Characterization of vitellogenin in the shrimp Metapenaeus ensis: expression studies and hormonal regulation of MeVg1 transcription in vitro.
During gonad maturation, female shrimp accumulate the major egg yolk protein vitellin (Vn) in premolt stage, and the process of molting and reproduction is synchronized. Using a polyclonal anti-Vn antibody, immunopositive signals could be detected in the ovary and among the proteins secreted by the hepatopancreas by Western blot. In the ovary, Vn immunoreactivity was located in the posterior lobe. Hepatopancreas proteins with sizes identical to ovary vitellogenin (Vg) subunits (i.e., 78 and 157 kDa) were immunoreactive to the Vn antibody and these proteins included amino acid sequences identical to parts of the MeVg1 precursor. A major 7.8 kb MeVg1 transcript, was detected in the ovary. In the hepatopancreas, the transcripts were primarily small (<2.3 kb) and while the 7.8 kb transcript which constitutes <50% of the total Vg mRNA. MeVg1 transcript could be detected in the hepatopancreas of juvenile females with a maximum level during late intermolt and early premolt. To study the effect of different hormones on expression of MeVg1, explant cultures of hepatopancreas and ovary were developed. Although several hormones (20-hydroxyecdysone, estradiol (ES), farnesoic acid (FA), juvenile hormone (JH) III, methyl farnesoate, and progesterone (PG)) apparently stimulated MeVg1 gene expression, only FA consistently stimulated MeVg1 expression by the hepatopancreas explants, while both FA and 20-hydroxyecdysone were stimulated ovarian explants. In summary, (i) Vg transcripts can be detected in both reproductive and nonreproductive females; (ii) the presence of large quantities of smaller Vg transcripts and the absence of a large Vg precursor from the hepatopancreas suggests that smaller MeVg1 transcripts provide an important contribution to Vg synthesis in shrimp. Our results suggest that there is differential processing of the MeVg1 precursor in the ovary and hepatopancreas of shrimp.
Tiu
Shirley H K
SH
Department of Zoology, The University of Hong Kong, Pokfulam, Hong Kong.
Hui
Jerome H L
JH
He
Jian-Guo
JG
Tobe
Stephen S
SS
Chan
Siu-Ming
SM
D017398:Alternative Splicing / Q000235:genetics; D000818:Animals; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D012739:Gonadal Steroid Hormones / Q000502:physiology*; D043143:Hepatopancreas / Q000378:metabolism / Q000502:physiology; D009928:Organ Specificity / Q000235:genetics; D010053:Ovary / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D033561:Penaeidae / Q000737:chemistry* / Q000235:genetics* / Q000502:physiology; D011499:Protein Processing, Post-Translational / Q000235:genetics; D014158:Transcription, Genetic; D014819:Vitellogenins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*
D012739:Gonadal Steroid Hormones; D014819:Vitellogenins