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Biology subjects

Gustafson, K.

Publications and source records attributed to Gustafson, K..

3 recordsLinked to original sources

Mapping the Fascicular Morphology and Organization of the Human Sciatic Nerve via High-Resolution MicroCT Imaging

ObjectiveImplanted neuroprostheses can restore standing and walking after spinal cord injury and somatosensation after limb loss. Yet current approaches often fail to reliably activate hamstring muscles crucial for upright stability and mobility or to target afferent fibers for sensory restoration. We developed a novel methodology using high-resolution micro-computed tomography (microCT) to visualize and track fascicle groups innervating distinct hamstring muscles along the human sciatic nerve. This approach provides a framework for mapping fascicular topography in complex neural pathways to optimize standing neuroprostheses. MethodsBilateral sciatic nerves were dissected and excised from an embalmed human cadaver, annotated with branch names, and stained with phosphotungstic acid before undergoing microCT scanning at 11.4 m isotropic resolution. Images were segmented with a 3D U-Net convolutional neural network. Segmentation results were used to quantify morphological metrics and track fascicular organization along [~]25 cm of the nerve. MicroCT reconstructions were validated against histological cross sections. ResultsGross dissection revealed matched proximal-to-distal branching between left and right sciatic nerves: branch to long head of the biceps femoris (lhBF), branch to hamstring part of the adductor magnus and semimembranosus (HAM/SM), and branch to semitendinosus (ST). All branches originated medially and followed an inferomedial trajectory. Branch-free lengths of the sciatic exhibited asymmetry, especially between the lumbosacral roots to the first branch (5.5 cm left vs. 1.5 cm right) and lhBF to the HAM/SM branch (9.0 cm left vs. 16.5 cm right). MicroCT analysis revealed bilateral symmetry in fascicle diameters ([~]0.4 mm) and total fascicle counts ([~]84) but asymmetry in hamstring-innervating fascicle counts (left [~]7, right: [~]9). The 3D fascicular maps revealed that hamstring fascicles were located in the anteromedial portion of the sciatic nerve cross section and remained separate for distances up to 15.9 cm proximal to their branching points. SignificanceOur microCT-based approach enables efficient, high-resolution 3D mapping of fascicular organization within large, complex peripheral nerves like the sciatic nerve, overcoming previous technical limitations. This methodology informs development of neuroprostheses with improved hamstring muscle activation for enhanced standing function.

bioengineering↗

Mitochondrial oxidation of the carbohydrate fuel driven by pyruvate dehydrogenase robustly enhances stemness of older and geriatric Intestinal Stem Cells

Background and AimsAging impairs Intestinal Stem Cell (ISC) function and attenuates their regenerative capacity. Although the transcriptional landscape governing ISC fate during aging has been described, almost nothing is known about how metabolite handling regulates ISC renewal and maintains stemness. We investigated how mitochondrial metabolism of glucose and fatty acid-derived carbons, regulated by the gatekeeper, pyruvate dehydrogenase (PDH) rescues ISC stemness in older and geriatric mice and humans. MethodsProximal small intestinal organoids (enteroids) generated from pinch biopsy specimens obtained from young (21-25y) and older individuals (64-75y), and GFP-sorted single ISCs from Lgr5-EGFP mice (2-24 months) were used to examine hallmarks of ISC stemness. Mitochondrial morphology was evaluated using transmission electron microscopy. Mitochondrial oxygen consumption rate (OCR), ATP (mitoATP), and glycolytic ATP production were measured in the presence of full and single metabolic substrates (pyruvate, glutamate, and fatty acids) in whole cell and isolated mitochondria using the high throughput Seahorse XF technology. Carbon flux through TCA cycle was determined by 13C6-glucose tracing and measuring 13C enrichment in TCA cycle intermediates using liquid chromatography mass spectrometry. ResultsAge induced decline in ISC stemness is driven by a dramatic decrease in PDH activity that shuttles pyruvate away from the TCA cycle. Restoring PDH activity by inhibition of pyruvate dehydrogenase kinase 4 (PDK4) drives glucose-derived carbon entry into TCA cycle and subsequently increases mitochondrial OCR and mitoATP, collectively rescuing the decline in stemness in aging ISCs. The observed shift in fuel preference from fatty acids to glucose is unaltered by PDK4 inhibition. ConclusionPDH upregulation rescues age-induced decline in ISC stemness in humans and mice via directing glucose derived carbons to TCA cycle and increasing mitoATP production.

cell biology↗

Supplier-origin gut microbiomes affect host body weight and select autism-related behaviors

Autism spectrum disorders (ASD) are complex human neurodiversities increasing in prevalence within the human population. In search of therapeutics to improve quality-of-life for ASD patients, the gut microbiome (GM) has become a promising target as a growing body of work supports roles for the complex community of microorganisms in influencing host behavior via the gut-brain-axis. However, whether naturally-occurring microbial diversity within the host GM affects these behaviors is often overlooked. Here we applied a model of population-level differences in the GM to a classic ASD model - the BTBR T+ Itpr3tf/J mouse - to assess how complex GMs affect host behavior. Leveraging the naturally occurring differences between supplier-origin GMs, our data demonstrate that differing, complex GMs selectively effect host ASD-related behavior - especially neonatal ultrasonic communication - and reveal a male-specific effect on behavior not typically observed in this strain. We then identified that the body weight of BTBR mice is influenced by the postnatal GM which was potentially mediated by microbiome-dependent effects on energy harvest in the gut. These data provide insight into how variability within the GM affects host behavior and growth, thereby emphasizing the need to incorporate naturally occurring diversity within the host GM as an experimental factor in biomedical research.

microbiology↗