Molecular cloning and characterization of the crustacean hyperglycemic hormone cDNA from Litopenaeus schmitti. Functional analysis by double-stranded RNA interference technique.
The crustacean hyperglycemic hormone (CHH) plays an important role in the regulation of hemolymph glucose levels, but it is also involved in other functions such as growth, molting and reproduction. In the present study we describe the first CHH family gene isolated from the Atlantic Ocean shrimp Litopenaeus schmitti. Sequence analysis of the amplified cDNA fragment revealed a high nucleotide sequence identity with other CHHs. Northern blot analysis showed that the isolated CHH mRNA from L. schmitti is present in the eyestalk but not in muscle or stomach. We also investigated the ability of dsRNA to inhibit the CHH function in shrimps in vivo. Injection of CHH dsRNA into the abdominal hemolymph sinuses resulted in undetectable CHH mRNA levels within 24 h and a corresponding decrease in hemolymph glucose levels, suggesting that functional gene silencing had occurred. These findings are the first evidence that dsRNA technique is operative in adult shrimps in vivo.
Lugo
Juana M
JM
Aquatic Biotechnology Department, Animal Biotechnology Division, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
Morera
Yuliet
Y
Rodríguez
Tania
T
Huberman
Alberto
A
Ramos
Laida
L
Estrada
Mario P
MP
D000818:Animals; D060829:Arthropod Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis; D005786:Gene Expression Regulation; D007447:Invertebrate Hormones; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D009928:Organ Specificity / Q000235:genetics; D033561:Penaeidae / Q000187:drug effects / Q000235:genetics*; D034622:RNA Interference; D012330:RNA, Double-Stranded / Q000494:pharmacology*; D016415:Sequence Alignment
D060829:Arthropod Proteins; D018076:DNA, Complementary; D007447:Invertebrate Hormones; D009419:Nerve Tissue Proteins; D012330:RNA, Double-Stranded; C015947:hyperglycemic hormone, crustacean