The Role Of Endocannabinoids In The Attachment Of Green Mussle,Perna Viridis

Cannabinoids are compounds that modulate neurotransmitter,immune,and other functions by acting on cannabinoid receptors and modulating neurotransmitter release.Some small molecules synthesized and secreted in the body can act on cannabinoid receptors,also known as endocannabinoid.Endogenous cannabinoids have been found in invertebrates.At present,AEA,2-AG and other endogenous cannabinoids have been extracted from Mytilus edulis and Hydra vulgaris.Moreover,studies in Dreissena polymorpha show that AEA and its analogues can inhibit the attachment of mussels in the Dreissena polymorpha,and this process is reversible.It suggests that endocannabinoids may be involved in the regulation of Dreissena polymorpha attachment.As one of the main fouling shellfish in the coastal area of China,the attachment mechanism of Perna viridis is really less studied.Based on the results of this study,the role of endocannabinoids in the attachment of Perna viridis was preliminarily discussed.The main results obtained are as follows:(1)Effects of AEA,2-AG,OEA,PEA and AA on the adhesion of Perna viridis shows that AEA,2-AG and AA had some inhibitory effects on the adhesion of Perna viridis.Experiments In vitro showed that AM1241,HU308 and 2-AG degrading enzyme inhibitor JZL184 had some inhibitory effects on the adhesion of Perna viridis.These results suggest that AEA and 2-AG may regulate the attachment of Perna viridis by cannabinoid receptors.Effect of CB1 receptor antagonist SR141716A and AM251 on AEA have no obviously competitive inhibition in the attachment of Perna viridis.The effects of AEA and 2-AG on the expression of adhesion proteins in Perna viridis showed that AEA might affect the expression of FP1V-1,which could inhibit the adhesion of Perna viridis.While 2-AG may affect the expression of FP1V-1,FP1V-2,FP2,FP4V-1 and FP4V-2,which could inhibit the adhesion of Perna viridis.(2)The LC-MS/MS results showed that Perna viridis do contain four kinds of endo-genous AEA,2-AG,OEA,PEA.Moreover,the content of 2-AG in unattached mussles was higher than that in attached mussles,indicating that 2-AG might be involved in the regulation of unattachment in Perna viridis.(3)In the annotation of the transcriptome Unigene,a CB1-like receptor is annotated in the Nr database which proved that there are cannabinoid receptors in Perna viridis.The results of transcriptome showed that the content of 2-AG in unatta-ched mussles was higher than that in attached.Differential expression analysis was performed between the two groups.The results showed that On the one hand,2-AG may act on the TRP ion channel on the presynaptic membrane,which can reduce the concentration of Ca2+ in cell,cause the DSE phenomenon,and lead to the reduce of NO.While NO was reported to be involved in the attachment of Bivalves.On the other hand,2-AG may act on the CB receptor on the mitochon-drial membrane,affecting the energy metabolism in the cell,thus inhibiting the attachment of Perna viridis.