Brennan, C. D. | bioRxiv Science

bioRxiv2025

Hindlimb Immobilization Impairs Neuromuscular Junction Transmission in Young Rats

BackgroundImmobilization, bed rest, or illness rapidly lead to weakness out of proportion to muscle atrophy. Although the contribution of muscle wasting to weakness is well described, the role of neuromuscular junction (NMJ) dysfunction in early disuse-related weakness is not well understood. ObjectiveWe investigated whether short-term unilateral hindlimb immobilization (HLI) in rats impairs NMJ transmission and contributes to muscle weakness out of proportion to atrophy. MethodsFour-month-old male Fischer-344/Brown Norway rats underwent 10 days of unilateral HLI (n=6) or remained mobile (n=6). Neuromuscular excitability and transmission were assessed using compound muscle action potentials (CMAP), repetitive nerve stimulation (RNS), and single-fiber electromyography (SFEMG). Muscle contractility testing quantified tetanic torque, and post-mortem analysis measured muscle mass. ResultsTen days of HLI reduced gastrocnemius, soleus, and plantaris muscle mass by [~]20-35%. Plantarflexion peak tetanic torque normalized to body weight declined by 23%, and torque-time integral was reduced by 36%, indicating disproportionate functional loss (muscle size versus contractile output) and supporting underlying neural impairment. CMAP amplitude decreased from 69 mV to 53.23 mV (a 22.9% reduction; p = 0.0109), indicating a loss of summated neuromuscular excitability. Furthermore, both RNS and SFEMG indicated features consistent with NMJ transmission defects. RNS revealed CMAP decrement from [~]0% pre-HLI to -9.15% at 40 Hz stimulation and -8.63% at 50 Hz post-HLI. Similarly, SFEMG confirmed marked NMJ transmission defects, with jitter increasing 103% and blocking increased from <1% to >11% of fibers. ConclusionsOur findings suggest that short-term immobilization produces rapid and pronounced impairments in NMJ transmission that contribute to weakness beyond the degree of muscle atrophy. These findings identify the NMJ as an early and vulnerable site of disuse-induced dysfunction and highlight the potential for synaptic-targeted therapies to preserve muscle performance during immobilization and recovery.

physiology↗

bioRxiv2023

A leptin-responsive hypothalamic circuit inputs to the circadian feeding network

Salient cues, such as the rising sun or the availability of food, play a crucial role in entraining biological clocks, allowing for effective behavioral adaptation and ultimately, survival. While the light-dependent entrainment of the central circadian pacemaker (suprachiasmatic nucleus, SCN) is relatively well defined, the molecular and neural mechanisms underlying entrainment associated with food availability remains elusive. Using single nucleus RNA sequencing during scheduled feeding (SF), we identified a leptin receptor (LepR) expressing neuron population in the dorsomedial hypothalamus (DMH) that upregulates circadian entrainment genes and exhibits rhythmic calcium activity prior to an anticipated meal. We found that disrupting DMHLepR neuron activity had a profound impact on both molecular and behavioral food entrainment. Specifically, silencing DMHLepR neurons, mis-timed exogenous leptin administration, or mis-timed chemogenetic stimulation of these neurons all interfered with the development of food entrainment. In a state of energy abundance, repetitive activation of DMHLepR neurons led to the partitioning of a secondary bout of circadian locomotor activity that was in phase with the stimulation and dependent on an intact SCN. Lastly, we discovered that a subpopulation of DMHLepR neurons project to the SCN with the capacity to influence the phase of the circadian clock. This leptin regulated circuit serves as a point of integration between the metabolic and circadian systems, facilitating the anticipation of meal times.

neuroscience↗