Characterization of two vitellogenin cDNAs from a Pandalus shrimp ( Pandalopsis japonica): Expression in hepatopancreas is down-regulated by endosulfan exposure

• Published in: Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology Vol. 157; no. 1; pp. 102 - 112
• Main Authors: Jeon, Jeong-Min, Lee, Sun-Ok, Kim, Kyoung Sun, Baek, Hae-Ja, Kim, Suam, Kim, Il-Kyu, Mykles, Donald L., Kim, Hyun-Woo
• Format: Journal Article
• Language: English
• Published: England Elsevier Inc 01.09.2010
• Subjects: Amino Acid Sequence; Animals; Arthropod; Base Sequence; cDNA; Crustacean; Differential display; DNA sequence; DNA, Complementary - genetics; Down-Regulation - drug effects; Endocrine disruptor; Endosulfan; Endosulfan - pharmacology; Evolution, Molecular; Female; Gene expression; Gene Expression Regulation - drug effects; Hepatopancreas; Hepatopancreas - drug effects; Hepatopancreas - metabolism; Humans; Male; Molecular Sequence Data; mRNA; Ovary; Pandalidae - drug effects; Pandalidae - genetics; Pandalidae - physiology; Pandalopsis japonica; Pesticide; Phylogeny; Reproduction; Reproduction - drug effects; Sequence Alignment; Shrimp; Tissue distribution; Vitellin; Vitellogenin; Vitellogenins - chemistry; Vitellogenins - genetics; Endosulfan; Arthropod; DNA sequence; Amino acid sequence; Pandalopsis japonica; mRNA; Endocrine disruptor; Gene expression; Vitellogenin; Vitellin; Ovary; Reproduction; Pesticide; Crustacean; Hepatopancreas; cDNA; Differential display; Tissue distribution; Shrimp
• ISSN: 1096-4959; 1879-1107
• DOI: 10.1016/j.cbpb.2010.05.006

Endosulfan is a neurotoxic organochlorine insecticide of the cyclodiene family of pesticides that inhibits molting and reproduction in aquatic crustaceans. In order to determine the molecular mechanism of endosulfan as an endocrine disrupting chemical (EDC), differential display RT-PCR (DDRT-PCR) was used to isolate genes in the shrimp, Pandalopsis japonica, affected by endosulfan exposure. PCR screening of cDNA from the hepatopancreas from control and endosulfan-exposed animals, using 120 sets of random primers, yielded partial cDNAs encoding two vitellogenin-like proteins (Pj-Vg1 and -Vg2). Complete sequences were obtained using a combination of RT-PCR and RACE-PCR. Pj-Vg1 (7883 bp) encoded a protein composed of 2533 amino acid residues (283.27 kDa estimated mass), whereas Pj-Vg2 (7792 bp) encoded a protein composed of 2537 amino acids residues (284.87 kDa estimated mass). Alignment of the Pj-Vgs with those of other vitellogenins identified a conserved subtilisin cleavage site (RQKR) and the lipoprotein N-terminal (vitellin), DUF1081, and von Willebrand factor type D domains, indicating both genes encoded functional proteins. Phylogenetic analysis showed that Pj-Vg1 and -Vg2 were most similar to Pandalus hypsinotus Vg. Both Pj-Vg1 and -Vg2 were expressed primarily in the hepatopancreas, although the Pj-Vg2 transcript was also detected in the ovary. The effects of the 3-day endosulfan exposure (2.5 µg/L and 25 µg/L) on Vg expression in the hepatopancreas were determined by quantitative RT-PCR. Expression of both transcripts was significantly inhibited at 25 µg/L suggesting that Pj-Vgs can be used as indicator for endosulfan exposure.