Multiusuary equipment conceeded by process: 2010/05426-8-bioimaging CCD system

• Grant number: 17/22262-8
• Support Opportunities: Multi-user Equipment Program
• Start date: February 01, 2018
• End date: January 31, 2025
• Field of knowledge: Biological Sciences - Biophysics - Radiology and Photobiology
• Principal Investigator: Vadim Viviani
• Grantee: Vadim Viviani
• Host Institution: Centro de Ciências e Tecnologias para a Sustentabilidade (CCTS). Universidade Federal de São Carlos (UFSCAR). Sorocaba , SP, Brazil
• Associated research grant: 10/05426-8 - Arthropod bioluminescence: biological diversity in Brazilian biomes, biochemical origin, structural/functional evolution of luciferases, molecular differentiation of lanterns, biotechnological, environmental and educational applications, AP.TEM
• EMU web page: https://ppgbma.wixsite.com/ppgbma-ufscar-sor/equipamentos-multiusuarios
• Type of equipment: Processos Biológicos - Caracterização - Leitores de fluorescência (imageamento, placas); Caracterização de Materiais - Imageamento - Fluorescência (in vivo)
• Manufacturer: Berthold
• Model: Nightowl

Abstract

Bioluminescence, the emission of cold light by living organisms, has been the subject of many multidisciplinary studies involving physics, chemistry and biology, resulting in important scientific discoveries and biotechnological applications in the last century. It can be considered as one of the signatures of life, and for this reason has been used as an excellent bioindicator at the molecular, cellular, organismic and biospheric levels. In the terrestrial environment it occurs among bacteria, fungi, annelids, mollusks, and especially among the arthropods. Among the insects, the bioluminescent beetles system is well known, whereas the bioluminescent Diptera systems remain largely unknown. Brazil hosts the greatest biodiversity of bioluminescent organisms in the world, including beetles, fungi and marine species. However, with the exception of surveys and studies of bioluminescent beetles Atlantic rain forest, and more recently of fungi, little is known about the bioluminescent biodiversity of the Cerrado biome and Amazon forest which are severely threatened by agricultural progress. In the last decade the cDNAs encoding various luciferases from Brazilian beetles with different properties of bioluminescence (colors, kinetic, pH-sensitivity, thermal stability) were cloned, expressed, purified and investigated under the structural/functional and evolutionary aspects by our research group, revealing important determinants of bioluminescence spectra, pH-sensitivity and oxigenase activity. The resultant knowledge is already allowing to engineer these proteins to improve their biotechnological applicability, and some of these luciferases have already acquired effective commercial application. However, several questions remain open regarding bioluminescent systems of insects, including: (1) the origin and structural/functional evolution of luciferases of Beetles and Diptera; (2) the identity of the luciferases, luciferins and accessory proteins in bioluminescent Diptera; (3) the biochemical and anatomical origins of the bioluminescent apparatus in diptera and Beetles, and their possible evolutionary relationships. In addition, despite the undoubted utility of beetles luciferases and other organisms for bioanalytical purposes, in Brazil we still do not use the unique broad repertoire of recombinant luciferases derived from our fauna for bioanalytical purposes, using imported reagents. Thus, this thematic proposal seeks to achieve broad multidisciplinary study of the terrestrial bioluminescence (especially of insects in Brazil), involving: (i) survey of the biodiversity of bioluminescent arthropods in Cerrado biomes, Amazon Forest, with the investigation of their bioluminescence properties, molecular evolution and phylogeny, and application as environmental bio-indicator; (II) to investigate the bioluminescent system of Diptera, especially the molecular identity of luciferases, luciferins and accessory protein Orfelia fultonii of the Palearctic region; (III) clone and investigate the structure, function and molecular evolution of luciferases and accessory proteins of bioluminescent beetles coming from our fauna and Diptera; (IV) to investigate the anatomical and metabolic origin of bioluminescence in the three families of bioluminescent beetles and Diptera through an approach involving cell biology and functional genomics, and (V) to investigate the applicability of the investigated and optimized luciferases by our research group in the development of biotech, environmental and educational applications (enzyme and cell luminescent biosensors, luminescent bioassays and bio-imaging, educational kits). (AU)