The influence of juvenile hormone analogue, fenoxycarb on the midgut remodeling in Bombyx mori (L., 1758) (Lepidoptera: Bombycidae) during larval-pupal metamorphosis

Yıl 2011, Cilt: 35 Sayı: 2, 179 - 194, 30.06.2011

Ebru Goncu Osman Parlak

Öz

During larval-pupal metamorphosis, midgut tissues of holometabolous insects undergo remodeling and this remodeling process involves two metamorphic events. Degeneration of larval midgut occurs via programmed cell death and pupal midgut develops from regenerative (stem) cells. All metamorphic events in insects are regulated by ecdysone and juvenile hormone. Molecular mechanisms of ecdysone are well known but action mechanism of juvenile hormone is not well understood. Fenoxycarb, 0-ethyl N-(2-(4-pheoxyphenoxy)-ethyl) carbamate has been previously shown one of the most potent juvenile hormone analogue against a variety of insect species and widely used for investigations the mechanisms of juvenile hormone. In this study, we aimed to analyze the metamorphic events in the midgut of Bombyx mori Linnaeus, 1758 and investigate the effects of fenoxycarb on this period. For this purpose, we applied two different doses of fenoxycarb (1ng/10µl and 10ng/10µl) topically on day 0 of fifth instar larvae. We followed the progression of remodeling processes with morphological observations and analyzed programmed cell death specific proteins, caspase 3 for apoptosis and acid phosphatase for autophagic cell death in control and experiment groups. Our results indicated that, programmed cell death of larval midgut has morphologic characteristics of apoptosis and caspase activation occurs during this period and juvenile hormone analogue fenoxycarb inhibits or delays remodeling process in dose dependent manner.

Anahtar Kelimeler

Apoptosis, autophagic cell death, Bombyx mori, fenoxycarb, midgut

Destekleyen Kurum

The research was supported by Ege University

Proje Numarası

2009 Science 021

Bombyx mori’ L., 1758 (Lepidoptera: Bombycidae) de larval-pupal metamorfoz süresince orta barsaktaki yeniden şekillenme sürecine juvenil hormon analoğu fenoxycarb’ın etkisi

Öz

Larval-pupal metamorfoz boyunca, tam başkalaşım geçiren böceklerin orta barsakları yeniden şekillenme sürecine girer ve bu süreç 2 metamorfik olayı içerir. Larval orta barsağın dejenerasyonu programlanmış hücre ölümü ile meydana gelirken pupal orta barsak rejeneratif (kök) hücrelerden oluşur. Böceklerdeki tüm metamorfik olaylar ekdizon ve juvenil hormon tarafından kontrol edilir. Ekdizonun moleküler mekanizması gayet iyi bilinmekle birlikte juvenil hormonun moleküler mekanizması henüz tam olarak anlaşılamamıştır. Fenoksikarb, O-ethyl N-(2-(4-pheoxyphenoxy)-ethyl) karbamat’ın pek çok böcek türünde etkili juvenil hormon analoglarından birisi olduğu ortaya konulmuştur ve juvenil hormonun mekanizmasına ilişkin çalışmalarda sıklıkla kullanılmaktadır. Bu çalışmada, Bombyx mori Linnaeus, 1758’nin orta barsağındaki metamorfik olayların analiz edilmesi ve bu süreçte fenoksikarb’ın etkisinin araştırılması amaçlanmıştır. Bu amaçla, beşinci larval döneminin 0. gününde iki farklı dozda fenoksikarb (1ng/10µl and 10ng/10µl) topikal olarak uygulanmıştır. Yeniden şekillenme sürecinin ilerleyişini morfolojik gözlemler ve programlanmış hücre ölümüne spesifik proteinlerin - apoptosis için kaspaz 3 ve otofajik hücre ölümü için asit fosfataz- kontrol ve deney gruplarında analizi ile takip edilmiştir. Sonuçlar larval orta barsakta programlanmış hücre ölümünün apoptozisin morfolojik karakterlerine sahip olduğunu, bu period boyunca kaspaz aktivasyonunun meydana geldiğini ve juvenil hormon analoğu fenoksikarb’ın doza bağlı olarak yeniden şekillenme sürecini geciktirdiğini veya engellediğini göstermiştir.

Anahtar Kelimeler

Apoptozis, otofajik hücre ölümü Bombyx mori, fenoksikarb, orta barsak

Proje Numarası

2009 Science 021

Kaynakça

• Adachi, T., M. Tomita & K. Yoshizato, 2005. Synthesis of prolyl 4-hydroxylase a subunit and type IV collagen in hemocytic granular cells of silkworm, Bombyx mori: involvement of type IV collagen in self-defense reaction and metamorphosis. Matrix Biology, 24: 136 - 154.
• Ahmad, M., S. M. Srinivasula, L. Wang, G. Litwack, T. Fernandes- Alnemri & E. S. Alnemri, 1997. Spodoptera frugiperda caspase-1, a novel insect death protease that cleaves the nuclear immunophilin FKBP46, is the target of the baculovirus antiapoptotic protein p35. The Journal of Biological Chemistry, 272: 1421–1424.
• Aidells, B., R. A. Lockshin & A. M. Cullin, 1971. Breakdown of the silk glands of Galleria mellonella - acid phosphatase in involuting glands. Journal of Insect Physiology, 17: 857-869.
• Anonymous, 1991. The Pesticide Manual: A World Compendium. Ninth Edition. Unwin Brothers Limited. The British Crop Protection Council, Great Britain, 1141 pp.
• Bergmeyer, H. U., 1974. Reagents for enzymatic analysis. pp 495-496. In: Methods of Enzymatic Analysis Vol I. (Eds: Bergmeyer HU, Gawehn K) Verlag Chemie, Weinheim.
• Bowen, E. D., T. A. Ryder & C. Dark, 1976 The effects of starvation on the planarian worm Polycelis tenuis Iijima. Cell Tissue Research, 169: 193–209.
• Bradford, M. M., 1976. Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Analytical Biochemistry, 72: 248-254.
• Cavalcante, V. M. & C. Cruz-Landim, 2004. Electrophoretic protein pattern and acid phosphatase activity in the midgut extracts of Apis mellifera L. (Hymenoptera: Apidae) during metamorphosis. Neotropical Entomology, 33 (2): 169-171.
• Csikós, G. & M. M. Sass, 1997. Changes of acid phosphatase content and activity in the fat body and the hemolymph of the flesh fly Neobellieria (Sarcophaga) bullata during metamorphosis. Archives of Insect Biochemistry and Physiology, 34: 369–391.
• Dedos, S. G. & H. Fugo, 1999. Induction of dauer larvae by application of fenoxycarb early in the 5th Instar of the silkworm, Bombyx mori. Journal of Insect Physiology, 45 (8): 769-775.
• Dedos, S. G., F. Szurdoki, A. Székács, T. Shiotsuki, B. D. Hammock, J. Shimada & H. Fugo, 2002. Fenoxycarb levels and their effects on general and juvenile hormone esterase activity in the hemolymph of the silkworm, Bombyx mori. Pesticide Biochemistry and Physiology, 73 (3): 174-187. Extension Toxicology Network, 1993. (Web page: http://ace.orst.edu/cgi-bin/mfs/01/pips/ methopr. (Data accessed: 10.03.2010)
• Goncu, E. & O. Parlak, 2008. Some autophagic and apoptotic features of programmed cell death in the anterior silk glands of the silkworm, Bombyx mori. Autophagy, 4 (8): 1069-1072.
• Halaby, R., M. L. Martinez,R. Lockshin & Z. Zakeri, 1998. 20-hydroxyecdysone induces apoptosis in the labial gland of Manduca sexta. Journal of Research on the Lepidoptera, 37: 3-10.
• Hakim, R. S., K. M. Baldwin & M. Loeb, 2001. The role of stem cells in midgut growth and regeneration. In Vitro Cellular & Developmental Biology – Animal, 37: 338-342.
• Hiruma, K., T. Shinoda , F. Malone & L. M. Riddiford,. 1999. Juvenile hormone modulates 20-hydroxyecdysone-inducible ecdysone receptor and ultraspiracle gene expression in the tobacco hornworm, Manduca sexta. Developmental Genes and Evolution, 209 (1):18-30.
• Jiang, C., E. H. Baehrecke, & C. S. Thummel, 1997. Steroid regulated programmed cell death during Drosophila metamorphosis Development, 124 (22): 4673-83.
• Kamimura, M., 1995. Effects of juvenile hormone analogue, fenoxycarb, on larval growth of the silkworm, Bombyx mori (Lepidoptera: Bombycidae). Applied Entomology and Zoology, 30: 487-489.
• Kamimura, M. & M. Kiuchi, 1998. Effects of a juvenile hormone analog, fenoxycarb, on 5th stadium of the larvae of the silkworm, Bombyx mori (Lepidoptera:Bombycidae). Applied Entomology and Zoology, 33 (2): 333-338.
• Lee, C. Y., B. A. K. Cooksey & E. H. Baehrecke, 2002 Steroid regulation of midgut cell death during Drosophila development. Developmental Biology, 250: 101-111.
• Lehane, M. & P. Billingsley, 1996. Structure and Ultrastructure of the Insect Midgut. In: Biology of the Insect Midgut (Eds: M. Lehane & P. Billingsley) Springer; First Edition, 504 pp.
• Leonardi, M. G., S. Cappellozza, P. Ianne, L. Cappellozza, P. Parentis & B. Giordina, 1996. Effects of the topical application an insect growth regulator (fenoxycarb) on some physiological parameters in the fifth instar larvae of the silkworm Bombyx mori. Comparative Biochemistry and Physiology, 113B (2): 361-365.
• Lockshin, R. A. & J. Beaulaton, 1974. Programmed cell death. Life Sciences, 15: 1549-1566.
• Lockshin, R. A. & C. M. Williams, 1965a. Programmed cell death. I. Cytology of degeneration in the intersegmental muscles of the pernye silkmoth. Journal of Insect Physiology, 11: 123-133.
• Lockshin, R. A. & C. M. Williams, 1965b. Programmed cell death. V. Cytolytic enzymes in relation to the breakdown of the intersegmental muscles of silkmoths. Journal of Insect Physiology, 11: 833-844.
• Lockshin, R. A. & Z. Zakari, 2004. Apoptosis, autophagy and more. The International Journal of Biochemistry & Cell Biology, 36: 2405–2419.
• Masner, P., M. Angst & S. Dorn, 1987. Fenoxycarb, an insect growth regulator with juvenile hormone activity: A candidate for Heliothis virescens (F.) control on cotton. Pesticide Science, 18: 89-94.
• Matolcsy, G., M. Nadasy & V. Andriska, 1988. Pesticide Chemistry: Studies in Environmental Science, Elsevier. New York, 800 pp.
• Micchelli, C.A. & N. Perrimon, 2006. Evidence that stem cells reside in the adult Drosophila midgut epithelium. Nature, 439: 475–479.
• Monconduit H. & B. Mauchamp, 1999. Fenoxycarb, a potent contaminant of the silkworm Bombyx mori L., does not influence its juvenile hormone titer. Archives of Insect Biochemistry and Physiology, 40: 141-149.
• Miyamoto, J., M. Hirano,Y. Takimoto & M. Hatakoshi, 1993. Insect growth regulators for pest control with emphasis on juvenile hormone analogs: present status and future prospects. Pest Control With Enhanced Environmental Safety, 524: 144-168.
• Nardi, J. B., C. Gao & M. R. Kanost, 2001. The extracellular matrix protein lacunin is expressed by a subset of hemocytes involved in basal lamina morphogenesis. Journal of Insect Physiology, 47: 997–1006.
• Parthasarathy, P. & S. R. Palli, 2007. Developmental and hormonal regulation of midgut remodeling in a Lepidopteran insect Heliothis virescens. Mechanism of Development, 124: 23–34.
• Riddiford, L. M., 1975. Juvenile hormone-induced delay of metamorphosis of the viscera of the cecropia silkworm. The Biological Bulletin, 142: 310-325.
• Riddiford, L. M., K. Hiruma, Q. Lan & B. Zhou, 1999. Regulation and role of nuclear receptors during larval molting and metamorphosis of Lepidoptera. American Zoologist, 39: 736-746.
• Sahota, T. S., 1975. Effect of juvenile hormone on acid phosphatases in the degenerating flight muscles of the douglas- fire beetle, Dendroctonus pseudotsugae. Journal of Insect Physiology, 21: 471–478.
• Sakurai, S., M. Kaya & S. Satake, 1998. Hemolymph ecdysteroid titer and ecdysteroid-dependent developmental events in the last-larval stadium of the silkworm, Bombyx mori: Role of low ecdysteroid titer in larval-pupal metamorphosis and reappraisal of the head critical period. Journal of Insect Physiology, 44: 867-881.
• Saraste, A. & K. Pulkki, 2000. Morphologic and biochemical hallmarks of apoptosis. Cardiovascular Research, 45 (3): 528-537.
• Sass, M., L. Kőműves, G. Csikós & J. Kovács, 1989. Changes in the activities of lysosomal enzymes in the fat body and midgut of two Lepidopteran insects (Mamestra brassicae and Pieris brassicae) during metamorphosis. Comparative Biochemistry and Physiology, 92 (3): 285-289.
• Shinohara, Y., N. Ishii, S. Takahashiand & T. Okazaki, 2008. Appearance of apoptotic cells and granular cells in the silkworm midgut lumen during larval-pupal ecdysis. Zoological Science, 25: 139–145.
• Takaki, K. & S. Sakurai, 2003. Regulation of prothoracic gland ecdysteroidogenic Activity leading to pupal metamorphosis. Insect Biochemistry and Molecular Biology, 33(12): 1189-99.
• Tettamanti, G., A. Grimaldi, M. Casartelli, E. Ambrosetti, B. Ponti, T. Congiu, R. Ferrarese, M. L. Rivas-Pena, F. Pennacchio & M. Eguileor, 2007. Programmed cell death and stem cell differentiation are responsible for midgut replacement in Heliothis virescens during prepupal instar. Cell and Tissue Research, 330 (2): 345-59.
• Tsujimoto, Y. & S. Shimizu, 2005. Another way to die: autophagic programmed cell death. Cell Death and Differentation, 12: 1528-34.
• Uwo, M.F., K. Ui-Tei, P. Park & M. Takeda, 2002. Replacement of midgut epithelium in the greater wax moth, Galleria mellonella, during larval–pupal molt. Cell and Tissue Research, 308: 319–331.
• Vilaplana, L., N. Pascual, N. Perera & X. Belle´ s, 2007. Molecular characterization of an inhibitor of apoptosis in the egyptian armyworm, Spodoptera littoralis, and midgut cell death during metamorphosis. Insect Biochemistry and Molecular Biology, 37: 1241–1248
• Wigglesworth, V. B., 1972. Digestion and Nutrition, 476–552. In: The Principles of Insect Physiology. Chapman & Hall, London, 827 pp.
• Wu, Y., R. Parthasarathy, H. Bai & S. R. Palli, 2006. Mechanisms of midgut remodeling: juvenile hormone analog methoprene blocks midgut metamorphosis by modulating ecdysone action. Mechanism of Development, 123: 530-547.
• Zakeri, Z., D. Quaglino, T. Latham, K. Woo & R. A. Lockshin, 1996. Programmed cell death in the tobacco hornworm, Manduca sexta: alterations in protein Synthesis. Microscope Research and Technique, 34: 192-201.