title: Differential effects of silencing crustacean hyperglycemic hormone gene expression on the metabolic profiles of the muscle and hepatopancreas in the crayfish Procambarus clarkii.
abstract: In order to functionally characterize the metabolic roles of crustacean hyperglycemic hormone (CHH), gene expression of CHH in the crayfish (Procambarus clarkii) was knocked down by in vivo injection of CHH double-stranded RNA (dsRNA), followed by metabolomic analysis of 2 CHH target tissues (the muscle and hepatopancreas) using nuclear magnetic resonance spectroscopy. Compared to the levels in untreated and saline-injected (SAI) animals, levels of CHH transcript, but not those of molt-inhibiting hormone (a CHH-family peptide), in the eyestalk ganglia of CHH dsRNA-injected (DSI) animals were significantly decreased at 24, 48, and 72 hour post injection (hpi), with concomitant changes in levels of CHH peptide in the sinus gland (a neurohemal organ) and hemolymph. Green fluorescence protein (GFP) dsRNA failed to affect levels of CHH transcript in the eyestalk ganglia of GFP DSI animals. Number of metabolites whose levels were significantly changed by CHH dsRNA was 149 and 181 in the muscle and 24 and 12 in the hepatopancreas, at 24 and 48 hpi, respectively. Principal component analysis of these metabolites show that metabolic effects of silencing CHH gene expression were more pronounced in the muscle (with the cluster of CHH DSI group clearly being separated from that of SAI group at 24 hpi) than in the hepatopancreas. Moreover, pathway analysis of the metabolites closely related to carbohydrate and energy metabolism indicate that, for CHH DSI animals at 24 hpi, metabolic profile of the muscle was characterized by reduced synthesis of NAD+ and adenine ribonucleotides, diminished levels of ATP, lower rate of utilization of carbohydrates through glycolysis, and a partially rescued TCA cycle, whereas that of the hepatopancreas by unaffected levels of ATP, lower rate of utilization of carbohydrates, and increased levels of ketone bodies. The combined results of metabolic changes in response to silenced CHH gene expression reveal that metabolic functions of CHH on the muscle and hepatopancreas are more diverse than previously thought and are differential between the two tissues.
authors.affiliation: Department of Biology, National Changhua University of Education, Changhua, Taiwan.
authors.forename: Wenfeng
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Aquaculture, National Kaohsiung Marine University, Kaohsiung, Taiwan.
authors.forename: Kuo-Hsun
authors.initials: KH
authors.lastname: Chiu
authors.affiliation: Department of Biology, National Changhua University of Education, Changhua, Taiwan.
authors.forename: Yi-Chun
authors.initials: YC
authors.lastname: Tien
authors.affiliation: Department of Biology, National Changhua University of Education, Changhua, Taiwan.
authors.forename: Shih-Fu
authors.initials: SF
authors.lastname: Tsai
authors.affiliation: Department of Medical Laboratory, Taoyuan Armed Forces General Hospital, Taoyuan, Taiwan.
authors.forename: Li-Jane
authors.initials: LJ
authors.lastname: Shih
authors.affiliation: Department of Biology, National Changhua University of Education, Changhua, Taiwan.
authors.forename: Chien-Hsun
authors.initials: CH
authors.lastname: Lee
authors.affiliation: Sorbonne Universités, UPMC Université Paris 06, UMR 7144 CNRS, Equipe ABICE, Station Biologique de Roscoff, Roscoff, France.|CNRS, UMR 7144, Adaptation et Diversité en Milieu Marin, Station Biologique de Roscoff, Roscoff, France.
authors.forename: Jean-Yves
authors.initials: JY
authors.lastname: Toullec
authors.affiliation: Department of Biology, National Changhua University of Education, Changhua, Taiwan.
authors.forename: Chi-Ying
authors.identifier: http://orcid.org/0000-0002-9591-1370
authors.initials: CY
authors.lastname: Lee
mesh_terms: D000818:Animals; D060829:Arthropod Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D003400:Astacoidea / Q000235:genetics / Q000378:metabolism*; D004734:Energy Metabolism; D005786:Gene Expression Regulation; D020868:Gene Silencing; D006458:Hemolymph / Q000378:metabolism; D043143:Hepatopancreas / Q000378:metabolism*; D007447:Invertebrate Hormones / Q000037:antagonists & inhibitors* / Q000235:genetics; D055442:Metabolome; D009132:Muscles / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics; D012330:RNA, Double-Stranded / Q000235:genetics
chemical_list: D060829:Arthropod Proteins; D007447:Invertebrate Hormones; D009419:Nerve Tissue Proteins; D012330:RNA, Double-Stranded; C015947:hyperglycemic hormone, crustacean