跳至主要内容

Identification and Characterization of the Lipid Transport System in the Tarantula Grammostola rosea

Read full paper at:
http://www.scirp.org/journal/PaperInformation.aspx?PaperID=52876#.VKtJrMnQrzE


Lipids due to its hydrophobic nature are transported in the hemolymph mainly by lipoproteic fractions. In the present work we studied the lipoproteic fractions present in the hemolymph of the spider Grammostola rosea. Through ultracentrifugation two lipoproteic fractions are isolated, one of high density named Gr-HDL (δ = 1.13 - 1.15 g/ml) and the other of very high density called Gr-VHDL (δ = 1.24 - 1.27 g/ml) Gr-VHDL in hemolymph is majority in relation to Gr-HDL. In this sense Gr-VHDL fraction has 98.6% of hemolymphatic proteins, and 89.3% of lipids presents in the hemolymph. Both lipoproteic fractions possess phospholipids such as majority lipids (phosphatidylcholine and phosphatidylethanolamine) and 18:1, 16:0, 18:2 and 18:0 as the major fatty acids. In order to confirm the role played by lipoproteic fractions in vitro assays with different 14C-lipid were performed. It was observed that Gr-VHDL takes up mainly free fatty acids and triacylglycerols unlike that observed for Gr-HDL in relation to phosphatidylcholine. Through electrophoresis it was observed that Gr-VHDL has three proteins: a predominant band of 68 kDa and two others of 99 and 121 kDa. Gr-HDL displayed a predominant band of 93 kDa, and other minority of 249 kDa. In conclusion, this study reports lipid characterization of the lipoproteic fractions present in the hemolymph of the tarantula, G. rosea. The role of each lipoproteic fraction in relation to lipid uptake is sustained by in vitro assays. Similarities and differences are found when it is compared to lipoproteins of only the three species of spiders studied.
Cite this paper
Laino, A. , Cunningham, M. , Suárez, G. and Garcia, C. (2015) Identification and Characterization of the Lipid Transport System in the Tarantula Grammostola rosea. Open Journal of Animal Sciences, 5, 9-20. doi: 10.4236/ojas.2015.51002.
 

[1] Arrese, E.L., Gazard, J.L., Flowers, M.T., Soulages, J.L. and Wells, M.A. (2001) Diacylglycerol Transport in the Insect Fat Body: Evidence of Involvement of Lipid Droplets and the Cytosolic Fraction. Journal of Lipid Research, 42, 225-234.
[2] Blacklock, B.J. and Ryan, R.O. (1994) Hemolymph Lipid Transport. Insect Biochemistry and Molecular Biology, 24, 855-873. http://dx.doi.org/10.1016/0965-1748(94)90015-9
[3] Chino, H. (1985) Lipid Transport: Biochemistry of Hemolymph Lipophorin. In: Kerkut, G.A. and Gilbert, L.I., Eds., Comprehensive Insect Physiology, Biochemistry and Pharmacology, Pergamon Press, New York, Vol. 10, 115-135.
[4] Gonzalez, M.S., Rimoldi, O. and Brenner, R. (1995) Studies on Very-High-Density Lipoprotein of Triatoma infestans Hemolymph in Relation to Its Function as Free Fatty Acid Carrier. Comparative Biochemistry and Physiology B, 110, 767-775. http://dx.doi.org/10.1016/0305-0491(94)00202-6
[5] Soulages, J.L. and Wells, M.A. (1994) Effect of Diacylglycerol Content on Some Physicochemical Properties of the Insect Lipoprotein, Lipophorin. Correlation with the Binding of Apolipophorin-III. Biochemistry, 33, 2356-2362. http://dx.doi.org/10.1021/bi00175a002
[6] Chang, E.S. and O’Connor, J.M. (1983) Metabolism and Transport of Carbohydrates and Lipid. In: Mantel, L.H., Ed., The Biology of Crustacea, Academic Press, New York, 263-278.
[7] García, F., Cunningham, M., González Baró, M.R., Garda, H.A. and Pollero, R.J. (2002) Effect of Fenitrothion on the Physical Properties of Crustacean Lipoproteins. Lipids, 37, 673-679.
http://dx.doi.org/10.1007/s11745-002-0948-5
[8] García, F., González Baró, M.R. and Pollero, R.J. (2002) Transfer of Lipids between Hemolymph and Hepatopancreas in the Shrimp Macrobrachium borellii. Lipids, 37, 581-585.
http://dx.doi.org/10.1007/s11745-002-0936-9
[9] Lee, R.F. (1991) Lipoproteins from the Hemolymph and Ovaries of Marine Invertebrates. In: Gilles, R., Ed., Advances in Comparative and Environmental Physiology, Springer-Verlag, London, Vol. 7. 187-208.
http://dx.doi.org/10.1007/978-3-642-75897-3_3
[10] Lee, R.F. and Puppione, D.L. (1978) Serum Lipoproteins in the Spiny Lobster, Panulirus interruptus. Comparative Biochemistry and Physiology B, 59, 239-243.
[11] Cunningham, M., Pollero, R.J. and Gonzalez, A. (1994) Lipid Circulation in Spiders. Transport of Phospholipids, Free Acids and Triacylglycerols as the Major Lipid Classes by a High-Density Lipoprotein Fraction Isolated from Plasma of Polybetes pythagoricus. Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 109, 333-338. http://dx.doi.org/10.1016/0305-0491(94)90017-5
[12] Cunningham, M., Gonzalez, A. and Pollero, R.J. (2000) Characterization of Lipoproteins Isolated from the Hemolymph of the Spider Latrodectus mirabilis. Journal of Arachnology, 28, 49-55.
http://dx.doi.org/10.1636/0161-8202(2000)028[0049:COLIFT]2.0.CO;2
[13] Cunningham, M., García, F. and Pollero, R.J. (2007) Arachnid Lipoproteins: Comparative Aspects. Comparative Biochemistry and Physiology Part C, 146, 79-87.
[14] Cunningham, M. and Pollero, R.J. (1996) Characterization of Lipoprotein Fractions with High Content of Hemocyanin in the Hemolymphatic Plasma of Polybetes pythagoricus. Journal of Experimental Zoology, 274, 275-280. http://dx.doi.org/10.1002/(SICI)1097-010X(19960401)274:5<275::AID-JEZ2>3.0.CO;2-M
[15] Haunerland, N.H. and Bowers, W.S. (1987) Lipoproteins in the Hemolimph of the Tarantula, Eurypelma californicum. Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 86, 571-574.
http://dx.doi.org/10.1016/0305-0491(87)90450-0
[16] Stratakis, E., Fragkiadakis, G. and Tentes, I. (1993) Purification and Properties of the Fatty Acid-Binding VHDL from the Hemolymph of the Spider Eurypelma californicum. Journal of Experimental Zoology, 267, 483-492. http://dx.doi.org/10.1002/jez.1402670503
[17] Bridges, C.R. (2001) Modulation of Haemocyanin Oxygen Affinity: Properties and Physiological Implications in a Changing World. Journal of Experimental Biology, 204, 1021-1032.
[18] Cunningham, M., Gomez, C. and Pollero, R.J. (1999) Lipid Binding Capacity of Spider Hemocyanin. Journal of Experimental Zoology, 284, 368-373.
http://dx.doi.org/10.1002/(SICI)1097-010X(19990901)284:4<368::AID-JEZ2>3.0.CO;2-I
[19] Decker, H. and Rimke, T. (1998) Tarantula Hemocyanin Shows Phenoloxidase Activity. Journal of Biological Chemistry, 273, 25889-25892. http://dx.doi.org/10.1074/jbc.273.40.25889
[20] Decker, H. and Jaenicke, E. (2004) Recent Findings on Phenoloxidase Activity and Antimicrobial Activity of Hemocyanins. Developmental and Comparative Immunology, 28, 673-687.
http://dx.doi.org/10.1016/j.dci.2003.11.007
[21] Jaenicke, E. and Decker, H. (2004) Functional Changes in the Family of Type 3 Copper Proteins in Evolution. European Journal of Chemical Biology, 5, 163-176.
[22] Terwilliger, N.B. (1998) Functional Adaptations of Oxygen-Transport Proteins. Journal of Experimental Biology, 201, 1085-1098.
[23] Zatta, P. (1981) Protein-Lipid Interactions in Carcinus maenas (Crustacea) Hemocyanin. Comparative Biochemistry and Physiology Part B: Comparative Biochemistry, 69, 731-735.
http://dx.doi.org/10.1016/0305-0491(81)90377-1
[24] Laino, A., Cunningham, M., García, F. and Heras, H. (2009) First Insight into the Lipid Uptake, Storage and Mobilization in Arachnids: Role of Midgut Diverticula and Lipoproteins. Journal of Insect Physiology, 55, 1118-1124. http://dx.doi.org/10.1016/j.jinsphys.2009.08.005
[25] Laino, A., Cunningham, M., Heras, H. and García, F. (2011) In Vitro Lipid Transfer between Lipoproteins and Midgut-Diverticula in the Spider Polybetes pythagoricus. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 160, 181-186. http://dx.doi.org/10.1016/j.cbpb.2011.08.003
[26] Baumann, T., Kuhn-Nentwig, L., Largiadèr, C.R. and Nentwig, W. (2010) Expression of Defensins in Non-Infected Araneomorph Spiders. Cellular and Molecular Life Sciences, 67, 2643-2651.
http://dx.doi.org/10.1007/s00018-010-0354-2
[27] Garcia, F., Cunningham, M., Soulages, J.L., Garda, H.A. and Pollero, R.J. (2006) Structural Characterization of the Lipovitellin from the Shrimp Macrobrachium borellii. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 145, 365-370. http://dx.doi.org/10.1016/j.cbpb.2006.08.014
[28] Lowry, O.H., Rosenbrough, N.J., Farr, A.L. and Randall, R. (1951) Protein Measurement with the Folin Phenol Reagent. Journal of Biological Chemistry, 193, 265-275.
[29] Laemmli, U.K. (1970) Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature, 227, 680-685. http://dx.doi.org/10.1038/227680a0
[30] Garín, C.F., Heras, H. and Pollero, R.J. (1996) Lipoproteins of the Egg Perivitellin Fluid of Pomacea canaliculata Snails (Mollusca: Gastropoda). Journal of Experimental Zoology, 276, 307-314.
http://dx.doi.org/10.1002/(SICI)1097-010X(19961201)276:5<307::AID-JEZ1>3.0.CO;2-S
[31] Folch, J., Lees, M. and Sloane-Stanley, G.H. (1957) A Simple Method for the Isolation and Purification of Total Lipids from Animal Tissues. Journal of Biological Chemistry, 226, 497-509.
[32] Ackman, R.G., McLeod, C. and Banerjee, A.K. (1990) An Overview of Analyses by Chromarod-Iatroscan TLC-FID. Journal of Planar Chromatography, 3, 450-490.
[33] Morrison, W.R. and Smith, L.M. (1992) Preparation of Fatty Acid Methyl Esters and Dimethylacetals from Lipid with Boron Fluoride-Methanol. Journal of Lipid Research, 5, 600-608.
[34] Garcia, F., González Baró, M.R. and Pollero, R.J. (2005) Effect of Fenitrothion on the Acylglyceride Exchang in Crustacean Lipoproteins. Pesticide Biochemistry and Physiology, 82, 177-184.
http://dx.doi.org/10.1016/j.pestbp.2004.11.005
[35] Chino, H., Downer, R.G., Wyatt, G. and Gilbert, L.I. (1981) Lipophorins, a Major Class of Lipoproteins of Insect Hemolimph. Insect Biochemistry, 11, 491. http://dx.doi.org/10.1016/0020-1790(81)90085-8
[36] Chapman, M.J. (1983) A Broad Perspective on Invertebrate and Lower Vertebrate Apolipoproteins: Crustaceans, Insects. Fish, Amphibians, and Reptiles. In: Lewis, L.A. and Naito, H.K., Eds., Handbook of Electrophoresis, Vol. 4, CRC Press, Boca Raton, 285-310.
[37] Lamy, J., Lamy, J., Weill, J., Markl, J., Schneider, H.J. and Linton, S.M. (1979) Hemocyanins in Spiders. VII. Inmunologycal Comparison of the Subunits of Eurypelma californicum. Hoppe-Seyler’s Zeitschrift für Physiologische Chemie, 360, 889-895. http://dx.doi.org/10.1515/bchm2.1979.360.2.889
[38] Markl, J. (1986) Evolution and Function of Structurally Diverse Subunits in the Respiratory Protein Hemocyanin from Arthropods. Biological Bulletin, 171, 90-115.
[39] Schneider, H.J., Markl, J., Schartau, W. and Linzen, B. (1977) Hemocyanins in Spiders. IV. Subunits Heterogeneity of Eurypelma californicum (Dugesiella) Hemocyanin, and Separation of Polypeptide Chains. Hoppe-Seyler’s Zeitschrift für Physiologische Chemie, 358, 1133-1141.
http://dx.doi.org/10.1515/bchm2.1977.358.2.1133
[40] Cunningham, M., Garcia, F., Garda, H. and Pollero, R. (2006) Hemocyanin Lipid Uptake in Polybetes pythagoricus Is Altered by Fenitrothion. Pesticide Biochemistry and Physiology, 86, 57-62.
http://dx.doi.org/10.1016/j.pestbp.2005.11.006
[41] Heras, H. and Pollero, R.J. (1990) Occurrence of Plasma Lipoproteins in Octopods. Partial Characterization and Interorgan Transport of Lipids. Journal of Experimental Marine Biology and Ecology, 140, 29-38.
http://dx.doi.org/10.1016/0022-0981(90)90078-Q
[42] Pollero, R.J., González Baró, M.R. and Garín, C.F. (1992) Lipid Transport in Snails. Partial Characterization of a High-Density Lipoprotein Isolated from Ampullaria canaliculata Plasma. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 102, 717-720.
[43] Hall, M., van Heusden, M.C. and Söderhäll, K. (1995) Identification of the Major Lipoproteins in Crayfish Hemolymph as Proteins Involved in Immune Recognition and Clotting. Biochemical and Biophysical Research Communications, 216, 939-946. http://dx.doi.org/10.1006/bbrc.1995.2711
[44] Laino, A., Cunningham, M., Heras, H. and Garcia, F. (2011) Isolation and Characterization of Two Vitellins from Eggs of the Spider Polybetes pythagoricus (Araneae: Sparassidae). Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 158, 142-148.
http://dx.doi.org/10.1016/j.cbpb.2010.11.001
[45] Laino, A., Cunningham, M., Costa, F.G. and Garcia, F. (2013) Energy Sources from the Eggs of the Wolf Spider Schizocosa malitiosa: Isolation and Characterization of Lipovitellins. Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology, 165, 172-180.
http://dx.doi.org/10.1016/j.cbpb.2013.04.004
[46] Schartau, W. and Leidescher, T. (1983) Composition of the Hemolymph of the Tarantula Eurypelma californicum. Journal of Comparative Physiology, 152, 73-77. http://dx.doi.org/10.1007/BF00689730
[47] Pollero, R.J., Huca, G. and Brenner, R.R. (1985) Role of Hemocytes and Plasma on Lipid Transport in the Freshwater Mollusc Diplodon delodontus. Comparative Biochemistry and Physiology Part A: Physiology, 82, 339-343. http://dx.doi.org/10.1016/0300-9629(85)90865-5            eww150105lx
标签：

评论

发表评论

此博客中的热门博文

Does Immigration Promote the Investment of the Monopolistic Firm?

In the present paper, we examine the effect of increasing uncertainty of immigrants’ growth on the optimal timing of investment of a firm that has a monopolistic power over the labor market. It is revealed that when the uncertainty of immigrants’ growth is more than a threshold level, increasing uncertainty of immigrants’ growth accelerates the optimal timing of firms’ investment and enhances the economic growth, even if the uncertainty of immigrants’ growth is formulated by the geometric Brownian motion, which is in sharp contrast to the standard result that an increase in the uncertainty postpones the optimal timing. With an increase in the immigrants over the past ten years, workforces in the host countries have been growing significantly to the extent that the immigrants represent 70% of the increase in the workforce in Europe, and 47% in the United States as OECD indicates. In the present paper, we attempted to investigate the effect of increased uncertainty caused by the growi...
发表评论
阅读全文

Education Policy Implementation: A Mechanism for Enhancing Primary Education Development in Zanzibar

Education is one of the fundamental rights of individuals; therefore, the government of a country needs to develop and strengthen educational policy and quality as well as to ensure that everyone has equal access to basic education. The improvement of access and quality of education in the world is becoming as an essential factor in development, whereas the basic education (primary school), is acknowledged as a foundation of the higher educational development for every country. To fulfill this goal, governments introduce several policies and procedures; however, it requires some reforms and participation from the politician, policymakers, and other stakeholders to re-examine educational policy so that it can lead to multiplication and betterment of the reforms. Educational reforms actually focus on accountability. A positive educational development and reform is very challenging and needs more effort and strategy on how to use and utilize the resources effectively as such it can achie...
发表评论
阅读全文