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Intestinal microvillar membranes and their secretory vesicles in Lepidoptera Spodoptera frugiperda and Diptera Musca domestica

Grant number: 14/14183-2
Support Opportunities:Scholarships in Brazil - Post-Doctoral
Effective date (Start): December 01, 2014
Effective date (End): June 30, 2017
Field of knowledge:Biological Sciences - Biochemistry - Enzymology
Principal Investigator:Clelia Ferreira Terra
Grantee:Felipe Jun Fuzita
Host Institution: Instituto de Química (IQ). Universidade de São Paulo (USP). São Paulo , SP, Brazil
Associated research grant:11/51685-8 - Insect digestion: a molecular, cellular, physiological and evolutionary approach, AP.TEM
Associated scholarship(s):16/09511-6 - Glycan structural determination of glycoproteins from the midgut of Spodoptera frugiperda, BE.EP.PD


In mammals, proteins going to the intestinal apical membrane are present in lipid rafts. The lipid rafts are structures rich in cholesterol and glycosphingolipids, more resistant to certain detergents than the rest of the membrane and can be purified by density gradient centrifugation. The incorporation of enzymes in the rafts can alter their activities.The composition and properties of insect intestinal microvillar membranes are poorly known. The amount and variety of proteins in this membrane increase in more derived insect orders and differences can also be seen in distinct parts of the intestinal tract. In Lepidoptera some parts of the intestine secrete enzymes by a microapocrine mechanism, where small vesicles migrate through the cell microvilli and, before or after being fused with the microvillar membrane at the tips of the microvilli, are discharged into the lumen by a pinching- off process. These vesicles can be purified from the ectoperitrophic fluid, a fluid bathing the microvilli.We intend to isolate lipid rafts and total microvilar membranes from different parts of the intestine of S. frugiperda and M. domestica and the microapocrine vesicles from S. frugiperda. The preparations will have the enzyme and protein composition analyzed and compared.Desired skills: Experience in small animal dissection, sub-cellular fractionation, density gradient centrifugation, analysis of protein by mass spectrometry

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
FUZITA, FELIPE J.; PIMENTA, DANIEL C.; PALMISANO, GIUSEPPE; TERRA, WALTER R.; FERREIRA, CLELIA. Detergent-resistant domains in Spodoptera frugiperda midgut microvillar membranes and their relation to microapocrine secretion. COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, v. 235, p. 8-18, . (17/08103-4, 14/14183-2, 14/06863-3)
FUZITA, FELIPE J.; PALMISANO, GIUSEPPE; PIMENTA, DANIEL C.; TERRA, WALTER R.; FERREIRA, CLELIA. A proteomic approach to identify digestive enzymes, their exocytic and microapocrine secretory routes and their compartmentalization in the midgut of Spodoptera frugiperda. COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, v. 257, . (14/06863-3, 17/08103-4, 14/14183-2)
FUZITA, FELIPE J.; CHANDLER, KEVIN B.; HASERICK, JOHN R.; FERREIRA, CLELIA; TERRA, WALTER R.; COSTELLO, CATHERINE E.. Glycoproteins in the Midgut Microvilar Membrane of Spodoptera frugiperda (Lepidoptera: Noctuidae). FASEB JOURNAL, v. 31, p. 2-pg., . (11/51685-8, 14/14183-2, 16/09511-6)

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