Acute effects of a ketone monoester, whey protein, or their coingestion on mTOR trafficking and protein-protein colocalization in human skeletal muscle.

We recently demonstrated that acute oral ketone monoester intake induces a stimulation of postprandial myofibrillar protein synthesis rates comparable to that elicited following the ingestion of 10 g whey protein or their coingestion. The present investigation aimed to determine the acute effects of ingesting a ketone monoester, whey protein, or their coingestion on mechanistic target of rapamycin (mTOR)-related protein-protein colocalization and intracellular trafficking in human skeletal muscle. In a randomized, double-blind, parallel group design, 36 healthy recreationally active young males (age: 24.2 ± 4.1 yr) ingested either: 1) 0.36 g·kg-1 bodyweight of the ketone monoester (R)-3-hydroxybutyl (R)-3-hydroxybutyrate (KET), 2) 10 g whey protein (PRO), or 3) the combination of both (KET + PRO). Muscle biopsies were obtained in the overnight postabsorptive state (basal conditions), and at 120 and 300 min in the postprandial period for immunofluorescence assessment of protein translocation and colocalization of mTOR-related signaling molecules. All treatments resulted in a significant (Interaction: P < 0.0001) decrease in tuberous sclerosis complex 2 (TSC2)-Ras homolog enriched in brain (Rheb) colocalization at 120 min versus basal; however, the decrease was sustained at 300 min versus basal (P < 0.0001) only in KET + PRO. PRO and KET + PRO increased (Interaction: P < 0.0001) mTOR-Rheb colocalization at 120 min versus basal; however, KET + PRO resulted in a sustained increase in mTOR-Rheb colocalization at 300 min that was greater than KET and PRO. Treatment intake increased mTOR-wheat germ agglutinin (WGA) colocalization at 120 and 300 min (Time: P = 0.0031), suggesting translocation toward the fiber periphery. These findings demonstrate that ketone monoester intake can influence the spatial mechanisms involved in the regulation of mTORC1 in human skeletal muscle.NEW & NOTEWORTHY We explored the effects of a ketone monoester (KET), whey protein (PRO), or their coingestion (KET + PRO) on mTOR-related protein-protein colocalization and intracellular trafficking in human muscle. All treatments decreased TSC2-Rheb colocalization at 120 minutes; however, KET + PRO sustained the decrease at 300 min. Only PRO and KET + PRO increased mTOR-Rheb colocalization; however, the increase at 300 min was greater in KET + PRO. Treatment intake increased mTOR-WGA colocalization, suggesting translocation to the fiber periphery. Ketone bodies influence the spatial regulation of mTOR.

The J Domain of Sacsin Disrupts Intermediate Filament Assembly.

Autosomal Recessive Spastic Ataxia of the Charlevoix Saguenay (ARSACS) is caused by mutation in the SACS gene resulting in loss of function of the protein sacsin. A key feature is the formation of abnormal bundles of neurofilaments (NF) in neurons and vimentin intermediate filaments (IF) in cultured fibroblasts, suggesting a role of sacsin in IF homeostasis. Sacsin contains a J domain (SacsJ) homologous to Hsp40, that can interact with Hsp70 chaperones. The SacsJ domain resolved NF bundles in cultured Sacs-/- neurons. Having studied the mechanism using NF assembled in vitro from purified NF proteins, we report that the SacsJ domain interacts with NF proteins to disassemble NFL filaments, and to inhibit their initial assembly. A cell-penetrating peptide derived from this domain, SacsJ-myc-TAT was efficient in disassembling NF bundles in cultured Sacs-/- motor neurons, restoring the NF network; however, there was some loss of vimentin IF and NF in cultured Sacs+/+ fibroblasts and motor neurons, respectively. These results suggest that sacsin through its SacsJ domain is a key regulator of NF and vimentin IF networks in cells.

The influence of awareness on implicit visuomotor adaptation.

It is well documented that reaches are adapted when reaching with a visuomotor distortion (i.e., rotated cursor feedback). Less clear is the influence of awareness on visuomotor adaptation, where awareness encompasses knowledge of the changes in one's reaches and the visuomotor distortion itself. In the current experiment, we asked if awareness governs the magnitude of implicit (i.e., unconscious) visuomotor adaptation achieved, independent of how the distortion is introduced (i.e., abruptly vs. gradually introduced visuomotor distortion), and hence initial errors experienced. Participants were divided into two groups that differed with respect to how the visuomotor distortion was introduced (i.e., Abrupt vs. Gradual Groups) and reached in a virtual environment where a cursor on the screen misrepresented the position of their hand. Participants completed three blocks of 150 reach training trials in the following order: aligned cursor feedback (baseline), rotated cursor feedback (adaptation) and aligned cursor feedback (washout). For the Abrupt Group, the cursor was immediately rotated 45° clockwise (CW) relative to hand motion in the adaptation block, whereas in the Gradual Group, the 45° cursor rotation was gradually introduced over adaptation trials. Following reach training, participants' awareness of changes in their reaches and the visuomotor distortion were established based on a drawing task, where participants drew the path their hand took to get the cursor on target, as well as a post-experiment questionnaire. Participants were subsequently divided into the following 3 groups: Abrupt-Aware (n = 16), Gradual-Aware (n = 11) and Gradual-Unaware (n = 14). Results revealed that errors differed for the Gradual-Unaware Group at the end of adaptation training compared to the Gradual-Aware Group and at the start of the washout block compared to the Abrupt-Aware Group. Errors in the two aware groups did not differ from each other. These results suggest that awareness may lead to reduced implicit adaptation, regardless of the size of initial errors experienced.