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LUNA, M. S. A. Comparative proteomic analysis of differencitial expression induced by noradrenergic innervation in venom gland and accessory gland of Bothrops jararaca snake. 2013. 170 p. Ph. D. thesis (Cell Biology and Tissue) – Instituto de Ciências Biomédicas, Universidade de São Paulo, São Paulo, 2013. Venom gland of viperid snakes have a basal-central lumen where the venom produced by secretory cells is stored. When the venom is depleted from the lumen, secretory cells are activated and more venom is produced. In previous studies, we have shown that venom extraction promotes changes in the protein synthesis of venom gland. In this thesis, we performed a proteomic analysis of venom gland and accessory gland during venom production cycle and verify the influence of noradrenergic innervation on protein synthesis of the venom gland. We identified exclusive proteins in each stage. Among protein species of the cytoplasm, we verify that the number of exclusive protein species identified as actin, vimentin, tropomyosin, and proteasome subunit alpha type-1, thioredoxin, and 40S ribosomal increased after venom extraction. Among protein species of the endoplasmic reticulum, we verify that the number of species of protein identified as disulfide isomerase and glucose regulated protein 78 kDa increased after venom extraction. Regarding secreted proteins, specifically about venom toxins, we showed for the first time that many toxins are present in quiescent stage. SVSP are more abundant in quiescent stage and others toxins CTL-like such as GPIb-BP and coagulation factor IX/X were present only in this stage. Other toxins such as SVMP, LAAO, PLA2, PLI, Bj46a, and disintegrin are more abundant in activated stages of venom gland, indicating the production and secretion of toxins is not synchronized. Noradrenergic innervation regulates positively the synthesis of vimentin and proteasome subunit alpha and negatively the synthesis of myosin light chain and ATP-synthase mitochondrial-like. Considering toxins, noradrenergic innervation regulates negatively the synthesis of CTL-like such as GPIb-BP, coagulation factor IX/X, PLA2 and SVMP-PII and positively the synthesis of Bj46a and SVMP-PI. In the accessory gland, the synthesis of nuclear protein histone H2 increased after venom extraction and the synthesis of proteins HSP90 (endoplasmic reticulum) and DPPIV (membrane) decreased after venom extraction. Surprisingly, we showed for the first time the presence of toxins in accessory gland such as CTLs, SVSP, SVMP, PLA2, LAAO and in a less abundance, disintegrin, GC, PN identified only in accessory gland that not underwent venom extraction. We also identified the presence of enzyme inhibitors such as PLI, Bj46a and the precursor of the tripeptide pEKW. It is important to show that from SVMP class, only SVMP-PII and SVMPI-PIII were identified and mainly in accessory gland 4 days after venom extraction. Similar to venom gland, the syntheses and secretion of new toxins are asynchronized in accessory gland. The proteomic composition of gland of snake venom apparatus on different stages of venom production cycle, and the participation noradrenergic innervation in this process will contribute to better understand the mechanisms involved in venom gland activation and consequently, venom production. These data will also contribute to the studies on snake toxins and their diversities.

Análise proteômica comparativa da expressão diferencial de proteínas induzida pela ativação da inervação noradrenérgica em glândulas de veneno e acessória da serpente Bothrops jararaca.