Neuronal insulin signaling and resistance: a balancing act of kinases and phosphatases.

Insulin signaling cascade in peripheral insulin-sensitive tissues regulates whole-body glucose metabolism. Any deregulation in this pathway leads to insulin resistance, ultimately leading to metabolic diseases like type 1 diabetes, type 2 diabetes, and obesity. Insulin signaling in the brain has also been studied for many decades and associated with many primary functions like maintenance of synaptic plasticity, regulation of cognition, and circadian rhythm. Importantly, neuronal insulin signaling has also been associated with the regulation of neuronal glucose uptake. Any impairment in neuronal insulin signaling affecting neuronal glucose uptake has been associated with neurodegenerative disorders like Alzheimer's disease, the process now being termed as type 3 diabetes. Since the criticality lies in proper signaling cascade, determining important points of deregulation is important. In this review, we have discussed some critical points of such deregulation, dividing them into two classes of enzymes: kinases and phosphatases. We have highlighted their individual roles in neuronal insulin signaling, along with their possible implications in neuronal insulin resistance. Future strategies targeting these nodes in neuronal insulin signaling might be helpful in exploring potential therapeutic opportunities to overcome neuronal insulin resistance and related neurodegenerative diseases.

PP1γ regulates neuronal insulin signaling and aggravates insulin resistance leading to AD-like phenotypes.

BACKGROUND: PP1γ is one of the isoforms of catalytic subunit of a Ser/Thr phosphatase PP1. The role of PP1γ in cellular regulation is largely unknown. The present study investigated the role of PP1γ in regulating neuronal insulin signaling and insulin resistance in neuronal cells. PP1 was inhibited in mouse neuroblastoma cells (N2a) and human neuroblastoma cells (SH-SY5Y). The expression of PP1α and PP1γ was determined in insulin resistant N2a, SH-SY5Y cells and in high-fat-diet-fed-diabetic mice whole-brain-lysates. PP1α and PP1γ were silenced by siRNA in N2a and SH-SY5Y cells and effect was tested on AKT isoforms, AS160 and GSK3 isoforms using western immunoblot, GLUT4 translocation by confocal microscopy and glucose uptake by fluorescence-based assay. RESULTS: Results showed that, in one hand PP1γ, and not PP1α, regulates neuronal insulin signaling and insulin resistance by regulating phosphorylation of AKT2 via AKT2-AS160-GLUT4 axis. On the other hand, PP1γ regulates phosphorylation of GSK3ß via AKT2 while phosphorylation of GSK3α via MLK3. Imbalance in this regulation results into AD-like phenotype. CONCLUSION: PP1γ acts as a linker, regulating two pathophysiological conditions, neuronal insulin resistance and AD. Video Abstract.

PP2Cα aggravates neuronal insulin resistance leading to AD-like phenotype in vitro.

Neuronal insulin resistance is a major risk for development of Alzheimer's Disease (AD). Studies already reported few kinases participating in neuronal insulin signaling connected with progression of AD pathogenesis, yet complete information is missing. α isoform of Protein Phosphatase-2C (PP2C) is a Ser/Thr phosphatase, only known in 3T3-L1 adipocytes as a positive regulator of insulin signaling. However, many aspects of its function in neuronal insulin signaling and insulin resistance are unidentified. Recently, we reported that PP2Cα positively regulates neuronal glucose uptake possibly by a mechanism of dephosphorylation of IRS-1 at Ser522 and by inactivating AMPK, exacerbating hyperinsulinemia mediated neuronal insulin resistance. Since PP2Cα affected neuronal insulin signaling and AD is connected to neuronal insulin resistance, in the present study, we studied the role of PP2Cα in regulating activities of both isoforms of GSK3α and GSK3ß (one of the leading kinases for AD progression). The results led us to test the role of PP2Cα on AD hallmarks. Silencing of PP2Cα caused hyperphosphorylation of a potential kinase Tau, leading to NFT formation and increased Aß deposition. Our study thereby demonstrates escalation of hyperinsulinemia mediated neuronal insulin resistance leading to AD-like pathogenesis by PP2Cα in vitro and hints a novel molecule, PP2Cα, linking AD pathogenesis.

Cellular and Molecular Regulation of Exercise-A Neuronal Perspective.

The beneficial effects of exercise on the proper functioning of the body have been firmly established. Multi-systemic metabolic regulation of exercise is the consequence of multitudinous changes that occur at the cellular level. The exercise responsome comprises all molecular entities including exerkines, miRNA species, growth factors, signaling proteins that are elevated and activated by physical exercise. Exerkines are secretory molecules released by organs such as skeletal muscle, adipose tissue, liver, and gut as a function of acute/chronic exercise. Exerkines such as FNDC5/irisin, Cathepsin B, Adiponectin, and IL-6 circulate through the bloodstream, cross the blood-brain barrier, and modulate the expression of important signaling molecules such as AMPK, SIRT1, PGC1α, BDNF, IGF-1, and VEGF which further contribute to improved energy metabolism, glucose homeostasis, insulin sensitivity, neurogenesis, synaptic plasticity, and overall well-being of the body and brain. These molecules are also responsible for neuroprotective adaptations that exercise confers on the brain and potentially ameliorate neurodegeneration. This review aims to detail important cellular and molecular species that directly or indirectly mediate exercise-induced benefits in the body, with an emphasis on the central nervous system.

PP2Cα positively regulates neuronal insulin signalling and aggravates neuronal insulin resistance.

PP2Cα is one of the newly identified isoforms of metal-dependent protein phosphatases (PPM). The role of this phosphatase in neuronal insulin signalling is completely unknown. In the present study, we show insulin-mediated rapid upregulation of a protein of the insulin signalling cascade, PP2Cα, in mouse N2a cells and human SH-SY5Y cells. By contrast, such PP2Cα upregulation is not observed in insulin-resistant conditions despite insulin stimulation. Here, we report that, under insulin-sensitive and insulin-resistant conditions, the translation of PP2Cα was regulated by insulin through c-Jun N-terminal kinase. PP2Cα in turn dephosphorylated a novel inhibitory site of insulin receptor substrate-1 at Ser522 and AMP-activated protein kinase, hence positively regulating neuronal insulin signalling and insulin resistance.

Ser/Thr phosphatases: One of the key regulators of insulin signaling.

Protein phosphorylation is an important post-translational modification that regulates several cellular processes including insulin signaling. The evidences so far have already portrayed the importance of balanced actions of kinases and phosphatases in regulating the insulin signaling cascade. Therefore, elucidating the role of both kinases and phosphatases are equally important. Unfortunately, the role of phosphatases is less studied as compared to kinases. Since brain responds to insulin and insulin signaling is reported to be crucial for many neuronal processes, it is important to understand the role of neuronal insulin signaling regulators. Ser/Thr phosphatases seem to play significant roles in regulating neuronal insulin signaling. Therefore, in this review, we discussed the involvement of Ser/Thr phosphatases in regulating insulin signaling and insulin resistance in neuronal system at the backdrop of the same phosphatases in peripheral insulin sensitive tissues.

Review article: irritable bowel syndrome: natural history, bowel habit stability and overlap with other gastrointestinal disorders.

Symptoms of irritable bowel syndrome (IBS) characteristically fluctuate over time. We aimed to review the natural history of IBS and IBS subgroups including bowel habit disturbances, and the overlap of IBS with other gastrointestinal disorders. The community incidence of IBS is approximately 67 per 1000 person years. The prevalence of IBS is stable over time because symptoms fluctuate and there is a portion who experience resolution of their GI symptoms similar in number to those developing new-onset IBS. The proportion who report resolution of symptoms varies amongst population-based studies from 17% to 55%. There is evidence of substantial movement between subtypes of IBS. For example in a clinical trial cohort, only one in four patients retained their baseline classification throughout the study periods, two in three moved between IBS-C (constipation) and IBS-M (mixed), while over half switched between IBS-D (diarrhoea) and IBS-M. The least stable group was IBS-M. There are very limited data on drivers of bowel habit change in IBS. There are emerging evidence fluctuations in intestinal immune activity might account for symptom variability over time. It is of clinical importance to recognise the substantial overlap of IBS symptoms with other gastrointestinal syndromes including gastro-oesophageal reflux disease. This is important to ensure the correct clinical diagnosis of IBS is made and patients are not over investigated. Knowledge of the natural history, stability of subgroups and overlap of IBS with other gastrointestinal conditions should be considered in therapeutic decision making.

Fully Nonlinear ${SP}_{3}$ Approximation Based Fluorescence Optical Tomography.

In fluorescence optical tomography, many works in the literature focus on the linear reconstruction problem to obtain the fluorescent yield or the linearized reconstruction problem to obtain the absorption coefficient. The nonlinear reconstruction problem, to reconstruct the fluorophore absorption coefficient, is of interest in imaging studies as it presents the possibility of better reconstructions owing to a more appropriate model. Accurate and computationally efficient forward models are also critical in the reconstruction process. The approximation to the radiative transfer equation (RTE) is gaining importance for tomographic reconstructions owing to its computational advantages over the full RTE while being more accurate and applicable than the commonly used diffusion approximation. This paper presents Gauss-Newton-based fully nonlinear reconstruction for the approximated fluorescence optical tomography problem with respect to shape as well as the conventional finite-element method-based representations. The contribution of this paper is the Frechet derivative calculations for this problem and demonstration of reconstructions in both representations. For the shape reconstructions, radial-basis-function represented level-set-based shape representations are used. We present reconstructions for tumor-mimicking test objects in scattering and absorption dominant settings, respectively, for moderately noisy data sets in order to demonstrate the viability of the formulation. Comparisons are presented between the nonlinear and linearized reconstruction schemes in an element wise setting to illustrate the benefits of using the former especially for absorption dominant media.

Towards crossbar nanoarray structure via microcontact printing.

The method for patterning arrays of multiwalled carbon nanotubes (MWCNT's) in symmetric patterns to form junctions has been demonstrated. This has been achieved by incorporating the technique of microcontact printing using poly-dimethylsiloxane (PDMS) molds. Relief structures in the order of a few micrometers were fabricated that enabled the transfer of continuous horizontal arrays of MWCNT's in aqueous suspension in a controlled manner. The MWCNT's were patterned onto silicon microelectrode substrates with metallic gold electrodes. These were fabricated using standard photolithography techniques. The silicon substrates served as a base platform with suitable measurement microelectrodes for electrically characterizing the crossbar junction arrays. Using a dual alignment and stamping process, PDMS molds were inked alternatively with p-type and n-type suspensions of MWCNT's and transferred in a grid-like manner onto the base platform. Parallel alignment of the MWCNT's was achieved due to the geometry of the mold relief structures. This step-by-step assembly resulted in the formation of crossbar MWCNT array structures. Each of these crosspoints in the individual junction can function as an addressable crossbar nanodevice. The functionality of this circuit was demonstrated through the current-voltage (I-V) characteristics. Using these high-density crossarray circuit patterns, addressable nanostructures that form the building blocks of highly integrated device arrays can be built.