Introduction: Contradictory reports of the myosin heavy chain (MHC) composition of adult human suprahyoid muscles leave unresolved the extent to which these muscles express developmental and unconventional MHC. Methods: By immunohistochemistry, separation sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)-Coomassie, separation SDS-PAGE-Western blot, and mRNA PCR, we tested for conventional MHCI, MHCIIA, MHCIIX, developmental MHC embryonic and MHC neonatal, and unconventional MHC alpha-cardiac, MHC extraocular, and MHC slow tonic in adult human anterior digastric (AD), geniohyoid (GH), and mylohyoid (MH) muscles. Results: By separation SDS-PAGE-Coomassie and Western blot, only conventional MHC are present. By immunohistochemistry all muscle fibers are positive for MHCI, MHCIIA, or MHCIIX, and fewer than 4 fibers/mm2 are positive for developmental or unconventional MHC. By PCR, mRNA of MHCI and MHCIIA dominate, with sporadically detectable MHC alpha-cardiac and without detectable mRNA of other developmental and unconventional MHC. Conclusions: We conclude that human suprahyoid muscles AD, GH, and MH are composed almost exclusively of conventional MHC isoforms.

Thoracic paravertebral sympathetic postganglionic neurons (tSPNs) comprise the final integrative output of the distributed sympathetic nervous system controlling vascular and thermoregulatory systems. Considered a non-integrating relay, what little is known of tSPN intrinsic excitability has been determined by sharp microelectrodes with presumed impalement injury. We thus undertook the first electrophysiological characterization of tSPN cellular properties using whole-cell recordings and coupled results with a conductance-based model to explore the principles governing their excitability in adult mice of both sexes. Recorded membrane resistance and time constant values were an order of magnitude greater than values previously obtained, leading to a demonstrable capacity for synaptic integration in driving recruitment. Variation in membrane resistivity was the primary determinant controlling cell excitability with vastly lower currents required for tSPN recruitment. Unlike previous microelectrode recordings in mouse which observed inability to sustain firing, all tSPNs were capable of repetitive firing. Computational modeling demonstrated that observed differences are explained by introduction of a microelectrode impalement injury conductance. Overall, tSPNs largely linearly encoded injected current magnitudes over a broad frequency range with distinct subpopulations differentiable based on repetitive firing signatures. Thus, whole-cell recordings reveal tSPNs have more dramatically amplified excitability than previously thought, with greater intrinsic capacity for synaptic integration and with the ability for maintained firing to support sustained actions on vasomotor tone and thermoregulatory function. Rather than acting as a relay, these studies support a more responsive role and possible intrinsic capacity for tSPNs to drive sympathetic autonomic function.

Objective Uncertain biological consequences of titanium-magnet (Ti-mag) tongue implants constrain application of the Tongue Drive System (TDS), a brain-tongue-computer interface for individuals with severe physical impairment. Here we describe oromotor function and tongue tissue response following Ti-Mag implantation and explantation in the miniature pig, an animal model with a tongue similar in size to humans. Design A 1.8 × 6.2 mm Ti-mag tracer was implanted into the anterior tongue in five Yucatan minipigs. X-rays were taken immediately and >six days after implantation to evaluate tracer migration. In three minipigs, the tracer was explanted?>16 days after implantation. Twenty-five days post-explantation, tongue tissue was harvested and processed for histological and immunohistochemical (IHC) markers of healing. In two minipigs tissue markers of healing were evaluated post-mortem following >12 days implantation. Drink cycle rate (DCR) was characterized to determine the impact of procedures on oromotor function. Results Neither implantation (N = 5) nor explantation (N = 3) changed DCR. X-rays revealed minimal tracer migration (N = 4, 0–4 mm). By histology and IHC a robust capsule was present two weeks post-implantation with limited fibrosis. Explantation produced localized fibrosis and limited muscle remodeling. Conclusions These findings suggest the safety of Ti-mag anterior tongue implants for assistive technologies in humans.

Objective: The Tongue Drive System (TDS) is a new wearable assistive technology (AT), developed to translate voluntary tongue movements to user-defined computer commands by tracking the position of a titanium-encased magnetic tracer (Ti-Mag) implanted into the tongue. TDS application, however, is constrained by limited information on biological consequence and safety of device implantation into the tongue body. Here we implant a stainless-steel pellet in the rat tongue and assay pellet migration, tongue lick function, and tongue histology to test the safety and biocompatibility of unanchored tongue implants. Design: Water consumption, weight and lick behavior were measured before and for >24 days after implantation of a stainless-steel spherical pellet (0.5 mm) into the anterior tongue body of twelve adult male rats. X-rays were obtained weekly to assess pellet migration. Pellet location and tissue reaction to implantation were determined by post-mortem dissection and histology of the anterior tongue. Results: By dissection pellets were distributed across the transverse plane of the tongue. Measures of water consumption, weight, and lick behavior were unchanged by implantation except for a decrease in consumption immediately post-implantation in some animals. By X-ray, there was no migration of the implant, a finding supported by pellet encapsulation demonstrated histologically. Measures of lick behavior were minimally impacted by implantation. Conclusion: A smooth spherical stainless-steel implant in the anterior tongue of the rat does not migrate, is encapsulated and does not substantially impact lick behavior. These findings support the implantation of small tracers in the anterior tongue in humans for operating wearable assistive technologies.

Introduction Equivocal decline of tongue muscle performance with age is compatible with resistance of the tongue to sarcopenia, the loss of muscle volume and function that typically occurs with aging. To test this possibility we characterized anatomical and molecular indices of sarcopenia in the macaque tongue muscle styloglossus (SG). Methods We quantified myosin heavy chain (MHC), muscle fiber MHC phenotype and size and total and phosphorylated growth- and atrophy-related proteins by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), immunoblot and immunohistochemistry (IHC) in the SG in twenty-four macaque monkeys (Macaca rhesus, age range 9 months to 31 years) categorized into Young ( <  8 years of age), Middle-aged (15–21 years of age) and Old ( >  22 years of age) groups. Results In Young, Middle and Old age groups, by SDS-PAGE MHCI comprised ~ 1/3 and MHCII ~ 2/3 of total MHC. MHCI relative frequency was lower and MHCII higher in Middle versus Young (p = 0.0099) and Middle versus Old (p = 0.052). Relative frequencies of MHC fiber phenotype were not different by age but were different by phenotype (rates 233, 641 and 111 per 1000 fibers for MHCI, MHCII and MHCI-II respectively, p = 0.03). Few or no fibers were positive for developmental MHC. Mean cross-sectional area (CSA) was not different among the three age groups for MHCII and MHCI-II; however MHCI fibers tended to be larger in Middle versus Old and Young (mean = 2257 μm 2 ,1917 μm 2 (p = 0.05) and 1704 μm 2 (p = 0.06), respectively). For each age group, mean CSA increased across MHC phenotype (lowest mean CSA for MHCI and highest mean CSA for MHCII). Spearman analysis demonstrated age-related increases in total p70 ribosomal protein S6 kinase (P70), phosphorylated P70 421/424 , phosphorylated P38 mitogen-activated protein kinase and muscle atrophy F-Box, a trend to age-related decrease in total extracellular signal-regulated kinase (ERK), and no age-related change in total protein kinase B (Akt/PKB), phosphorylated Akt, phosphorylated ERK, phosphorylated c-Jun N-terminal kinase (JNK46) and phosphorylated P70 389 . Conclusion Common anatomical and molecular indices of sarcopenia are absent in our sample of macaque SG. Relative frequencies of MHCII protein and phenotype are preserved with age. Although MAFbx expression increases with age, this is not associated with fiber atrophy, perhaps reflecting compensatory growth signaling by p70. The resistant nature of the styloglossus muscle to sarcopenia may be related to routine activation of tongue muscles in respiration and swallowing and the preservation of hypoglossal motoneuron number with age.