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A busca pela longevidade acompanha o homem desde os primórdios, por meio de histórias fantásticas como as que rodeavam a alquimia. Com o avanço científico, essa busca adquiriu um novo caráter e passou a incluir também a investigação de genes que estivessem associados a uma maior expectativa de vida. Nesse contexto, estudos investigando o gene Mth da mosca de fruta Drosophila melanogaster mostraram que este, quando mutado, pode prolongar a vida do organismo. O gene Mth codifica um receptor transmembrana para os peptídeos endóge - nos Stunted A e B, e se acopla à proteína G. Receptores acoplados à proteína G estão relacionados a diversas atividades biológicas como neurotransmissão, fisiologia hormonal, resposta a drogas e transdução de estímulos. Desse modo, tal receptor atua recebendo e repassando informações para a célula, e assim, interferindo em sua longevidade. Estudos como esses fazem com que, aos poucos, nosso entendimento sobre a expectativa de vida torne-se mais amplo e detalhado, o que também traz a longevidade para mais perto da nossa realidade.

The study was partially presented as a poster in the 53rd Annual Meeting of the European Association for the Study of Diabetes, Lisbon 2017., The role of insulin-degrading enzyme (IDE), a metalloprotease with high affinity for insulin, in insulin clearance remains poorly understood. OBJECTIVE: This study aimed to clarify whether IDE is a major mediator of insulin clearance, and to define its role in the etiology of hepatic insulin resistance., [Methods] We generated mice with liver-specific deletion of Ide (L-IDE-KO) and assessed insulin clearance and action., [Results] L-IDE-KO mice exhibited higher (~20%) fasting and non-fasting plasma glucose levels, glucose intolerance and insulin resistance. This phenotype was associated with ~30% lower plasma membrane insulin receptor levels in liver, as well as ~55% reduction in insulin-stimulated phosphorylation of the insulin receptor, and its downstream signaling molecules, AKT1 and AKT2 (reduced by ~40%). In addition, FoxO1 was aberrantly distributed in cellular nuclei, in parallel with up-regulation of the gluconeogenic genes Pck1 and G6pc. Surprisingly, L-IDE-KO mice showed similar plasma insulin levels and hepatic insulin clearance as control mice, despite reduced phosphorylation of the carcinoembryonic antigen-related cell adhesion molecule 1, which upon its insulin-stimulated phosphorylation, promotes receptor-mediated insulin uptake to be degraded., [Conclusion] IDE is not a rate-limiting regulator of plasma insulin levels in vivo., This work was supported by grants from the Ministerio de Economía, Industria y Competitividad: SAF2014-58702-C2-1-R and SAF2016-77871-C2-1-R to ICC; SAF2014-58702-C2-2-R and SAF2016-77871-C2-2-R to GP; supported by the EFSD European Research Programme on New Targets for Type 2 Diabetes supported by an educational research grant from MSD to ICC and GP; the National Institutes of Health: R01-DK054254, R01-DK083850 and RO1-HL-112248 to SMN, and R01-GM115617 to MAL; and the American Diabetes Association: Career Development Award 7-11-CD-13 to MAL.

Sporadic Alzheimer's disease (sAD) is associated with energy metabolism deficiency and impairment of insulin receptor (IR) signaling in the brain. In this context, low doses of intracerebroventricular (icv) injection of streptozotocin (STZ) in rodents has been used as an experimental model of sAD which leads to an insulin-resistant brain state and neurodegeneration. However, the STZ effects on brain insulin signaling-related proteins it is not appropriately elucidated. The aim of this study was to evaluate the beginning and progression of alterations in the brain IR pathway of rats after 1, 3, 5, and 7 days of STZ injection and investigate intracellular signaling involved on STZ induced insulin resistance. We observed that STZ injection causes cognitive impairment in the animals, a temporal variation of the insulin signaling-related proteins and apoptosis cell death in the hippocampus. We also have shown that STZ causes insulin resistance and impairment on phosphoinositide 3-kinase (PI3K) activity in the Neuro-2a cells through protein kinase B (Akt) inactivation by S-nitrosylation, which could upregulate GSK3-β activity. STZ ability to cause an insulin-resistant neuron state involves NO production and ROS production which may play an important role in the mechanism linked to STZ-induced neurotoxicity. The icv injection of STZ model and STZ exposed Neuro-2a cells may be potential experimental models for assessing molecules related to the pathogenesis of sAD.

Summary The food a honey bee female larva receives determines whether she develops into a large long-lived fertile queen or a short-lived sterile worker. Through well-established nutrient sensing and growth promoting functions in metazoans, the insulin/insulin-like growth factor 1 signaling (IIS) pathway has become a focal topic in investigations on how differences in food environment can be translated into internal signals responsible for queen-worker determination. However, low expression levels of two insulin receptors (AmInRs) in honey bee larvae and the failure of one AmInR to influence caste differentiation are in potential conflict with such a classical growth promoting role of IIS in queen-worker development. In view of such an apparent contradiction, and the fact that binding partners and affinities of these two AmInRs have not been worked out, we performed a functional study on insulin-like peptide genes (AmILP1 and AmILP2) in honey bee larvae by using a double-stranded RNA (dsRNA)-mediated gene knockdown approach. We found that juvenile hormone (JH) levels were diminished by vAmILP1 dsRNA treatment, while the AmILP2 knockdown caused a reduction in ovary size. Blood sugar titers were not significantly affected by the treatments. From these results we conclude that AmILP2 transcript levels may influence specific organ development, such as the ovary and body mass, while more general traits of caste differentiation, such as mandibles, may require additional regulators. In addition, JH production may be regulated by AmILP1 expressed locally in the brain, similar to the function of certain ILPs in Drosophila.

[CAT] La panerola Blattella germanica és un insecte de metamorfosi incompleta (hemimetàbol) i anautògen. Aquesta és una estratègia reproductiva per la qual les femelles no es poden reproduir fins que no han menjat. D'alguna manera els senyals nutricionals han d'arribar als diferents teixits i cèl•lules per informar-los que s'ha assolit un estat nutricional adequat per iniciar els processos reproductius. Tant la via de TOR com la via del receptor d'insulina (InR) semblen formar part conjunta d'aquesta senyalització nutricional. A B. germanica la vitel•logènesi s'activa mitjançant l'hormona juvenil (HJ), que es fabrica i secreta per unes glàndules retrocerebrals anomenades corpora allata (CA). Quan falta l'aliment, els CA no sintetitzen HJ i aquesta no pot activar la síntesi de vitel•logenina (Vg) en el cos gras (CG), que no pot ser incorporada als ovaris, impedint que aquests es desenvolupin correctament. Prèviament havia estat clonada la seqüència de TOR de B. germanica (BgTOR). En aquest treball s'han clonat les de BgInR, BgS6K i de BgPepck. Gràcies al dsRNAi s'ha estudiat la funció del InR com a regulador del creixement, veient que no només afecta a la maduresa sexual de les femelles sinó també la seva mida quan s'injecta en estadis nimfals. La interferència del InR també produeix problemes en el correcte desenvolupament i desplegament de les ales i les tegmines. Així mateix, s'ha comprovat que la interferència del InR fa baixar els nivells dels trànscrits de HMG-S1, HMG-S2, HMG-R i epoxidasa en els CA, implicats en la síntesi d'HJ, així com de la Vg en CG , independentment de l'efecte de la HJ sobre aquesta. També s'ha vist que la interferència del InR produeix una major longevitat en les femelles, a causa que aquestes no es reprodueixen, evitant el desgast que això produeix. Alhora, l'aplicació de dsInR confereix certa protecció davant situacions de dejuni. Altres resultats suggereixen que l'activació d'S6K, tal com succeeix en altres organismes, es realitza per modificacions post-transcripcionals, probablement per fosforilació de la proteïna. S'ha començat a posar a punt un sistema de western-blot per usar-lo per a la detecció de l'estat de fosforilació de S6K i per tant del seu estat d'activació. Mitjançant la tècnica del RNAi s'ha comprovat que la interferència per S6K fa baixar els transcrits de HMG-S1, HMGS2, HMG-R i epoxidasa en els CA, implicats en la síntesi d'HJ, així com la Vg en CG, indicant que està implicat en la transmissió del senyal nutricional als teixits implicats en la vitel•logènesi. Finalment s'ha establert Pepck com a marcador de l'estat nutricional, ja que s'incrementa la seva expressió en situacions de dejuni. No s'ha observat, però, que augmenti quan es redueixen els nivells de S6K o InR mitjançant RNAi, la qual cosa ens indica que la influència de la situació de dejuni sobre la transcripció de PEPCK es pot realitzar per vies alternatives a S6K i InR, si bé sembla que necessita de la presència de InR funcional, [EN] The cockroach Blatella germanica is a hemimetabolous insect with anautogenous reproduction. TOR and Insulin Receptor (InR) pathways are responsible of the nutritional signaling and to control reproductive processes. In B. germanica the yolk protein synthesis is activated by juvenile hormone (JH), which is secreted by retrocerebrals glands known as corpora allata (CA). In starvation, CA are not able to synthesize JH and it can’t activate the synthesis of vitelogenin (Vg) in the fat body (FB), and it can’t be incorporated by the ovaries, preventing their proper development. In this work we have cloned the sequences of BgInR, BgS6K and BgPepck. Using RNAi, InR has been studied as a regulator of growth, affecting not only sexual maturity of females, but also their size when RNAi is injected at nymphs. InR interference also produces problems for the proper development and deployment of wings and elytra. It has also been observed that InR interference lowers the levels of HMG-S1, HMG-S2, HMG-R and epoxidase transcription in CA, involved on the synthesis of JH by CA, and consequently, reducing Vg levels. Independently of JH, InR has a direct effect on Vg expression. Besides, InR interference confers greater longevity in females and protection in front of fasting. We have also begun to implement western-blot technique to detect phosphorylation of S6K. The S6K interference reduce transcription levels of HMG-S1, HMG-S2, HMG-R and epoxidase in CA, and Vg in the CG , indicating that it is involved in nutritional signal transmission to the tissues involved in vitelogenesis. Finally, we established Pepck as a marker of the nutritional status, as its expression is increased in starvation. However, it has not been observed any increase when reducing the levels of S6K or InR using RNAi, indicating that the influence of starvation on Pepck transcription can be performed in alternative ways to S6K and InR, but needs the presence of functional InR.

Colon carcinoma is the most common tumor of the gastrointestinal tract. According to some investigators, insulin, epidermal growth factor (EGF) and insulin-like growth factor I (IGF-I) may be involved in the neoplastic proliferation. Insulin-binding and receptor tyrosine kinase activity were investigated in colon carcinomas and in normal colons. The insulin receptor concentration, as shown by binding assays, was 17.4 ± 4.3 fmol/J.Lg in normal colon and 29.69 ± 9.4 fmol/ J.Lg in colon carcinoma. Nevertheless, the insulin affinity of the receptor was similar in both groups (Kd ::: 1 nM). Both normal and neoplastic colon showed phosphorylation of the insulin receptor. The electrophoretic migration of the B-subunit of the insulin receptors purified from colon carcinomas was similar to that of normal colon and both tissues demonstrated an insulin-dependent autophosphorylation. The receptor tyrosine kinase activity was measured by the incorporation of[gamma32P]ATP into the B-subunit. The basal and the insulin-stimulated tyrosine kinase activities were significantly higher in colon carcinomas compared to normal colon tissues (2.2 and 1.6 times, respectively). Understanding the metabolism of neoplastic cells may contribute to the development of prevention strategies as well as new therapies. It is now necessary to study other steps of the insulin signal transduction pathway, such as insulin receptor substrate 1 phosphorylation.