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Bioelectrical and cytoskeletal patterns correlate with altered axial polarity in the follicular epithelium of the Drosophila mutant gurken

机译:生物电和细胞骨架模式与果蝇突变体gurken的卵泡上皮细胞的轴向极性改变相关。

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摘要

Comparison of wt and follicles. The dorsal side of wt S10B is defined by a thicker, columnar follicular epithelium (FE) and by an anterodorsal position of the oocyte nucleus (ON, red circle; cFC, centripetal follicle cells; mFC, mainbody follicle cells; pFC, posterior follicle cells). S10B lacks dorsoventral (d-v) polarity and is characterised by a uniform cuboidal, ventralised FE covering the oocyte (Ooc). While, in wt S10B, border cells (BC) are located close to the ON, in S10B, disrupted body-axis formation leads to undefined positioning of BC amongst the nurse cells (NC). The ON is often located at the posterior end of the Ooc in a typical protrusion. Transheterozygous combinations of alleles HF48 and 2B6 result in ventralised follicles of all vitellogenic stages (S8–14; bright-field image). In S12–14, wt-typical dorsal respiratory appendages are missing and a second micropylar structure appears at the posterior end. To visualise basal microfilaments (bMF) and microtubules (MT) in the FE, tangential optical sections using structured-illumination microscopy (SIM; focal plane: red line) were used. For analysis of V - and pH -patterns, median optical sections (SIM; focal plane: turquoise line) were used. Quantification of transversal ( ) and anteroposterior (a-p; ) gradients of V and pH , respectively, in the FE of S10B. Example of a follicle (SIM) where DiBAC-fluorescence intensities of FE (area marked in yellow) and FE (white) as well as of aFE (red) and pFE (blue) were measured using ImageJ (“mean grey value”). In wt follicles, the d-v axis was identified via the anterodorsal position of the ON, and the fluorescence intensities of the dorsal and ventral FE were quantified accordingly
机译:wt和卵泡的比较。 wt S10B的背侧由较厚的圆柱状卵泡上皮(FE)和卵母细胞核的前角位置定义(ON,红色圆圈; cFC,向心卵泡细胞; mFC,主体卵泡细胞; pFC,后卵泡细胞)。 S10B缺乏背腹(d-v)极性,其特征是覆盖卵母细胞(Ooc)的均匀长方体,腹侧FE。虽然在wt S10B中,边界单元(BC)的位置接近ON,但在S10B中,体轴形成的中断会导致BC在护士单元(NC)之间的定位不确定。 ON通常位于典型突起中Ooc的后端。等位基因HF48和2B6的超杂合组合导致所有卵黄形成期的腹面卵泡(S8-14;明场图像)。在S12-14中,典型的背侧呼吸附件缺失,第二个毛孔结构出现在后端。为了可视化FE中的基底微丝(bMF)和微管(MT),使用了结构照明显微镜(SIM;焦平面:红线)的切向光学切片。为了分析V和pH模式,使用了中值光学部分(SIM;焦平面:绿松石线)。在S10B的有限元分析中,分别对V和pH的横向()和前后(a-p;)梯度进行定量。滤泡(SIM)的示例,其中使用ImageJ(“平均灰度值”)测量了FE(黄色区域)和FE(白色)以及aFE(红色)和pFE(蓝色)的DiBAC荧光强度。在野生型卵泡中,d-v轴是通过ON的前体位置确定的,因此对背侧和腹侧FE的荧光强度进行了定量

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