Transfer of polyglutamine aggregates in neuronal cells occurs in tunneling nanotubes.

Huntington's disease (HD) is a dominantly inherited neurodegenerative disease caused by CAG expansion in the huntingtin gene, which adds a homopolymeric tract of polyglutamine (polyQ) to the encoded protein leading to the formation of toxic aggregates. Despite rapidly accumulating evidences supporting a role for intercellular transmission of protein aggregates, little is known about whether and how huntingtin (Htt) misfolding progresses through the brain. It has been recently reported that synthetic polyQ peptides and recombinant fragments of mutant Htt are readily internalized in cell cultures and able to seed polymerization of a reporter wild-type Htt. However, there is no direct evidence of aggregate transfer between cells and the mechanism has not been explored. By expressing recombinant fragments of mutant Htt in neuronal cells and in primary neurons, we found that aggregated fragments formed within one cell spontaneously transfer to neighbors in cell culture. We demonstrate that the intercellular spreading of the aggregates requires cell-cell contact and does not occur upon aggregate secretion. Interestingly, we found that the expression of mutant, but not wild-type Htt fragments, increases the number of tunneling nanotubes, which in turn provide an efficient mechanism of transfer.

The cell biology of prion-like spread of protein aggregates: mechanisms and implication in neurodegeneration.

The misfolding and aggregation of specific proteins is a common hallmark of many neurodegenerative disorders, including highly prevalent illnesses such as Alzheimer's and Parkinson's diseases, as well as rarer disorders such as Huntington's and prion diseases. Among these, only prion diseases are 'infectious'. By seeding misfolding of the PrP(C) (normal conformer prion protein) into PrP(Sc) (abnormal disease-specific conformation of prion protein), prions spread from the periphery of the body to the central nervous system and can also be transmitted between individuals of the same or different species. However, recent exciting data suggest that the transmissibility of misfolded proteins within the brain is a property that goes way beyond the rare prion diseases. Evidence indicates that non-prion aggregates [tau, α-syn (α-synuclein), Aß (amyloid-ß) and Htt (huntingtin) aggregates] can also move between cells and seed the misfolding of their normal conformers. These findings have enormous implications. On the one hand they question the therapeutical use of transplants, and on the other they indicate that it may be possible to bring these diseases to an early arrest by preventing cell-to-cell transmission. To better understand the prion-like spread of these protein aggregates it is essential to identify the underlying cellular and molecular factors. In the present review we analyse and discuss the evidence supporting prion-like spreading of amyloidogenic proteins, especially focusing on the cellular and molecular mechanisms and their significance.

Characterization of the role of dendritic cells in prion transfer to primary neurons.

TSEs (transmissible spongiform encephalopathies) are neurodegenerative diseases caused by pathogenic isoforms (PrPSc) of the host-encoded PrPc (cellular prion protein). After consumption of contaminated food, PrPSc deposits rapidly accumulate in lymphoid tissues before invasion of the CNS (central nervous system). However, the mechanisms of prion spreading from the periphery to the nervous system are still unclear. In the present study, we investigated the role of DCs (dendritic cells) in the spreading of prion infection to neuronal cells. First, we determined that BMDCs (bone-marrow-derived DCs) rapidly uptake PrPSc after exposure to infected brain homogenate. Next, we observed a progressive catabolism of the internalized prion aggregates. Similar experiments performed with BMDCs isolated from KO (knockout) mice or mice overexpressing PrP (tga20) indicate that both PrPSc uptake and catabolism are independent of PrPc expression in these cells. Finally, using co-cultures of prion-loaded BMDCs and cerebellar neurons, we characterized the transfer of the prion protein and the resulting infection of the neuronal cultures. Interestingly, the transfer of PrPSc was triggered by direct cell-cell contact. As a consequence, BMDCs retained the prion protein when cultured alone, and no transfer to the recipient neurons was observed when a filter separated the two cultures or when neurons were exposed to the BMDC-conditioned medium. Additionally, fixed BMDCs also failed to transfer prion infectivity to neurons, suggesting an active transport of prion aggregates, in accordance with a role of TNTs (tunnelling nanotubes) observed in the co-cultures.

Doppel and PrPC co-immunoprecipitate in detergent-resistant membrane domains of epithelial FRT cells.

Dpl (doppel) is a paralogue of the PrPC (cellular prion protein), whose misfolded conformer (the scrapie prion protein, PrPSc) is responsible for the onset of TSEs (transmissible spongiform encephalopathies) or prion diseases. It has been shown that the ectopic expression of Dpl in the brains of some lines of PrP-knockout mice provokes cerebellar ataxia, which can be rescued by the reintroduction of the PrP gene, suggesting a functional interaction between the two proteins. It is, however, still unclear where, and under which conditions, this event may occur. In the present study we addressed this issue by analysing the intracellular localization and the interaction between Dpl and PrPC in FRT (Fischer rat thyroid) cells stably expressing the two proteins separately or together. We show that both proteins localize prevalently on the basolateral surface of FRT cells, in both singly and doubly transfected clones. Interestingly we found that they associate with DRMs (detergent-resistant membranes) or lipid rafts, from where they can be co-immunoprecipitated in a cholesterol-dependent fashion. Although the interaction between Dpl and PrPC has been suggested before, our results provide the first clear evidence that this interaction occurs in rafts and is dependent on the integrity of these membrane microdomains. Furthermore, both Dpl and PrPC could be immunoprecipitated with flotillin-2, a raft protein involved in endocytosis and cell signalling events, suggesting that they share the same lipid environment.

Modelling picture naming in aphasia: The relationship between consistency and predictability of responses.

OBJECTIVE: Two aspects of aphasic picture naming were examined: response consistency, that is, the extent to which the accuracy of the response to the same stimulus is replicated in a successive examination, and response predictability, that is, the extent to which accuracy depends on the characteristics of each stimulus. METHODS: Thirty-eight aphasic participants were examined twice. The response pattern was the same across the 2 presentations (response stability) for 36 participants, who were classified into 3 groups according to the prevailing error-type (lexical-semantic, phonological, or a balance between the two error-types): Their item-consistency was quantified with Cohen's kappa. In each case the roles played by lexical frequency, precocity of acquisition and length of the target word, and visual complexity and image agreement of the stimulus picture were examined; the ability to predict response accuracy of a model simultaneously including these 5 variables was quantified by means of the McFadden index. Finally, the relationship between predictability (McFadden index) and consistency (Cohen's kappa) was analyzed. RESULTS: For 34 of 36 participants, consistency was higher than chance. Consistency was directly correlated to the prevalence of lexical-semantic errors. On regression analysis, the relationship between consistency and predictability was significant. CONCLUSIONS: Response consistency reflects the existence of a clear difficulty gradient within the items of a battery. The significant relationship between consistency and error type suggests that, in principle, lexical-semantic errors might be more predictable than phonological errors based on the characteristics of each stimulus. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

The role of Broca's area in speech perception: evidence from aphasia revisited.

Motor theories of speech perception have been re-vitalized as a consequence of the discovery of mirror neurons. Some authors have even promoted a strong version of the motor theory, arguing that the motor speech system is critical for perception. Part of the evidence that is cited in favor of this claim is the observation from the early 1980s that individuals with Broca's aphasia, and therefore inferred damage to Broca's area, can have deficits in speech sound discrimination. Here we re-examine this issue in 24 patients with radiologically confirmed lesions to Broca's area and various degrees of associated non-fluent speech production. Patients performed two same-different discrimination tasks involving pairs of CV syllables, one in which both CVs were presented auditorily, and the other in which one syllable was auditorily presented and the other visually presented as an orthographic form; word comprehension was also assessed using word-to-picture matching tasks in both auditory and visual forms. Discrimination performance on the all-auditory task was four standard deviations above chance, as measured using d', and was unrelated to the degree of non-fluency in the patients' speech production. Performance on the auditory-visual task, however, was worse than, and not correlated with, the all-auditory task. The auditory-visual task was related to the degree of speech non-fluency. Word comprehension was at ceiling for the auditory version (97% accuracy) and near ceiling for the orthographic version (90% accuracy). We conclude that the motor speech system is not necessary for speech perception as measured both by discrimination and comprehension paradigms, but may play a role in orthographic decoding or in auditory-visual matching of phonological forms.