From hepatopancreas to ovary: molecular characterization of a shrimp vitellogenin receptor involved in the processing of vitellogenin.
We report the first cloning and characterization of cDNA encoding a putative vitellogenin (Vg) receptor (VgR) from the shrimp, Penaeus monodon. The shrimp VgR cDNA is 6.8 kb; the deduced protein has 1943 amino acids with a molecular weight of 211 kDa. VgR is ovary specific and consists of conserved cysteine-rich domains, epidermal growth factor-like domains, and YWTD motifs similar to the low-density lipoprotein, very low-density lipoprotein, and VgR of insects and vertebrates. VgR expression level in the ovary is low during early vitellogenesis and increases to maximum levels in females with a gonadosomatic index of 3-4, presumably when needed for receptor-mediated endocytosis during the rapid phase of extraovarian Vg production by the hepatopancreas. A peptide from the C-terminal end of VgR was synthesized for antibody production. Anti-VgR antibody recognized an ovarian membrane protein, and the level of this protein was high when extraovarian production of Vg reached peak levels. By immunohistochemical analysis, VgR was detected strongly in the membranes of larger oocytes. VgR expression was knocked down after the shrimp were injected with VgR double-stranded RNA, leading to a decrease in VgR protein content in the ovary, but an increase in the hemolymph level of Vg. This study represents the first report of the functional analysis of a putative VgR in a crustacean.
Tiu
Shirley Hiu Kwan
SH
School of Biological Sciences, The University of Hong Kong, Hong Kong, People's Republic of China.
Benzie
John
J
Chan
Siu-Ming
SM
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000302:isolation & purification; D004527:Egg Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology; D005260:Female; D043143:Hepatopancreas / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D010053:Ovary / Q000737:chemistry / Q000378:metabolism*; D033561:Penaeidae / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D011956:Receptors, Cell Surface / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology; D017422:Sequence Analysis, DNA; D014018:Tissue Distribution; D014818:Vitellogenesis / Q000235:genetics; D014819:Vitellogenins / Q000096:biosynthesis*
D018076:DNA, Complementary; D004527:Egg Proteins; D011956:Receptors, Cell Surface; D014819:Vitellogenins; C030474:vitellogenin receptor