Defensive and offensive behaviours in a Kleefstra syndrome mouse model.

Kleefstra syndrome in humans is characterized by a general delay in development, intellectual disability and autistic features. The mouse model of this disease (Ehmt1±) expresses anxiety, autistic-like traits, and aberrant social interactions with non-cagemates. To investigate how Ehmt1± mice behave with unfamiliar conspecifics, we allowed adult, male animals to freely interact for 10 min in a neutral, novel environment within a host-visitor setting. In trials where the Ehmt1± mice were hosts, there were defensive and offensive behaviors. Our key finding was that Ehmt1± mice displayed defensive postures, attacking and biting; in contrast, wild-type (WT) interacting with other WT did not enact such behaviors. Further, if there was a fight between an Ehmt1± and a WT mouse, the Ehmt1± animal was the most aggressive and always initiated these behaviors.

Edible plant by-products as source of polyphenols: prebiotic effect and analytical methods.

Polyphenols with high chemical diversity are present in vegetables both in the edible parts and by-products. A large proportion of them remains unabsorbed along the gastrointestinal tract, being accumulated in the colon, where they are metabolized by the intestinal microbiota. These polyphenols have been found to have "prebiotic-like" effects. The edible plant industry generates tons of residues called by-products, which consist of unutilized plant tissues (peels, husks, calyxes and seeds). Their disposal requires special and costly treatments to avoid environmental complications. Reintroducing these by-products into the value chain using technological and biotechnological practices is highly appealing since many of them contain nutrients and bioactive compounds, such as polyphenols, with many health-promoting properties. Edible plant by-products as a source of polyphenols highlights the need for analytical methods. Analytical methods are becoming increasingly selective, sensitive and precise, but the great breakthrough lies in the pretreatment of the sample and in particular in the extraction methods. This review shows the importance of edible plant by-products as a source of polyphenols, due to their prebiotic effect, and to compile the most appropriate analytical methods for the determination of the total content of phenolic compounds as well as the detection and quantification of individual polyphenols.

A Real-Time PCR Assay to Detect and Quantify Root-Knot Nematodes from Soil Extracts.

Root-knot nematodes cause forking and stubbing of the growing carrot root tip, decreasing market value and reducing yield by up to 45%. Since crop damage by these nematodes depends on their initial population densities at planting, preplant detection of potentially low nematode numbers is critical for predicting future yield loss. The aim of this study was to overcome some of the drawbacks of the labor- and time-intensive process of root-knot nematode identification and quantification by developing and field testing a real-time PCR (qPCR) assay. Primers were designed targeting the root-knot nematode Meloidogyne incognita species complex, which includes M. incognita as well as the closely related Meloidogyne javanica and Meloidogyne arenaria. The qPCR assay successfully detected each species and showed little amplification for nontarget nematode groups except for the sister group Meloidogyne enterolobii, which is not known to occur in California. Predicted nematode densities related well with microscopic counts of nematodes from prepared solutions, as well as from solutions extracted from field soil. In a greenhouse experiment, the qPCR assay distinguished between low, medium, and high levels of M. incognita infection and qPCR predicted densities at planting were negatively related in linear models with final carrot fresh weight, length, and diameter. These results suggest that qPCR assays could be a valuable diagnostic tool to predict nematode infections and prevent crop losses.[Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Cytoglobin deficiency potentiates Crb1-mediated retinal degeneration in rd8 mice.

PURPOSE: The purpose of this study is to determine the effect of Cytoglobin (Cygb) deficiency on Crb1-related retinopathy. The Crb1 cell polarity complex is required for photoreceptor function and survival. Crb1-related retinopathies encompass a broad range of phenotypes which are not completely explained by the variability of Crb1 mutations. Genes thought to modify Crb1 function are therefore important targets of research. The biological function of Cygb involves oxygen delivery, scavenging of reactive oxygen species, and nitric oxide metabolism. However, the relationship of Cygb to diseases involving the Crb1 cell polarity complex is unknown. METHODS: Cygb knockout mice homozygous for the rd8 mutation (Cygb-/-rd8/rd8) were screened for ocular abnormalities and imaged using optical coherence tomography and fundus photography. Electroretinography was performed, as was histology and immunohistochemistry. Quantitative PCR was used to determine the effect of Cygb deficiency on transcription of Crb1 related cell polarity genes. RESULTS: Cygb-/-rd8/rd8 mice develop an abnormal retina with severe lamination abnormalities. The retina undergoes progressive degeneration with the ventral retina more severely affected than the dorsal retina. Cygb expression is in neurons of the retinal ganglion cell layer and inner nuclear layer. Immunohistochemical studies suggest that cell death predominates in the photoreceptors. Electroretinography amplitudes show reduced a- and b-waves, consistent with photoreceptor disease. Cygb deficient retinas had only modest transcriptional perturbations of Crb1-related cell polarity genes. Cygb-/- mice without the rd8 mutation did not exhibit obvious retinal abnormalities. CONCLUSIONS: Cygb is necessary for retinal lamination, maintenance of cell polarity, and photoreceptor survival in rd8 mice. These results are consistent with Cygb as a disease modifying gene in Crb1-related retinopathy. Further studies are necessary to investigate the role of Cygb in the human retina.

The Object Space Task reveals increased expression of cumulative memory in a mouse model of Kleefstra syndrome.

Kleefstra syndrome is a disorder caused by a mutation in the EHMT1 gene characterized in humans by general developmental delay, mild to severe intellectual disability and autism. Here, we characterized cumulative memory in the Ehmt1+/- mouse model using the Object Space Task. We combined conventional behavioral analysis with automated analysis by deep-learning networks, a session-based computational learning model, and a trial-based classifier. Ehmt1+/- mice showed more anxiety-like features and generally explored objects less, but the difference decreased over time. Interestingly, when analyzing memory-specific exploration, Ehmt1+/- show increased expression of cumulative memory, but a deficit in a more simple, control memory condition. Using our automatic classifier to differentiate between genotypes, we found that cumulative memory features are better suited for classification than general exploration differences. Thus, detailed behavioral classification with the Object Space Task produced a more detailed behavioral phenotype of the Ehmt1+/- mouse model.

Basal Forebrain Gating by Somatostatin Neurons Drives Prefrontal Cortical Activity.

The basal forebrain provides modulatory input to the cortex regulating brain states and cognitive processing. Somatostatin-expressing neurons constitute a heterogeneous GABAergic population known to functionally inhibit basal forebrain cortically projecting cells thus favoring sleep and cortical synchronization. However, it remains unclear if somatostatin cells can regulate population activity patterns in the basal forebrain and modulate cortical dynamics. Here, we demonstrate that somatostatin neurons regulate the corticopetal synaptic output of the basal forebrain impinging on cortical activity and behavior. Optogenetic inactivation of somatostatin neurons in vivo rapidly modified neural activity in the basal forebrain, with the consequent enhancement and desynchronization of activity in the prefrontal cortex, reflected in both neuronal spiking and network oscillations. Cortical activation was partially dependent on cholinergic transmission, suppressing slow waves and potentiating gamma oscillations. In addition, recruitment dynamics was cell type-specific, with interneurons showing similar temporal profiles, but stronger responses than pyramidal cells. Finally, optogenetic stimulation of quiescent animals during resting periods prompted locomotor activity, suggesting generalized cortical activation and increased arousal. Altogether, we provide physiological and behavioral evidence indicating that somatostatin neurons are pivotal in gating the synaptic output of the basal forebrain, thus indirectly controlling cortical operations via both cholinergic and non-cholinergic mechanisms.

Molecular mechanism of prion-like tau-induced neurodegeneration.

INTRODUCTION: Accumulation of hyperphosphorylated tau and the disruption of microtubules are correlated with synaptic loss and pathology of Alzheimer's disease (AD). Impaired cognitive function and pathology of AD is correlated with this lesion. This review looks at the mechanism of neurodegeneration, the prion-like behavior of tau in its interaction with normal MAPs in correlation with tau hyperphosphorylation. METHODS: We reviewed our work in the field as well as current literature that pertains to tau phosphorylation and the biological effects. RESULTS: Hyperphosphorylation of tau in AD, in vitro, in cells, or in animal models converts this protein into a prion-like protein that is able to propagate the altered conformation. DISCUSSION: These findings suggest that phosphorylation of tau is a critical event in neurodegeneration. The combination of phosphorylation sites can generate a gain of toxic function for tau. The mechanism of tau toxicity might involve not only the microtubule system but also interference with other cellular compartments such as the nucleus and the actin cytoskeleton.

Tau: More than a microtubule-binding protein in neurons.

Tau protein was discovered as a microtubule-associated protein nearly 50 years ago, and our understanding of tau has revolved around that role. Even with tau's rise to stardom as a central player in neurodegenerative disease, therapeutic efforts have largely been targeted toward cytoskeletal changes. While some studies hinted toward non-cytoskeletal roles for tau, it is only fairly recently that these ideas have begun to receive considerable attention. Many new binding partners for tau have been identified, including DNA, RNA, RNA-binding proteins, some receptors, and other tau molecules. The diversity of tau binding partners coupled with the discovery of tau other than axonal compartments such as nucleus, dendrites, and synapses have led to the proposal of novel functions for tau in roles such as nuclear stability, cell signaling, transcriptional processing, and protein synthesis. Tau self-assembly in particular has made an impact, leading to the hypothesis that a prion-like function of hyperphosphorylated tau is central to tauopathies. With tau emerging as a multifaceted protein that operates in many parts of the cell and with many molecular partners, the field of tau biology is primed for discoveries that can provide new perspectives on both the unique biochemistry of tau and the nature of devastating neurological diseases.

Composition and Functional Properties of the Edible Spear and By-Products from Asparagus officinalis L. and Their Potential Prebiotic Effect.

Asparagus is a healthy food appreciated for its organoleptic characteristics, nutritional composition and physiological properties. During its industrial processing, a large amount of by-products are generated, since only the apical part of the vegetable is considered edible and a large amount of by-products are generated that could be of nutritional interest. Therefore, the nutritional composition of the edible part and the two by-products of the plant (root and stem) was evaluated, including dietary fiber, inulin, low-molecular-weight carbohydrates, low-molecular-weight polyphenols and macromolecular polyphenols. The hydration properties, oil retention capacity, glucose retardation index and impact on bacterial growth of both probiotic bacteria and pathogenic strains were determined. All samples were high in fiber (>22 g/100 g dw), fructans (>1.5 g/100 g dw) and polyphenolic compounds (>3 g/100 g dw) and had good water-, oil- and glucose-binding capacity. In addition, they promoted the growth of probiotic strains but not pathogenic ones. The effects were more pronounced in the spear by-product samples and appear to be related to the components of dietary fiber. The results indicate that edible spear has potential beneficial effects on host health and microbiota when ingested as part of a healthy diet, while the by-products could be used as supplements and/or as natural ingredients in fiber-enriched foods that require emulsification and are intended to achieve a prebiotic effect.

UV-C Treatment Impact on the Availability of Water-Soluble Carbohydrates, Polyphenols, and Antioxidant Capacity of an Algerian Underutilized Date Fruit (Phoenix dactylifera L.).

Underutilized dates are considered as a socioeconomically important fruit for local and global communities, such as Degla-Beida, a common date fruit variety. The aim of this research was to elucidate, for the first time, the efficiency of UV-C light treatment (over different irradiation durations 5, 10, 20, and 40 min) in the enhancement of soluble carbohydrates and phenolic compounds, and to evaluate its effect on the antioxidant capacity. Furthermore, the content of dietary fiber was analyzed: insoluble dietary fiber (11.89 g/100 g); soluble dietary fiber (5.15 g/100 g); and total dietary fiber (17.06 g/100 g). The techno-functional properties were also determined: swelling capacity (3.94 mL/g); oil holding capacity (7.38 g/g); water holding capacity (9.30 g/g); and bulk density (1.81 g/mL). All were carried out to study the potential of exploiting this underutilized fruit for other applications as for feed or food. The results suggest that UV-C technology changes minimally the total water-soluble carbohydrate content; however, this preservation technology can affect the availability of different soluble carbohydrates depending on the irradiation time (IT), increasing the high molecular weight polysaccharides with IT up to 20 min, and some oligosaccharides with IT up to 5 min. The polyphenolic content determined by HPLC-QTOF was increased when the samples were submitted to UV-C reaching the maximum at 20 min (111.62 mg/100 g) and then to decrease in those submitted to IT of 40 min (12.05 mg/100 g). Regarding antioxidant capacity in the UV-C treated samples, FRAP decreased and EC50 on DPPH increased when IT was increased, while ORAC was hardly maintained. In addition, considering UV-C radiation associated with preservation and the studied date fruit as a rich source of dietary fiber with adequate techno-functional properties, this study presents valuable information for its potential use as a new food ingredient.

CBD lengthens sleep but shortens ripples and leads to intact simple but worse cumulative memory.

Cannabidiol (CBD) is on the rise as over-the-counter medication to treat sleep disturbances, anxiety, pain, and epilepsy due to its action on the excitatory/inhibitory balance in the brain. However, it remains unclear if CBD also leads to adverse effects on memory via changes of sleep macro- and microarchitecture. To investigate the effect of CBD on sleep and memory consolidation, we performed two experiments using the object space task testing for both simple and cumulative memory in rats. We show that oral CBD administration extended the sleep period but changed the properties of rest and non-REM sleep oscillations (delta, spindle, ripples). Specifically, CBD also led to less long (>100 ms) ripples and, consequently, worse cumulative memory consolidation. In contrast, simple memories were not affected. In sum, we can confirm the beneficial effect of CBD on sleep; however, this comes with changes in oscillations that negatively impact memory consolidation.

Increased cortical plasticity leads to memory interference and enhanced hippocampal-cortical interactions.

Our brain is continuously challenged by daily experiences. Thus, how to avoid systematic erasing of previously encoded memories? While it has been proposed that a dual-learning system with 'slow' learning in the cortex and 'fast' learning in the hippocampus could protect previous knowledge from interference, this has never been observed in the living organism. Here, we report that increasing plasticity via the viral-induced overexpression of RGS14414 in the prelimbic cortex leads to better one-trial memory, but that this comes at the price of increased interference in semantic-like memory. Indeed, electrophysiological recordings showed that this manipulation also resulted in shorter NonREM-sleep bouts, smaller delta-waves and decreased neuronal firing rates. In contrast, hippocampal-cortical interactions in form of theta coherence during wake and REM-sleep as well as oscillatory coupling during NonREM-sleep were enhanced. Thus, we provide the first experimental evidence for the long-standing and unproven fundamental idea that high thresholds for plasticity in the cortex protect preexisting memories and modulating these thresholds affects both memory encoding and consolidation mechanisms.

Nutritional composition of green asparagus (Asparagus officinalis L.), edible part and by-products, and assessment of their effect on the growth of human gut-associated bacteria.

Asparagus is considered a healthy food with a high content of bioactive compounds. In this study, the proximate and mineral composition, non-digestible carbohydrates and bioactive compounds of edible spear, spear by-product and root have been evaluated. Their activity on the growth of human gut-associated bacteria has been studied. The results support the high nutritional and functional value of the asparagus, including its by-products, highlighting the potential of the non-edible parts to be used as prebiotics. A remarkable content in xylose, inulin, flavonoids and saponins has been found. It has been shown that the spear by-product can be selectively used to promote the growth of commensal or probiotic lactobacilli and bifidobacteria strains. It has been confirmed that any part of the asparagus has a potential future as a healthy food or as health-promoting ingredients, however more work is required to identify the compounds able to modulate the human gut microbiota.

Pathological Human Tau Induces Alterations in the Brain Insulin Signaling Cascade.

The process of neurodegeneration in Alzheimer's disease has been associated with a disruption of insulin signaling cascade in neurons, and to insulin resistance. T2DM correlates with Alzheimer's disease, but mechanisms of interaction are unknown. We have developed a mouse model of tau induced neurodegeneration expressing pseudo-phosphorylated tau [Pathological Human Tau (PH-Tau)] in neurons. This model (PH-Tau-Tg) recapitulated cognitive decline and neurodegeneration observed in AD. In this study we examined if expression of PH-Tau could affect neuronal excitability and insulin receptor signaling. Neuronal excitability was investigated using intracerebral recordings of extracellular field potentials from prefrontal cortex after insulin and kainic acid (KA) injection. Analysis of baseline recordings indicated an increased excitability of PH-Tau-Tg as evidenced by higher spectrum densities (PSDs) of high frequencies brain waves. Injection of insulin (1IU, s.c) led to a decrease of fast ripples PSDs, more pronounced in PH-Tau-Tg mice than controls. Subsequent injection of kainic acid (KA, 5 mg/kg, s.c) led to significant increase in firing rate, amplitude of extracellular field potentials and PSDs of high frequency brain waves in control mice only. To further investigate the role of insulin in PH-Tau-Tg mice, we subjected mice to a glucose tolerance test. We found that PH-Tau-Tg mice were significantly hyperglycemic prior to glucose injection. Interestingly, the PH-Tau-Tg mice showed a moderate increase at 30 min due to the higher baseline, indicating a low sensitivity of insulin receptor in these mice. This is consistent with increased levels of activated insulin receptors in the brain and the inhibitory effect of insulin on ictal activity post KA injection in PH-Tau-Tg mice. We suggest that these mice have reduced insulin sensitivity (hyperglycemia) and as a compensatory mechanism there is overactivation/expression of insulin receptor in the brain rendering neuronal circuits resistant to seizure induction after injection of insulin. These data indicate that insulin signal transduction pathway is altered in PH-Tau-Tg mice, and that injection of exogenous insulin reduces hypersynchronous bursting activity of field potentials recorded from cortical neuronal circuits. We propose that the appearance of abnormal tau might potentiate the toxic environment by interfering with the insulin signaling cascade in the brain of patients with Alzheimer's disease.

Optogenetic Suppression of Lateral Septum Somatostatin Neurons Enhances Hippocampus Cholinergic Theta Oscillations and Local Synchrony.

The septal complex regulates both motivated and innate behaviors, chiefly by the action of its diverse population of long-range projection neurons. A small population of somatostatin-expressing GABAergic cells in the lateral septum projects deep into subcortical regions, yet on its way it also targets neighboring medial septum neurons that profusely innervate cortical targets by ascending synaptic pathways. Here, we used optogenetic stimulation and extracellular recordings in acutely anesthetized transgenic mice to show that lateral septum somatostatin neurons can disinhibit the cholinergic septo-hippocampal pathway, thus enhancing the amplitude and synchrony of theta oscillations while depressing sharp-wave ripple episodes in the dorsal hippocampus. These results suggest that septal somatostatin cells can recruit ascending cholinergic pathways to promote hippocampal theta oscillations.

Importin-Mediated Pathological Tau Nuclear Translocation Causes Disruption of the Nuclear Lamina, TDP-43 Mislocalization and Cell Death.

Tau is a cytosolic protein that has also been observed in the nucleus, where it has multiple proposed functions that are regulated by phosphorylation. However, the mechanism underlying the nuclear import of tau is unclear, as is the contribution of nuclear tau to the pathology of tauopathies. We have previously generated a pathological form of tau, PH-tau (pseudophosphorylation mutants S199E, T212E, T231E, and S262E) that mimics AD pathological behavior in cells, Drosophila, and a mouse model. Here, we demonstrated that PH-tau translocates into the nucleus of transiently transfected HEK-293 cells, but wildtype tau does not. We identified a putative importin binding site in the tau sequence, and showed that disruption of this site prevents tau from entering the nucleus. We further showed that this nuclear translocation is prevented by inhibitors of both importin-α and importin-ß. In addition, expression of PH-tau resulted in an enlarged population of dying cells, which is prevented by blocking its entry into the nucleus. PH-tau-expressing cells also exhibited disruption of the nuclear lamina and mislocalization of TDP-43 to the cytoplasm. We found that PH-tau does not bundle microtubules, and this effect is independent of nuclear translocation. These results demonstrate that tau translocates into the nucleus through the importin-α/ß pathway, and that PH-tau exhibits toxicity after its nuclear translocation. We propose a model where hyperphosphorylated tau not only disrupts the microtubule network, but also translocates into the nucleus and interferes with cellular functions, such as nucleocytoplasmic transport, inducing mislocalization of proteins like TDP-43 and, ultimately, cell death.

Pasteurization Modifies the Sensorial Attributes and Nutritional Profile of Orange Pulp By-Product.

After orange processing, different by-products are generated, i.e., peels, seeds and pulps. The pulp is highly perishable, being an unstable food matrix that needs a preservation process to be stored and used again in the food production chain. Pasteurization is the technique of choice before aseptically packaging and storing under refrigerated conditions. In this study, the effect of pasteurization has been evaluated on the chemical, functional and sensorial profiles. Ash content decreased (p < 0.05) after the thermal treatment. Indeed, magnesium, calcium and zinc diminished, although copper was found to be higher (p < 0.05) in the pasteurized product. Total dietary fiber decreased (p < 0.05), but soluble dietary fiber raised (p < 0.05) due to hydrolysis caused by pasteurization. SDF:IDF ratio, hydration properties, and fat binding capacity were improved. Total soluble phenolic compounds remained similar but FRAP and DPPH scavenging activity decreased (p < 0.05) in the pasteurized by-product. Regarding the sensorial profile, pasteurization produced darkening, appearance of a cooked smell and an increase in bitterness. Therefore, pasteurization deteriorates the sensorial profile being able to change the attributes of an added-pasteurized-pulp juice; however, it is a good choice to preserve the orange pulp by-product to formulate food products different from juices or other beverages.

Optimization of Conventional Extraction Parameters for Recovering Phenolic Compounds from Potato (Solanum tuberosum L.) Peels and Their Application as an Antioxidant in Yogurt Formulation.

The aim of this work was to optimize the conventional parameters for the extraction of phenolic compounds from potato (Solanum tuberosum L.) peels (PP). A central composite design (CCD) was used to establish the impacts of ethanol concentration (%), extraction time (min), and liquid/solid ratio (mL/g). The optimal experimental conditions that maximized extraction were ethanol at a concentration of 80% (v/v) for a time of 150 min with a ratio of 1 g/30 mL. Under optimal conditions, the total phenolic content (TPC) and the total flavonoid content (TFC) were 204.41 ± 8.64 mg GAE/100 g DW and 21.47 ± 0.76 mg QE/100 g DW, respectively. The PP extract had a potent antioxidant capacity tested by phosphomolybdate and DPPH assays with IC50 of 10.65 ± 0.21 and 179.75 ± 3.18 µg/mL, respectively. Furthermore, by fortifying yogurt with PP as a natural ingredient, an improvement ofits physical, nutritional, antioxidant, and sensorial qualities was attempted in this study. The yogurts formulated with PP revealed significantly higher (p ≤ 0.05) TPC, TFC, and antioxidant capacity in comparison with the control sample. In addition, the sensory evaluation showed that the yogurts enriched with PP were preferred over the control yogurt. The results indicate that PP can be considered an interesting byproduct since it can improve the nutritional, bioactive, and sensorial profile of yogurt, highlighting that PP, due to its high phenol content, can substantially improve the antioxidant effect of the new formulated yogurt.

Influence of high-pressure processing on nutritional composition and bioactive compounds of Phaseolus coccineus L.

The influence of high-pressure processing (HPP) prior to cooking on nutritional composition and bioactive compounds content of four varieties of Phaseolus coccineus L. was studied. Cooking and HPP+C increased the protein content. However, minerals, total carbohydrates, ciceritol and α-galactosides were reduced. Fat was not affected by cooking but decreased after HPP+C. For dietary fiber, the behavior observed was different depending on the sample and the treatment applied. HPP+C could be considered a good processing technology to retain the advantageous lower myo-inositol phosphates isoforms and supply prebiotic oligosaccharides. The trypsin inhibitors activity was lower in the cooked and HPP+C samples; however, there were no significant differences between both thermal treatments. Thus, HPP+C reduced cooking time and preserving or improving the nutritional composition of the beans and their bioactive compounds content.

Phosphorylation of tau at Thr212, Thr231, and Ser262 combined causes neurodegeneration.

Abnormal hyperphosphorylation of the microtubule-associated protein Tau is a hallmark of Alzheimer disease and related diseases called tauopathies. As yet, the exact mechanism by which this pathology causes neurodegeneration is not understood. The present study provides direct evidence that Tau abnormal hyperphosphorylation causes its aggregation, breakdown of the microtubule network, and cell death and identifies phosphorylation sites involved in neurotoxicity. We generated pseudophosphorylated Tau proteins by mutating Ser/Thr to Glu and, as controls, to Ala. These mutations involved one, two, or three pathological phosphorylation sites by site-directed mutagenesis using as backbones the wild type or FTDP-17 mutant R406W Tau. Pseudophosphorylated and corresponding control Tau proteins were expressed transiently in PC12 and CHO cells. We found that a single phosphorylation site alone had little influence on the biological activity of Tau, except Thr(212), which, upon mutation to Glu in the R406W background, induced Tau aggregation in cells, suggesting phosphorylation at this site along with a modification on the C-terminal of the protein facilitates self-assembly of Tau. The expression of R406W Tau pseudophosphorylated at Thr(212), Thr(231), and Ser(262) triggered caspase-3 activation in as much as 85% of the transfected cells, whereas the corresponding value for wild type pseudophosphorylated Tau was 30%. Cells transfected with pseudophosphorylated Tau became TUNEL-positive.