| In this study, the full-length MIH c DNA named Ec MIH was cloned by rapid amplification of c DNA ends(RACE) from the eyestalk ganglia of the ridgetail white prawn E. carinicauda.The full-length of MIH c DNA was 1218 bp containing a 420 bp 5’untranslated region(5’UTR), a438 bp 3’untranslated region(3’UTR) and a 360 bp open reading frame(ORF) encoding 120 amino acids which consist of a 41-residue signal peptide and a 79-residue mature peptide, with the molecμLar mass of 13.71 k Da and an estimated p I of 8.02. Sequence analysis indicated that the mih was homology with that of Marsupenaeus japonicas and Macrobrachium rosenbergii,were 87% and 86% identity respectively. Tissue expression analysis showed that MIH-like m RNA expressed at the highest level in eyestalk while extremely low in ovary, but not expressed in liver, muscle, gill and blood cells. In the ovarian and larval development, the expression profiles of mih showed different pattern. With the ovarian development, the highest level of mih m RNA expression was found in ovary at mature stage and significant decrease of mih m RNA expression occurred at stage I, II, III and V. In the larval development, the expression of the mih was the highest level at larval I stage and the lest level at larval V stage and extremely significantly among different stages(P<0.01). The results suggested that mih gene played inhibitory roles in ovarian development and participated in larval molting in E. carinicauda.To study the roles of the Gonad-inhibiting hormone(GIH) in ovarian larval development,the GIH c DNA of E. carinicauda was first cloned by rapid amplification(RACE) method, which was named gih. The full-length c DNA sequence of gih was 1009 bp, which contained a 523bp3’untranslated region(3’UTR), a 128 bp 5’untranslated region(5’UTR), and one 333 bp open reading frame(ORF) that encoded 111 amino acids with the molecular mass of 12.708 k Da and an estimated p I of 8.01. GIH was composed of signal peptide and mature peptide, its N-terminal contained 32 amino acid composition of signal peptide and mature peptide was composed of 79 amino acids. Blast analysis indicated that the homologys of gih with the GIHs of Rimicaris kairei was 81% and phylogenetic tree analysis showed that gih was in the same category as GIH of Rimicaris kairei. The tissue expression analysis revealed that GIH expressed in eyestalk ganglia, ventral ganglion, obvious, heart, intestine, blood and stomach. In the ovarian and larval development, the expression profiles of GIH showed different pattern. With the ovarian development, The highest level of GIH m RNA expression was found in ovary at mature stage and significant decrease of GIH m RNA expression occurred at stage I, II, III and V. In the larval development, the expression of the GIH was the lest level at larval IV stage and extremely significantly among different stages(P<0.01). The results suggested that GIH gene played inhibitory roles in ovarian development and participated in larval molting in E. carinicauda.To study the roles of the hyperglycemic hormone(CHH) in ovarian and larval development,the CHH c DNA of E. carinicauda was first cloned by rapid amplification(RACE) method,which was named chh. The full-length c DNA sequence of chh was 710 bp, which contained a 85 bp 3’untranslated region(3’UTR), a 167 bp 5’untranslated region(5’UTR), and one 426 bp openreading frame(ORF) that encoded 142 amino acids with the molecular mass of 15.9k Da and an estimated p I of 8.26. chh gene by signal peptide, mature peptide and related peptide precursor.Signal peptide cut site between Ala and Trp, and consists of 27 amino acids, mature peptide from74 amino acids and related peptide precursor consists of 33 amino acids. The second base cutting locus for KR, mature peptide on 12 th CHH family Ⅱ peculiar to the type of peptide Gly residues. Sequence analysis indicated that the chh was homology with that of M. japonicas and M. rosenbergii, were 82% and 81% identity respectively. Tissue expression analysis showed that chh m RNA expressed at the highest level in eyestalk while extremely low in blood, In the ovarian and larval development, the expression profiles of chh showed different pattern. With the ovarian development, The highest level of chh m RNA expression was found in ovary at III stage and significant decrease of chh m RNA expression occurred at stage I, II, IV and V. the expression of the chh was the lest level in ovary at IV stage and extremely significantly among different stages(P<0.01). In the larval development, the expression of the chh was the lest level at larval IV stage and extremely significantly among different stages(P<0.01), and Larval stage I to V below hepatopancreatic. The results suggested that chh gene played promoting roles in ovarian development and participated in larval molting in E. carinicauda.pH, salinity and NH4 Cl is mariculture shrimp crab physiological ecology of important environment factor, can directly affect the survival, growth, molting, and reproduction of aquatic creatures and immune activity. This study found that, the gene has obvious difference in time.After challenged with p H 6.5, 24 h and 48 h eyestalk and ventral ganglion MIH-like gene expression level than the control group were significantly increased(P<0.05); After challenged with 4 mg/L NH4 Cl, 12 h and 24 h eyestalk and ventral ganglion MIH-like gene expression level than the control group were significantly increased(P<0.05); After challenged with salinity 39,12 h and 24 h eyestalk and ventral ganglion MIH-like gene expression level than the control group were significantly increased(P<0.05); Results show that the MIH involved in stress response after the environmental stresses. |
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