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Dong Y, Huang L, Liu L. Comparative analysis of testicular fusion in Spodoptera litura (cutworm) and Bombyx mori (silkworm): Histological and transcriptomic insights. Gen Comp Endocrinol 2024; 356:114562. [PMID: 38848820 DOI: 10.1016/j.ygcen.2024.114562] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 01/31/2024] [Revised: 05/30/2024] [Accepted: 05/31/2024] [Indexed: 06/09/2024]
Abstract
Spodoptera litura commonly known as the cutworm, is among the most destructive lepidopteran pests affecting over 120 plants species. The powerful destructive nature of this lepidopteran is attributable to its high reproductive capacity. The testicular fusion that occurs during metamorphosis from larvae to pupa in S.litura positively influences the reproductive success of the offspring. In contrast, Bombyx mori, the silkworm, retains separate testes throughout its life and does not undergo this fusion process. Microscopic examination reveals that during testicular fusion in S.litura, the peritoneal sheath becomes thinner and more translucent, whereas in B.mori, the analogous region thickens. The outer basement membrane in S.litura exhibits fractures, discontinuity, and uneven thickness accompanied by a significant presence of cellular secretions, large cell size, increased vesicles, liquid droplets, and a proliferation of rough endoplasmic reticulum and mitochondria. In contrast, the testicular peritoneal sheath of B.mori at comparable developmental stage exhibits minimal change. Comparative transcriptomic analysis of the testicular peritoneal sheath reveals a substantial difference in gene expression between the two species. The disparity in differential expressed genes (DEGs) is linked to an enrichment of numerous transcription factors, intracellular signaling pathways involving Ca2+ and GTPase, as well as intracellular protein transport and signaling pathways. Meanwhile, structural proteins including actin, chitin-binding proteins, membrane protein fractions, cell adhesion, extracellular matrix proteins are predominantly identified. Moreover, the study highlights the enrichment of endopeptidases, serine proteases, proteolytic enzymes and matrix metalloproteins, which may play a role in the degradation of the outer membrane. Five transcription factors-Slforkhead, Slproline, Slcyclic, Slsilk, and SlD-ETS were identified, and their expression pattern were confirmed by qRT-PCR. they are candidates for participating in the regulation of testicular fusion. Our findings underscore significant morphological and trancriptomic variation in the testicular peritoneal sheath of S.litura compared to the silkworm, with substantial changes at the transcriptomic level coinciding with testicular fusion. The research provides valuable clues for understanding the complex mechanisms underlying this unique phenomenon in insects.
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Affiliation(s)
- Yaqun Dong
- Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Guangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, China
| | - Lihua Huang
- Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Guangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, China
| | - Lin Liu
- Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Guangzhou Key Laboratory of Insect Development Regulation and Application Research, Institute of Insect Science and Technology, School of Life Sciences, South China Normal University, Guangzhou 510631, China.
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Dorn DC, Dorn A. Stem cell autotomy and niche interaction in different systems. World J Stem Cells 2015; 7:922-944. [PMID: 26240680 PMCID: PMC4515436 DOI: 10.4252/wjsc.v7.i6.922] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 07/20/2014] [Accepted: 05/27/2015] [Indexed: 02/06/2023] Open
Abstract
The best known cases of cell autotomy are the formation of erythrocytes and thrombocytes (platelets) from progenitor cells that reside in special niches. Recently, autotomy of stem cells and its enigmatic interaction with the niche has been reported from male germline stem cells (GSCs) in several insect species. First described in lepidopterans, the silkmoth, followed by the gipsy moth and consecutively in hemipterans, foremost the milkweed bug. In both, moths and the milkweed bug, GSCs form finger-like projections toward the niche, the apical cells (homologs of the hub cells in Drosophila). Whereas in the milkweed bug the projection terminals remain at the surface of the niche cells, in the gipsy moth they protrude deeply into the singular niche cell. In both cases, the projections undergo serial retrograde fragmentation with progressing signs of autophagy. In the gipsy moth, the autotomized vesicles are phagocytized and digested by the niche cell. In the milkweed bug the autotomized vesicles accumulate at the niche surface and disintegrate. Autotomy and sprouting of new projections appears to occur continuously. The significance of the GSC-niche interactions, however, remains enigmatic. Our concept on the signaling relationship between stem cell-niche in general and GSC and niche (hub cells and cyst stem cells) in particular has been greatly shaped by Drosophila melanogaster. In comparing the interactions of GSCs with their niche in Drosophila with those in species exhibiting GSC autotomy it is obvious that additional or alternative modes of stem cell-niche communication exist. Thus, essential signaling pathways, including niche-stem cell adhesion (E-cadherin) and the direction of asymmetrical GSC division - as they were found in Drosophila - can hardly be translated into the systems where GSC autotomy was reported. It is shown here that the serial autotomy of GSC projections shows remarkable similarities with Wallerian axonal destruction, developmental axon pruning and dying-back degeneration in neurodegenerative diseases. Especially the hypothesis of an existing evolutionary conserved “autodestruction program” in axons that might also be active in GSC projections appears attractive. Investigations on the underlying signaling pathways have to be carried out. There are two other well known cases of programmed cell autotomy: the enucleation of erythroblasts in the process of erythrocyte maturation and the segregation of thousands of thrombocytes (platelets) from one megakaryocyte. Both progenitor cell types - erythroblasts and megakaryocytes - are associated with a niche in the bone marrow, erythroblasts with a macrophage, which they surround, and the megakaryocytes with the endothelial cells of sinusoids and their extracellular matrix. Although the regulatory mechanisms may be specific in each case, there is one aspect that connects all described processes of programmed cell autotomy and neuronal autodestruction: apoptotic pathways play always a prominent role. Studies on the role of male GSC autotomy in stem cell-niche interaction have just started but are expected to reveal hitherto unknown ways of signal exchange. Spermatogenesis in mammals advance our understanding of insect spermatogenesis. Mammal and insect spermatogenesis share some broad principles, but a comparison of the signaling pathways is difficult. We have intimate knowledge from Drosophila, but of almost no other insect, and we have only limited knowledge from mammals. The discovery of stem cell autotomy as part of the interaction with the niche promises new general insights into the complicated stem cell-niche interdependence.
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Iga M, Blais C, Smagghe G. Study on ecdysteroid levels and gene expression of enzymes related to ecdysteroid biosynthesis in the larval testis of Spodoptera littoralis. ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY 2013; 82:14-28. [PMID: 23007959 DOI: 10.1002/arch.21068] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/01/2023]
Abstract
We investigated here the ecdysteroid titers and the expression of six genes coding for known enzymes of the ecdysteroid biosynthesis in the testes of last instar larvae of the pest cotton leafworm, Spodoptera littoralis. We showed that the timing of the ecdysteroid profile was the same in testes and in hemolymph, with a small peak at day 2 and a large one at day 4 after ecdysis. Ecdysone and 20-hydroxyecdysone (20E) were detected in both tissues. 20E was the major ecdysteroid in testes and in hemolymph from day 4. Interestingly, the gene expression of the steroidogenetic enzymes, Neverland, and the five cytochrome P450 enzymes encoded by the Halloween genes was confirmed in the testes, and varied during the instar. However, from the data obtained so far, we cannot conclude that the measured ecdysteroids in the testes result from the activity of the genes under study. Indeed, it is suggested that the ecdysone produced centrally in the prothoracic glands, could have been transformed into 20E in the testes, where Sl-shade is well expressed.
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Affiliation(s)
- Masatoshi Iga
- Department of Crop Protection, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium
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Dorn DC, Dorn A. Structural characterization and primary in vitro cell culture of locust male germline stem cells and their niche. Stem Cell Res 2010; 6:112-28. [PMID: 21256099 DOI: 10.1016/j.scr.2010.11.002] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 06/18/2010] [Revised: 10/17/2010] [Accepted: 11/12/2010] [Indexed: 11/19/2022] Open
Abstract
The establishment of in vitro culture systems to expand stem cells and to elucidate the niche/stem cell interaction is among the most sought-after culture systems of our time. To further investigate niche/stem cell interactions, we evaluated in vitro cultures of isolated intact male germline-niche complexes (i.e., apical complexes), complexes with empty niche spaces, and completely empty niches (i.e., isolated apical cells) from the testes of Locusta migratoria and the interaction of these complexes with isolated germline stem cells, spermatogonia (of transit-amplifying stages), cyst progenitor cells, cyst progenitor cell-like cells, cyst cells, and follicle envelope cells. The structural characteristics of these cell types allow the identification of the different cell types in primary cultures, which we studied in detail by light and electron microscopy. In intact testes germline stem cells strongly adhere to their niche (the apical cell), but emigrate from their niche and form filopodia if the apical complex is put into culture with "standard media." The lively movements of the long filopodia of isolated germline stem cells and spermatogonia may be indicative of their search for specific signals to home to their niche. All other incubated cell types (except for follicle envelope cells) expressed rhizopodia and lobopodia. Nevertheless isolated germline stem cells in culture do not migrate to empty niche spaces of nearby apical cells. This could indicate that apical cells lose their germline stem cell attracting ability in vitro, although apical cells devoid of germline stem cells either by emigration of germline stem cells or by mechanical removal of germline stem cells are capable of surviving in vitro up to 56 days, forming many small lobopodia and performing amoeboid movements. We hypothesize that the breakdown of the apical complex in vitro with standard media interrupts the signaling between the germline stem cells and the niche (and conceivably the cyst progenitor cells) which directs the typical behavior of the male regenerative center. Previously we demonstrated the necessity of the apical cell for the survival of the germline stem cell. From these studies we are now able to culture viable isolated germline stem cells and all cells of its niche complex, although DNA synthesis stops after Day 1 in culture. This enables us to examine the effects of supplements to our standard medium on the interaction of the germline stem cell with its niche, the apical cell. The supplements we evaluated included conditioned medium, tissues, organs, and hemolymph of male locusts, insect hormones, mammalian growth factors, Ca(2+) ion, and a Ca(2+) ionophore. Although biological effects on the germline stem cell and apical cell could be detected with the additives, none of these supplements restored the in vivo behavior of the incubated cell types. We conclude that the strong adhesion between germline stem cells and apical cells in vivo is actively maintained by peripheral factors that reach the apical complex via hemolymph, since a hemolymph-testis barrier does not exist. The in vitro culture model introduced in this study provides a platform to scan for possible regulatory factors that play a key role in a feedback loop that keeps germline stem cell division and sperm disposal in equilibrium.
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Affiliation(s)
- David C Dorn
- Department of Hematology, Hemostasis, Oncology, and Stem-Cell Transplantation, Hannover Medical School, 30625 Hannover, Germany.
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Zahn J, Doormann P, Dorn A, Dorn DC. Apoptosis of male germ-line stem cells after laser ablation of their niche. Stem Cell Res 2007; 1:75-85. [PMID: 19383387 DOI: 10.1016/j.scr.2007.09.005] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 05/03/2007] [Revised: 08/27/2007] [Accepted: 09/24/2007] [Indexed: 01/25/2023] Open
Abstract
Male germ-line stem cells (GSCs) and their niche-the apical cells or hub cells-display a unique feature at the apices of insect testicular follicles. In the locust, Locusta migratoria, the niche consists of only one large apical cell surrounded by about 60 GSCs. The apical cell can be readily identified in the intact follicle. Using laser ablation it is feasible to destroy the apical cell exclusively without injuring neighboring GSCs or any other cells. The most immediate effect on GSCs is the loss of their structural polarity. Beginning about 3 h after laser treatment chromatin starts to clump and condense in individual GSCs, and some show the first signs of cellular breakdown. These symptoms intensify during the 96-h observation period after laser ablation of the apical cell. TUNEL staining and electron microscopic observations confirm an apoptotic cell death of the GSCs. Laser ablation of individual GSCs had no effect on neighboring GSCs or the apical cell. Destroyed apical cells were not replaced during the observation period. Mitotic divisions of GSCs ceased after about 24 h after apical cell ablation. It is speculated that it might be a general principle in stem cell-niche relationships that stem cells undergo apoptosis when the niche is dysfunctional. This could be a control mechanism to prevent tumor growth of orphaned GSCs.
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Affiliation(s)
- Johannes Zahn
- Institut für Zoologie, Johannes Gutenberg-Universität, Mainz, Germany
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Vafopoulou X, Steel CGH. Testis ecdysiotropic peptides in Rhodnius prolixus: biological activity and distribution in the nervous system and testis. JOURNAL OF INSECT PHYSIOLOGY 2005; 51:1227-39. [PMID: 16139295 DOI: 10.1016/j.jinsphys.2005.06.013] [Citation(s) in RCA: 13] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Received: 03/06/2005] [Revised: 06/26/2005] [Accepted: 06/28/2005] [Indexed: 05/04/2023]
Abstract
In Rhodnius prolixus, testes from both pharate adult and adult males are shown to produce and release ecdysteroids in vitro. Proteinaceous brain extracts from these stages caused stimulation of ecdysteroid production by testes of unfed adults. Therefore, the brain of Rhodnius contains peptides with testis ecdysiotropic activity. The Lymantria testis ecdysiotropin (LTE) also stimulated the in vitro production of ecdysteroids by unfed adult testis but had no stimulatory effect on prothoracic glands. Western blot analysis of brain peptides using anti-LTE revealed the presence of several medium to small size immunoreactive peptides. Two of these peptides with sizes of 16.8 and 11.0 kDa were present only during pharate adult development and the adult stage. Immunohistochemical analysis using confocal laser scanning microscopy revealed abundant LTE-immunoreactive material in cytoplasmic granules of specific neurosecretory cells in the brain and suboesophageal ganglion and the epithelium of the testis sheath. Clusters of two cytologically distinct cell types were seen within the medial neurosecretory cells (MNC) and also a pair of neurons in the posterior protocerebrum. Feeding in both larvae and adult males resulted in massive release of LTE-immunoreactive material from the MNC cells, suggesting a role of LTE-related peptides in both larval-adult development and in male reproductive development. Release from the MNC cells of LTE-immunoreactive material exhibited a clear daily cycling during larval-adult development, which was synchronous with the rhythms of release of prothoracicotropic hormone and bombyxin reported previously. The testis sheath exhibited intense immunofluorescence in pharate adults and unfed adults, which disappeared following a blood meal. It is concluded that LTE-related peptides are developmentally regulated in several locations and may act as ecdysiotropins in Rhodnius. Those in the MNC cells are very probably classical hormones, i.e. are transported to their target sites via the insect haemolymph.
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Affiliation(s)
- Xanthe Vafopoulou
- Biology Department, York University, 4700 Keele St., Toronto, Ontario, Canada M3J 1P3
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Maas U, Dorn A. No evidence of androgenic hormone from the testes of the glowworm, Lampyris noctiluca. Gen Comp Endocrinol 2005; 143:40-50. [PMID: 15993103 DOI: 10.1016/j.ygcen.2005.02.025] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/26/2004] [Revised: 02/22/2005] [Accepted: 02/27/2005] [Indexed: 11/16/2022]
Abstract
The widely accepted concept, stating that insects have no true sex hormones, and that primary as well as secondary sex characteristics are controlled by the genetic inventory of each single cell, is challenged by the report of Naisse, J. [1966a. Contrôle endocrinien de la différenciation sexuelle chez l'Insecte Lampyris noctiluca (Coléoptère Malacoderme Lampyride). I. Rôle androgène des testicules, Arch. Biol. Liège, 77, 139-201] on the discovery of an androgenic hormone in the glowworm, Lampyris noctiluca. This case is of special interest, since it may point to an ancestral mode of sex differentiation in arthropods, considering that androgenic hormones have been discovered and characterized in crustaceans. With the intention to further characterize the androgenic hormone in the glowworm, and to establish a bioassay, we tried to repeat Naisses's transplantation experiments, according to which, androgen producing testes implanted into female larvae should masculinize the female's gonads and all other female features of the sexually strongly dimorphic pupae and beetles. We found, however, that larval development of the glowworm proceeded differently than reported by Naisse, and that sexing of larvae was not possible. Therefore, "blind" transplantations had to be performed. The results of our experiments showed, however, unequivocally, that an androgenic hormone, allegedly synthesized by the apical tissue of larval testes, was not involved in sex differentiation. We found, that in transplants, where testes and ovaries were even located closely to each other, both matured and formed spermatozoa in the testes and vitellogenic oocytes in the ovaries. Masculinization of ovaries was never observed, and the sex of the recipient was always in accordance with the sex of its own gonads. We therefore conclude that Lampyris noctiluca does not synthesize an androgenic hormone in the larval testes, and that sex differentiation is probably regulated as in other insect species. (The apical testis tissue of the glowworm was previously shown to represent progenitors of spermatogonial cyst cells [Balles, S., Maas, U., Sehn, E., Dorn, A., 2002. Testis differentiation in the glowworm, Lampyris noctiluca, with special reference to the apical tissue. J. Morphol. 251, 22-37].).
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Affiliation(s)
- Ulrike Maas
- Institute of Zoology, Johannes Gutenberg University, D-55099 Mainz, Germany
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SCHMIDT ESTHERD, SEHN ELISABETH, DORN AUGUST. Differentiation and ultrastructure of the spermatogonial cyst cells in the milkweed bug,Oncopeltus fasciatus. INVERTEBR REPROD DEV 2002. [DOI: 10.1080/07924259.2002.9652773] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
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Balles S, Maas U, Sehn E, Dorn A. Testis differentiation in the glowworm, Lampyris noctiluca, with special reference to the apical tissue. J Morphol 2002; 251:22-37. [PMID: 11746466 DOI: 10.1002/jmor.1072] [Citation(s) in RCA: 6] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/10/2022]
Abstract
The gonads of Lampyris noctiluca are sexually undifferentiated during the first larval instars. They consist of many gonadal follicles that include the germ stem cells enclosed by the somatic cells of the follicle wall. Follicle wall cells are more numerous at the follicle apices than at the distal parts, but different cell types cannot be distinguished. In male larvae, the appearance of apical follicle tissue, derived from follicle wall cells, marks the onset of testis differentiation. When maximally expressed, the apical tissue occupies about the upper half of the testis follicles and can be observed in larvae of the fifth and sixth instar. The apical tissue is characterized by its "light" appearance (due to poor stainability) caused by the small number cellular organelles, especially a paucity of free ribosomes. Maximal expression of the apical tissue must be very brief, since in most examined fifth and sixth instar larvae the apical tissue is partly or mostly translocated into the center of the upper half of the follicles and spermatogonia then occupy the apical follicle tips. During and after translocation apical cells form projections that grow around clusters of spermatogonia (spermatocysts). Thus, the apical cells transform into spermatocyst envelope cells. They retain their "light" appearance but undergo dramatic subcellular differentiation: smooth ER becomes extremely prominent, forming stacks and whorls of parallel cisternae. Golgi complexes are also conspicuous. The cellular organization suggests secretory activity. The possibility of ecdysteroid production and its function is discussed. The spermatocyst envelope cells persist into the pupal stage. When spermiohistogenesis takes place in cysts, cyst envelope cells show signs of regression. At all stages of testis development apical cells and their derivatives, the spermatocyst envelope cells, phagocytize degenerating spermatogonia. Although this is an important task of these cells, the impressive formation of sER in the cyst envelope cells is indicative of an additional, as yet unknown, function.
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Affiliation(s)
- S Balles
- Institut für Zoologie, Johannes Gutenberg-Universität, D-55099 Mainz, Germany
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Loeb MJ, De Loof A, Gelman DB, Hakim RS, Jaffe H, Kochansky JP, Meola SM, Schoofs L, Steel C, Vafopoulou X, Wagner RM, Woods CW. Testis ecdysiotropin, an insect gonadotropin that induces synthesis of ecdysteroid. ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY 2001; 47:181-188. [PMID: 11462222 DOI: 10.1002/arch.1050] [Citation(s) in RCA: 15] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/23/2023]
Abstract
Testes of lepidoptera synthesized ecdysteroid in a somewhat different temporal pattern than the prothoracic glands that release ecdysteroid to the hemolymph. Brain extracts from Heliothis virescens and Lymantria dispar induced testes to synthesize ecdysteroid, but did not affect prothoracic glands. The testis ecdysiotropin (LTE) was isolated from L. dispar pupal brains by a series of high-pressure chromatography steps. Its sequence was Ile-Ser-Asp-Phe-Asp-Glu-Tyr-Glu-Pro-Leu-Asn-Asp-Ala-Asp-Asn-Asn-Glu-Val-Leu-Asp-Phe-OH, of molecular mass 2,473 Daltons. The predominant signaling pathway for LTE was via G(i) protein, IP3, diacylglycerol and PKC; a modulating pathway, apparently mediated by an angiotensin II-like peptide, was controlled via G(s) protein, cAMP, and PKA. Testis ecdysteroid caused isolated testis sheaths to also synthesize a growth factor that induced development of the male genital tract. The growth factor appeared to be a glycoprotein similar to vertebrate alpha-1-glycoprotein. A polyclonal antibody to LTE indicated LTE-like peptide in L. dispar brain medial neurosecretory cells, the suboesophageal, and other ganglia, and also in its target organ, the testis sheath. LTE immunoreactivity was also seen in testis sheaths of Rhodnius prolixus. LTE-like immunoactivity was also detected in developing optic lobes, antennae, frontal ganglia, and elongating spermatids of developing L. dispar pupae. This may indicate that LTE has a role in development as well as stimulation of testis ecdysteroid synthesis. Published 2001 Wiley-Liss, Inc.
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Affiliation(s)
- M J Loeb
- Insect Biocontrol Laboratory, U.S. Department of Agriculture, Beltsville, Maryland 20705, USA.
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Gelman DB, Rojas MG, Kelly TJ, Hu JS, Bell RA. Ecdysteroid and free amino acid content of eggs of the Colorado potato beetle, Leptinotarsa decemlineata. ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY 2000; 44:172-182. [PMID: 10918312 DOI: 10.1002/1520-6327(200008)44:4<172::aid-arch4>3.0.co;2-u] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/23/2023]
Abstract
In order to identify components of the Colorado potato beetle (CPB) egg that may be required by Edovum puttleri, a parasitic wasp that parasitizes the CPB egg, to complete development, ecdysteroid and free amino acid content of CPB eggs were analyzed by reversed phase high pressure liquid chromatography followed by radioimmunoassay to identify ecdysteroids. Ecdysteroid titers were relatively low (<300 pg/egg) through day 2 post-oviposition and then increased sharply, reaching concentrations >2,500 pg/egg on day 3 post-oviposition. Ecdysone (E), 20-hydroxyecdysone (20E), and polar conjugates of E were prominent ecdysteroids present in eggs sampled on days 0 and 1 post-ecdysis, and E, 20E, three peaks containing more polar ecdysteroids (metabolic inactivation products), and polar conjugates of E were present in eggs sampled on day 2. Thus, at a time when parasitization of CPB eggs by E. puttleri is relatively high (0-48 h), physiologically-active ecdysteroids (20E and perhaps E are physiologically active) are present at concentrations between 50 and 200 pg/egg. Ecdysone and 20E reached their highest levels in day-3 eggs, indicating that ecdysteroid may direct physiological processes associated with the completion of CPB embryonic development. In day-4 eggs, the concentration of E and 20E fall dramatically and polar metabolites of E and/or 20E are now responsible for the high ecdysteroid content of the eggs. Interestingly, conjugates of E decrease to relatively low levels in day-3 eggs and are absent in day-4 eggs. Therefore, it is likely that the increase in E in day-3 eggs is due, in part, to the breakdown of polar conjugates of E. Nine amino acids were present in significant quantities in eggs sampled at various times between 0 and 48 h post-oviposition. These include histidine, glutamine, proline, asparagine, serine, glutamic acid, threonine, lysine, and tyrosine. The first three amino acids were present at concentrations that were approximately 2 to 6 times greater than the concentrations of the last six amino acids. Amounts of most of the free amino acids varied with the age of the eggs from which the extract was prepared, but in general, there was no correlation between the levels at times of maximum parasitization (0 and 30 h) and the levels at the less favored times of parasitization (16 and 48 h). This information should facilitate the development of diets for both parasites and predators of pest species of beetles. Arch. Insect Biochem. Physiol. 44:172-182, 2000. Published 2000 Wiley-Liss, Inc.
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Affiliation(s)
- D B Gelman
- Insect Biocontrol Laboratory, USDA, ARS, PSI, Beltsville, Maryland 20705, USA
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Gelman DB, Carpenter JE, Greany PD. Ecdysteroid levels/profiles of the parasitoid wasp, Diapetimorpha introita, reared on its host, Spodoptera frugiperda and on an artificial diet. JOURNAL OF INSECT PHYSIOLOGY 2000; 46:457-465. [PMID: 12770210 DOI: 10.1016/s0022-1910(99)00132-8] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 05/24/2023]
Abstract
Diapetimorpha introita is an ichneumonid ectoparasitoid of the fall armyworm, Spodoptera frugiperda. Since it has been reported that D. introita wasps reared on an artificial diet exhibit a significantly lower percentage of adult eclosion and fecundity than host-reared wasps, this study was undertaken to elucidate the factors responsible for the reduced viability observed in diet-reared wasps. A system of markers has been devised to track the development (from the initiation of cocooning through adult eclosion) of D. introita. Although wasps reared on artificial diet developed more slowly than did those reared on host pupae, both diet- and host-reared wasps passed through the same stages of development - the eyes enlarged and moved backward, the gut was purged and upon ecdysis the exarate pupa emerged. The thorax was the first to darken, followed by the head and then the abdomen. Pharate pupal formation occurred before gut purge. Two peaks of hemolymph ecdysteroids were observed, one in wasps in which gut purge was almost complete and the second in day-2 exarate pupae. Ecdysone and 20-hydroxyecdysone were the major ecdysteroids present in hemolymph sampled at these times. Small quantities of 20,26-dihydroxyecdysone, polar ecdysteroids and/or possibly 26-hydroxyecdysone were also present. In six stages of development, hemolymph ecdysteroid titers were significantly higher in host-reared than in diet-reared wasps (Eye 1, Eye 2, Gut Purge 2, Pharate Pupa, Head/Thorax Dark, and Abdomen Dark). Relatively high percentages of mortality were observed in diet-reared wasps in four of these stages and in two others which occurred in close proximity to one of the stages, the Abdomen Dark stage. Thus, insufficient ecdysteroid in the hemolymph may be responsible, in part, for the relatively high percentage of mortality that occurred in wasps reared on an artificial diet.
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Affiliation(s)
- D B. Gelman
- Insect Biocontrol Laboratory, USDA, ARS, PSI, Beltsville, MD, USA
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LOEB MARCIAJ, BELL ROBERTA, GELMAN DALEB, KOCHANSKY JAN, LUSBY WILLIAM, WAGNER RENEEM. Action cascade of an insect gonadotropin, testis ecdysiotropin, in male Lepidoptera. INVERTEBR REPROD DEV 1996. [DOI: 10.1080/07924259.1996.9672544] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
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Unique intertesticular tissue complex in larvae of Heliothis virescens (F.) (Lepidoptera : Noctuidae). ACTA ACUST UNITED AC 1995. [DOI: 10.1016/0020-7322(95)00005-o] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/15/2022]
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GILLOTT CEDRIC, ISMAIL PREETIM. In vitrosynthesis of ecdysteroid by the male accessory reproductive glands, testis and abdominal integument of the adult migratory grasshopper,Melanoplus sanguinipes. INVERTEBR REPROD DEV 1995. [DOI: 10.1080/07924259.1995.9672435] [Citation(s) in RCA: 21] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
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LOEB MJ. Development of Isolated Spermducts fromHeliothis virescens(Lepidoptera)in vitro. INVERTEBR REPROD DEV 1991. [DOI: 10.1080/07924259.1991.9672179] [Citation(s) in RCA: 9] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
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LOEB MJ. Growth and development of spermducts of the tobacco budworm mothHeliothis virescens, in vivoandin vitro. INVERTEBR REPROD DEV 1991. [DOI: 10.1080/07924259.1991.9672163] [Citation(s) in RCA: 18] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
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18
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DELBECQUE JEANPAUL, WEIDNER KARIN, HOFFMANN KLAUSH. Alternative sites for ecdysteroid production in insects. INVERTEBR REPROD DEV 1990. [DOI: 10.1080/07924259.1990.9672126] [Citation(s) in RCA: 61] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 10/18/2022]
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