A dopamine beta hydroxylase from Chlamys farreri and its induced mRNA expression in the haemocytes after LPS stimulation

The expression of dopa decarboxylase and dopamine beta hydroxylase and their responding to bacterial challenge during the ontogenesis of scallop Chlamys farreri

“…In the adult scallop, catecholaminergic nervous system could be activated by immune stimulation, resulting in the increase of CfDDC and CfDBH mRNA expression level in haemocytes [7,8]. In the present study, the temporal expression of CfDDC and CfDBH mRNA was determined in the trochophore, late D-hinged and veliger scallop larvae challenged by pathogenic bacteria V. anguillarum to get an insight into the response of the growing catecholaminergic nervous system to the immune stimulation.…”

“…For the measurement of CfDDC mRNA, a 110 bp product was amplified with the sense primer P1 and the antisense primer P2 (Table 1) from cDNA template [7]. The sense primer P3 and the antisense primer P4 (Table 1) were used to amplify a 139 bp product to quantify CfDBH mRNA [8]. Two CfEF-1a (elongation factor 1 alpha from scallop C. farreri) primers, sense primer P5 and antisense primer P6 (Table 1), were used to amplify an 86 bp fragment as an internal control to verify the successful reverse transcription and to calibrate the cDNA template.…”

“…The shells of older larvae (day 2 and up) were decalcified in 5% EDTA in PBS for 30 min, rinsed in PBS 3 Â 15 min, and blocked overnight in the blocking solution (10% normal goat serum, 0.25% bovine serum albumin, 1% TritonX-100 and 0.03% sodium azide in PBS). Then larvae were labeled by incubating them in anti-CfDDC or anti-CfDBH antibody (which were acquired via immunization 6-week old rats with CfDDC and CfDBH recombinant protein and confirmed to be specific by the western blotting, diluted 1:500 in the blocking solution) for 1e3 days at 4 C [7,8]. These larvae were then washed (3 Â 1 h) in PBS-TX and incubated for 1 day in goat anti-rat IgG conjugated to Alexa 488 (Invitrogen) diluted 1:100 in PBS.…”

The immune system and its modulation mechanism in scallop

“…In adult scallops, dopamine, norepinephrine and epinephrine could be detected in the haemolymph [117]. Four crucial catecholamine metabolism enzymes have been identified recently from scallop, including phenylalanine hydroxylase (PAH) [135], DDC [123], dopamine beta hydroxylase (DBH) [135] and monoamine oxidase (MAO) [136]. Their mRNA transcripts were ubiquitously detectable in all the tested tissues including hemocytes, hepatopancreas, kidney, adductor muscle, gonad, gill and mantle.…”

The enkephalinergic nervous system and its immunomodulation on the developing immune system during the ontogenesis of oyster Crassostrea gigas

“…A complex neuroendocrine-immune regulatory network exists in all vertebrates and some of the invertebrates, and it optimizes the immune defense against the invasive pathogenic microorganisms and maintenances homeostasis of the host [1,2]. Enkephalinergic nervous system is one of the most important nervous systems in vertebrates, which interacts with the immune system to modulate immune response [3].…”