;Scherfer et al, 2006).…”

“…The control stock used in all experiments was w 1118 . Two fon null alleles, fon Δ17 and fon Δ24 (Bajzek et al, 2012) were used to remove fon and paired with the deficiency stock, Df(2L)Exel6043. Other mutant alleles used in experiments were Brkd J29 (a gift from Troy Littleton, Massachusetts Institute of Biology, Cambridge, MA, USA), hop Tum-l (Bloomington Drosophila Stock Center, BL-8492), Mhc S1 (a gift from Troy Littleton), and Tl 3 (BL-3338).…”

A Common Suite of Coagulation Proteins Function inDrosophilaMuscle Attachment

“…The following stocks were used to drive tissue-specific expression: tubP(aTub84B.PL)-Gal4 (BL-5138), 24B-Gal4 (BL-1767), sr-Gal4 and sr-Gal4, UAS-CD8-GFP (gifts from T. Volk), ppl-Gal4 (a gift from L. Dobens), and da-Gal4 (originally BL-37291 outcrossed 10 times to w 1118 to remove background lethals). The following fondue mutations were used: the null alleles fon D24 and fon D17 are deletions that remove only fon coding sequence (Bajzek et al 2012); hypomorphic allele w 1118 ; Mi{ET1}fon MB11923 /SM6a (fon MB ; BL-29262) (Bajzek et al 2012), fon RNAi [originally from R. Ueda; described in Scherfer et al (2006)], and w 1118 ; P{UASfon.GFP}28e [(fon-GFP; BL-43646) (Lindgren et al 2008)]. Additional alleles and/or stocks analyzed are as follows: Tig A1 and Tig X (Bunch et al 1998), fon D24 A; da-Gal4 (Bajzek et al 2012), UAS-Tig RNAi (BL-31570), UAS-Tsp RNAi (VDRC; v10072s), UAS-Lsp1g RNAi (BL-55389), UAS-Gelsolin RNAi (BL-31205; BL-41704), and P{PTT-un1} vkg G00454 (Morin et al 2001).…”

“…For one candidate, this abnormal pupal phenotype was caused by a Minos insertion into the fondue (fon) locus (Mi{ET1}fon MB11923 referred to hereafter as fon MB ). Interestingly, the Dushay group had previously reported that homozygous fon mutants exhibit longer or curved pupae that failed to eclose ( Figure 1A) (Scherfer et al 2006;Bajzek et al 2012). To further characterize the pupal morphology phenotype, we measured the axial ratio (length/width) in fon alleles compared to wild-type (WT) control pupae.…”

“…To further characterize the pupal morphology phenotype, we measured the axial ratio (length/width) in fon alleles compared to wild-type (WT) control pupae. In fon MB / fon MB or deletion mutants (fon D17 or fon D24 ) that remove portions of fon coding region (Bajzek et al 2012), pupae exhibited a greater axial ratio than WT individuals ( Figure 1B). This lethality and associated pupal morphological changes seemed unlikely to result solely from the role of Fon in hemolymph coagulation, as this phenomenon occurred in unwounded individuals, and other more severe clotting mutants (e.g., hemolectin) did not exhibit this same phenotype (Lesch et al 2007).…”