Molecular cloning and differential expression pattern of two structural variants of the crustacean hyperglycemic hormone family from the mud crab Scylla olivacea

Metabolic effects of parasitization by the barnacle Polyascus plana (Cirripedia: Rhizocephala: Sacculinidae) on a grapsid host, Metopograpsus thukuhar

“…The reagents (excepting the primary antibody), experimental protocol, and data processing for the ELISA followed the methods described by Lee et al (2001). The primary antibody used was the IgG fraction of an antibody (antiSco-CHH), which was raised against a C-terminal stretch (residues 59−71) of the mud crab Scylla olivacea CHH, Sco-CHH (Tsai et al 2008). Specificity of the primary antibody for the putative CHH in the sinus gland samples of M. thukuhar was assessed by 2 tests.…”

“…Specificity of the primary antibody for the putative CHH in the sinus gland samples of M. thukuhar was assessed by 2 tests. First, samples were assayed with the ELISA using the anti-Sco-CHH pre-adsorbed with the synthetic peptide that was used for raising the antibody (Tsai et al 2008). Second, instead of using the anti-Sco-CHH antibody, sinus gland samples were assayed using the IgG fraction of the pre-immune serum.…”

Hyperglycemic activity of the recombinant crustacean hyperglycemic hormone B1 isoform (CHH-B1) of the Pacific white shrimp Litopenaeus vannamei

“…Bands of ∼10 kDa were not detected in control samples. Since the Western blot analysis of culture supernatants showed that the expression of rCHH-B1 was similar among the induction conditions analyzed, the condition of 2% methanol for 24 h was selected [38]; Pt, P. trituberculatus (CHH-1, ACB46189; CHH-2, AIZ94611) [49]; Cs, C. sapidus (ES-CHH (CHH-1), AAS45136; CsPO-CHH (CsCHH-2, ABG67921, ABC61678) [8,18,53]; Gel, G. lateralis (EG-CHH-A, ABF48652; Po-CHH, ABF58091) [22,52]; Cam, C. maenas (SG-CHH, AAG29442; PO-CHH, AAG29435) [10]; Sco, S. olivacea (CHH, AAQ75760; CHH-L, ABP88270) [2,41]; Poi, P. ibericum (CHHXO, ABA70560; CHHPO, ABA70561) [38]; Mar, M. rosenbergii (CHH, AAF29534; CHH-L, AF372657) [3,29]; Prc, P. clarkii (CHH1, BAA89003; CHH1-L, AAL79193; CHH2, AFV95082; CHH2-L, AFV95078) [19,48]; Pj, P. japonica (CHH1ES, AFG16933; CHH1PO, AFG16932) [16]; Lv, L. vannamei (CHH-B2, AAN86057; CHH-B1, AY167045) [21,30]; Hyperglycemic activity tested in vivo: N, using native neuropeptides; R, recombinant neuropeptides; (+), with activity; (−) without activity; aa, number of amino acids in sequences; % I.D., identity percentage relative to the CHH-B1 sequence. CHH, CHH isoforms; CHH-L, CHH-like isoforms.…”

“…Most of the type I genes described so far encode two transcripts that, in most cases, share the signal peptide sequence, the CHH precursor-related peptide (CPRP) and the N-terminal region (exons I and II), but differ in the C-terminal region, which can be encoded by exon III or exon IV [4,10]. These alternative splicing isoforms have been found in various crustaceans, including Carcinus maenas [10], Macrobrachium rosenbergii [3], Pachygrapsus marmoratus, Potamon ibericum [38], Gecarcinus lateralis [22], Callinectes sapidus [8,53], Scylla olivacea [41], Pandalopsis japonica [16], Procambarus clarkii [19,47,48,50], Portunus trituberculatus [49], and Litopenaeus vannamei [21].…”

Insect ion transport peptides are derived from alternatively spliced genes and differentially expressed in the central and peripheral nervous system

“…Surprisingly, several variants of mRNAs have been found in the shore crab, most of which definitely did not lead to a translated product and are, thus, of unknown function (Dircksen et al, 2001), a finding recently corroborated by studies on other crabs. These studies stated that preferentially long POCHH-encoding mRNAs occur in several other tissues but no expressed peptides could be found (Chung and Zmora, 2008;Tsai et al, 2008).…”