The insulin-degrading enzyme: from molecular evolution and subcellular localization to new roles in microglial physiology

Abstract

under our supervision, within the doctoral programme 1 DOCTORADO EN INVESTIGACIÓN BIOMÉDICA, under the modality of INTERNATIONAL DOCTOR MENTION, after taking into account the following considerations 2 : The candidate has fulfilled all the requirements and the level of training necessary to aspire to the Doctor's Degree with International Mention by the University of Valladolid, by performing an original research project. She has significantly contributed to the design of the scientific questions to be solved, framed at the interface of several disciplines, has designed, executed and analysed the experiments, and has performed data presentations in different formats (writing published work and presenting at scientific meetings), contributing to the dissemination of her results to the scientific community.

Insulin-degrading enzyme (IDE) as a modulator of microglial phenotypes in the context of Alzheimer’s disease and brain aging

“…So far, our analysis questions that this regulatory role is directly related to “effector functions” as a protease, though it cannot be discarded in still unexplored situations. This new frame of ideas is coherent with previous works where microglia phenotypic switches were correlated with IDE expression [ 46 , 70 ]. However, attention should be paid, since IDE net expression but also traffic (export into extracellular vesicles) is modified by stimulus-induced switches in microglial phenotypes [ 22 ].…”

“…Leissring (University of California-Irvine, USA), was backcrossed with WT mice on a C57BL/6J background (Charles River, France), and the colony was maintained by crossing heterozygous animals. Mouse genotypes were evaluated by PCR, as previously described [ 22 ], and confirmed by protein expression by immunoblot [ 46 ] and Additional file 1 : Fig. S1.2).…”

“…FITC-labeled Aβ (FAM-Aβ) oligomers were prepared as previously described [ 46 ]. Treatments with FAM-Aβ oligomers (1 µM), prepared in RPMI 1640 medium without FBS, were added to primary microglial cultures grown in 6-well plates.…”

Dual role of Apolipoprotein D as long-term instructive factor and acute signal conditioning microglial secretory and phagocytic responses

“…Mouse primary microglial cells were isolated from individual newborn mice, sexed and cultured as previously described ( Corraliza-Gomez, 2021 ). Briefly, after 21 days in culture of primary mixed glial cells, astrocytes were detached and isolated microglia were re-exposed to the mixed glia-conditioned media during the last 24 h, thus containing astroglial pro-survival and proliferative factors.…”

“…Human ApoD (hApoD) was purified from breast cystic fluid as described ( Ruiz et al, 2013 ). Aβ oligomers were prepared following the protocol by Corraliza-Gomez (2021) . Briefly, after solubilization in 1,1,1,3,3,3-Hexafluoropropanol (HFIP; Sigma-Aldrich), aliquots were left for 2 h to evaporate and the peptide film was dissolved in DMSO as a 5 mM stock, which was sonicated and stored at −20°C.…”