Molecular Cloning and Expression Analysis ofOvary-Specific Transcript 1(Pm-OST1) of the Giant Tiger Shrimp,Penaeus monodon

Abstract

Isolation and characterization of genes specifically expressed in ovaries are necessary for understanding sex differentiation and ovarian development processes in the giant tiger shrimp, Penaeus monodon. In this study, a transcript that significantly matched the polehole precursor was further characterized by RACE-PCR. The sequence obtained was 5151 bp in length and contained a coding region of 5031 bp corresponding to 1677 amino acids. This transcript was only expressed in ovaries but not in testes of Juveniles (N = 10) and broodstock (N = 22) of P. monodon. A tissue distribution analysis further confirmed ovary-specific expression of this transcript (called P. monodon ovary-specific transcript 1, Pm-OST1) in female broodstock. Expression levels of Pm-OST 1 in ovaries of juvenile P. monodon upon 5-HT Injection (33.9+/-6.40 g; 50 microg/g body weight) were significantly higher at 12-72 hours post Injection (P<0.05). Quantitative real-time PCR Indicated that Pm-OST1 was comparably expressed throughout ovarian development in normal P. monodon broodstock (P>0.05). However, the expression level of Pm-OST1 was significantly higher in stage-III ovaries in eyestalk-ablated broodstock (P<0.05). Pm-OST1 was clearly localized in the ooplasm of previtellogenic and vitellogenic oocytes. Our results suggest that Pm-OST1 plays a functionally Important role in promoting the development of female germ cells and oocytes in P. monodon.

Isolation and characterization of genes functionally involved in ovarian development of the giant tiger shrimp Penaeus monodon by suppression subtractive hybridization (SSH)

“…Several reproduction-related genes that are up-regulated during the ovarian development of P. monodon , for example, Ovarian-Specific Transcript 1 ( Pm-OST1 ) and cyclin B ( PmCyB ), have been previously reported (Klinbunga et al , 2009; Visudtiphole et al , 2009). The down-regulation of keratinocyte-associated protein 2 , Ser/Thr Chk1 , DNA replication licensing factor mcm2 , PmSePM and egalitarian implied that lower levels of these gene products may be necessary for the development and final maturation of P. monodon oocytes.…”

“…EF-1 α was included as the positive control. Initially, nonquantitative RT-PCR (Klinbunga et al , 2009) was carried out using 100 ng of first-strand cDNA as the template, with varying concentrations of primers (0.05, 0.10, 0.15, 0.20, 0.25, 0.30 and 0.40 μM, respectively). Primer sequences are listed in Table 1.…”

Expression and polymorphism of farnesoic acid O-methyltransferase (FAMeT) and association between its SNPs and reproduction-related parameters of the giant tiger shrimp Penaeus monodon

“…Exogenous 5-HT administration clearly promoted expression of P. monodon Ovarian-Specific Transcript (Pm-OST1) in ovaries of subadults (5-month-old) at 12-78 hpi, with the peak level at 48 hpi (Klinbunga et al, 2009). In addition, its effects on promoting the expression of P. monodon adipose differentiation related protein (PmADRP) in ovaries of this shrimp at 48 hpi were also reported (Sittikankaew et al, 2010).…”

“…Knowledge on molecular mechanisms and functional involvement of reproduction-related genes and proteins in ovarian development of P. monodon is necessary for better understanding of the reproductive maturation of captive P. monodon to resolve the major constraint of this economically important species (Klinbunga et al, 2009;Preechaphol et al, 2007).…”

Improving the Reproductive Performance of Domesticated Giant Tiger Shrimp, Penaeus monodon

“…In addition domesticated P. monodon produce far fewer eggs and spawn less frequently than their wild-caught counterparts (Klinbunga et al, 2009, Coman et al, 2006, Hall, 2003, Peixoto et al, 2005. Peixoto et al (2005) noted that the ovaries of mature domesticated broodstock frequently contained high proportions of immature oocytes.…”