中国对虾中肠的超微结构与细胞化学研究

STUDIES ON ULTRASTRUCTURE AND CYTOCHEMISTRY OF MIDGUT INPENAEUS CHINENSIS

  • 摘要: (1)在透射电镜下观察中国对虾(Penaeus chinensis Osbeck)中肠肠壁的超微结构.结果显示:中国对虾中肠肠壁由内向外依次为上皮细胞层、结缔组织层、肌层和结缔组织层四层结构.中肠上皮有明、暗两种细胞,以暗细胞占多数,两种上皮细胞都具有从肠腔吸收营养物质和向结缔组织中转运营养物质的功能.(2)运用酶细胞化学技术,显示中肠组织细胞中的酸性磷酸酶(ACP)和酚氧化酶(PO)活性,电镜观察显示:ACP阳性反应出现在明细胞顶部的圆形小泡和明、暗两种细胞的溶酶体中,暗细胞顶部聚集着大量ACP阴性反应的酶原颗粒,酶原颗粒主要由暗细胞合成并分泌.PO颗粒出现在患病虾肠壁结缔组织内解体的颗粒细胞上和游离于结缔组织中.结缔组织除具有维持上皮细胞的形态、贮存营养物质的功能,还在对虾免疫反应中发挥作用.

     

    Abstract: The electronic microscope technique and the enzyme cyt ochemistry technique were used to studied the ultrastructure of the wall of the midgut of penaeus chinensis, and active site of phenoloxidasa (PO) and acid phosphatase (ACP) in the tissue cells of midgut well. Right midgut from penaeus chinensis were removed and making experiments using three methods: ①midgut tissue was cut into blocks of approximately 1mm 3, the blocks were fixed in 3% fresh glutaraldehyde solution and then were post-fixed in 1% osmic acid. Embeldded it in Epon821. Semit him sections of the midgut ware cut on the ULTRACUT E. Stained it with toluidine blue and examined under microscope. Ultrathin sections of the midgut were cut on the LKB-Nova ultramicrotome. ②The midgut tissue blocks were fixed in 2% glut araldehyde solutions, then the blocks cut into thin sections approximately 60nm and incubated in the ACP medium and post-fixed in 1% osmic acid, embedded it in Epon821 and cut it into ultrathin sections. ③the midgut tissue blocks of healt hy and virus-infectious penaeus chinensis were fixed in 1% Bakers formol calciu m-and cut it into thin sections approximately 100 nm, incubated it in the pheno l oxidase medius then deal the samples with common ultrasection technique. All abo ve ultrathin sections were observed under a Hitachi H-7000 electron microscope. The results showed: the epithelium of midgut includes light cells and da rk cells, mostly dark cells, both kinds of cells have the function of absorbing nutrient substances from the enterocolic cavity and transferring them to connect ive tissue. Cytochemistry reaction results showed: ACP positive reaction was sho wn in the rounded vesicles that are situated at the top of the light cells, also in the lysosomal bodies of both light and dark cells. The tops of the dark cell s were crowded with lots of ACP-negative zymogen granules that were mainly synt hesized and excreted by the dark cells. PO hardly have activity in health y wall of the midgut, while in the virus-infections penaeus chinensis, the PO- granules were found on the disassembled granular cells of the connective tissue and dissociate in the connective tissue. Besides maintaining the shapes of the epithelial cells and stoking nutriments, the connective itssue play role in the i mmune reaction of the penaeus chinensis.

     

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