中华皮肤科杂志 ›› 2002, Vol. 35 ›› Issue (5): 361-363.

• 论著 • 上一篇    下一篇

烟曲霉分生孢子表面形态改变及纤连蛋白受体分布状态

徐赤宇, 温海, 吴建华   

  1. 第二军医大学长征医院皮肤科, 上海200003
  • 收稿日期:2001-11-15 出版日期:2002-10-15 发布日期:2002-10-15
  • 基金资助:
    国家自然科学基金资助项目(39770002)

Change of Surface Shape and Distribution of Fibronectin Receptors on Aspergillus fumigatus Conidia

XU Chiyu, WEN Hai, WU Jianhua   

  1. Department of Dermatology, Chang Zheng Hospital, Second Military Medical University, Shanghai 200003, China
  • Received:2001-11-15 Online:2002-10-15 Published:2002-10-15

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摘要： 目的 观察烟曲霉分生孢子静止期及其在膨胀生发过程中表面形态的改变。观察纤连蛋白受体在孢子表面分布状态及其在膨胀生发过程中的变化。方法 取培养成熟的烟曲霉,磷酸盐缓冲液冲洗后,通过3.5～4.0μm的钢丝网过滤提取制备纯净分生孢子悬液(108细胞/mL).加入细胞培养液(RPMI1640+10%FBS),分别在0、2、4、6h取材固定,进行扫描电镜观察。另取定量分生孢子悬液(108细胞/mL)孵育在包被有定量纤连蛋白(50μg/mL)的微量测定细胞培养板中,37℃ 3h;用磷酸盐缓冲液冲洗3次,再分别加入兔抗人-纤连蛋白抗体和羊抗兔IgG-FITC孵育(37℃ 1h),胰酶消化后冲洗固定立即送共距焦免疫荧光电镜检查。结果 静止期分生孢子表面有特征性的小棘状层形式,即有许多刺猬毛状的突起分布在其表面。孢子膨胀生发过程中,刺猬毛样小棘状层结构逐渐降解脱落,大约4h后完全降解脱落。孢子变成由条索状纤维质构成的光滑细胞壁球体;通过共距焦免疫荧光电镜发现FITC均匀地分布粘附在分生孢子表面刺猬毛样棘状突起上。结论 静止期分生孢子的表面可表达出高水平纤连蛋白受体。带有放射荧光标记的纤连蛋白配体-受体均匀地分布在静止期分生孢子最外面小棘状层。孢子膨胀过程中表面小棘状层逐渐脱落。

关键词: 曲霉,烟, 孢子真菌, 受体,纤连蛋白

Abstract: Objective To observe the changes of the outer cell wall in resting phase and during swelling process of conidia of Aspergillus fumigatus,and the distribution of fibronectin receptors on conidia surface.Methods After being harvested with 3.5～4.0μm steel filters,the conidia were resuspended in RPMI 1640+10%FBS medium.The specimens were observed by the scanning electron microscopy after incubation for 0,2,4,6 hours at 37℃ respectively.The same amount of conidia was resuspendesd in PBS buffer,and put the suspension on 50μg/ml fibronectin coated microtitre plate,after incubation for 3hrs at 37℃,the conidia were washed three times with PBS.Then rabbit antifibronectin antibody and fluorescein isothiocyanate(FITC)conjugated goat anti-rabbit immunoglob-ulin antibody were added,respectively.After digesting with trypsin,the conidia were washed again with PBS and examined by immuno-confocal microscopy.Results Acharacteristic echinulate pattern of protrusions was shown in the cell wall surface of resting conidia.During swelling process,the echinulate layer of swollen conidia was gradually disintegrated,and almost completely shed 4hr later.Then,a smooth cell wall layer covered by some strands of fib-rillar material was shown.Binding of fibronectin to the surface of conidia was revealed by immuno-confocal microscopy.FITC was homogeneously adhered to the protrusions on the conidial outer cell wall layer.Conclusion In resting phase the conidia of A.fumigatus express high levels of fibronectin receptors on their surface,they are ho-mogeneously distributed over the external echinulate layer of the conidia,and shed gradually during the swelling process.

Key words: Spores,fungal, Receptors,fibronectin, Aspergillus fumigatus