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Volume 1, Issue 2, Pages 187-199 (August 2001) Two Zinc Finger Proteins, OMA-1 and OMA-2, Are Redundantly Required for Oocyte Maturation in C. elegans  Michelle R. Detwiler, Melanie Reuben, Xiumin Li, Eric Rogers, Rueyling Lin  Developmental Cell  Volume 1, Issue 2, Pages 187-199 (August 2001) DOI: 10.1016/S1534-5807(01)00026-0

Figure 1 Sequence Analysis of oma-1 and oma-2 Genes, Mutant Alleles, and Related Sequences (A) Alignment of OMA-1 and OMA-2 protein sequences with TIS11 fingers indicated by lines above. (B) Alignment of the two zinc fingers of OMA-1, OMA-2, five other C. elegans proteins, and mouse TIS11 (mTIS11). Black and gray backgrounds indicate residues identical and similar, respectively, to those in OMA-1. Arrows indicate the conserved C-C-C-H residues. Asterisks mark amino acids changed in four missense mutations (te36, te21, te22, and te50). (C) Schematic diagrams of oma-1 and oma-2 genomic sequences and positions of each mutation. Mutations are indicated by allele numbers, with nucleotide changes in small letters and amino acid changes in caps. Boxes are exons and lines are introns. Position 1 of the nucleotide sequence refers to the “a” of the initiating atg. M, ectopic initiator methionine; ss, splice site mutation; stop, nonsense mutation. Gray indicates zinc fingers Developmental Cell 2001 1, 187-199DOI: (10.1016/S1534-5807(01)00026-0)

Figure 2 Comparison of Gonad Morphology for Wild-Type, Oma, and Female Animals (A) Schematic diagram of a wild-type C. elegans adult hermaphrodite gonad arm. Germ cells proliferate mitotically in the distal region and then enter prophase of meiosis I. Distal germ nuclei (open circles) are in a syncitial cytoplasm, whereas proximal germ cells are enclosed by membranes. The −1 oocyte is the most proximal and most developed oocyte. (B–G) Light micrographs, all at the same magnification, of wild-type (B), oma-1(RNAi); oma-2(RNAi) (C), oma-1(te33);oma-2(te51) (D), and tra-2(q122gf)/+ (E), tra-2(q122gf)/+; oma-1(te33);oma-2(te51), mated with wild-type N2 males (F), tra-2(q122gf)/+;oma-1(te33); oma-2(te51), unmated (G) gonads. The portion of each gonad arm containing oocytes is indicated between the two arrowheads in (B) and (C). ut, uterus; sp, spermatheca. The scale bar represents 10 μm Developmental Cell 2001 1, 187-199DOI: (10.1016/S1534-5807(01)00026-0)

Figure 3 Expression of OMA-1 and OMA-2 Proteins (A) Extracts of bacteria expressing no fusion protein (lane 1), GST-OMA-1 (lane 2), or GST-OMA-2 (lane 3), and whole-worm extracts from wild-type N2 (lane 4), oma-1(te33) (lane 5), and oma-2(te51) (lane 6) were probed with antibodies to OMA-1, OMA-2, or actin. (B) Worm lysates from wild-type N2, glp-4(bn2), fem-3(q20sd), or fem-3(e2006) grown at either 15°C or 25°C probed with antibodies to OMA-1, OMA-2, or actin. (C) Dissected wild-type gonads stained with affinity-purified antibodies to either OMA-1 or OMA-2. The scale bar represents 10 μm Developmental Cell 2001 1, 187-199DOI: (10.1016/S1534-5807(01)00026-0)

Figure 4 Oocytes Are Arrested at Diakinesis in Oma Gonads Dissected gonads from wild-type (C), oma-1(te33);oma-2(te51) (D), lin-3(n1058) (E), fem-3(e1950) (F), and oma-1(RNAi);oma-2(RNAi) (G) were stained with DAPI. Wild-type (A) and Oma (B) gonads were also stained with the H3P antibody. Arrows in (A)–(D) point to the most distal oocytes with detectable H3P staining. sp, spermatheca. The scale bar represents 10 μm Developmental Cell 2001 1, 187-199DOI: (10.1016/S1534-5807(01)00026-0)

Figure 5 Comparison of Nuclear Envelope in Wild-Type and Oma Oocytes (A and B) Close up micrographs of live wild-type (A) and oma-1(te33);oma-2(te51) (B) proximal gonads. The arrows in (A), (D), and (E) point to nuclei in the −1 oocyte. Note that the −1 oocyte in (A) and (D) has initiated NEBD and that chromosomes in (E) have compacted further to form a tight cluster, whereas chromosomes in Oma oocytes remain scattered (F and G). (C and H) High magnification of wild-type (C) and Oma (H) −1 oocyte nuclear lamin staining. In wild-type, the lamin staining becomes wrinkled (C) then cytoplasmic (D) upon oocyte maturation. sp, spermatheca. The scale bar represents 10 μm Developmental Cell 2001 1, 187-199DOI: (10.1016/S1534-5807(01)00026-0)

Figure 6 Two Sperm-Dependent Events Do Not Occur in Oma Gonads Gonads dissected from wild-type (A, D, and G), oma-1(te33);oma-2(te51) (B, E, and H), or fem-3(e1950) (C, F, and I) are stained with MAPK-YT antibody (A–C), DAPI (D–F), or AIR-2 antibody (G–I). Brackets in (A) and (D) indicate the pachytene zone where MAP kinase is also activated to a low degree. Arrowheads in (A)–(C) point to weak MAPK-YT staining in distal oocytes. Arrows in (G)–(I) point to the position of −1 oocyte nuclei. Neither Oma nor Fem-3 gonads show chromosomal association of AIR-2. sp, spermatheca. The scale bar represents 10 μm Developmental Cell 2001 1, 187-199DOI: (10.1016/S1534-5807(01)00026-0)

Figure 7 wee-1.3(RNAi) Suppresses Oma Prophase Arrest Live micrographs (A and B) or DAPI staining (C and D) of wild-type (A and C) or oma-1(te33);oma-2(te51) (B and D) animals fed wee-1.3 dsRNA-producing bacteria. Brackets point to the uterus. One oocyte from each uterus is outlined with a dashed line. Arrows point to polyploid nuclei in the gonad arm, and arrowheads indicate nuclei remaining in diakinesis with six chromosomal bivalents. ut, uterus; sp, spermatheca. The scale bar represents 10 μm Developmental Cell 2001 1, 187-199DOI: (10.1016/S1534-5807(01)00026-0)