Table 1.
Gene-specific primers and expected amplicon sizes.
Fig 1.
Summary of BLAST searches in combined three-transcriptome dataset and unigene analyses in individual datasets (eyestalk, CNS, and ovary) of Macrobrachium rosenbergii.
(A) E-value distribution of BLASTx hits from all unigenes obtained within combined eyestalk, CNS, and ovary dataset. (B) Species distribution of BLASTx hits. (C) Comparison of unigenes present in individual datasets. (D) Pfam annotation of unigenes present in individual datasets.
Table 2.
Summary of transcripts encoding neuropeptides in M. rosenbergii eyestalk, CNS, and ovary transcriptomes.
Table 3.
Predicted mature peptides from the eyestalk, CNS, and ovary transcriptomes of M. rosenbergii.
Table 4.
Summary of transcripts encoding neuropeptide receptors in M. rosenbergii eyestalk, CNS, and ovary transcriptomes.
Fig 2.
Molecular characterization of Macrobrachium rosenbergii bursicon, CCAP, CLDH, eclosion hormone, and PDHs.
The schematic diagrams show the organization of putative neuropeptide precursors of M. rosenbergii, including signal peptides (blue), putative proteolytic cleavage sites (red), C-terminal glycine residues responsible for amidation (green), Cys residues and putative neuropeptides (yellow). Precursor sequence alignments are shown with site of putative neuropeptide denoted with a yellow bar. Conserved amino acids are shown in black shading while similar amino acids are shown with grey shading. Mro, is M. rosenbergii and for other species abbreviations see S1A Table.
Fig 3.
Molecular characterization of Macrobrachium rosenbergii DH45.
(A) Schematic diagram showing the organization of DH45 neuropeptide precursor of M. rosenbergii, including signal peptide (blue), putative proteolytic cleavage sites (red), C-terminal glycine residues responsible for amidation (green), and putative neuropeptide (yellow). Alignment of DH45, DH44, and ELH active peptides is shown. Mro, is M. rosenbergii and for other species abbreviations see S1A Table. Conserved amino acids are shown in black shading while similar amino acids are shown with grey shading. (B) Phylogenetic tree of DH45, DH44, and ELH active peptides based on maximum likelihood estimation. Mro-CHH peptide sequence is used as a 'non-related' peptide for confirmation of phylogenetic tree reliability. Scale bar represents the number of amino acid substitutions per site.
Fig 4.
Molecular characterization of Macrobrachium rosenbergii neuroparsin, NPF, SIFamide, and sulfakinin.
The schematic diagrams show the organization of putative neuropeptide precursors of the M. rosenbergii, including signal peptides (blue), putative proteolytic cleavage sites (red), C-terminal glycine residues responsible for amidation (green), Cys residues, and putative neuropeptides (yellow). Precursor sequence alignments are shown with site of putative neuropeptide denoted with a yellow bar. Conserved amino acids are shown in black shading while similar amino acids are shown with grey shading. Mro, is M. rosenbergii and for other species abbreviations see S1A Table.
Fig 5.
Molecular characterization of Macrobrachium rosenbergii CHH.
(A) The schematic diagrams show the organization of putative CHH neuropeptide precursors of the M. rosenbergii, including signal peptides (blue), putative proteolytic cleavage sites (red), C-terminal glycine residues responsible for amidation (green), Cys residues, and putative neuropeptides (yellow). Precursor sequence alignments are shown with site of putative neuropeptide denoted with a yellow bar. Conserved amino acids are shown in black shading while similar amino acids are shown with grey shading. Mro, is M. rosenbergii and for other species abbreviations see S1A Table. (B) LC-MS/MS spectra showing fragment matching CHH precursor-related peptide (CPRP) which is indicated by red underline in (A).
Fig 6.
Tissue-specific expression of Macrobrachium rosenbergii neuropeptide genes, using RT-PCR.
Expression of 11 neuropeptide genes using gene-specific primers, as well as the β-actin gene. PCR used cDNA derived from 13 tissues of female (including 4 stages of ovarian tissue) and 1 tissue of male (testis). Negative control represents no cDNA in PCR. CCAP, Crustacean cardioactive peptide; CLDH, Calcitonin-like diuretic hormone; EH, Eclosion hormone; NPF, Neuropeptide F; SK, Sulfakinin; SIF, SIFamide; NP, Neuroparsin.