Annual Report 2006

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D evelopmental Biology Department Expression of a nanos homologue in eggs The silkworm has no recognizable germ plasm, a morphologically distinguishable egg cytoplasm that contains germ cell determinant(s), and its germ cells appear first on the ventral side of the embryo, not on the posterior pole as in During the search of germ cell specific genes in we found that transcripts of a homologue were dispersed along the ventral midline of the egg, the region where germ cell formation occurs in the future (Fig. 1a). In later stages of embryogenesis, the transcript was observed specifically in the germ cells (Fig. 1b). The localization of the transcripts may be interpreted as the presence of germ plasm in the ventral midline of the egg. However, its composition and localization mechanisms seem to be different from those of suggesting that has developed a specific mechanism(s) of germ cell formation during the evolution. Application of the tetracyclin controlled binary gene expression system (tTA/TRE system) to the sawfly, One of the powerful tools to analyze gene function in non-model organisms is the technique to regulate gene expression. We demonstrated that a transgenesis-based binary gene expression system worked in (Fig. 2). Embryos bearing both tetracyclinecontrolled transactivator (tTA) and a marker EGFP gene driven by the tTA response element (TRE-EGFP) expressed EGFP, while the embryos bearing either the tTA or the TRE-EGFP did not express EGFP. The EGFP expression in embryos bearing both tTA and TRE-EGFP was suppressed when tetracycline was orally applied to the parents at a concentration of 100 µg/ml. Fig. 2 The tTA/TRE binary expression system works in the sawfly, The embryo bearing tTA/TRE -EGFP expresses the marker EGFP (left panel, middle embryo). The expression was suppressed by application of tetracycline (right panel, middle embryo). Fig. 1 Expression pattern of a homologue during the early development (a) An egg at cleavage stage. Ventral view. Anterior is to the left. (b) An embryo after blastokinesis. The transcript is specifically observed in the gonad (arrow).
Expression patterns of two novel genes isolated from regenerating individuals of an oligochaete annelid, The spatiotemporal expression profiles of two novel genes, and which were isolated from regenerating were examined by RT-PCR and whole mount hybridization (WISH). For both genes, the expression was scarcely detectable in intact worms, but was dramatically activated following amputation. The expression levels were highest at the blastema formation stage (around 12-24 hours after amputation). The results suggest that these genes play important roles in annelid regeneration (Fig. 3). Ppet genes have a function on differentiation / proliferation of ES cells and also early development in the mouse We have previously reported that mouse ES cells were divided into three subpopulations according to the expression levels of platelet endothelial cell adhesion molecule 1 (PECAM-1) and stage-specific embryonic antigen (SSEA)-1. Quantitative RT-PCR and chimera formation revealed that the expression level of PECAM-1 and SSEA-1 were positively correlated with pluripotency of ES cell subpopulations. In order to identify novel regulatory factors in ES cell differentiation, we have performed comparison of gene expression profiles by oligo-DNA array analysis between three subpopulations. In this fiscal year, we focused on uncharacterized 23 genes (Ppet: PECAM-1 Positive ES cell-derived Transcripts) and attempt to elucidate the functions of them in ES cells and early embryos by small interfering RNA (siRNA) mediated gene knockdown. When fluorescein-labeled Ppet siRNAs were transfected into ES cells, plating efficiency of these ES cells showed no significant difference between fluorescein negative and positive cells in all groups. Akaline phosphatase (AL-P, a marker for undifferentiated ES cells) positive colonies were decreased by knockdown of Ppet 002, 005, 010, 015, 021 and 023 gene as well as Oct3/4 (Fig. 4). In contrast, AL-P positive colonies were increased by knockdown of Ppet 001, 003, 004 and 019 genes. Next, we performed knockdown of Ppet gene in early mouse embryos. The embryos that injected Ppet015, 019 and 021 siRNA exhibited developmental retardation and degradation. In cases of Ppet 019 and 021 gene knockdown, total cell numbers of blastocysts were reduced without morphological abnormality. Conversely, Ppet 010, 022 and 023 gene knockdown caused developmental facilitation. In cases of Ppet 022 Fig. 3 Expression patterns of and RT-PCR was performed using the total RNAs from intact and regenerating at 3-96 hours after amputation (A). The number of PCR cycles used are indicated at the right of each panel. WISH analysis was performed using DIG-labeled antisense riboprobes against regenerating fragments at 24 (B-D) or 36 hours after amputation (E). Lateral views, the anterior is to the left. Arrowheads indicate the base of the blastema. e, esophagus; g, gut. Scale bar = 200 µminB.
Page 1 and 2: 2006
Page 3 and 4: Teruo Ishige, President The Nationa
Page 5 and 6: List of Publication Original paper
Page 7 and 8: Topics of Research in This Year Pos
Page 9 and 10: studies. To construct a fully satur
Page 11 and 12: Fig. 2 Geographical distribution of
Page 13 and 14: (ABR0015, ABR0257, ABR0417, ABR0495
Page 15 and 16: Mulberry latex defense mulberry tre
Page 17 and 18: Fig. 2 Growth inhibitory effects of
Page 19 and 20: Fig. 3 Change of the levels of luci
Page 21 and 22: Next, we examined the effect of tra
Page 23 and 24: G enome Research Department Introdu
Page 25 and 26: The Rice Annotation Project Databas
Page 27 and 28: industrial and agricultural concern
Page 29 and 30: One of the main problems involving
Page 31 and 32: Fig. 3 Mapping of QTL for rachis in
Page 33 and 34: two genes, XOO2263 and XOO4134 cont
Page 35 and 36: G enebank Genebank Project The NIAS
Page 37 and 38: using dormant winter buds. Also, we
Page 39: Insect and Animal Sciences Division
Page 43 and 44: Fig. 5 PGC and EBC fused cells PGCs
Page 45 and 46: cells. The epidermal cells maintain
Page 47 and 48: Animal and cell culture models for
Page 49 and 50: terminus, respectively. A signal se
Page 51 and 52: Fig. 2 Trehalose content of body pa
Page 53 and 54: Fig. 6 Birth weight of 11 adult SNT
Page 55 and 56: The role of glucose as a metabolic
Page 57 and 58: analysis using microsatellite marke
Page 59 and 60: Fig. 2 Recorded Electromyograms (le
Page 61 and 62: Table 1 Menu of Commercial silkworm
Page 63 and 64: Fig. 3 Expression of GFP gene in th
Page 65 and 66: also surveyed in the genetic stocks
Page 67 and 68: esearch were located in or near gen
Page 69 and 70: elatives results in severe reductio
Page 71 and 72: and circadian clocks may control th
Page 73 and 74: B iochemistry Department Structural
Page 75 and 76: Proteomic analysis Based on the pro
Page 77 and 78: photoautotrophic medium (Fig. 1B).
Page 79 and 80: Fig. 4 Blast resistance of OsWRKY45
Page 81 and 82: phosphatase activity by SIPK depend
Page 83 and 84: the endosperm in co-operation with
Page 85 and 86: isolated genes in transgenic Anal
Page 87 and 88: Fig. 1 Gene structures and mutation
Page 89 and 90: cytochimera among the mulberries (
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15 G. Hariraj, Kinoshita Haruo, T.
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49 Kameda Tsunenori, Teramoto Hidet
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85 Matsuyama Shuichi, Ohkura Satosh
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120 Ozawa Kenjiro, Kawahigashi Hiro
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155 Taguchi-Shiobara Fumio, Yamamot
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192 Ueno Osamu, Agarie Sakae, (2005
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8 Murakami Ritsuko, Koyama Akio, Sh
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20 Todoroki Junichi, Kaneko Hiroyuk
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Executive Members and Research Staf
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Developmental Biology Department Di
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Research Leader Hayasaka Shoji Rese
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Mutation Genetics Laboratory Head N
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thousands of yen TOTAL BUDGET OPERA
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Annual Report 2006 (Apr. 2005Mar. 2
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Annual Report 2006

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