Caenorhabditis elegans exoskeleton collagen COL‐19: An adult‐specific marker for collagen modification and assembly, and the analysis of organismal morphology

Abstract

The integral role that collagens play in the morphogenesis of the nematode exoskeleton or cuticle makes them a useful marker in the examination of the collagen synthesizing machinery. In this study, a green fluorescent protein-collagen fusion has been constructed by using the Caenorhabditis elegans adult-specific, hypodermally synthesized collagen COL-19. In wild-type nematodes, this collagen marker localized to the circumferential annular rings and the lateral trilaminar alae of the cuticle. Crosses carried out between a COL-19::GFP integrated strain and several morphologically mutant strains, including blister, dumpy, long, small, squat, and roller revealed significant COL-19 disruption that was predominantly strain-specific and provided a structural basis for the associated phenotypes. Disruption was most notable in the cuticle overlying the lateral seam cell syncytium, and confirmed the presence of two distinct forms of hypodermis, namely the circumferentially contracting lateral seam cells and the laterally contracting ventral-dorsal hypodermis. The effect of a single aberrant collagen being sufficient to mediate widespread collagen disruption was exemplified by the collagen mutant strain dpy-5 and its disrupted COL-19::GFP and DPY-7 collagen expression patterns. Through the disrupted pattern of COL-19 and DPY-7 in a thioredoxin mutant, dpy-11, and through RNA interference of a dual oxidase enzyme and a vesicular transport protein, we also show the efficacy of the COL-19::GFP strain as a marker for aberrant cuticle collagen synthesis and, thus, for the identification of factors involved in the construction of collagenous extracellular matrices. Developmental Dynamics 226:523-539, 2003.

Multilocus Patterns of Polymorphism and Selection Across the X Chromosome of Caenorhabditis remanei

“…and col-19 encodes a collagen protein, which is associated with the cuticle (Thein et al 2003). The function of the C. elegans orthologs of the other genes is largely unknown.…”

Ultraviolet-A triggers photoaging in model nematode Caenorhabditis elegans in a DAF-16 dependent pathway

“…3 Differential expression of clk-1 and eat-2 upon UV-A exposure in C. elegans. The expression of clk-1 was downregulated during the course of exposure whereas eat-2 was downregulated consistently 2010), and it acts like an adult specific marker (Thein et al 2003;Li and Paik 2011). Further, it deciphers the accumulation of collagen inside the organism.…”

“…Since the model host lacks a proper epidermal layer as that of humans, the preliminary aim was to confirm whether UV-A can cause any damage in the nematode. For this, transgenic strain of collagen tagged with GFP, col-19::GFP strain (TP12), was exposed to UV-A for 2, 4, and 6 h. The col-19 gene is an adult specific marker and is also essential for the normal structure of the alae in adult C. elegans (Thein et al 2003). In this strain, the col-19 gene is fused with GFP and localized in the cuticle.…”

Regulation of Fertility, Survival, and Cuticle Collagen Function by the Caenorhabditis elegans eaf-1 and ell-1 Genes

“…To test this hypothesis, we examined the cuticle structure using the TP12 transgenic worm line, which expresses the cuticle collagen col-19 gene fused to GFP. This results in COL-19-GFP being incorporated into the collagen structure during cuticle formation (40). Both ell-1 RNAi treatment and eaf-1(tm3976) dramatically decreased GFP expression in the cuticle (Fig.…”