DISTRIBUTION OF HEMOCYANIN IN FENNEROPENAEUS CHINENSIS TISSUES

Hemocyanin,a multifunctional copper-containing respiratory protein,makes up the major part in the hemolymph of shrimp.It was reported that the expression of hemocyanin gene has a tissue specialty,however the distribution of hemocyanin in Fenneropenaeus chinensis has not been previously reported.In this paper,the distribution of hemocyanin in F.chinensis was studied by immunohistochemistry.Monoclonal antibodies are highly specific molecular probes,which allow identification based on molecular level.A monoclonal antibody which was against two subunits(73and 75kDa)of F.chinensis hemocyanin was screened out using Western-blotting.Using this Mab as the primary antibody,paraffin method and immunohistochemistry were employed to observe distribution of hemocyanin in gill,heart,stomach,intestine,lymphoid organ,ovary and hepatopancreas of F.chinensis.Immunohistochemistry results showed that positive signal was observed in all of sampled tissues.Strong positive result was found in blood vessel of gill and lacunaes among myocardial bundles of heart;Whereas positive signals presented in lymph cavities and blood sinus of lymphatic organ,hepatopancreatic tubules and blood sinus in connective tissues of mesenteron;Weak positive result could be observed in blood sinus among oocyte,blood sinus in connective tissues of the inside stomach circular muscle.No positive signal was observed in negative control.In conclusion,hemocyanin was mostly from the hemolymph rather than fundamental tissue,and also found where hemolymph can not reach.hemocyanin was widely distributed in various tissues of F.chinensis,and was found abundant in blood vessel or lacunae where plenty of hemolymph moved through.