FACIT collagen (1alpha-chain) is expressed by hemocytes and epidermis during the inflammatory response of the ascidian Ciona intestinalis.
Based on previous cloning and sequencing study, real-time PCR and in situ hybridization assays of the inflamed body wall of LPS-injected Ciona intestinalis showed the enhanced gene expression of a collagen with FACIT structural features (Ci-type IX-Col 1alpha-chain). By using specific antibodies raised against an opportunely chosen Ci-type IX-Col synthetic peptide, the fibroblast property of hemocytes challenged in vitro with LPS (at 4h) was displayed by flow cytometry, while immunocytochemistry identified hemocytes with large granules (morula cells) as collagen-producing cells. Hemocyte lysate supernatant analyzed in immunoblotting contained a 60 kDa band identifiable as 1alpha-chain-Ci-type IX-Col. Observations of body wall sections (immunohistochemistry method) supported the role of hemocytes and showed that epidermis expressed Ci-type IX-Col 1alpha-chain in the time course of the inflammatory reaction (within 24h). Transcript and protein were mainly found in the epidermis that outlined the proximal side of the tunic matrix (at 24h after LPS injection), in cells associated with the epidermis at 4 and 192 h. In conclusion, the C. intestinalis inflammatory response to LPS challenge appeared to be composed of a complex reaction set, and for the first time we showed in ascidians a granulation tissue with FACIT-collagen production that could participate in inflammation and wound healing. Like in vertebrates, C. intestinalis acute inflammatory reactions result in a regulated pattern of tissue repair with collagen expression during remodelling. Ci-type IX-Col could be involved in a network of non-fibril-forming collagens that participates in the organization of extracellular matrix and defense responses.
Department of Animal Biology, University of Palermo, Palermo, Italy.
Aiti
A
Vizzini
Margherita
M
Pergolizzi
Mirella
M
Vazzana
Giuseppina
G
Salerno
Caterina
C
Di Sano
Pasquale
P
Macaluso
Vincenzo
V
Arizza
Daniela
D
Parrinello
Matteo
M
Cammarata
Nicolò
N
Parrinello
D000818:Animals; D002938:Ciona intestinalis; D024081:Collagen Type IX / Q000096:biosynthesis* / Q000276:immunology* / Q000378:metabolism; D000078404:Epidermal Cells; D004817:Epidermis / Q000276:immunology* / Q000378:metabolism; D004926:Escherichia coli; D005109:Extracellular Matrix / Q000378:metabolism; D020869:Gene Expression Profiling; D006434:Hemocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D007249:Inflammation; D008070:Lipopolysaccharides / Q000494:pharmacology; D019899:Paracrine Communication; D011347:Procollagen / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects
D024081:Collagen Type IX; D008070:Lipopolysaccharides; D011347:Procollagen