Henry C B, Kleinstein E, Shum W, DeFouw D O
Department of Anatomy, Cell Biology and Injury Sciences, UMDNJ-New Jersey Medical School, Newark, USA.
Anat Rec. 1995 Mar;241(3):411-6. doi: 10.1002/ar.1092410316.
The chorioallantoic membrane (CAM) of the chick embryo expands during embryogenesis to meet the increased oxygen demands during growth and differentiation. Temporal and spatial glycosylation patterns of CAM ectodermal and endodermal proteins likely contribute to differentiation of the functional attributes of the CAM.
Using lectins for light and electron microscopic observations, we studied the patterns of glycoconjugate expression on the ectoderm and endoderm of the chorioallantoic membrane (CAM) of the chick at days 4.5, 5.0, 5.5, 6.0, and 10 of morphogenesis. For light microscopy, samples of unfixed CAM were incubated with the following FITC lectins: Con A, DBA, GSA-I, GSA-II, PNA, SBA, UEA-I, and WGA.
All lectins, except GSA-I and -II, gave positive results. The positive lectins, labeled with HRP, served to ultrastructurally localize PNA, SBA, and WGA, but not DBA binding to the luminal surface of the endoderm. UEA-I and Con A bound similarly except on day 10 when UEA-I no longer bound. On the ectodermal surface, only WGA bound at all times studied. PNA and SBA binding were present from days 5.0 to 6.0 but absent at days 4.5 and 10. DBA binding occurred through day 5.0 but was absent thereafter. UEA-I bound to the ectoderm at days 4.5, 5.0, and 10 but not days 5.5 and 6.0. Con A bound only on days 5.0 and 10.
That the ultrastructurally similar ectoderm and endoderm of the CAM display functional differences conforms to the hypothesis that differential expression of glycoconjugate microdomains likely contributes to such functional specialization.
鸡胚的绒毛尿囊膜(CAM)在胚胎发育过程中会扩张,以满足生长和分化过程中增加的氧气需求。CAM外胚层和内胚层蛋白的时空糖基化模式可能有助于CAM功能属性的分化。
我们使用凝集素进行光镜和电镜观察,研究了鸡胚绒毛尿囊膜(CAM)在形态发生的第4.5、5.0、5.5、6.0和10天,外胚层和内胚层上糖缀合物的表达模式。对于光镜检查,将未固定的CAM样本与以下异硫氰酸荧光素(FITC)凝集素孵育:刀豆球蛋白A(Con A)、大豆凝集素(DBA)、花生凝集素(GSA-I)、花生凝集素(GSA-II)、豌豆凝集素(PNA)、大豆凝集素(SBA)、荆豆凝集素I(UEA-I)和小麦胚凝集素(WGA)。
除GSA-I和GSA-II外,所有凝集素均给出阳性结果。用辣根过氧化物酶(HRP)标记的阳性凝集素用于超微结构定位PNA、SBA和WGA,但未用于定位与内胚层腔表面结合的DBA。除了在第10天UEA-I不再结合外,UEA-I和Con A的结合情况相似。在外胚层表面,在所有研究时间点只有WGA结合。PNA和SBA的结合在第5.0至6.0天存在,但在第4.5天和第10天不存在。DBA的结合持续到第5.0天,但此后不存在。UEA-I在第4.5、5.0和10天与外胚层结合,但在第5.5天和第6.0天不结合。Con A仅在第5.0天和第10天结合。
CAM超微结构相似的外胚层和内胚层表现出功能差异,这符合糖缀合物微结构域的差异表达可能有助于这种功能特化的假设。