摘要
胰岛素作为一种具有多种生物学效应的激素,除了在人们所熟知的蛋白质、脂肪、糖原的代谢中发挥重要的调节作用外,在调控细胞生长、增殖,保证正常的生长发育等方面也有重要的作用。近10年来,胰岛素信号传导途径对细胞大小和数目调控的研究取得了突破性的进展,弄清楚了整个信号传导途径中的大多数基因,关键分子的功能及其磷酸化调控。整个胰岛素信号传导途径是在线虫、果蝇和老鼠等模式动物中发现的,是一条相对保守的信号传导途径。
家蚕是新兴的遗传与发育研究的模式昆虫,是除果蝇以外最好的昆虫遗传学研究材料。利用家蚕研究昆虫生理和发育生物学可以克服以果蝇作为模式生物的一些缺点:家蚕个体大小适中,发育周期相对较短,繁殖能力较强,容易饲养,便于进行解剖、培养、生理测定等实验操作。家蚕中的胰岛素类似物是家蚕素(bombyxin);虽然家蚕基因组框架草图已经完成,但是很少有家蚕bombyxin的分子作用机制和胰岛素信号途径的相关报道。脂肪体是昆虫的营养代谢中心和能量贮存中心,胰岛素信号可能既快速地调控昆虫脂肪体的生理功能,又长期缓慢地调控其器官发育。
为了进一步研究家蚕胰岛素信号传导途径对脂肪体生理功能和器官发育的分子机理,本毕业论文已得出以下结论:
1家蚕胰岛素信号传导途径中上游基因IRS、PI3K110、PI3K60、PTEN、PDK和Akt同人、果蝇、伊蚊和赤拟谷盗的对应基因比对表明,它们编码的氨基酸序列和昆虫相对应的序列相似性很高,和人的序列比较相似性相对较低。其中,家蚕和其它昆虫的PI3K60和PTEN具有相同的蛋白质结构域,和人PI3K60和PTEN的蛋白质结构域有较大的差别,昆虫之间的也有较大差异:而Akt和PDK在这几个基因中最为保守。
2检测家蚕中胰岛素信号通路中上游基因InR、IRS、PI3K110、PI3K60、PTEN、PDK和Akt在脑、神经节、生殖腺、马氏管、中肠、丝腺和脂肪体组织中分布及表达水平发现,相对于内参基因rp49,这些基因表达的绝对量都很低,但这些基因在脂肪体和脑中表达量相对较高。1)除了Akt外,其它几个基因均在脂肪体中有较高水平的表达;2)脑中Akt表达水平较高,其它几个基因也有较高水平的表达;3)在丝腺中所有基因的表达水平都很低。
3检测从3龄1天到成虫2天,家蚕脂肪体中InR、IRS、PI3K110、PI3K60、PTEN、PDK和Akt的表达水平发现,InR、IRS、PTEN和PI3K110的表达量都分别在3龄末期、4龄末期和5龄末期出现了明显的峰值,而Akt、PDK和PI3K60在预蛹之前的表达量都非常低,没有显著的变化。
4 5龄1天的家蚕在受到饥饿刺激后,脂肪体中InR、IRS和PDK都有大幅度升高,PI3K110、PI3K60和Akt的表达水平都上升了1倍,PTEN的表达量基本没有变化。5龄5天的家蚕受到饥饿刺激后,所有基因的表达水平都没有非常显著的变化。这说明5龄1天家蚕脂肪体中胰岛素信号传导途径中上游基因的表达水平对于饥饿刺激更加敏感。
Insulin,as a multiple-function hormone,plays an important role in the regulation of protein、lipid and carbohydrate metabolism.Insulin is also essential to modulate cell growth and proliferation and to ensure normal body growth and development.During the last decade,many progresses have been made in understanding how insulin signaling pathway regulates cell size and cell number,including gene identification,elucidating the physiological function and phosphorylation regulation of key molecules involved in the pathway.Most of the insulin signaling pathway was explored in the model animals such as the worm(Caenorhabditis elegans),the fruitfly(Drosophila melanogaster),and the mouse(Mus musculus).The insulin signaling pathway is comparatively conserved during the evolution in the animal kingdom.
The silkworm(Bombyx mori) is a new model insect in the studies of genetics and developmental biology.Except for Drosophila melanogaster,Bombyx mori is the best model for insect research. The silkworm has several advantages in the studies of insect physiology and developmental biology, for example:a suitable body size、a relatively short lifespan、a strong reproductive ability and conveniences for maintenance、dissection、culture、bioassay and other experimental operations.The homologue of insulin in B.mori is bombyxin.Although the draft map of silkworm genome has been completed,studies on the molecular mechanism of bombyxin and insulin signaling pathway in B. mori is very rare.Fatbody is the center for nutrient metabolism and energy storage in insects.The insulin signaling might be important to regulate its physiological function in a short-term and rapid manner and its organ development in a long-term and slow manner.
To further understand how insulin signaling regulates fatbody function and development in B. mori,several conclusions might be drawn below based on the experiments conducted in this master thesis:
1.First,the amino acid sequences in the upstream genes of the insulin signaling pathway(IRS, PI3K110,PI3K60,PTEN,PDK and Akt) in B.mori have very high similarity to those of D. melanogaster、Aedes Aegypti and Tribolium castaneum,and comparatively low similarity to those of human.The protein domains of PI3K60 and PTEN are the same in insects,but are slightly different from those of human.Akt and PDK are the mostly conserved genes in this pathway.
2.Second,compared with the control gene rp49,the absolute expression levels of the upstream genes of the insulin signaling(InR,IRS,PI3K110,PI3K60,PTEN,PDK and Akt) are low in all tissues,including brain,nerve cord,testis and ovary,malpighian tube,midgut,silk gland,and fat body.These genes have the highest expression levels in brain and fat body.1) Except for Akt, expression levels of the other genes are comparatively high in fat body;2) Akt is highly expressed in brain,and other genes are also comparatively highly expressed in this tissue;3) All genes have the lowest expression levels in the silk gland.
3.Third,from day 1 of the third instar to day 2 of the adults,the expression levels of InR,IRS, PTEN and PI3K110 in the fat body reach a peak at the end of each larval stage.In contrast, before pupariation,the expression levels of Akt,PDK and PI3K60 in the fat body are always low and have no significant variation.
4.The expression levels of InR,IRS and PDK increase dramatically after 24 hours of starvation on day 1 of the fifth instar,PI3K110,PI3K60 and Akt increase～1-fold,but PTEN has no significant change.However,on day 5 of the fifth instar,the expression levels of all upstream genes in the insulin signaling pathway do not change dramatically after 24 hours of starvation.It indicates that the expression levels of those genes in the fat body are much more sensitive to starvation on day 1 than day 5 of fifth instar.
引文
[1] I Claeys, G Simonet, J Poels,L.T. Van, L Vercammen, L.A. De, B.J. Vanden.Insulin-Related Peptides and Their Conserved Signal Transduction Pathway[ J ].Peptides. 2002 ,23(4):807-816.
[2] L.C. Cantley.The Phosphoinositide 3-kinase Pathway[ J ]. Science. 2002, 296(5573):1655-1657
[3] H Stocker, M Andjelkovic, Oldham S Oldham,M Laffargue, M.P. Wymann, B.A.Hemmings, E Hafen. Living with Lethal PIP3 Levels: Viability of Flies Lacking PTEN Restored by a PH Domain Mutation in Akt/PKB[ J ]. Science. 2002,295(5562) :2088-2091
[4] T Maehama, J.E. Dixon.The Tumor Suppressor, PTEN/MMAC1, Dephosphorylates the Lipid Second Messenger, Phosphatidylinositol 3,4,5-trisphosphate[ J ]. J Biol Chem. 1998 ,273(22):13375-13378
[5] J.M. Edward, S Kei, J.A. Laura, R Leah, S Natalia, M Rodolfo, R.A. Dario. Role that Phosphorylation of GSK3 Plays in Insulin and Wnt Signalling Defined by Knockin Analysis[ J ].The EMBO Journal. 2005,24(8):1571-1583
[6] A.C. Gingras, B Raught, S.P. Gygi, A Niedzwiecka, M Miron, S.K. Burley, R.D.Polakiewicz, C.A. Wyslouch, R Aebersold, N Sonenberg. Hierarchical Phosphorylation of the Translation Inhibitor 4EBP1[ J ]. Genes Dev.2001 ,15(21):2852-2864
[7] B.M. Burgering, G.J. Kops. Cell Cycle and Death Control: Long Live Forkheads[ J ].Trends Biochem Sci. 2002 ,27(7):352-360
[8] P.F. Dijkers, R.H. Medema, C Pals, L Banerji, N.S. Thomas, E.W. Lam, B.M.Burgering, J.A. Raaijmakers, J.W. Lammers, L Koenderman, P.J. Coffer. Forkhead Transcription Factor FKHR-L1 Modulates Cytokine-dependent Transcriptional Regulation of p27(KIPl) [ J ]. Mol Cell Biol. 2000 , 20(24):9138-9148.
[9] M Holzenberger, J Dupont, B Ducos, P Leneuve, A Geloen, P.C. Even, P Cervera,B.Y. Le. IGF-1 Receptor Regulates Lifespan and Resistance to Oxidative Stress in Mice[ J ]. Nature. 2003 ,421(6919):182-187.
[10] M Bluher, R Paschke.Analysis of the Relationship between PPAR-gamma 2 Gene Variants and Severe Insulin Resistance in Obese Patients with Impaired Glucose Tolerance. Exp Clin Endocrinol Diabetes[ J ]. 2003, 111(2):85-90
[11]岳国华,朱尚权.胰岛素受体结构与功能研究概况[J].生物化学与生物物理进展.1992,19(1):1-5
[12]T Kjeldsen,F.C.Wiberg,A.S.Andersen.Chimeric Receptors Indicate that Phenylalanine 39 is a Major Contributor to Insulin Specificity of the Insulin Receptor[J].J Biol Chem.1994,269(52):32942-32946
[13]M Taousi,T.R.Levy,P Roach,S.Ⅰ.Taylor,P Gorden.Structural Basis by Which a Recessive Mutation in the α-subunit of the Insulin Receptor Affects Insulin Binding[J].J Biol Chem,1994,269(21):14912-14918
[14]A Ullrich,J Schlessinger.Signal Transduction by Receptors with Tyrosine Kinase Activity[J].Cell.1990,61(2):203-212
[15]Ⅰ Leconte,E Clauser.Two Sequences Flanking the Major Autophosphorylation Site of the Insulin Receptor are Essential for Tyrosine Kinase Activation[J].Biochem J.1995,306(Pt2):465-472
[16]Ⅰ Leconte,C Auzan,A Debant,B Rossi,E Clauser.N-linked Oligosaccharide Chains of the Insulin Receptor Beta Subunit are Essential for Transmembrane Signaling[J].J Biol Chem.1992,267(24):17415-17423
[17]X.J.Sun,P Rothenberg,C.R.Kahn,J.M.Backer,E Araki,P.A.Wilden,D.A.Cahill,B.J.Goldstein,M.F.White.Structure of the Insulin Receptor Substrate IRS-1Defines a Unique signal Transduction Protein[J].Nature.1991,352(6330):73-77
[18]X.J.Sun,S Pons,L.M.Wang,Y Zhang,L Yenush,D Burks,M.G.Myers,E Glasheen,N.G.Copeland,N.A.Jenkins,J.H.Pierce,M.F.White.Role of IRS-2 in Insulin and Cytokine Signaling[J].Nature.1995,377(2):173-177
[19]B.E.Lavan,Ⅴ.R.Fantin,E.T.Chang,W.S.Lane,S.R.Keller,G.E.Lienhard,A Novel 160-kDa Phosphotyrosine Protein in Insulin-treated Embryonic Kidney Cells is a New Member of the Insulin Receptor Substrate Family[J].J Biol Chem.1997,272(34):21403-21407
[20]B.E.Lavan,W.S.Lane,G.E.Lienhard.The 60Dk Phosphotyrosine Protein in Insulin-treated Adipocytes is a New Member of the Insulin Receptor Substrate Family[J].J Biol Chem.1997,272(17):11439-11443
[21]P.S.Dhe,E.A.Ottinger,R.T.Nolte,M.J.Eck,S.E.Shoelson.Crystal Structure of the Pleckstrin Homology-phosphotyrosine Binding(PH-PTB) Targeting Region of Insulin Receptor Substrate 1[J].Proc Natl Acad Sci U S A.1999,96(15):8378-8381
[22] B Giovannone, M.L. Scaldaferri, M Federici,0 Porzio, D Lauro, A Fusco,P Sbraccia, P Borboni, R Lauro, G Sesti. Insulin Receptor Substrate(IRS) Transduction System:Distinction and Overlapping Signaling Potential[ J ].Diabetes Metab Res Rev. 2000,16(6):434-441
[23] M Anai, H Ono, M Funaki, Y Fukushima, K Inukai, T Ogihara, H Sakoda, Y Onishi, Y Yazaki, M Kikuchi, Y Oka, T Asano. Different Subcellular Distribution and Regulation of Expression of Insulin Receptor Substrate(IRS)-3 from Those of IRS-1 and IRS-2[ J ].J Biol Chem,1998,273(45):29686-29692
[24] M Sugimoto, N Yamaguchi, K Kawai.Characterization of Gamma-GTP in a Human Pancreatic Cancer Cell Line [ J ]. Gastroenterol Jpn. 1984, 19(3):227-231
[25] S Djordjevic, P.C. Driscoll.Structural Insight into Substrate Specificity and Regulatory Mechanisms of Phosphoinositide 3-kinases[ J ]. Trends Biochem Sci .2002 ,27(8):426-432
[26] T Balla. Phosphoinositide-derived Messengers in Endocrine Signaling[ J ]. Journal of Endocrinology. 2006, 188: 135-153
[27] B Vanhaesebroeck, S.J. Leevers, K Ahmadi, J Timms, R Katso, P.C. Driscoll, R Woscholski, P.J. Parker, M.D. Waterfield. Synthesis and Function of 3-phosphorylated Inositol Lipids[ J ]. Annu Rev Biochem. 2001 ,70 :535-602
[28] MP Wymann , L P irola. Structure and Function of Phosphoinositide 3-kinases[ J ].Biochim Biophys Acta. 1998 ,1436(1-2) :127-150
[29] J Li,C Yen,K Podsypanina,S Bose, S.I. Wang, J Puc, C Miliaresis, L Roders, C.R.Mc,S.H. Bigner,B.C. Giovanella,M Ittmann,B Tycko, H Hibshoosh, M.H. Wigler,R Parsons. PTEN ,a Putative Protein Tyrosine Phosphatase Gene Mutated in Human Brain ,Breast and Prostate Cancer [J ].Science .1997 ,275(5308):1943-1947.
[30] A Besson, S.M. Robbins, V.W. Yong. PTEN/MMAC1/TEP1 in Signal Transduction and Tumorigenesis[ J ] . Eur J Biochem. 1999 , 263(3): 605-611
[31] K.M. Yamada, M Araki.Tumor suppressor PTEN: Modulator of Cell Signaling ,Growth, Migration and Apoptosis [ J ]. J Cell Sci. 2001 , 114(Pt13):2375-2382.
[32] L.D. Mayo, J.E. Dixon, D.L. Durden, N.K. Tonks, D.B. Donner.PTEN Protects p53 from Mdm2 and Sensitizes Cancer Cells to Chemotherapy[ J ] . J Biol Chem.2002 ,277(7):5484-5489.
[33] F Vazquez, S.R. Grossman, Y Takahashi, M.V. Rokas, N Nakamura, W.R. Sellers. Phosphorylation of the PTEN Tail Acts as an Inhibitory Switch by Preventing its Recruitment into a Protein Complex[J].J Biol Chem.2001,276(52):48627-48630.
[34]P.F.Jones,T Jakubowicz,F.J.Pitossi,F Maurer,B.A.Hemmings.Molecular Cloning and Identification of a Serine/threonine Protein Kinase of the Second-messenger Subfamily[J].Proc Natl Acad Sci U S A.1991,88(10):4171-4175
[35]A Bellacosa,J.R.Testa,S.P.Staal,P.N.Tsichlis.A Retroviral Oncogene akt,Encoding a Serine-threonine Kinase Containing an SH2-1ike Region[J].Science.1991,254(5029):274-277
[36]R.J.Haslam,H.B.Koide,B.A.Hemminhd.Pleckstrin Domain Homology[J].Nature.1993,363(6427):309-310
[37]T.F.Franke,S.I.Yang,T.O.Chan,A Kazlauskas,D.K.Morrison,D.R.Kaplan,P.N.Tsichlis.The Protein Kinase Encoded by the Akt Proto-oncogene is a Target of the PDGF-activated Phosphatidylinositol 3-kinase[J].Cell 1995,81(5):727-736.
[38]N.N.Ahmed,T.F.Franke,A Bellacosa,K Datta,P.M.Gonzalez,T Taguchi,J.R.Testa,P.N.Tsichlis.The Proteins Encoded by c-akt and v-akt Differ in Posttranslational Modification,subcellular localization and oncogenic potential[J].Oncogene.1993,8(7):1957-1963
[39]A Toker,A.C.Newton.Akt/protein Kinase B is Regulated by Autophosphorylation at the Hypothetical PDK-2 Site[J].J Biol Chem.2000,275(12):8271-8274
[40]H Dudek,S.R.Datta,T.F.Franke,M.J.Bimbaum,R Yao,G.M.Cooper,R.A.Segal,D.R,Kaplan,M.E.Greenberg.Regulation of Neuronal Survival by the Serine-threonine Protein Kinase Akt[J].Science.1997,275(5300):661-665.
[41]E.M.Eves,W Xiong,A Bellacosa.Akt,a Target of Phosphatidylinositol 32kinase,Inhibits Apoptosis in a Differentiating Neuroal Cell Line[J].Mol Cell Biol.1998,18:2143-2152.
[42]L.C.Cantley,B.G.Neel.New insight into tumor suppression:PTEN Suppresses Tumor Formation by Restraining the Phosphoinositide 3-kinase/AKT Pathway[J].Proc Natl Acad Sci USA.1999,96(8):4240-4245.
[43]A.D.Kohn,S.A.Summers,M.J.Birnbaum,R.A.Roth.Expression of a Constitutively Active Akt ser/thr Kinase in 3T3-L1 Adipocytes Stimulates Glucose Uptake and Glucose Transporter 4 Translocation[J].J Bio Chem.1996,271(49):31372-31378.
[44]C.E.Stewart,P Rotwein.Growth,Differentiation,and Survival:Multiple Physiological Functions for Insulin-like Growth Factors[J].Physiol Rev.1996,76(4):1005-1026
[45]D.J.Burks,M.J.Font,M Schubert,D.J.Withers,M.G.Myers,H.H.Towery.IRS-2pathways integrate female reproduction and energy homeostasis.Nature.2000,407(6802):377-382
[46]T.P.Neufeld.Shrinkage Control:Regulation of Insulin-Mediated Growth by FOXO Transcription Factors[J].J Biol.2003,2(3):18
[47]P Coffer.OutFOXing the Grim Reaper:Novel Mechanisms Regulating Longevity by Forkhead Transcription Factors[J].Sci Stke.2003,201:PE39
[48]A Antebi.Inside Insulin Signaling,Communication is Key to Long Life[J].Sci Aging Knowledge environ.2004,23:PE25
[49]M Barbieri,M Bonafe,C Franceschi,G Paolisso.Insulin / IGF-I-signaling Pathway:an Evolutionarily Conserved Mechanism of Longevity from Yeast to Humans[J].Am J Physiol Endocrinol Metab,2003,285(5):E1064-E1071
[50]M Tatar,A Bartke,A Antebi.The Endocrine Regulation of Aging by Insulin-like Signals[J].Science.2003,299(5611):1346-1351.
[51]A Richardson,F Liu,M.L.Adamo,R.H.Van,J.F.Nelson.The Role of Insulin and Insulin-like Growth Factor-Ⅰ in Mammalian Ageing[J].Best Pract Res Clin EndocrinolMetab.2004,18(3):393-406.
[52]M Holzenberger,J Dupont,B Ducos,P Leneuve,A Geloen,P.C.Even,P Cervera,B.Y.Le.IGF-1 Receptor Regulates Lifespan and Resistance to Oxidative Stress in Mice[J].Nature.2003,421(6919):182-187
[53]沈方,张晓明,刘伟国,朱晞。胰岛素与胰岛素受体在学习记忆中作用的研究进展[J].神经解剖学杂志.2006,22:359-361
[54]W Kern,A Peters,S.B.Fruehwald,E Deininger,J Born,H.L.Fehm.Improving Influence of Insulin on Cognitive Functions in Humans[J].Neuroendocrinology.2001,74(4):270-280
[55]Q Wu,R.B.Mark.Signaling and Function of Insulin-like Peptides in Insects[J].Annu.Rev.Entomol.2006,51:1-24
[56]H Nagasawa,F Guo,X.C.Zhong,B.Y.Xia,Z.S.Wang,X.J.Qui,D.Y.Wei,E.I. Chen, J.Z. Wang, A Suzuki, A Isogai, Y Hori, S Tamura, H Ishizaki. Large-scale Purification of Prothoracicotropic Hormone of the Silkworm (Bombyx mori) 。 Sci Sin. 1980,23(8):1053-1060
[57] H Nagasawa, H Kataoka, Y Hori, A Isogai, S Tamura, A Suzuki, F Guo, X.C.Zhong, A Mizoguchi, M Fujishita. Isolation and some Characterization of the Prothoracicotropic Hormone from Bombyx mori[ J ]. Gen Comp Endocrinol. 1984,53(1):143-152
[58] H Kondo, M Ino, A Suzuki, H Ishizaki, M Iwami. Multiple Gene Copies for Bombyxin, an Insulin-related Peptide of the Silkmoth Bombyx mori: Structural Signs for Gene Rearrangement and Duplication Responsible for Generation of Multiple Molecular Forms of Bombyxin[ J ]. J. Mol. Biol. 1996,259, 926-937
[59] M Iwami, I Furuya, H Kataoka. Bombyxin-related peptides: cDNA structure and expression in the brain of the hornworm Agrius convolvuli [corrected] [ J ]. Insect Biochem Mol Biol. 1996,26(1):25-32
[60] K.M. Kimura, M Iwami, A Kawakami, H Nagasawa, A Suzuki, H Ishizaki.Structure and expression of bombyxin-related peptide genes of the moth Samia cynthia ricini[ J ]. Gen Comp Endocrinol. 1992, 86(2):257-268
[61] M Iwami, A Tanaka, N Hano, S Sakurai. Bombyxin gene expression in tissues other than brain detected by reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization [ J ].Experientia. 1996, 52(9):882-7
[62] S Satake, M Masumura, H Ishizaki, K Nagata, H Kataoka, A Suzuki, A Mizoguchi. Bombyxin, an Insulin-related Peptide of Insects, Reduces the Major Storage Carbohydrates in the Silkworm Bombyx mori[ J ]. Comp Biochem Physiol B Biochem Mol Biol. 1997,118(2):349-357
[63] H.F. Nijhout, L.W. Grunert. Bombyxin is a Growth Factor for Wing Imaginal Disks in Lepidoptera[ J ]. Proc Natl Acad Sci U S A. 2002, 99(24): 15446-15450
[64] F Goltzene, F Holder, M Charlet, M Meister, T Oka. Immunocytochemical Localization of Bombyx-PTTH-like Molecules in Neurosecretory Cells of the Brain of the Migratory Locust, Locusta migratoria. Cell Tissue Res. 1992 ,269(1):133-140.
[65] C Cao, M.R. Brown.Localization of an Insulin-like Peptide in Brains of Two Flies.Cell Tissue Res. 2001, 304(2):317-321
[66] P Meneses, L.A. De, M Ortiz. A Protein Extract from Drosophila melanogaster with Insulin-like Activity[ J ]. Comp Biochem Physiol A. 1975, 51(2):483-485
[67] Y Ruan, C Chen, Y Cao, R.S. Garofalo. The Drosophila Insulin Receptor Contains a Novel Carboxyl-terminal Extension likely to Play an Important Role in Signal Transduction[ J ]. J Biol Chem. 1995, 270 (9):4236-4243
[68] R Fernandez, D Tabarini, N Azpiazu, M Frasch, J Schlessinger. The Drosophila Insulin Receptor Homolog: a GeneEssential for Embryonic Development Encodes two Receptor Isoforms with Different Signaling Potential[ J ]. EMBO J. 1995,14(14):3373-3384
[69] T Yamaguchi, R Fernandez, R.A. Roth. Comparison of the Signaling Abilities of the Drosophila and Human Insulin Receptors in Mammalian Cells[ J ] .Biochemistry. 1995,34 :4962-4968
[70] W Brogiolo, H Stocker, T Ikeya, F Rintelen, R Fernandez, E Hafen. An Evolutionarily Conserved Function of the Drosophila Insulin Receptor and Insulin-like Peptides in Growth Control[ J ]. Curr Biol. 2001,11(4) :213-221
[71] A.M. Garofalo, E.J. Strait, L.C. Johnson, H.R. La, E.A. Lazarus, G.A. Navratil, M Okabayashi, J.T. Scoville, T.S. Taylor, A.D. Turnbull. Sustained Stabilization of the Resistive-wall Mode by Plasma Rotation in the DIII-D Tokamak[ J ]. Phys Rev Lett. 2002,89(23) :235001
[72] J Song, L Wu, Z Chen, R.A. Kohanski, L Pick. Axons Guided by Insulin Receptor in Drosophila Visual System[ J ]. Science. 2003 ,300(5618):502-505
[73] R Bohni, E.J. Riesgo, S Oldham, W Brogiolo, H Stocker, B.F. Andruss, K Beckingham, E Hafen. Autonomous Control of Cell and Organ Size by CHICO, a Drosophila homolog of Vertebrate IRSl-4[ J ]. Cell. 1999 ,97 (7):865-875
[74] E.J. Rulifson, S.K. Kim, R Nusse. Ablation of Insulin-producing Neurons in Flies: Growth and Diabetic Phenotypes[ J ]. Science. 2002,296 (5570): 1118-1120
[75] H Huang, C.J. Potter, W Tao, D.M. Li, W Brogiolo, E Hafen, H Sun, T Xu. PTEN Affects Cell Size, Cell Proliferation and Apoptosis during Drosophila Eye Development[ J ]. Development. 1999 ,126 (23):5365-5372
[76] J.S. Britton, W.K. Lockwood, L Li, S.M. Cohen, B.A. Edgar. Drosophila's Insulin/PI3-kinase Pathway Coordinates Cellular Metabolism with Nutritional Conditions[ J ]. Dev Cell. 2002,2 (2):239-249
[77] M Iwami, I Furuya, H Kataoka. Bombyxin-related Peptides: cDNA Structure and Expression in the Brain of the Horn worm Agrius convolvuli [ J ]. Insect Biochem Mol Biol. 1996,26(1) :25-32
[78] G Fullbright, E.R. Lacy, E.E. Bullesbach. Bombyxin: an Insect Neurohormone Targets the Ovaries in Lepidoptera. SAAS Bull Biochem Biotechnol[ J ]. 1997,10:37-41
[79] K Nagata, H Hatanaka, H Kataoka, H Nagasawa, A Isogai, H Ishizaki, A Suzuki, F Inagaki. Identification of the Receptor-recognition Surface of Bombyxin-II, an Insulin-like Peptide of the Silkmoth Bombyx mori: Critical Importance of the B-chain Central Part[ J ]. J Mol Biol. 1995 ,253 (5):759-770
[80] M.A. Riehle, M.R. Brown. Insulin Receptor Expression during Development and a Reproductive Cycle in the Ovary of the Mosquito Aedes aegypti[ J ]. Cell Tissue Res. 2002 ,308(3):409-420
[81] M Tatar, A Kopelman, D Epstein, M.P. Tu, C.M. Yin, R.S. Garofalo. A mutant Drosophila Insulin Receptor Homolog that Extends Life-span and Impairs Neuroendocrine Function[ J ]. Science. 2001,292(5514) :41-43
[82] D.J. Clancy, D Gems, L.G. Harshman, S Oldham, H Stocker, E Hafen, S.J. Leevers,L Partridge. Extension of Life-span by Loss of CHICO, a Drosophila Insulin Recep tor Substrate Protein[ J ]. Science. 2001, 292 (5514): 104-106.
[83] P.E. Caldwell, M Walkiewicz, M Stern. Ras Activity in the Drosophila Prothoracic Gland Regulates Body Size and Developmental Rate Via Ecdysone Release[ J ].Curr Biol. 2005 ,15(20): 1785-1795
[84] J Colombani, L Bianchini, S Layalle, E Pondeville, V.C. Dauphin, C Antoniewski,C Carre, S Noselli, P Leopold. Antagonistic actions of ecdysone and insulins determine final size in Drosophila[ J ]. Science.2005, 310(5748):667-670
[85] T Kozlova, C.S. Thummel. Steroid Regulation of Postembryonic Development and Reproduction in Drosophila[J]. Trends Endocrinol Metab. 2000 ,11(7):276-80
[86] J.W. Truman, L.M. Riddiford. Endocrine Insights into the Evolution of Metamorphosis in Insects[ J ]. Annu Rev Entomol. 2002, 47:467-500
[87] B.D. Drummond, A.C. Spradling. Stem Cells and their Progeny Respond to Nutritional Changes during Drosophila Oogenesis[ J ]. Dev Biol. 2001,231(1):265-278
[88] M.A. Riehle, M.R. Brown. Insulin Stimulates Ecdysteroid Production through a Conserved Signaling Cascade in the Mosquito Aedes aegypti[ J ]. Insect Biochem Mol Biol. 1999,29(10):855-860
[89] H Nagasawa, H Kataoka, A Isogai, S Tamura, A Suzuki, A Mizoguchi, Y Fujiwara,A Suzuki, S.Y. Takahashi, H Ishizaki. Amino Ccid Sequence of a Prothoracicotropic Hormone of the Silkworm Bombyx mori[ J ]. Proc Natl Acad Sci .1986 ,83(16):5840-5843
[90] M.P. Tu, CM. Yin, M Tatar. Impaired Ovarian Ecdysone Synthesis of Drosophila Melanogaster Insulin Receptor Mutants[ J ]. Aging Cell. 2002,1(2): 158-160
[91] G Maniere, I Rondot, E.E. Bullesbach, F Gautron, E Vanhems, J.P. Delbecque.Control of Ovarian Steroidogenesis by Insulin-like Peptides in the Blowfly (Phormia regina) [ J ]. J Endocrinol. 2004, 181(1): 147-156
[92] Caldwell, M Wakliewicz, M Stern.Ras Activity in the Drosophila Prothoracic Gland Regulates Body Size and Developmental Rate via Ecdysone Release[ J ].Curr Biol. 2005 ,15(20): 1785-1795
[93] C Mirth, J.W. Truman, L.M. Riddiford. The Role of the Prothoracic Gland in Determining Critical Weight for Metamorphosis in Drosophila melanogaster[ J ].Curr Biol. 2005 ,15(20):1796-1807
[94] K.L. Kelly, I Merida, E.H. Wong, C.D. Di, J.M. Mato.A Phospho-oligosaccharide Mimics the Effect of Insulin to Inhibit Isoproterenol-dependent Phosphorylation of Phospholipid Methyltransferase in Isolated Adipocytes[ J ]. J Biol Chem.1987 ,262(31):15285-15290
[95] L.M. Riddiford. Hormone Receptors and the Regulation of Insect Metamorphosis [ J ]. Receptor. 1993, 3(3):203-209
[96] P Moshitzky, I Fleischmann, N Chaimov, P Saudan, S Klauser, E Kubli, S.W.Applebaum. Sex-peptide Activates Juvenile Hormone Biosynthesis in the Drosophila melanogaster Corpus Allatum[ J ]. Arch Insect Biochem Physiol. 1996,32 (3-4):363-374
[97] M.P. Tu, CM. Yin, M Tatar. Mutations in Insulin Signaling Pathway Alter Juvenile Hormone Synthesis in Drosophila melanogaster[ J ]. Gen Comp Endocrinol.2005 ,142(3):347-356
[98] J.S. Britton, B.A. Edgar. Environmental Control of the Cell Cycle in Drosophila:Nutrition Activates Mitotic and Endoreplicative Cells by Distinct Mechanisms[ J ].Development. 1998,125(11):2149-2158
[99] B.A. Edgar, W.T. Orr. Endoreplication Cell Cycles: more for less [ J ]. Cell. 2001,105(3):297-306
[100] J.K. King, C.S. Thummel, Developmental Biology. Less Steroids Make Bigger Flies[ J ]. Science. 2005 ,310(5748):630-631
[101] A.W. Shingleton.Body-size Regulation: Combining Genetics and Physiology [ J ].Curr Biol. 2005, 15(20):R825-827
[102] M.H. Orme, S.J. Leevers. Flies on Steroids: the Interplay between Ecdysone and Insulin Signaling[ J ]. Cell Metab. 2005 ,2(5):277-278
[103] L.E. Canavoso, Z.E. Jouni, K.J. Karnas, J.E. Pennington, M.A. Wells. Fat Metabolism in Insects[ J ]. Annu Rev Nutr. 2001,21 :23-46