DOCSLIB.ORG

Original Article Proteomic Analysis of the Peritrophic Matrix from The

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
Original Article Proteomic Analysis of the Peritrophic Matrix from The 56 Biomed Environ Sci, 2016; 29(1): 56-65 Original Article Proteomic Analysis of the Peritrophic Matrix from the Midgut of Third Instar Larvae, Musca domestica* WANG Yu1,2, XIU Jiang Fan1, CHENG Jin Zhi1, LUO Man1, ZHAO Peng1, SHANG Xiao Li1, WANG Tao1, and WU Jian Wei1,# 1. Department of Parasitology, Guizhou Medical University, Guiyang 550004, Guizhou, China; 2. Guizhou Provincial Center for Disease Control and Prevention, Guiyang 550004, Guizhou, China Abstract Objective To better comprehend the molecular structure and physiological function of the housefly larval peritrophic matrix (PM), a mass spectrometry approach was used to investigate the PM protein composition. Methods The PM was dissected from the midgut of the third instar larvae, and protein extracted from the PM was evaluated using SDS-PAGE. A 1D-PAGE lane containing all protein bands was cut from top to bottom, the proteins in-gel trypsinised and analysed via shotgun liquid chromatography- tandem mass spectrometry (LC-MS/MS). Results In total, 374 proteins, with molecular weights varying from 8.225 kD to 996.065 kD and isoelectric points ranging from 3.83 to 11.24 were successfully identified, most identified proteins were mainly related to immunity, digestion, nutrient metabolism and PM structure. Furthermore, many of these proteins were functionally associated with pattern binding, polysaccharide binding, structural constituent of peritrophic membrane and chitin binding, according to Gene Ontology annotation. Conclusion The PM protein composition, which provides a basis for further functional investigations of the identified proteins, will be useful for understanding the housefly larval gut immune system and may help to identify potential targets and exploit new bioinsecticides. Key words: Musca domestica; Peritrophic matrix; Proteome Biomed Environ Sci, 2016; 29(1): 56-65 doi: 10.3967/bes2016.006 ISSN: 0895-3988 www.besjournal.com (full text) CN: 11-2816/Q Copyright ©2016 by China CDC INTRODUCTION physiology, as it protects the midgut epithelium from abrasion by food particles and toxins, serves as the n many insects, the midgut epithelium is first biophysical barrier that alters the temporal generally lined with an extracellular, kinetics of host immune responses to pathogens I semi-permeable structure referred to as the ingested during feeding, and increases digestive peritrophic matrix (PM), PM and mucous secretions efficiency by compartmentalizing of digestive are somewhat similar, but there are big differences processes[1-6]. The insect PM is composed of chitin too. The PM is essential for insect digestive and glycoproteins, exhibiting characteristic *This work was supported by National Natural Science Foundation of China (No. 81360254); National Sci-Tech Support Plan of China under grant (No.2011BAC06B12); and the Science and Technology Program of Guizhou Province Agricultural research Project [No. NY (2014)3054]. #Correspondence should be addressed to WU Jian Wei, Professor, PhD, Tel: 86-851-86825253, E-mail: wjw@ gmc.edu.cn Biographical note of the first author: WANG Yu, male, born in 1987, PhD candidate, majoring in insect molecular biology. Proteomic analysis of the peritrophic matrix 57 chitin-binding activity[7]. PM proteins are classified evidence suggests that the housefly gut is a primary into four categories on the basis of extractability site of pathogen replication after oral infection[25-26]. under different conditions, and most of these Therefore, the importance of the PM as a crucial proteins play significant roles in the functions of the component of the local intestinal immune system PM[7]. Studies of the red flour beetle Tribolium merits further research. However, few studies have castaneum have demonstrated that specific investigated the housefly’ PM, and therefore, the individual PM proteins may regulate PM available information with which to understand its permeability, and that a gradient of PM barrier biological function is very limited. For a better function is essential for survival[8]. The PM not only understanding of how the PM performs these plays important roles in facilitating food digestion functions, a thorough revision of the molecular and protecting the gut epithelium, but also may act architecture of the housefly PM is required. as a significant structural target for insect In this study, we attempted to achieve a control[9-10]. comprehensive identification of proteins in the PM The housefly (Musca domestica L.; Diptera: of the housefly, Musca domestica. Muscidae) is a major domestic, medical, and veterinary pest that causes more than 100 human MATERIALS AND METHODS and animal diseases, including bacterial, protozoan, [11-12] helminthic, viral, and rickettsial infections . Housefly Larval Rearing Conditions and Isolation of Although insecticides have been widely used to Larval PM control insect pests, the housefly has shown a remarkable and rapid ability to evolve House flies were reared at the Department of [13-14] Parasitology, Guizhou Medical University (Guiyang, resistance . Therefore, effective novel strategies [27] of housefly control are vital for limiting the spread of China) . Larvae were raised in a climate-controlled disease, the evolution of resistance, and the room at 25 °C with a relative humidity of 75%-85% economic losses associated with reduced and were provided, medium comprising wheat bran production[15]. Because bacteria are both nutritional (500 g), heat-inactivated yeast (30 g), and water and developmental requirement of housefly (1500 mL) until pupation. After eclosion, adult flies larvae[16], adult flies associate with microbe-rich were fed water, sugar, and milk powder. Flies were animal waste or septic substrates throughout their maintained at 25 °C under a 12 h light /12 h dark life cycle[17], but rarely show signs of disease, cycle (LD12:12). indicating the presence of efficient systems for gut The PM was isolated from the midgut of the defense in conjunction with biophysical barriers third instar larvae. The larval midgut was dissected, against microbes[18]. Bacteria do not pass through and the PM allowed slid to out when the end of the the PM and are sequestered therein by size midgut was cut transversely, the PM was exclusion, in addition, some bacterial species appear subsequently washed with a 0.75% NaCl solution to be immobilized within the PM by an unknown until no food debris remained. Approximately 200 mechanism[19]. PMs were pooled and stored at -80 °C until further Housefly larvae play a vital role in ecosystems as use. decomposers of organic waste[14]. Adaptation to Sample Preparation and Gel Electrophoresis different ecological environments may have led to the evolution of a housefly defense response. The PM was homogenized on ice for 5 min, and Therefore, a comparison of the PM proteome from total proteins were extracted with 20 μL of lysis Musca domestica with those from species exposed buffer (2.5% SDS, 10% glycerin, 5% to different niches and microorganisms, such as β-mercaptoethanol, and 50 mmol/L Tris-HCl, pH 8.8) Anopheles and Drosophila, may be very significant, per mg of sample weight[24]. PMs were disrupted and will provide insights into housefly survival after incubation with the lysis buffer, mixed several maintenance while in close contact with many times during a 1 h incubation at 4 °C, and pathogens[12,20]. The housefly niches is unique centrifuged at 13,000 rpm and 4 °C for 20 min, after relative to that of other insects (e.g., Anopheles which the supernatant was recovered and gambiae[21], Helicoverpa armigera[22] and Glossina recentrifuged. Protein concentrations were morsitans morsitans[23], Bombyx mori[9,24]), for which measured according to the Bradford method[28]. the PM composition have been reported. Increasing Each sample was subsequently boiled for 10 min and 58 Biomed Environ Sci, 2016; 29(1): 56-65 centrifuged at 13,000 rpm and 4 °C for 10 min, after C18 column (75 µm × 15 cm, C18, 3 µm, 120 Å) with a which the proteins in the supernatant were spray tip. Data acquisition was performed with a separated by one-dimensional sodium dodecyl triple time-of-flight (TOF) 5600 system (AB SCIEX) sulfate-polyacrylamide gel electrophoresis (SDS- fitted with a nanospray III source (AB SCIEX), with a PAGE) with a 5% stacking gel and 12% resolving gel, pulled quartz tip as the emitter (New Objective, according to the standard method. The gels were Woburn, MA, USA). Data were acquired using an ion then stained with Coomassie Brilliant Blue R250 spray voltage of 2.5 kV, curtain gas of 30 PSI, (Sigma-Aldrich, St. Louis, MO, USA). nebulizer gas of 5 PSI, and an interface heater temperature of 150 °C. For information dependent In-gel Digestion acquisition (IDA), survey scans were acquired over a A 1D-PAGE lane containing all protein bands was 250 ms period, and as many as 35 product ion scans manually cut from top to bottom, excised gel slices were collected if they exceeded a threshold of 150 were washed twice with ultrapure water and counts per second (counts/s) with a 2+ to 5+ destained in 25 mmol/L ammonium bicarbonate/ charge-state. The total cycle time was fixed at 2.5 s. 50% acetonitrile (pH 8.0) at room temperature for A rolling collision energy setting was applied to all 30 min. Gel slices were subsequently dehydrated for precursor ions for collision-induced dissociation (CID). 30 min in 50% acetonitrile and 100% acetonitrile, Dynamic exclusion was set for ½ half of the peak respectively. The acetonitrile was then removed and width (18 s), and the precursor was accordingly the gel slices were reduced for 1 h at 37 °C with 1 refreshed from the exclusion list. mmol/L dithiothreitol in 25 mmol/L ammonium Based on the combined MS and MS/MS spectra, bicarbonate, followed by alkylation with 50 mmol/L proteins were successfully identified at a 95% or iodoacetamide in 25 mmol/L ammonium higher confidence interval, using their scores in the bicarbonate for 30 min at room temperature.
Load more

Recommended publications
  • Arachnida, Solifugae) with Special Focus on Functional Analyses and Phylogenetic Interpretations
    HISTOLOGY AND ULTRASTRUCTURE OF SOLIFUGES Comparative studies of organ systems of solifuges (Arachnida, Solifugae) with special focus on functional analyses and phylogenetic interpretations HISTOLOGIE UND ULTRASTRUKTUR DER SOLIFUGEN Vergleichende Studien an Organsystemen der Solifugen (Arachnida, Solifugae) mit Schwerpunkt auf funktionellen Analysen und phylogenetischen Interpretationen I N A U G U R A L D I S S E R T A T I O N zur Erlangung des akademischen Grades doctor rerum naturalium (Dr. rer. nat.) an der Mathematisch-Naturwissenschaftlichen Fakultät der Ernst-Moritz-Arndt-Universität Greifswald vorgelegt von Anja Elisabeth Klann geboren am 28.November 1976 in Bremen Greifswald, den 04.06.2009 Dekan ........................................................................................................Prof. Dr. Klaus Fesser Prof. Dr. Dr. h.c. Gerd Alberti Erster Gutachter .......................................................................................... Zweiter Gutachter ........................................................................................Prof. Dr. Romano Dallai Tag der Promotion ........................................................................................15.09.2009 Content Summary ..........................................................................................1 Zusammenfassung ..........................................................................5 Acknowledgments ..........................................................................9 1. Introduction ............................................................................
    [Show full text]
  • Genetic Evidence for a Protective Role of the Peritrophic Matrix Against Intestinal Bacterial Infection in Drosophila Melanogaster
    Genetic evidence for a protective role of the peritrophic matrix against intestinal bacterial infection in Drosophila melanogaster Takayuki Kuraishi1, Olivier Binggeli, Onya Opota, Nicolas Buchon, and Bruno Lemaitre1 Global Health Institute, École Polytechnique Fédérale de Lausanne, 1015 Lausanne, Switzerland Edited by Alexander S. Raikhel, University of California, Riverside, CA, and approved August 17, 2011 (received for review April 14, 2011) The peritrophic matrix (PM) forms a layer composed of chitin and of the cytokine Upd3 from damaged enterocytes, which then glycoproteins that lines the insect intestinal lumen. This physical activates the JAK/STAT pathway in intestinal stem cells to pro- barrier plays a role analogous to that of mucous secretions of the mote both their division and differentiation, establishing a ho- vertebrate digestive tract and is thought to protect the midgut meostatic regulatory loop (9, 10). Interestingly, both Imd pathway epithelium from abrasive food particles and microbes. Almost activity and epithelium renewal are also stimulated at a basal level nothing is known about PM functions in Drosophila, and its function by the indigenous gut microbiota (10). as an immune barrier has never been addressed by a genetic ap- The peritrophic matrix (PM) forms a layer composed of chitin proach. Here we show that the Drosocrystallin (Dcy) protein, a pu- and glycoproteins that lines the insect midgut lumen (11, 12). tative component of the eye lens of Drosophila, contributes to adult Although structurally different, it plays a role analogous to that of PM formation. A loss-of-function mutation in the dcy gene results mucous secretions of the vertebrate digestive tract and is thought in a reduction of PM width and an increase of its permeability.
    [Show full text]
  • Vsgovercomesanearlybarriertos
    COMMENTARY VSG overcomes an early barrier to survival of African trypanosomes in tsetse flies COMMENTARY Shaden Kamhawia,1 and Iliano V. Coutinho-Abreua The widespread expansion of vector-borne diseases is susceptible to infection compared with adults that a testament to their success. According to the World produce a mature PM (13). Importantly, the mecha- Health Organization, over half the global population is nism by which trypanosomes traverse the PM to en- at risk for contracting a vector-borne disease, and over able gut colonization in susceptible tsetse had been a million deaths are annually attributed to diseases unknown until now. In PNAS, Aksoy et al. (14) share transmitted by insects (1). An absence of preventative their discovery of how trypanosomes disrupt the vaccines combined with the rising resistance to insec- PM of the tsetse fly and implicate the variant sur- ticides has led to a surge in efforts to develop alter- face glycoprotein (VSG) of the blood stream form nate approaches toward vector control. One such (BSF)—famed for its critical part in escaping the im- approach is the interruption of pathogen transmission. mune system of the mammalian host (15)—in its Understanding the molecular basis of parasite–vector disruption, revealing a dual role for VSG in the life interactions can identify critical steps in pathogen de- cycle of trypanosomes. velopment to be targeted for disruption. After millions Infected blood ingested by a tsetse fly contains a of years adapting to their vectors, pathogens have large number of slender and a few stumpy BSF try- evolved complex and innovative survival strategies panosomes, both expressing a similar coat of VSG aimed at overcoming host defenses.
    [Show full text]
  • Proteases As Insecticidal Agents
    Toxins 2010, 2, 935-953; doi:10.3390/toxins2050935 OPEN ACCESS toxins ISSN 2072-6651 www.mdpi.com/journal/toxins Review Proteases as Insecticidal Agents Robert L. Harrison 1 and Bryony C. Bonning 2,* 1 Invasive Insect Biocontrol and Behavior Laboratory, USDA Agricultural Research Service, Plant Sciences Institute, 10300 Baltimore Avenue, Beltsville, Maryland 20705, USA; E-Mail: [email protected] 2 Department of Entomology, Iowa State University, 418 Science II, Ames, IA 50011-3222, USA * Author to whom correspondence should be addressed; E-Mail: [email protected]; Tel.: +01-515-294-1989; Fax: +01-515-294-5957. Received: 12 April 2010; in revised form: 26 April 2010 / Accepted: 30 April 2010 / Published: 5 May 2010 Abstract: Proteases from a variety of sources (viruses, bacteria, fungi, plants, and insects) have toxicity towards insects. Some of these insecticidal proteases evolved as venom components, herbivore resistance factors, or microbial pathogenicity factors, while other proteases play roles in insect development or digestion, but exert an insecticidal effect when over-expressed from genetically engineered plants or microbial pathogens. Many of these proteases are cysteine proteases, although insect-toxic metalloproteases and serine proteases have also been examined. The sites of protease toxic activity range from the insect midgut to the hemocoel (body cavity) to the cuticle. This review discusses these insecticidal proteases along with their evaluation and use as potential pesticides. Keywords: insecticides; basement membrane; cuticle; peritrophic matrix; plant defense; microbial defense 1. Toxic Proteins as Insecticidal Agents The use of classical chemical insecticides to control agricultural pests poses hazards to human health, other non-target species including beneficial insects such as pollinators, and the environment.
    [Show full text]
  • Downloaded the Hidden Markov Model (HMM) Containing the CBM 14 Family from the PFAM Database (
    International Journal of Molecular Sciences Article Identification of Peritrophins and Antiviral Effect of Bm01504 against BmNPV in the Silkworm, Bombyx mori Xu-Le Zha 1, Xin-Bo Yu 1, Hong-Yan Zhang 1, Han Wang 1, Xian-Zhi Huang 2, Yi-Hong Shen 1,* and Cheng Lu 1,* 1 State Key Laboratory of Silkworm Genome Biology, Southwest University, Beibei, Chongqing 400715, China; [email protected] (X.-L.Z.); [email protected] (X.-B.Y.); [email protected] (H.-Y.Z.); [email protected] (H.W.) 2 Science and Technology Department, Southwest University, Chongqing 400715, China; [email protected] * Correspondence: [email protected] (Y.-H.S.); [email protected] (C.L.); Tel.: +86-138-8360-7000 (Y.-H.S.); +86-23-6825-0346 (C.L.) Received: 11 October 2020; Accepted: 23 October 2020; Published: 27 October 2020 Abstract: The insect midgut secretes a semi-permeable, acellular peritrophic membrane (PM) that maintains intestinal structure, promotes digestion, and protects the midgut from food particles and pathogenic microorganisms. Peritrophin is an important PM protein (PMP) in the PM. Here, we identified 11 peritrophins with 1–16 chitin binding domains (CBDs) comprising 50–56 amino acid residues. Multiple CBDs in the same peritrophin clustered together, rather than by species. The CBD contained six highly conserved cysteine residues, with the key feature of amino acids between them being CX11-15CX5CX9-14CX11-12CX6-7C. Peritrophins with 2 and 4 CBDs (Bm09641 and Bm01504, respectively), and with 1, 8, and 16 CBDs (Bm11851, Bm00185, and Bm01491, respectively) were mainly expressed in the anterior midgut, and throughout the midgut, respectively.
    [Show full text]
  • Larval Peritrophic Matrix Yu-Bo Lin1,2†, Jing-Jing Rong1,2†, Xun-Fan Wei3, Zhuo-Xiao Sui3, Jinhua Xiao3 and Da-Wei Huang3*
    Lin et al. Proteome Science (2021) 19:7 https://doi.org/10.1186/s12953-021-00175-x RESEARCH Open Access Proteomics and ultrastructural analysis of Hermetia illucens (Diptera: Stratiomyidae) larval peritrophic matrix Yu-Bo Lin1,2†, Jing-Jing Rong1,2†, Xun-Fan Wei3, Zhuo-Xiao Sui3, Jinhua Xiao3 and Da-Wei Huang3* Abstract Background: The black soldier fly (Hermetia illucens) has significant economic potential. The larvae can be used in financially viable waste management systems, as they are voracious feeders able to efficiently convert low-quality waste into valuable biomass. However, most studies on H. illucens in recent decades have focused on optimizing their breeding and bioconversion conditions, while information on their biology is limited. Methods: About 200 fifth instar well-fed larvae were sacrificed in this work. The liquid chromatography-tandem mass spectrometry and scanning electron microscopy were employed in this study to perform a proteomic and ultrastructural analysis of the peritrophic matrix (PM) of H. illucens larvae. Results: A total of 565 proteins were identified in the PM samples of H. illucen, of which 177 proteins were predicted to contain signal peptides, bioinformatics analysis and manual curation determined 88 proteins may be associated with the PM, with functions in digestion, immunity, PM modulation, and others. The ultrastructure of the H. illucens larval PM observed by scanning electron microscopy shows a unique diamond-shaped chitin grid texture. Conclusions: It is the first and most comprehensive proteomics research about the PM of H. illucens larvae to date. All the proteins identified in this work has been discussed in details, except several unnamed or uncharacterized proteins, which should not be ignored and need further study.
    [Show full text]
  • Bulletin Number / Numéro 4 Entomological Society of Canada December / Décembre 2011 Société D’Entomologie Du Canada
    ............................................................ ............................................................ Volume 43 Bulletin Number / numéro 4 Entomological Society of Canada December / décembre 2011 Société d’entomologie du Canada Published quarterly by the Entomological Society of Canada Publication trimestrielle par la Société d’entomologie du Canada ........................................................ .......................................................................................................................................................... ............................................................................................................................................................ ......................................................................................................................................................................................................................... ........................................................................................... ............................................................... .......................................................................................................................................................................................... List of contents / Table des matières Volume 43(4), December / décembre 2011 Up front / Avant-propos ..............................................................................................................169 Moth balls / Boules à mites...................................................................................................172
    [Show full text]
  • Proteases As Insecticidal Agents Robert L
    Entomology Publications Entomology 2010 Proteases as Insecticidal Agents Robert L. Harrison United States Department of Agriculture Bryony C. Bonning Iowa State University, [email protected] Follow this and additional works at: http://lib.dr.iastate.edu/ent_pubs Part of the Entomology Commons The ompc lete bibliographic information for this item can be found at http://lib.dr.iastate.edu/ ent_pubs/2. For information on how to cite this item, please visit http://lib.dr.iastate.edu/ howtocite.html. This Article is brought to you for free and open access by the Entomology at Iowa State University Digital Repository. It has been accepted for inclusion in Entomology Publications by an authorized administrator of Iowa State University Digital Repository. For more information, please contact [email protected]. Proteases as Insecticidal Agents Abstract Proteases from a variety of sources (viruses, bacteria, fungi, plants, and insects) have toxicity towards insects. Some of these insecticidal proteases evolved as venom components, herbivore resistance factors, or microbial pathogenicity factors, while other proteases play roles in insect development or digestion, but exert an insecticidal effect when over-expressed from genetically engineered plants or microbial pathogens. Many of these proteases are cysteine proteases, although insect-toxic metalloproteases and serine proteases have also been examined. The sites of protease toxic activity range from the insect midgut to the hemocoel (body cavity) to the cuticle. This review discusses these insecticidal proteases along with their evaluation and use as potential pesticides. Keywords insecticides, basement membrane, cuticle, peritrophic matrix, plant defense, microbial defense Disciplines Entomology Comments This article is from Toxins 2, no.
    [Show full text]
  • Volatile DMNT Directly Protects Plants Against Plutella Xylostella By
    RESEARCH ARTICLE Volatile DMNT directly protects plants against Plutella xylostella by disrupting the peritrophic matrix barrier in insect midgut Chen Chen1†, Hongyi Chen1†, Shijie Huang1†, Taoshan Jiang1, Chuanhong Wang1, Zhen Tao1, Chen He1, Qingfeng Tang2, Peijin Li1* 1The National Key Engineering Lab of Crop Stress Resistance Breeding, the School of Life Sciences, Anhui Agricultural University, Hefei, China; 2Key Laboratory of Biology and Sustainable Management of Plant Diseases and Pests of Anhui Higher Education Institutes, the School of Plant Protection, Anhui Agricultural University, Hefei, China Abstract Insect pests negatively affect crop quality and yield; identifying new methods to protect crops against insects therefore has important agricultural applications. Our analysis of transgenic Arabidopsis thaliana plants showed that overexpression of pentacyclic triterpene synthase 1, encoding the key biosynthetic enzyme for the natural plant product (3E)-4,8-dimethyl- 1,3,7-nonatriene (DMNT), led to a significant resistance against a major insect pest, Plutella xylostella. DMNT treatment severely damaged the peritrophic matrix (PM), a physical barrier isolating food and pathogens from the midgut wall cells. DMNT repressed the expression of PxMucin in midgut cells, and knocking down PxMucin resulted in PM rupture and P. xylostella death. A 16S RNA survey revealed that DMNT significantly disrupted midgut microbiota *For correspondence: populations and that midgut microbes were essential for DMNT-induced killing. Therefore, we [email protected] propose that the midgut microbiota assists DMNT in killing P. xylostella. These findings may †These authors contributed provide a novel approach for plant protection against P. xylostella. equally to this work Competing interests: The authors declare that no Introduction competing interests exist.
    [Show full text]
  • Abstracts Content
    Abstracts Content Keynotes 1 Oral Presentations 11 Behavioral Biology 12 Developmental Biology 32 Ecology 46 Evolutionary Biology 60 Morphology 86 Neurobiology 100 Physiology 114 Zoological Systematics 128 Keynotes Poster Presentations 142 Behavioral Biology 143 Developmental Biology 160 Ecology 166 Evolutionary Biology 173 Morphology 193 Neurobiology 211 Physiology 248 Zoological Systematics 258 Satellite Symposia 266 Satellite Symposium I 267 Arthropod Neuro Network (ANN) Satellite Symposium II 278 Epigenetics in Animal Physiology Satellite Symposium III 281 Insect Biotechnology Satellite Symposium IV 299 Animal Phylogeny - Past, Present and Future Author Index 311 1 Keynotes KN-01 KN-02 Oral Presentations Insect Biotechnology C. elegans in an Evolutionary and Ecological Context: Vulva Development and Natural Pathogens 1,2 Poster A. Vilcinskas Presentations 1Institute of Phytopathology and Applied Zoology, Justus-Liebig- M.- A. Félix1 University of Gießen, Applied Entomology, Gießen, Germany 1Institut de Biologie de l’Ecole Normale Supérieure, CNRS-ENS-Inserm, Satellite Symposia 2Fraunhofer Institute of Molecular Biology and Applied Ecology, Paris, France Department Bioresources, Gießen, Germany Biological processes are generally studied in the laboratory under one Insect Biotechnology, alternatively called yellow biotechnology, has environmental condition and in one reference genetic background. We been defined as the use of biotechnology-based approaches to exploit try to widen this horizon to answer questions on the relationship between insects or insect-derived molecules, cells, organs or microorganisms genetic and phenotypic evolution, by placing a paradigmatic model for development of products or services for mediacl, agricultural or system in developmental biology, C. elegans vulval cell fate patterning, in industrial applications. This ermerging field is developed within the its evolutionary context.
    [Show full text]
  • Gut Microbiota Is Essential in PGRP-LA Regulated Immune
    Gao et al. Parasites Vectors (2020) 13:3 https://doi.org/10.1186/s13071-019-3876-y Parasites & Vectors RESEARCH Open Access Gut microbiota is essential in PGRP-LA regulated immune protection against Plasmodium berghei infection Li Gao1,2, Xiumei Song1,2 and Jingwen Wang1,2* Abstract Background: Malaria remains to be one of the deadliest infectious diseases and imposes substantial fnancial and social costs in the world. Mosquitoes rely on the immune system to control parasite infection. Peptidoglycan recog- nition proteins (PGRPs), a family of pattern-recognition receptors (PRR), are responsible for initiating and regulating immune signaling pathways. PGRP-LA is involved in the regulation of immune defense against the Plasmodium para- site, however, the underlying mechanism needs to be further elucidated. Methods: The spatial and temporal expression patterns of pgrp-la in Anopheles stephensi were analyzed by qPCR. The function of PGRP-LA was examined using a dsRNA-based RNA interference strategy. Western blot and periodic acid schif (PAS) staining were used to assess the structural integrity of peritrophic matrix (PM). Results: The expression of pgrp-la in An. stephensi was induced in the midgut in response to the rapid proliferat- ing gut microbiota post-blood meal. Knocking down of pgrp-la led to the downregulation of immune efectors that control gut microbiota growth. The decreased expression of these immune genes also facilitated P. berghei infection. However, such dsLA treatment did not infuence the structural integrity of PM. When gut microbiota was removed by antibiotic treatment, the regulation of PGRP-LA on immune efectors was abolished and the knock down of pgrp-la failed to increase susceptibility of mosquitoes to parasite infection.
    [Show full text]
  • The Chitinase Pfcht1 from the Human Malaria Parasite Plasmodium Falciparum Lacks Proenzyme and Chitin-Binding Domains and Displays Unique Substrate Preferences
    The chitinase PfCHT1 from the human malaria parasite Plasmodium falciparum lacks proenzyme and chitin-binding domains and displays unique substrate preferences Joseph M. Vinetz*†, Sanat K. Dave*, Charles A. Specht‡, Kenneth A. Brameld§,BoXu¶, Rhian Haywardʈ, and David A. Fidockʈ *WHO Center for Tropical Diseases, University of Texas Medical Branch, Galveston, TX 77555; ‡Department of Molecular and Cell Biology, Goldman School of Dental Medicine, Boston University, Boston, MA 02118; §Department of Pharmaceutical Chemistry, University of California, San Francisco, CA 94143; ¶Department of Human Biological Chemistry and Genetics, University of Texas Medical Branch, Galveston, TX 77555; and ʈLaboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892 Communicated by Phillips W. Robbins, Boston University, Boston, MA, September 30, 1999 (received for review August 2, 1999) Within hours after the ingestion of a blood meal, the mosquito (6). The PMs of the Plasmodium vectors Anopheles gambiae midgut epithelium synthesizes a chitinous sac, the peritrophic (which carries human malaria parasites) and A. aegypti (which matrix. Plasmodium ookinetes traverse the peritrophic matrix carries avian malaria parasites) are composed of chitin, a while escaping the mosquito midgut. Chitinases (EC 3.2.1.14) are ␤-1,4-linked polymer of GlcNAc, with intercalated proteins critical for parasite invasion of the midgut: the presence of the including trypsins and peritrophins (7–9). P. gallinaceum ooki- chitinase inhibitor, allosamidin, in an infectious blood meal pre- netes secrete active chitinase (refs. 10 and 11; J.V., J. Valenzuela, vents oocyst development. A chitinase gene, PgCHT1, recently has L. Aravind, J. Ribeiro, and D. Kaslow, unpublished data).
    [Show full text]