Biochemistry and physiology of arachnids


Team of work

Cunningham, Mónica Liliana
Principal Investigator
monilicu@yahoo.com.ar

García, Carlos Fernando
Principal Investigator
cfgarcia1123@yahoo.com.ar
Laino, Aldana
Investigator
aldana_laino@hotmail.com
Romero, Sofía
Research Fellow
sofiaromero321@msn.com
Suarez, Gustavo
Thesis Students
gustavosuarez964@hotmail.com
   


Collaborators from other institutions

International

Costa, Fernando G. 
Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay.

Sotelo-Mundo, Rogerio
Centro de Investigación en Alimentación y Desarrollo (CIAD) y Universidad de Sonora (UNISON). Hermosillo, Sonora, Mexico.

Trabalon Marie
Departamento de Biología, Universidad de Rennes. Rennes, Francia.

National

Barneche Jorge
Centro de Estudios Parasitológicos y de Vectores (CEPAVE), CONICET-UNLP. La Plata.

González, Alda
Centro de Estudios Parasitológicos y de Vectores (CEPAVE), CONICET-UNLP. La Plata.

Mattoni, Camilo. 
Laboratorio de Biología Reproductiva y Evolución, Instituto de Diversidad y Ecología Animal (CONICET-UNC).

Ojanguren-Affilastro, Andrés.
Museo Argentino de Ciencias Naturales, Bernardino Rivadavia (CABA), Argentina.


Line overview

Arachnids are one of the arthropod groups that have greater diversity; there are over 100 families of spiders distributed all over the world with more than 42,000 species described. In Argentina, 48 families are represented with approximately 1,000 species with an important ecological role. The general objective of the present line of research is to advance on the knowledge about physiological, biochemical and biological aspects of these organisms, mainly in relation to the role of not only hemolymph but also vitellin lipoproteins; enzymes, and immunologic and reproductive processes.

Sub-projects:

  1. Identification and characterization of vitellogenin in the spiders Grammostola rosea and Polybetes

The knowledge and understanding of the vitellogenin functionality are aspects of great interest, since they can determine the survival of the embryos and, therefore, of the species. However, the lack of knowledge of vitellogenins in arachnids impedes any speculation related to the subject. One of the objectives of the project is to identify and characterize the lipoproteins present in the tarantula G. rosea and P. pythagoricus in reproductive and non-reproductive stages in order to determine the complete functioning of the lipoprotein system.

Specific objectives:

  • To isolate and identify the different lipoproteins: hemolymph, vitellogenin and of egg of the 2 model spider.
  • To characterize such lipoprotein proteins regarding its protein composition in dissociating and native conditions, employing different electrophoretic techniques,
  • To characterize lipid composition of the different lipoproteins, as well as the fatty acid composition, utilizing chromatography techniques.
  • To correlate protein and lipid compositions of lipoprotein with their different functions.
  • To generate polyclonal antibodies against vitellus lipoprotein.
  • To determine the existing relations among the different lipoproteins present in hemolymph vitellogenin), in ovary, and in egg by means of their immunologic identities.
  1. Comparative analysis of lipids and fatty acids present in different organs of several species of scorpions.

 Lipids generally have three fundamental functions in the organisms: energy reserve, majority components of membranes, and signaling. Within arthropods most data regarding the way of mobilization and storage of lipids are found in insects and crustaceans. A few works were carried out in scorpions. This order has a remarkable importance as model in biochemistry of arthropods, since they are characterized by understanding very primitive animals with little variation throughout the time.

Specific objectives:

  • To isolate the different organs involved in the dynamics of lipids: hepatopancreas, muscle, hemolymph.
  • To characterize comparatively the lipids and fatty acids of three species of scorpions: Timogenes elegans, Timogenes dorbignyi, and Brachistosternus ferrugineus, focusing on the main organs/tissues involved in the dynamics of lipids.
  1. Functional and structural characterization of phospho-kinase Arginine Kinase (AK) of Polybetes pythagoricus.

In collaboration with the group of work of Dr. Rogerio Sotelo-Mundo from Centro de Investigación en Alimentación y Desarrollo (CIAD) y Universidad de Sonora (UNISON) (Hermosillo, México), we performed structural studies, particularly of Arginine Kinase (AK). This enzyme has a particular importance since it is not only a phospho-kinase involved in the energetic metabolism of invertebrates, but also responsible for the production of Ig-E in humans, mediating in allergenic processes.

Specific objectives:

  • To obtain the amino acid sequence of the Arginine-kinase enzyme.
  • To overexpress the protein in E. coli.
  • To carry out activity measurements through coupled enzymatic assays.
  • To perform functional and structural characterization of this enzyme through the production of crystals and titration isothermal measurements.
  • To study its allergenic capacity through immunological techniques.
  1. Comparative analysis of the dynamics of lipids in Spiders Araneomorphae and Mygalomorphae. Differences and similarities among Infraorders

Lipids generally have three main functions in the organisms: energy reserve, majority components in membranes and signaling. With arthropods most data related to the way of mobilization and storage of lipids are found in insects and crustaceans. Literature is scarce and confusing regarding arachnids. We know the different species of arachnids have differences and similarities in their lipid dynamics, which are subject to phylogenetic relationships and life habits, with fatty acids patterns in the different species of spiders and immunological identity among lipophorins of different species of spiders and among the hemocyanins of several species of studied spiders.

Specific objectives:

  • To study the lipid dynamics in different species of arachnids (representatives of the Infraorders Araneomorpha and Mygalomorphae), its storage, hemolymph transport and their presence in different organs.
  • To determine quali- and quantitatively the lipid composition of the intestinal diverticulum, hemolymph and muscle of the different studied species.
  • To determine quali- and quantitatively the fatty acid composition of the intestinal diverticulum, hemolymph and muscle of the different studied species.
  • To determine the immunologic identity of the different proteins present in the hemolymph.
  • To correlate the possible differences and similarities taking into consideration the phylogenetic relationships.
  1. Effect of insecticides on enzymes involved in detoxification mechanisms of spiders present in agroecosystems.

In agroecosystems spiders are considered natural enemies of harmful insects, being the reason why the study of the communities of these insects in agricultural environments has gained importance in the last years. The aim of the present line of research is to study the effect of insecticides commonly used in diverse crops on the expression/activity of different enzymes involved in the detoxification mechanisms.

Specific objectives:

  • To determine the lethal dose of 50 of cypermethrin and other insecticides used in crops.
  • To evaluate the effects of these insecticides on the activity of different enzymes such as: acetylcholinesterase, superoxide-dismutase, catalase and glutathione S-transferase using colorimetric techniques.
  • To evaluate the effects of the insecticides on the differential expression of the enzymes using real time-PCR.
  1. Characterization of hemocyte, hemolymph and antimicrobial peptides in the spider Polybetes pythagoricus. Role of hemocyanin in the immune system of arachnids.

Given the abundance of Hc in the hemolymph of invertebrates, the absence of a real phenoloxidase (PO) enzyme in chelicerates, the diverse immune functions described in Hcs of diverse organisms, and our recent results that indicate Hc of the spider Polybetes pythagoricus has PO activity induced with SDS, we propose that maybe this organism can use Hc as a defense molecule in the event of an infection. In this way, Hc of Polybetes pythagoricus could be precursor of antimicrobial peptides (AMPs); it could also show antiviral properties due to a hemolytic and/or agglutinating behavior or the synthesis of reactive oxygen species. Therefore, hemocytes could have certain peptides also acting in the defense system.

Specific objectives:

  • To generate an immunological challenge and analyze from now on the possible peptides and their characteristics not only in hemolymph but also in hemocytes of the spider Polybetes pythagoricus.
  • To identify, sequence, and synthesize synthetic peptides from peptides generated by the immunological challenge, and then to perform antimicrobial assays.
  • To analyze the possible role of hemocyanin in the defense processes against microorganisms.
  • To evaluate if hemocyanin has the capacity of agglutinating bacteria and/or erythrocytes, and if it can generate antimicrobial peptide by partial proteolysis.

Publications

 

  • Acute toxicity of organophosphate fenitrothion on biomarkers in prawn Palaemonetes argentinus (Crustacea: Palaemonidae).
    Lavarias, S; García, C. F.
    2015. Environmental Monitoring and Assessment. , Berlin: SPRINGER, N° 65, p. 187
    -197 ISSN 0167-6369

  • Analysis of lipid and fatty acid composition of three species of scorpions with relation to different organs.
    Laino, A; Mattoni, C; Ojanguren, A; Cunningham, M; García, F.
    2015. Comparative Biochemistry and Physiology. part b, Biochemistry & Molecular Biology: ELSEVIER SCIENCE INC, vol. 190, p. 27
    -36. ISSN 1096-4959

  • Characterization of Phenoloxidase Activity From Spider Polybetes pythagoricus Hemocyanin.
    Laino, A; Lavarías, S; Suarez, G; Lino, A; Cunningham, M.
    2015. Journal of Experimental Zoology part a- Ecological Genetics and Physiology. , New York: WILEY-LISS, DIV JOHN WILEY & SONS INC, vol. 323, p. 547-555. ISSN 1932-5223

  • Identification and characterization of the Lipid transport system in the tarantula Grammostola rosea.
    Laino, A; Cunningham, M; Suarez, G; García, C. F.
    2015.
    Open Journal of Animal Science. : Scientific Research, vol. 5, p. 9
    -20. ISSN 2161-7597

  • Protein characterization and fatty acid composition of VHDL subfraction II of the spider Polybetes pythagoricus.
    Laino, A; García, F; Cunningham, M.
    2015.
    Biocell. , Mendoza: INST HISTOL EMBRIOL-CONICET, vol. 39, p. 33
    -40. ISSN 0327-9545

  • Energy sources from the eggs of the wolf spider Schizocosa malitiosa: Isolation and characterization of lipovitellins. 
    Laino, A; Cunningham, M; Costa, F; Garcia, C. F. 
    2013. Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology: ELSEVIER SCIENCE INC. vol. 165, p. 172-180. ISSN 1096-4959
  • Study of biochemical biomarkers in freshwater prawn Macrobrachium borellii (Crustacea: Palaemonidae) exposed to organophosphate fenitrothion. 
    Lavarias, S; García, F; Crespo, R; Pedrini, N; Heras, H.
    2013. Ecotoxicology and Environmental Safety. San Diego: Academic Press Inc Elsevier Science. vol. 96, p. 10-16. ISSN 0147-6513
  • The multi-component venom of the spider Cupiennius salei: A bioanalytical investigation applying different strategies. 
    Trachsel, C; Siegemund, D; Kämpfer, U; Kopp, L; Bühr, C; Grossmann, J; Lüthi, C; Cunningham, M; Nentwig, W; Kuhn-Nentwig, L; Schürch, S; Schaller, J. 
    2012. Febs journal. Londres: Wiley-Blackwell Publishing, Inc, vol. 279, p. 2683-2694


  • Vitellogenin and Lipovitellin from the Macrobrachium borellii as hydrocarbon pollution biomarker. 
    García, C. F; Heras, F. 
    2012. Marine Pollution Bulletin. Amsterdam: Pergamon-ElSevier Science Ltd, vol. 64, p. 1631-1636


  • Vitellogenin levels in hemolymph, ovary and hepatopancreas of the freshwater crayfish Cherax quadricarinatus (Decapoda: Parastacidae) during the reproductive cycle.
    Ferre, L; Medesani, D; García, C. F; Grodzielski , M; Rodríguez, E.
    2012. Revista de Biología Tropical, vol. 60, p. 253-261


  • In vitro lipid transfer between lipoproteins and midgut-diverticula in the spider Polybetes pythagoricus.
    Laino, A; Cunningham, M. L; Heras, H; García, F. 
    2011. Comp. Biochem Physiol B 160: 181-186.
  • Isolation and characterization of two vitellins from eggs of the spider Polybetes pythagoricus (Araneae: Sparassidae). 
    Laino, A; Cunningham, M; Heras, H; García, F. 
    2011. Comparative Biochemistry and Physiology, Part B 158: 142-148
  • Structure and stability of crustacean lipovitellin: Influence of lipid content and composition.
    García, C. F; Cunningham, M; Soulages, J. L; Heras, H; Garda, H. A. 
    2010. Comparative biochemistry and physiology. part b, biochemistry & molecular biology. Elsevier Science Inc. vol. 155, n° 2, p. 126-131
  • First insight into the lipid uptake, storage and mobilization in arachnids. Role of midgut diverticula and lipoproteins. 
    Laino, A; Cunningham, M; Garcia, F; Heras, H. 
    2009. Journal of Insect Physiology 55: 1118-1124
  • Embryo lipoproteins and yolk lipovitellin consumption during embryogenesis in Macrobrachium borellii (Crustacea: Palaemonidae). 
    García, C. F; Cunningham, M; Garda, H. A; Heras, H. 
    2008. Comparative biochemistry and physiology. part b, biochemistry & molecular biology, vol. 151, p. 317-322
  • Effect of the water-soluble fraction of petroleum on microsomal lipid metabolism of machrobrachium borellii (arthropoda: crustacea). 
    Lavarias, S. M. L; García, F; Pollero, R; Heras, H.  
    2007. Aquatic toxicology. Elsevier. vol. 82, p. 265-271
  • Arachnid lipoproteins: Comparative aspects. Comparative biochemistry and physiology. Comparative pharmacology and toxicology.
    Cunningham, M; García, F; Pollero, R. 
    2007. Elsevier Sci, Oxford. vol. 146, p. 79-87

Book - Book chapters

  • The freshwater prawns M borellii and P argentinus (Crustacea: Palaemonidae) as valid indicators of organophospathe pesticide exposure. A biochemical assasement.
    Garcia, C. F and Sotelo-Mundo, R.
    2017. “Ecotoxicology and Genotoxicology: non-traditional Aquatic Models” ISBN 978-1-78262-781-4 en Books Editorial Thomas Graham House, Science Park, Milton Road, Cambridge CB4 OWF, UK. Pag 196-2011. Enero 2017


  • Pid Synthesis and Transport in Shrimps.
    Garcia, C. F; Gonzalez-Baró, M. 
    2013. Hauppauge, NY: Nova Science Publishers, p. 71-94. ISBN 978-1-62417-317-2




INIBIOLP - Instituto de Investigaciones Bioquímicas de La Plata "Profesor Doctor Rodolfo R. Brenner"
Consejo Nacional de Investigaciones Científicas y Técnicas - Facultad de Ciencias Médicas - U.N.L.P.
Avenida 60 y 120 - C.P. 1900 La Plata - Prov. de Buenos Aires - Argentina. Tel. 54-221-4824894 - Fax. 54-221-4258988 - E-mail :