Biology subjects

Friedman, M. A.

Publications and source records attributed to Friedman, M. A..

5 recordsLinked to original sources

bioRxiv2023

Genetic Diversity Modulates The Physical And Transcriptomic Response Of Skeletal Muscle To Simulated Microgravity

Developments in long-term space exploration necessitate advancements in countermeasures against microgravity-induced skeletal muscle loss. Astronaut data shows considerable variation in muscle loss in response to microgravity. Previous experiments suggest that genetic background influences the skeletal muscle response to unloading, but no in-depth analysis of genetic expression was performed. Here, we placed eight inbred founder strains of the diversity outbred mice (129S1/SvImJ, A/J, C57BL/6J, CAST/EiJ, NOD/ShiLtJ, NZO/HILtJ, PWK/PhJ, and WSB/EiJ) in simulated microgravity (SM) via hindlimb unloading for three weeks. Body weight, muscle morphology, muscle strength, protein synthesis marker expression, and RNA expression were collected. A/J and CAST/EiJ mice were most susceptible to SM-induced muscle loss, whereas NOD/ShiLtJ mice were the most protected. In response to SM, A/J and CAST/EiJ mice experienced reductions in body weight, muscle mass, muscle volume, and muscle cross-sectional area. A/J mice had the highest number of differentially expressed genes (68) and associated gene ontologies (328). Downregulation of immunological gene ontologies and genes encoding anabolic immune factors suggest that immune dysregulation contributes to the response of A/J mice to SM. Several muscle properties showed significant interactions between SM and mouse strain and a high degree of heritability. These data imply that genetic background plays a role in the degree of muscle loss in SM and that more individualized programs should be developed for astronauts to protect their skeletal muscles against microgravity on long term missions.

bioengineering↗

bioRxiv2023

Connexin 43 and Cell Culture Substrate Differentially Regulate OCY454 Osteocytic Differentiation and Signaling to Primary Bone Cells

Connexin 43 (Cx43), the predominate gap junction protein in bone, is essential for intercellular communication and skeletal homeostasis. Previous work suggests osteocyte-specific deletion of Cx43 leads to increased bone formation and resorption, however the cell-autonomous role of osteocytic Cx43 in promoting increased bone remodeling is unknown. Recent studies using 3D culture substrates in OCY454 cells suggest 3D cultures may offer increased bone remodeling factor expression and secretion, such as sclerostin and RANKL. In this study, we compared culturing OCY454 osteocytes on 3D Alvetex scaffolds to traditional 2D tissue culture, both with (WT) and without Cx43 (Cx43 KO). Conditioned media from OCY454 cell cultures was used to determine soluble signaling to differentiate primary bone marrow stromal cells into osteoblasts and osteoclasts. OCY454 cells cultured on 3D portrayed a mature osteocytic phenotype, relative to cells on 2D, shown by increased osteocytic gene expression and reduced cell proliferation. In contrast, OCY454 differentiation based on these same markers was not affected by Cx43 deficiency in 3D. Interestingly, increased sclerostin secretion was found in 3D cultured WT cells compared to Cx43 KO cells. Conditioned media from Cx43 KO cells promoted increased osteoblastogenesis and increased osteoclastogenesis, with maximal effects from 3D cultured Cx43 KO cells. These results suggest Cx43 deficiency promotes increased bone remodeling in a cell autonomous manner with minimal changes in osteocyte differentiation. Finally, 3D cultures appear better suited to study mechanisms from Cx43-deficient OCY454 osteocytes in vitro due to their ability to promote osteocyte differentiation, limit proliferation, and increase bone remodeling factor secretion. New and Noteworthy3D cell culture of OCY454 cells promoted increased differentiation compared to traditional 2D culture. While Cx43 deficiency did not affect OCY454 differentiation, it resulted in increased signaling, promoting osteoblastogenesis and osteoclastogenesis. Our results suggest Cx43 deficiency promotes increased bone remodeling in a cell autonomous manner with minimal changes in osteocyte differentiation. Also, 3D cultures appear better suited to study mechanisms in Cx43-deficient OCY454 osteocytes.

cancer biology↗

bioRxiv2022

Structural and functional analysis of the human Cone-rod homeobox transcription factor

The cone-rod homeobox (CRX) protein is a critical K50 homeodomain transcription factor responsible for the differentiation and maintenance of photoreceptor neurons in the vertebrate retina. Mutant alleles in the human gene encoding CRX result in a variety of distinct blinding retinopathies, including retinitis pigmentosa, cone-rod dystrophy, and Leber congenital amaurosis. Despite the success of using in vitro biochemistry, animal models, and genomics approaches to study this clinically relevant transcription factor over the past 24 years since its initial characterization, there are no high-resolution structures in the published literature for the CRX protein. In this study, we use bioinformatic approaches and small-angle x-ray scattering (SAXS) structural analysis to further understand the biochemical complexity of the human CRX homeodomain (CRX-HD). We find that the CRX-HD is a compact, globular monomer in solution that can specifically bind functional cis-regulatory elements encoded upstream of retina -specific genes. This study presents the first structural analysis of CRX, paving the way for a new approach to studying the biochemistry of this protein and its disease-causing mutant protein variants.

biochemistry↗

bioRxiv2020

Calcium- and Phosphorus-Supplemented Diet Increases Bone Volume After Four Weeks of High-Speed Treadmill Exercise in Adult Mice

Exercise has long-lasting benefits to bone health. Bone mass and the ability to exercise both decline with age, making it ideal to exercise earlier in life and to maximize gains in bone mass. Increasing strain on bone and frequency of loading during exercise can increase bone formation rate and cross-sectional area. Combining a short-term exercise program with a calcium- and phosphorus-supplemented diet increases cortical bone tissue mineral content (TMC) and area more than exercise alone in adult mice. It was hypothesized that combining high-speed running with a mineral-supplemented diet would lead to greater cortical TMC and area than high-speed running on a standard diet and low-speed running on a supplemented diet after 4 weeks. Male, 15-week old mice were assigned to 7 groups - a baseline group, non-exercised groups fed a control or supplemented diet, low-speed exercised groups fed a control or supplemented diet, and high-speed exercised groups fed a control or supplemented diet. Exercise consisted of 4 weeks of daily treadmill running for 20 min/day at 12 m/min or 20 m/min for low- and high-speed exercise, respectively. High-speed exercised mice had significantly lower body weight and lower tibial length after 4 weeks. Cortical TMC and area were significantly higher in high-speed exercised mice on the supplemented diet than high-speed exercised mice on the control diet. Trabecular bone volume (BV) and bone density were significantly higher in all groups on the supplemented diet than groups on the control diet, regardless of exercise. For mice on the control diet, non-exercised mice had significantly lower trabecular BV than baseline, while both speeds of exercise prevented this decline. There were few effects of exercise or diet on mechanical properties. For mice on the control diet, exercise significantly decreased serum PINP/CTX ratio on day 9 which may be preventing exercise from increasing bone mass or strength after only 4 weeks. For non-exercised mice on the supplemented diet, the serum PINP/CTX ratio on day 30 was significantly greater than for exercised mice, suggesting the supplemented diet may also lead to significantly greater bone mass in non-exercised mice if these interventions were extended beyond 4 weeks. Increasing treadmill speed can lower body weight while maintaining cortical and trabecular bone mass. A mineral-supplemented diet increases cortical and trabecular bone mass with high-speed exercise.

physiology↗

bioRxiv2020

Genetic Variability affects the Skeletal Response to Unloading

Mechanical unloading decreases bone volume and strength. In humans and mice, bone mineral density is highly heritable, and in mice the response to changes in loading varies with genetic background. Thus, genetic variability may affect the response of bone to unloading. As a first step to identify genes involved in bone response to unloading, we evaluated the effects of unloading in eight inbred mouse strains: C57BL/6J, PWK/PhJ, WSB/EiJ, A/J, 129S1/SvImJ, NOD/ShiLtJ, NZO/HlLtJ, and CAST/EiJ. C57BL/6J and NOD/ShiLtJ mice had the greatest unloading induced loss of diaphyseal cortical bone volume and strength. NZO/HlLtJ mice had the greatest metaphyseal trabecular bone loss, and C57BL/6J, WSB/EiJ, NOD/ShiLtJ, and CAST/EiJ mice had the greatest epiphyseal trabecular loss. Bone loss in the epiphyses displayed the highest heritability. With immobilization, mineral:matrix was reduced, and carbonate:phosphate and crystallinity were increased. A/J mice displayed the greatest unloading induced loss of mineral:matrix. Changes in gene expression in response to unloading were greatest in NOD/ShiLtJ and CAST/EiJ mice. The most upregulated genes in response to unloading were associated with increased collagen synthesis and extracellular matrix formation. Our results demonstrate a strong differential response to unloading as a function of strain. Diversity outbred (DO) mice are a high-resolution mapping population derived from these eight inbred founder strains. These results suggest DO mice will be highly suited for examining the genetic basis of the skeletal response to unloading.

genetics↗