Isoprenoids Control Germ Cell Migration Downstream of HMGCoA Reductase

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Isoprenoids Control Germ Cell Migration Downstream of HMGCoA Reductase Ana C Santos, Ruth Lehmann Developmental Cell Volume 6, Issue 2, Pages 283-293 (February 2004) DOI: 10.1016/S1534-5807(04)00023-1

Figure 1 The Isoprenoid/Cholesterol Biosynthetic Pathway in the Fly There are two main branches downstream of HMGCoAr, the cholesterol biosynthetic branch (shaded in green) and the isoprenoid biosynthetic branch (Moebius et al., 2000). The isoprenoids/cholesterol pathway (also called mevalonate pathway) produces many other molecules (terpenoids, Vitamins K and E, ubiquinone) that are not shown for simplicity. Dashed, gray arrows represent enzymes absent in the fly genome (see Results). Genes with germ cell migration defects are indicated in red. Developmental Cell 2004 6, 283-293DOI: (10.1016/S1534-5807(04)00023-1)

Figure 2 fpps and qm Are Highly Expressed in the Mesoderm RNA expression profile of several genes in the mevalonate pathway. Anterior is to the left and dorsal is up in all panels (except for B4 which is a frontal view). All embryos are lateral views except (A2), which is a top view. (A) Most genes in the pathway are broadly expressed in the embryo throughout development but show increased expression in tissues such as (A1) pharynx and esophagus (arrow) (CG33009/mevalonate kinase), (A2) ring gland (arrow) (CG10268/phosphomevalonate kinase), and (A3) esophagus (arrow) (CG8239/diphosphomevalonate decarboxylase). (B) Hmgcr RNA expression. Hmgcr is expressed in the mesoderm at stage 10 (B1) and 11 (B2) and gets restricted to the SGPs (arrowheads) at stage 12 (B3) and in coalesced gonads (B4). (C) fpps RNA expression profile. fpps is expressed throughout the mesoderm at stage 10 and 11 (C1 and C2, respectively). As the germ band retracts (C3) mesodermal expression is no longer predominant and after germ band retraction fpps is mostly expressed in the CNS (C4) and ring gland (not shown). (D) qm RNA expression profile. qm is expressed in the visceral mesoderm throughout development, from stage 10 (D1) and 11 (D2) to stage 13 (D3). At the end of embryogenesis, qm is highly expressed in the CNS (D4). Developmental Cell 2004 6, 283-293DOI: (10.1016/S1534-5807(04)00023-1)

Figure 3 fpps and qm Mutant Embryos Display Germ Cell Migration Defects Embryos of the indicated genotypes were labeled with α-Vasa to mark the germline. Dorsal view, anterior to the left in all panels. All embryos are at stage 14. (A) Wild-type embryo, (B) Hmgcrclb18 embryo showing several germ cells that failed to migrate into the gonad (arrow) and few germ cells that correctly migrated and formed gonads (white oval), (C) fppsk06103embryo showing several lost germ cells (arrow), (D) qmL14.4 embryo showing some lost germ cells (arrow), (E) fppsM−Z− showing many lost germ cells (arrow) and very few germ cells in gonads (white oval), and (F) fpps;qm embryo showing many lost germ cells (arrow) and no germ cells in the gonads (white oval). Developmental Cell 2004 6, 283-293DOI: (10.1016/S1534-5807(04)00023-1)

Figure 4 Mutations in Either fpps or qm Enhance Weak Hmgcr Phenotype Embryos of the indicated genotypes were labeled with α-Vasa to mark the germline. Dorsal view, anterior to the left in all panels. All embryos are at stage 13–14. (A) Hmgcr01152 embryo showing few lost germ cells. (B) fpps;Hmgcr embryo showing many germ cells that did not undergo normal germ cell migration and (C) qm;Hmgcr embryo showing several lost germ cells. Developmental Cell 2004 6, 283-293DOI: (10.1016/S1534-5807(04)00023-1)

Figure 5 fpps and qm Are Sufficient to Attract Germ Cells to the Nervous System Embryos of the indicated genotypes were labeled with α-Vasa to mark the germline except (B), which shows Hmgcr RNA expression. Dorsal view, anterior to the left in all panels. All embryos are at stage 13–14. (A) elavGal4;UAS-Hmgcr showing several germ cells being attracted to the nervous system (arrow). (B) High level of Hmgcr RNA expression in the nervous system in elavGal4;UAS-Hmgcr (compare to Figures 2B3 and 2B4). (C) elavGal4;UAS-fpps and (D) elavGal4;UAS-qm. (C and D) Few germ cells close to the CNS (arrows). (E) Summary of fpps, qm, and Hmgcr misexpression phenotypes. Percentage of embryos having 0 to 2, 3 to 4, or more than 5 germ cells ectopically located in the nervous system. N = total number of embryos analyzed at stage 13 for each genotype. Developmental Cell 2004 6, 283-293DOI: (10.1016/S1534-5807(04)00023-1)

Figure 6 Geranylgeranyl Transferase Type I Mutant Embryos Display Germ Cell Migration Defects Embryos of the indicated genotypes were labeled with α-Vasa to mark the germline. Dorsal view, anterior to the left in all panels. All embryos are at stage 14. (A) β GGT-Is−483 and (B) β GGT-Ixs−2554 show some lost germ cells (arrow). Developmental Cell 2004 6, 283-293DOI: (10.1016/S1534-5807(04)00023-1)