Comparative proteomics analysis of teleost intermuscular bones and ribs provides insight into their development.
Intermuscular bones (IBs) and ribs both are a part of skeletal system in teleosts, but with different developing process. The chemical composition of fish IBs and ribs as well as the underlying mechanism about their development have not been investigated. In the present study, histological structures showed that one bone cavity containing osteoclasts were existed in ribs, but not in IBs of Megalobrama amblycephala. We constructed the first proteomics map for fish bones including IBs and ribs, and identified the differentially expressed proteins between IBs and ribs through iTRAQ LC-MS/MS proteomic analysis.



The proteins extracted from IBs and ribs at 1- to 2-year old M. amblycephala were quantified 2,342 proteins, with 1,451 proteins annotated with GO annotation in biological processes, molecular function and cellular component. A number of bone related proteins as well as pathways were identified in the study. A total of 93 and 154 differently expressed proteins were identified in comparison groups of 1-IB-vs-1-Rib and 2-IB-vs-2-Rib, which indicated the obvious differences of chemical composition between these two bone tissues. The two proteins (vitronectin b precursor and matrix metalloproteinase-2) related to osteoclasts differentiation were significantly up-regulated in ribs compared with IBs (P < 0.05), which was in accordance with the results from histological structures. In comparison groups of 1-IB-vs-2-IB and 1-Rib-vs-2-Rib, 33 and 51 differently expressed proteins were identified and the function annotation results showed that these proteins were involved in regulating bone development and differentiation. Subsequently, 11 and 13 candidate proteins in comparison group of 1-IB-vs-1-Rib and 1-IB-vs-2-IB related to bone development were validated by MRM assays.



Our present study suggested the different key proteins involved in the composition of fish ribs and IBs as well as their growth development. These findings could provide important clues towards further understanding of fish skeletal system and the roles of proteins playing in regulating diverse biological processes in fish.
College of Fisheries, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education/Key Lab of Freshwater Animal Breeding, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei, 430070, China.|Collaborative Innovation Center for Healthy Freshwater Aquaculture of Hubei Province, Wuhan, 430070, China.
Chun-Hong
CH
Nie
College of Fisheries, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education/Key Lab of Freshwater Animal Breeding, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei, 430070, China.|Collaborative Innovation Center for Healthy Freshwater Aquaculture of Hubei Province, Wuhan, 430070, China.
Shi-Ming
SM
Wan
Department for Fisheries, Beekeeping, Game management and Special Zoology, Faculty of Agriculture, University of Zagreb, Zagreb, Croatia.
Tea
T
Tomljanovic
Department for Fisheries, Beekeeping, Game management and Special Zoology, Faculty of Agriculture, University of Zagreb, Zagreb, Croatia.
Tomislav
T
Treer
Department of Bioscience Technology, Chung Yuan Christian University, Chung-Li, Taiwan.
Chung-Der
CD
Hsiao
College of Fisheries, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education/Key Lab of Freshwater Animal Breeding, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei, 430070, China.
Wei-Min
WM
Wang
College of Fisheries, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education/Key Lab of Freshwater Animal Breeding, Ministry of Agriculture, Huazhong Agricultural University, Wuhan, Hubei, 430070, China. gaozexia@hotmail.com.|Collaborative Innovation Center for Healthy Freshwater Aquaculture of Hubei Province, Wuhan, 430070, China. gaozexia@hotmail.com.|Hubei Provincial Engineering Laboratory for Pond Aquaculture, Wuhan, 430070, Hubei, China. gaozexia@hotmail.com.
Ze-Xia
ZX
Gao
D000818:Animals; D001846:Bone Development; D003530:Cyprinidae / Q000254:growth & development* / Q000378:metabolism*; D029941:Fish Proteins / Q000378:metabolism; D009132:Muscles / Q000378:metabolism*; D040901:Proteomics; D012272:Ribs / Q000254:growth & development* / Q000378:metabolism*
D029941:Fish Proteins
Growth and differentiation; Intermuscular bones; MRM validation; Megalobrama amblycephala; Ribs; iTRAQ proteomics