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. 2020 Jul 31;9:e58619. doi: 10.7554/eLife.58619

Figure 2. INX-8(T239I D24N) compromises gap junction formation between soma and germline.

(A) Somatic INX-8 in wild-type (WT) gonads localizes to puncta representing soma-germline gap junctions where DTC or sheath cells overlie germ cells. (B) In WT distal gonad arms, gap junction puncta aggregate in small clusters associated with each germ cell; in inx-8(rf) distal arms, gap junction puncta localize more apically, where sheath dips down between germ cells, outlining the cell (honeycomb pattern). Localization of INX-21 and INX-22 define the two different classes of germline gap junction hemichannels, both of which include INX-14 subunits. (C) At the loop region, WT gap junction puncta form large aggregates (arrows) which are absent in inx-8(rf). (D) In WT proximal arms, gap junction formation with INX-22-containing hemichannels is extensive; in inx-8(rf), INX-22 and INX-8(T239I D24N) are strongly expressed but show little evidence of gap junction formation. ‘–1’ indicates the most proximal oocyte; inx-8(rf) hermaphrodites frequently show undersized oocytes at this position as well (<). (E) Heterozygous +/inx-8(rf) shows little effect on gap junction localization in an emb-8(hc69ts) background at 22°C. Antibodies specific to INX-8, INX-21, or INX-22 were used as indicated. DAPI-stained DNA in blue. Bar, 50 μm.

Figure 2.

Figure 2—figure supplement 1. INX-8(rf)::GFP can be endocytosed by oocytes.

Figure 2—figure supplement 1.

(A) inx-8(+)::gfp expressed in the somatic gonad sheath is endocytosed during ovulation and appears as distinct fluorescent puncta in early embryos. (B) inx-8(rf)::gfp expressed on an extrachromosomal array in inx-8(0) inx-9(0) animals is endocytosed by ovulating oocytes. (C) inx-8::gfp expressed by the DTC in inx-8/9 (0); Ex[inx-8(DTC+,Sh-)] is not observed in early embryos. Bar, 20 μm.