26 results on '"Butterfield, Natalie C."'
Search Results
2. Osteocyte transcriptome mapping identifies a molecular landscape controlling skeletal homeostasis and susceptibility to skeletal disease.
- Author
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Youlten SE, Kemp JP, Logan JG, Ghirardello EJ, Sergio CM, Dack MRG, Guilfoyle SE, Leitch VD, Butterfield NC, Komla-Ebri D, Chai RC, Corr AP, Smith JT, Mohanty ST, Morris JA, McDonald MM, Quinn JMW, McGlade AR, Bartonicek N, Jansson M, Hatzikotoulas K, Irving MD, Beleza-Meireles A, Rivadeneira F, Duncan E, Richards JB, Adams DJ, Lelliott CJ, Brink R, Phan TG, Eisman JA, Evans DM, Zeggini E, Baldock PA, Bassett JHD, Williams GR, and Croucher PI
- Subjects
- Age Factors, Animals, Bone Diseases metabolism, Bone and Bones metabolism, Computational Biology, Female, Humans, Male, Mice, Mice, Knockout, Osteocytes cytology, Osteoporosis genetics, Sequence Analysis, RNA, Sex Factors, Bone Diseases genetics, Homeostasis, Osteocytes metabolism, Transcriptome
- Abstract
Osteocytes are master regulators of the skeleton. We mapped the transcriptome of osteocytes from different skeletal sites, across age and sexes in mice to reveal genes and molecular programs that control this complex cellular-network. We define an osteocyte transcriptome signature of 1239 genes that distinguishes osteocytes from other cells. 77% have no previously known role in the skeleton and are enriched for genes regulating neuronal network formation, suggesting this programme is important in osteocyte communication. We evaluated 19 skeletal parameters in 733 knockout mouse lines and reveal 26 osteocyte transcriptome signature genes that control bone structure and function. We showed osteocyte transcriptome signature genes are enriched for human orthologs that cause monogenic skeletal disorders (P = 2.4 × 10
-22 ) and are associated with the polygenic diseases osteoporosis (P = 1.8 × 10-13 ) and osteoarthritis (P = 1.6 × 10-7 ). Thus, we reveal the molecular landscape that regulates osteocyte network formation and function and establish the importance of osteocytes in human skeletal disease.- Published
- 2021
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3. Osteoclasts recycle via osteomorphs during RANKL-stimulated bone resorption.
- Author
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McDonald MM, Khoo WH, Ng PY, Xiao Y, Zamerli J, Thatcher P, Kyaw W, Pathmanandavel K, Grootveld AK, Moran I, Butt D, Nguyen A, Corr A, Warren S, Biro M, Butterfield NC, Guilfoyle SE, Komla-Ebri D, Dack MRG, Dewhurst HF, Logan JG, Li Y, Mohanty ST, Byrne N, Terry RL, Simic MK, Chai R, Quinn JMW, Youlten SE, Pettitt JA, Abi-Hanna D, Jain R, Weninger W, Lundberg M, Sun S, Ebetino FH, Timpson P, Lee WM, Baldock PA, Rogers MJ, Brink R, Williams GR, Bassett JHD, Kemp JP, Pavlos NJ, Croucher PI, and Phan TG
- Published
- 2021
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4. Osteoclasts recycle via osteomorphs during RANKL-stimulated bone resorption.
- Author
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McDonald MM, Khoo WH, Ng PY, Xiao Y, Zamerli J, Thatcher P, Kyaw W, Pathmanandavel K, Grootveld AK, Moran I, Butt D, Nguyen A, Corr A, Warren S, Biro M, Butterfield NC, Guilfoyle SE, Komla-Ebri D, Dack MRG, Dewhurst HF, Logan JG, Li Y, Mohanty ST, Byrne N, Terry RL, Simic MK, Chai R, Quinn JMW, Youlten SE, Pettitt JA, Abi-Hanna D, Jain R, Weninger W, Lundberg M, Sun S, Ebetino FH, Timpson P, Lee WM, Baldock PA, Rogers MJ, Brink R, Williams GR, Bassett JHD, Kemp JP, Pavlos NJ, Croucher PI, and Phan TG
- Subjects
- Animals, Apoptosis, Bone Resorption metabolism, Cell Fusion, Cells, Cultured, Humans, Macrophages cytology, Mice, Osteochondrodysplasias drug therapy, Osteochondrodysplasias genetics, Osteochondrodysplasias metabolism, Osteochondrodysplasias pathology, Osteoclasts metabolism, Signal Transduction, Bone Resorption pathology, Osteoclasts pathology, RANK Ligand metabolism
- Abstract
Osteoclasts are large multinucleated bone-resorbing cells formed by the fusion of monocyte/macrophage-derived precursors that are thought to undergo apoptosis once resorption is complete. Here, by intravital imaging, we reveal that RANKL-stimulated osteoclasts have an alternative cell fate in which they fission into daughter cells called osteomorphs. Inhibiting RANKL blocked this cellular recycling and resulted in osteomorph accumulation. Single-cell RNA sequencing showed that osteomorphs are transcriptionally distinct from osteoclasts and macrophages and express a number of non-canonical osteoclast genes that are associated with structural and functional bone phenotypes when deleted in mice. Furthermore, genetic variation in human orthologs of osteomorph genes causes monogenic skeletal disorders and associates with bone mineral density, a polygenetic skeletal trait. Thus, osteoclasts recycle via osteomorphs, a cell type involved in the regulation of bone resorption that may be targeted for the treatment of skeletal diseases., Competing Interests: Declaration of interests F.H.E. and S.S. are employees and shareholders of BioVinc LLC, who has licensed patents for fluorescent probe compositions used in this work. P.I.C. has grant funding from Amgen. Other authors have no competing financial interests., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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5. A molecular quantitative trait locus map for osteoarthritis.
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Steinberg J, Southam L, Roumeliotis TI, Clark MJ, Jayasuriya RL, Swift D, Shah KM, Butterfield NC, Brooks RA, McCaskie AW, Bassett JHD, Williams GR, Choudhary JS, Wilkinson JM, and Zeggini E
- Subjects
- Gene Expression Profiling, Gene Expression Regulation, Genome-Wide Association Study, Humans, Phenotype, Transcription Factors genetics, Transcriptome, Genetic Predisposition to Disease genetics, Osteoarthritis genetics, Quantitative Trait Loci genetics
- Abstract
Osteoarthritis causes pain and functional disability for over 500 million people worldwide. To develop disease-stratifying tools and modifying therapies, we need a better understanding of the molecular basis of the disease in relevant tissue and cell types. Here, we study primary cartilage and synovium from 115 patients with osteoarthritis to construct a deep molecular signature map of the disease. By integrating genetics with transcriptomics and proteomics, we discover molecular trait loci in each tissue type and omics level, identify likely effector genes for osteoarthritis-associated genetic signals and highlight high-value targets for drug development and repurposing. These findings provide insights into disease aetiopathology, and offer translational opportunities in response to the global clinical challenge of osteoarthritis.
- Published
- 2021
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6. Accelerating functional gene discovery in osteoarthritis.
- Author
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Butterfield NC, Curry KF, Steinberg J, Dewhurst H, Komla-Ebri D, Mannan NS, Adoum AT, Leitch VD, Logan JG, Waung JA, Ghirardello E, Southam L, Youlten SE, Wilkinson JM, McAninch EA, Vancollie VE, Kussy F, White JK, Lelliott CJ, Adams DJ, Jacques R, Bianco AC, Boyde A, Zeggini E, Croucher PI, Williams GR, and Bassett JHD
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- Animals, Bone and Bones pathology, CRISPR-Cas Systems, Cartilage pathology, Clustered Regularly Interspaced Short Palindromic Repeats, Disease Models, Animal, Drug Discovery, Gene Editing, Gonadotropin-Releasing Hormone genetics, Iodide Peroxidase, Mice, Mice, Knockout, Osteoarthritis pathology, Osteoarthritis surgery, Paired Box Transcription Factors genetics, Phenotype, Iodothyronine Deiodinase Type II, Genetic Association Studies, Genetic Predisposition to Disease genetics, Osteoarthritis genetics
- Abstract
Osteoarthritis causes debilitating pain and disability, resulting in a considerable socioeconomic burden, yet no drugs are available that prevent disease onset or progression. Here, we develop, validate and use rapid-throughput imaging techniques to identify abnormal joint phenotypes in randomly selected mutant mice generated by the International Knockout Mouse Consortium. We identify 14 genes with functional involvement in osteoarthritis pathogenesis, including the homeobox gene Pitx1, and functionally characterize 6 candidate human osteoarthritis genes in mouse models. We demonstrate sensitivity of the methods by identifying age-related degenerative joint damage in wild-type mice. Finally, we phenotype previously generated mutant mice with an osteoarthritis-associated polymorphism in the Dio2 gene by CRISPR/Cas9 genome editing and demonstrate a protective role in disease onset with public health implications. We hope this expanding resource of mutant mice will accelerate functional gene discovery in osteoarthritis and offer drug discovery opportunities for this common, incapacitating chronic disease.
- Published
- 2021
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7. Response to Letter to the Editor: "IGSF1 Deficiency Results in Human and Murine Somatotrope Neurosecretory Hyperfunction".
- Author
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Joustra SD, Roelfsema F, van Trotsenburg ASP, Schneider HJ, Kosilek RP, Kroon HM, Logan JG, Butterfield NC, Zhou X, Toufaily C, Bak B, Turgeon MO, Brûlé E, Steyn FJ, Gurnell M, Koulouri O, Le Tissier P, Fontanaud P, Bassett JHD, Williams GR, Oostdijk W, Wit JM, Pereira AM, Biermasz NR, Bernard DJ, and Schoenmakers N
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- Animals, Humans, Mice, Immunoglobulins, Membrane Proteins
- Published
- 2020
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8. IGSF1 Deficiency Results in Human and Murine Somatotrope Neurosecretory Hyperfunction.
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Joustra SD, Roelfsema F, van Trotsenburg ASP, Schneider HJ, Kosilek RP, Kroon HM, Logan JG, Butterfield NC, Zhou X, Toufaily C, Bak B, Turgeon MO, Brûlé E, Steyn FJ, Gurnell M, Koulouri O, Le Tissier P, Fontanaud P, Duncan Bassett JH, Williams GR, Oostdijk W, Wit JM, Pereira AM, Biermasz NR, Bernard DJ, and Schoenmakers N
- Subjects
- Adult, Aged, Aged, 80 and over, Animals, Growth Hormone biosynthesis, Humans, Immunoglobulins deficiency, Insulin-Like Growth Factor I analysis, Intercellular Signaling Peptides and Proteins deficiency, Male, Membrane Proteins deficiency, Mice, Middle Aged, Immunoglobulins physiology, Intercellular Signaling Peptides and Proteins physiology, Membrane Proteins physiology, Neurosecretion physiology, Somatotrophs physiology
- Abstract
Context: The X-linked immunoglobulin superfamily, member 1 (IGSF1), gene is highly expressed in the hypothalamus and in pituitary cells of the POU1F1 lineage. Human loss-of-function mutations in IGSF1 cause central hypothyroidism, hypoprolactinemia, and macroorchidism. Additionally, most affected adults exhibit higher than average IGF-1 levels and anecdotal reports describe acromegaloid features in older subjects. However, somatotrope function has not yet been formally evaluated in this condition., Objective: We aimed to evaluate the role of IGSF1 in human and murine somatotrope function., Patients, Design, and Setting: We evaluated 21 adult males harboring hemizygous IGSF1 loss-of-function mutations for features of GH excess, in an academic clinical setting., Main Outcome Measures: We compared biochemical and tissue markers of GH excess in patients and controls, including 24-hour GH profile studies in 7 patients. Parallel studies were undertaken in male Igsf1-deficient mice and wild-type littermates., Results: IGSF1-deficient adult male patients demonstrated acromegaloid facial features with increased head circumference as well as increased finger soft-tissue thickness. Median serum IGF-1 concentrations were elevated, and 24-hour GH profile studies confirmed 2- to 3-fold increased median basal, pulsatile, and total GH secretion. Male Igsf1-deficient mice also demonstrated features of GH excess with increased lean mass, organ size, and skeletal dimensions and elevated mean circulating IGF-1 and pituitary GH levels., Conclusions: We demonstrate somatotrope neurosecretory hyperfunction in IGSF1-deficient humans and mice. These observations define a hitherto uncharacterized role for IGSF1 in somatotropes and indicate that patients with IGSF1 mutations should be evaluated for long-term consequences of increased GH exposure., (© Endocrine Society 2019.)
- Published
- 2020
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9. PYY is a negative regulator of bone mass and strength.
- Author
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Leitch VD, Brassill MJ, Rahman S, Butterfield NC, Ma P, Logan JG, Boyde A, Evans H, Croucher PI, Batterham RL, Williams GR, and Bassett JHD
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- Animals, Bone Density, Bone Development, Bone Resorption pathology, Bone Resorption physiopathology, Calcification, Physiologic, Cortical Bone blood supply, Cortical Bone ultrastructure, Female, Femur diagnostic imaging, Femur ultrastructure, Male, Mice, Inbred C57BL, Mice, Knockout, Organ Size, Osteoclasts pathology, Porosity, Bone and Bones anatomy & histology, Bone and Bones physiology, Peptide YY metabolism
- Abstract
Objective: Bone loss in anorexia nervosa and following bariatric surgery is associated with an elevated circulating concentration of the gastrointestinal, anorexigenic hormone, peptide YY (PYY). Selective deletion of the PYY receptor Y1R in osteoblasts or Y2R in the hypothalamus results in high bone mass, but deletion of PYY in mice has resulted in conflicting skeletal phenotypes leading to uncertainty regarding its role in the regulation of bone mass. As PYY analogs are under development for treatment of obesity, we aimed to clarify the relationship between PYY and bone mass., Methods: The skeletal phenotype of Pyy knockout (KO) mice was investigated during growth (postnatal day P14) and adulthood (P70 and P186) using X-ray microradiography, micro-CT, back-scattered electron scanning electron microscopy (BSE-SEM), histomorphometry and biomechanical testing., Results: Bones from juvenile and Pyy KO mice were longer (P < 0.001), with decreased bone mineral content (P < 0.001). Whereas, bones from adult Pyy KO mice had increased bone mineral content (P < 0.05) with increased mineralisation of both cortical (P < 0.001) and trabecular (P < 0.001) compartments. Long bones from adult Pyy KO mice were stronger (maximum load P < 0.001), with increased stiffness (P < 0.01) and toughness (P < 0.05) compared to wild-type (WT) control mice despite increased cortical vascularity and porosity (P < 0.001). The increased bone mass and strength in Pyy KO mice resulted from increases in trabecular (P < 0.01) and cortical bone formation (P < 0.05)., Conclusions: These findings demonstrate that PYY acts as a negative regulator of osteoblastic bone formation, implicating increased PYY levels in the pathogenesis of bone loss during anorexia or following bariatric surgery., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2019
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10. Slc20a2, Encoding the Phosphate Transporter PiT2, Is an Important Genetic Determinant of Bone Quality and Strength.
- Author
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Beck-Cormier S, Lelliott CJ, Logan JG, Lafont DT, Merametdjian L, Leitch VD, Butterfield NC, Protheroe HJ, Croucher PI, Baldock PA, Gaultier-Lintia A, Maugars Y, Nicolas G, Banse C, Normant S, Magne N, Gérardin E, Bon N, Sourice S, Guicheux J, Beck L, Williams GR, and Bassett JHD
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- Animals, Animals, Newborn, Bone Development, Bone Resorption physiopathology, Bone and Bones diagnostic imaging, Calcification, Physiologic, Calcinosis diagnostic imaging, Calcinosis genetics, Cells, Cultured, Chondrocytes metabolism, Humans, Incisor ultrastructure, Mice, Inbred C57BL, Mice, Knockout, Osteoblasts metabolism, Phenotype, Skull diagnostic imaging, Sodium-Phosphate Cotransporter Proteins, Type III deficiency, Tooth growth & development, X-Ray Microtomography, Bone and Bones physiology, Sodium-Phosphate Cotransporter Proteins, Type III genetics
- Abstract
Osteoporosis is characterized by low bone mineral density (BMD) and fragility fracture and affects over 200 million people worldwide. Bone quality describes the material properties that contribute to strength independently of BMD, and its quantitative analysis is a major priority in osteoporosis research. Tissue mineralization is a fundamental process requiring calcium and phosphate transporters. Here we identify impaired bone quality and strength in Slc20a2
-/- mice lacking the phosphate transporter SLC20A2. Juveniles had abnormal endochondral and intramembranous ossification, decreased mineral accrual, and short stature. Adults exhibited only small reductions in bone mass and mineralization but a profound impairment of bone strength. Bone quality was severely impaired in Slc20a2-/- mice: yield load (-2.3 SD), maximum load (-1.7 SD), and stiffness (-2.7 SD) were all below values predicted from their bone mineral content as determined in a cohort of 320 wild-type controls. These studies identify Slc20a2 as a physiological regulator of tissue mineralization and highlight its critical role in the determination of bone quality and strength. © 2019 The Authors. Journal of Bone and Mineral Research Published by Wiley Periodicals Inc., (© 2019 The Authors. Journal of Bone and Mineral Research Published by Wiley Periodicals Inc.)- Published
- 2019
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11. Genetic and Pharmacological Targeting of Transcriptional Repression in Resistance to Thyroid Hormone Alpha.
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Freudenthal B, Shetty S, Butterfield NC, Logan JG, Han CR, Zhu X, Astapova I, Hollenberg AN, Cheng SY, Bassett JHD, and Williams GR
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- Animals, Bone Density, Bone Development, Calcification, Physiologic, Mice, Mice, Inbred C57BL, Thyroid Hormone Resistance Syndrome drug therapy, Thyroxine pharmacology, Vorinostat pharmacology, Nuclear Receptor Co-Repressor 1 physiology, Thyroid Hormone Receptors alpha physiology, Thyroid Hormone Resistance Syndrome genetics
- Abstract
Background: Thyroid hormones act in bone and cartilage via thyroid hormone receptor alpha (TRα). In the absence of triiodothyronine (T3), TRα interacts with co-repressors, including nuclear receptor co-repressor-1 (NCoR1), which recruit histone deacetylases (HDACs) and mediate transcriptional repression. Dominant-negative mutations of TRα cause resistance to thyroid hormone alpha (RTHα; OMIM 614450), characterized by excessive repression of T3 target genes leading to delayed skeletal development, growth retardation, and bone dysplasia. Treatment with thyroxine has been of limited benefit, even in mildly affected individuals, and there is a need for new therapeutic strategies. It was hypothesized that (i) the skeletal manifestations of RTHα are mediated by the persistent TRα/NCoR1/HDAC repressor complex containing mutant TRα, and (ii) treatment with the HDAC inhibitor suberoylanilide hydroxamic acid (SAHA) would ameliorate these manifestations. Methods: The skeletal phenotypes of (i) Thra1
PV/+ mice, a well characterized model of RTHα; (ii) Ncor1ΔID/ΔID mice, which express an NCoR1 mutant that fails to interact with TRα; and (iii) Thra1PV/+ Ncor1ΔID/ΔID double-mutant adult mice were determined. Wild-type, Thra1PV/+ , Ncor1ΔID/ΔID , and Thra1PV/+ Ncor1ΔID/ΔID double-mutant mice were also treated with SAHA to determine whether HDAC inhibition results in amelioration of skeletal abnormalities. Results: Thra1PV/+ mice had a severe skeletal dysplasia, characterized by short stature, abnormal bone morphology, and increased bone mineral content. Despite normal bone length, Ncor1ΔID/ΔID mice displayed increased cortical bone mass, mineralization, and strength. Thra1PV/+ Ncor1ΔID/ΔID double-mutant mice displayed only a small improvement of skeletal abnormalities compared to Thra1PV/+ mice. Treatment with SAHA to inhibit histone deacetylation had no beneficial or detrimental effects on bone structure, mineralization, or strength in wild-type or mutant mice. Conclusions: These studies indicate treatment with SAHA is unlikely to improve the skeletal manifestations of RTHα. Nevertheless, the findings (i) confirm that TRα1 has a critical role in the regulation of skeletal development and adult bone mass, (ii) suggest a physiological role for alternative co-repressors that interact with TR in skeletal cells, and (iii) demonstrate a novel role for NCoR1 in the regulation of adult bone mass and strength.- Published
- 2019
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12. Author Correction: An atlas of genetic influences on osteoporosis in humans and mice.
- Author
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Morris JA, Kemp JP, Youlten SE, Laurent L, Logan JG, Chai RC, Vulpescu NA, Forgetta V, Kleinman A, Mohanty ST, Sergio CM, Quinn J, Nguyen-Yamamoto L, Luco AL, Vijay J, Simon MM, Pramatarova A, Medina-Gomez C, Trajanoska K, Ghirardello EJ, Butterfield NC, Curry KF, Leitch VD, Sparkes PC, Adoum AT, Mannan NS, Komla-Ebri DSK, Pollard AS, Dewhurst HF, Hassall TAD, Beltejar MG, Adams DJ, Vaillancourt SM, Kaptoge S, Baldock P, Cooper C, Reeve J, Ntzani EE, Evangelou E, Ohlsson C, Karasik D, Rivadeneira F, Kiel DP, Tobias JH, Gregson CL, Harvey NC, Grundberg E, Goltzman D, Adams DJ, Lelliott CJ, Hinds DA, Ackert-Bicknell CL, Hsu YH, Maurano MT, Croucher PI, Williams GR, Bassett JHD, Evans DM, and Richards JB
- Abstract
In the version of this article initially published, in Fig. 5a, the data in the right column of 'DAAM2 gRNA1' were incorrectly plotted as circles indicating 'untreated' rather than as squares indicating 'treated'. The error has been corrected in the HTML and PDF versions of the article.
- Published
- 2019
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13. An atlas of genetic influences on osteoporosis in humans and mice.
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Morris JA, Kemp JP, Youlten SE, Laurent L, Logan JG, Chai RC, Vulpescu NA, Forgetta V, Kleinman A, Mohanty ST, Sergio CM, Quinn J, Nguyen-Yamamoto L, Luco AL, Vijay J, Simon MM, Pramatarova A, Medina-Gomez C, Trajanoska K, Ghirardello EJ, Butterfield NC, Curry KF, Leitch VD, Sparkes PC, Adoum AT, Mannan NS, Komla-Ebri DSK, Pollard AS, Dewhurst HF, Hassall TAD, Beltejar MG, Adams DJ, Vaillancourt SM, Kaptoge S, Baldock P, Cooper C, Reeve J, Ntzani EE, Evangelou E, Ohlsson C, Karasik D, Rivadeneira F, Kiel DP, Tobias JH, Gregson CL, Harvey NC, Grundberg E, Goltzman D, Adams DJ, Lelliott CJ, Hinds DA, Ackert-Bicknell CL, Hsu YH, Maurano MT, Croucher PI, Williams GR, Bassett JHD, Evans DM, and Richards JB
- Subjects
- Adult, Aged, Animals, Female, Fractures, Bone genetics, Genome-Wide Association Study methods, Humans, Male, Mice, Mice, Knockout, Middle Aged, Phenotype, Polymorphism, Single Nucleotide genetics, Bone Density genetics, Genetic Predisposition to Disease genetics, Osteoporosis genetics
- Abstract
Osteoporosis is a common aging-related disease diagnosed primarily using bone mineral density (BMD). We assessed genetic determinants of BMD as estimated by heel quantitative ultrasound in 426,824 individuals, identifying 518 genome-wide significant loci (301 novel), explaining 20% of its variance. We identified 13 bone fracture loci, all associated with estimated BMD (eBMD), in ~1.2 million individuals. We then identified target genes enriched for genes known to influence bone density and strength (maximum odds ratio (OR) = 58, P = 1 × 10
-75 ) from cell-specific features, including chromatin conformation and accessible chromatin sites. We next performed rapid-throughput skeletal phenotyping of 126 knockout mice with disruptions in predicted target genes and found an increased abnormal skeletal phenotype frequency compared to 526 unselected lines (P < 0.0001). In-depth analysis of one gene, DAAM2, showed a disproportionate decrease in bone strength relative to mineralization. This genetic atlas provides evidence linking associated SNPs to causal genes, offers new insight into osteoporosis pathophysiology, and highlights opportunities for drug development.- Published
- 2019
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14. Type 2 deiodinase polymorphism causes ER stress and hypothyroidism in the brain.
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Jo S, Fonseca TL, Bocco BMLC, Fernandes GW, McAninch EA, Bolin AP, Da Conceição RR, Werneck-de-Castro JP, Ignacio DL, Egri P, Németh D, Fekete C, Bernardi MM, Leitch VD, Mannan NS, Curry KF, Butterfield NC, Bassett JHD, Williams GR, Gereben B, Ribeiro MO, and Bianco AC
- Subjects
- Amino Acid Substitution, Animals, Endoplasmic Reticulum enzymology, Endoplasmic Reticulum genetics, Golgi Apparatus enzymology, Golgi Apparatus genetics, HEK293 Cells, Humans, Mice, Mice, Transgenic, Mutation, Missense, Thyroxine therapeutic use, Triiodothyronine therapeutic use, Iodothyronine Deiodinase Type II, Brain enzymology, Brain pathology, Endoplasmic Reticulum Stress, Hypothyroidism drug therapy, Hypothyroidism enzymology, Hypothyroidism genetics, Hypothyroidism pathology, Iodide Peroxidase genetics, Iodide Peroxidase metabolism, Polymorphism, Genetic, Unfolded Protein Response
- Abstract
Levothyroxine (LT4) is a form of thyroid hormone used to treat hypothyroidism. In the brain, T4 is converted to the active form T3 by type 2 deiodinase (D2). Thus, it is intriguing that carriers of the Thr92Ala polymorphism in the D2 gene (DIO2) exhibit clinical improvement when liothyronine (LT3) is added to LT4 therapy. Here, we report that D2 is a cargo protein in ER Golgi intermediary compartment (ERGIC) vesicles, recycling between ER and Golgi. The Thr92-to-Ala substitution (Ala92-D2) caused ER stress and activated the unfolded protein response (UPR). Ala92-D2 accumulated in the trans-Golgi and generated less T3, which was restored by eliminating ER stress with the chemical chaperone 4-phenyl butyric acid (4-PBA). An Ala92-Dio2 polymorphism-carrying mouse exhibited UPR and hypothyroidism in distinct brain areas. The mouse refrained from physical activity, slept more, and required additional time to memorize objects. Enhancing T3 signaling in the brain with LT3 improved cognition, whereas restoring proteostasis with 4-PBA eliminated the Ala92-Dio2 phenotype. In contrast, primary hypothyroidism intensified the Ala92-Dio2 phenotype, with only partial response to LT4 therapy. Disruption of cellular proteostasis and reduced Ala92-D2 activity may explain the failure of LT4 therapy in carriers of Thr92Ala-DIO2.
- Published
- 2019
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15. Quantitative X-Ray Imaging of Mouse Bone by Faxitron.
- Author
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Butterfield NC, Logan JG, Waung J, Williams GR, and Bassett JHD
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- Animals, Bone and Bones physiology, Image Processing, Computer-Assisted instrumentation, Mice, Microradiography instrumentation, Models, Animal, Software, Bone Density, Bone and Bones diagnostic imaging, Image Processing, Computer-Assisted methods, Microradiography methods
- Abstract
This chapter describes the use of point projection digital microradiography for rapid imaging and quantitation of bone mineral content in mice.
- Published
- 2019
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16. Identification of 153 new loci associated with heel bone mineral density and functional involvement of GPC6 in osteoporosis.
- Author
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Kemp JP, Morris JA, Medina-Gomez C, Forgetta V, Warrington NM, Youlten SE, Zheng J, Gregson CL, Grundberg E, Trajanoska K, Logan JG, Pollard AS, Sparkes PC, Ghirardello EJ, Allen R, Leitch VD, Butterfield NC, Komla-Ebri D, Adoum AT, Curry KF, White JK, Kussy F, Greenlaw KM, Xu C, Harvey NC, Cooper C, Adams DJ, Greenwood CMT, Maurano MT, Kaptoge S, Rivadeneira F, Tobias JH, Croucher PI, Ackert-Bicknell CL, Bassett JHD, Williams GR, Richards JB, and Evans DM
- Subjects
- Animals, Disease Models, Animal, Female, Femur chemistry, Gene Expression Profiling, Glypicans deficiency, Glypicans genetics, Glypicans physiology, Growth Disorders genetics, Humans, Male, Mice, Mice, Knockout, Molecular Sequence Annotation, Osteoblasts metabolism, Osteochondrodysplasias congenital, Osteochondrodysplasias genetics, Osteoclasts metabolism, Osteocytes metabolism, Osteoporosis pathology, Phenotype, Bone Density genetics, Calcaneus pathology, Genome-Wide Association Study, Osteoporosis genetics, Polymorphism, Single Nucleotide
- Abstract
Osteoporosis is a common disease diagnosed primarily by measurement of bone mineral density (BMD). We undertook a genome-wide association study (GWAS) in 142,487 individuals from the UK Biobank to identify loci associated with BMD as estimated by quantitative ultrasound of the heel. We identified 307 conditionally independent single-nucleotide polymorphisms (SNPs) that attained genome-wide significance at 203 loci, explaining approximately 12% of the phenotypic variance. These included 153 previously unreported loci, and several rare variants with large effect sizes. To investigate the underlying mechanisms, we undertook (1) bioinformatic, functional genomic annotation and human osteoblast expression studies; (2) gene-function prediction; (3) skeletal phenotyping of 120 knockout mice with deletions of genes adjacent to lead independent SNPs; and (4) analysis of gene expression in mouse osteoblasts, osteocytes and osteoclasts. The results implicate GPC6 as a novel determinant of BMD, and also identify abnormal skeletal phenotypes in knockout mice associated with a further 100 prioritized genes.
- Published
- 2017
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17. Pitx1 determines characteristic hindlimb morphologies in cartilage micromass culture.
- Author
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Butterfield NC, Qian C, and Logan MPO
- Subjects
- Alcian Blue, Animals, Blotting, Western, Cartilage cytology, Cartilage embryology, Cells, Cultured, Chondrogenesis genetics, Forelimb cytology, Forelimb embryology, Forelimb metabolism, Hindlimb cytology, Hindlimb embryology, Limb Buds cytology, Limb Buds embryology, Limb Buds metabolism, Mice, Knockout, Mice, Transgenic, Paired Box Transcription Factors metabolism, Staining and Labeling methods, Body Patterning genetics, Cartilage metabolism, Gene Expression Regulation, Developmental, Hindlimb metabolism, Paired Box Transcription Factors genetics
- Abstract
The shapes of homologous skeletal elements in the vertebrate forelimb and hindlimb are distinct, with each element exquisitely adapted to their divergent functions. Many of the signals and signalling pathways responsible for patterning the developing limb bud are common to both forelimb and hindlimb. How disparate morphologies are generated from common signalling inputs during limb development remains poorly understood. We show that, similar to what has been shown in the chick, characteristic differences in mouse forelimb and hindlimb cartilage morphology are maintained when chondrogenesis proceeds in vitro away from the endogenous limb bud environment. Chondrogenic nodules that form in high-density micromass cultures derived from forelimb and hindlimb buds are consistently different in size and shape. We described analytical tools we have developed to quantify these differences in nodule morphology and demonstrate that characteristic hindlimb nodule morphology is lost in the absence of the hindlimb-restricted limb modifier gene Pitx1. Furthermore, we show that ectopic expression of Pitx1 in the forelimb is sufficient to generate nodule patterns characteristic of the hindlimb. We also demonstrate that hindlimb cells are less adhesive to the tissue culture substrate and, within the limb environment, to the extracellular matrix and to each other. These results reveal autonomously programmed differences in forelimb and hindlimb cartilage precursors of the limb skeleton are controlled, at least in part, by Pitx1 and suggest this has an important role in generating distinct limb-type morphologies. Our results demonstrate that the micromass culture system is ideally suited to study cues governing morphogenesis of limb skeletal elements in a simple and experimentally tractable in vitro system that reflects in vivo potential.
- Published
- 2017
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18. Tbx5 Buffers Inherent Left/Right Asymmetry Ensuring Symmetric Forelimb Formation.
- Author
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Sulaiman FA, Nishimoto S, Murphy GR, Kucharska A, Butterfield NC, Newbury-Ecob R, and Logan MP
- Subjects
- Abnormalities, Multiple genetics, Abnormalities, Multiple pathology, Animals, DNA-Binding Proteins genetics, Embryo, Mammalian, Gene Expression Regulation, Developmental, Heart Defects, Congenital genetics, Heart Defects, Congenital pathology, Heart Septal Defects, Atrial genetics, Heart Septal Defects, Atrial pathology, Humans, Limb Buds growth & development, Limb Deformities, Congenital pathology, Lower Extremity Deformities, Congenital genetics, Lower Extremity Deformities, Congenital pathology, Mice, Somites growth & development, Upper Extremity Deformities, Congenital genetics, Upper Extremity Deformities, Congenital pathology, Embryonic Development genetics, Forelimb growth & development, Limb Deformities, Congenital genetics, T-Box Domain Proteins genetics
- Abstract
The forelimbs and hindlimbs of vertebrates are bilaterally symmetric. The mechanisms that ensure symmetric limb formation are unknown but they can be disrupted in disease. In Holt-Oram Syndrome (HOS), caused by mutations in TBX5, affected individuals have left-biased upper/forelimb defects. We demonstrate a role for the transcription factor Tbx5 in ensuring the symmetric formation of the left and right forelimb. In our mouse model, bilateral hypomorphic levels of Tbx5 produces asymmetric forelimb defects that are consistently more severe in the left limb than the right, phenocopying the left-biased limb defects seen in HOS patients. In Tbx hypomorphic mutants maintained on an INV mutant background, with situs inversus, the laterality of defects is reversed. Our data demonstrate an early, inherent asymmetry in the left and right limb-forming regions and that threshold levels of Tbx5 are required to overcome this asymmetry to ensure symmetric forelimb formation., Competing Interests: The authors have declared that no competing interests exist.
- Published
- 2016
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19. Mutations in mouse Ift144 model the craniofacial, limb and rib defects in skeletal ciliopathies.
- Author
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Ashe A, Butterfield NC, Town L, Courtney AD, Cooper AN, Ferguson C, Barry R, Olsson F, Liem KF Jr, Parton RG, Wainwright BJ, Anderson KV, Whitelaw E, and Wicking C
- Subjects
- Abnormalities, Multiple embryology, Abnormalities, Multiple metabolism, Animals, Chromosome Mapping, Craniofacial Abnormalities embryology, Craniofacial Abnormalities metabolism, Cytoskeletal Proteins, Embryo, Mammalian, Fibroblast Growth Factors metabolism, Forelimb abnormalities, Forelimb metabolism, Hedgehog Proteins metabolism, Intercellular Signaling Peptides and Proteins metabolism, Intracellular Signaling Peptides and Proteins, Mice, Mutagenesis, Mutation, Missense, Phenotype, Polydactyly embryology, Polydactyly genetics, Polydactyly metabolism, Proteins chemistry, Ribs abnormalities, Signal Transduction, Abnormalities, Multiple genetics, Cilia physiology, Cilia ultrastructure, Craniofacial Abnormalities genetics, Proteins genetics
- Abstract
Mutations in components of the intraflagellar transport (IFT) machinery required for assembly and function of the primary cilium cause a subset of human ciliopathies characterized primarily by skeletal dysplasia. Recently, mutations in the IFT-A gene IFT144 have been described in patients with Sensenbrenner and Jeune syndromes, which are associated with short ribs and limbs, polydactyly and craniofacial defects. Here, we describe an N-ethyl-N-nitrosourea-derived mouse mutant with a hypomorphic missense mutation in the Ift144 gene. The mutant twinkle-toes (Ift144(twt)) phenocopies a number of the skeletal and craniofacial anomalies seen in patients with human skeletal ciliopathies. Like other IFT-A mouse mutants, Ift144 mutant embryos display a generalized ligand-independent expansion of hedgehog (Hh) signalling, in spite of defective ciliogenesis and an attenuation of the ability of mutant cells to respond to upstream stimulation of the pathway. This enhanced Hh signalling is consistent with cleft palate and polydactyly phenotypes in the Ift144(twt) mutant, although extensive rib branching, fusion and truncation phenotypes correlate with defects in early somite patterning and may reflect contributions from multiple signalling pathways. Analysis of embryos harbouring a second allele of Ift144 which represents a functional null, revealed a dose-dependent effect on limb outgrowth consistent with the short-limb phenotypes characteristic of these ciliopathies. This allelic series of mouse mutants provides a unique opportunity to uncover the underlying mechanistic basis of this intriguing subset of ciliopathies.
- Published
- 2012
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20. Inactivation of Patched1 in the mouse limb has novel inhibitory effects on the chondrogenic program.
- Author
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Bruce SJ, Butterfield NC, Metzis V, Town L, McGlinn E, and Wicking C
- Subjects
- Animals, Cell Death, Cell Differentiation, Cells, Cultured, Chondrocytes cytology, Chondrocytes metabolism, Female, Forelimb metabolism, Forelimb physiology, Hedgehog Proteins metabolism, Hindlimb metabolism, Hindlimb physiology, Homeodomain Proteins genetics, Ligands, Male, Mice, Mice, Transgenic, Molecular Imaging, Patched Receptors, Patched-1 Receptor, Peanut Agglutinin metabolism, Phenotype, Receptors, Cell Surface deficiency, Receptors, Cell Surface genetics, Staining and Labeling, Time Factors, Chondrogenesis, Extremities physiology, Receptors, Cell Surface metabolism
- Abstract
The bones of the vertebrate limb form by the process of endochondral ossification, whereby limb mesenchyme condenses to form an intermediate cartilage scaffold that is then replaced by bone. Although Indian hedgehog (IHH) is known to control hypertophic differentiation of chondrocytes during this process, the role of hedgehog signaling in the earlier stages of chondrogenesis is less clear. We have conditionally inactivated the hedgehog receptor Ptc1 in undifferentiated limb mesenchyme of the mouse limb using Prx1-Cre, thus inducing constitutively active ligand-independent hedgehog signaling. In addition to major patterning defects, we observed a marked disruption to the cartilage elements in the limbs of Prx1-Cre:Ptc1(c/c) embryos. Using an in vitro micromass culture system we show that this defect lies downstream of mesenchymal cell condensation and likely upstream of chondrocyte differentiation. Despite early increases in levels of chondrogenic genes, soon after mesenchymal condensation the stromal layer of Prx1-Cre:Ptc1(c/c)-derived micromass cultures is characterized by a loss of cell integrity, which is associated with increased cell death and a striking decrease in Alcian blue staining cartilage nodules. Furthermore, inhibition of the hedgehog pathway activation using cyclopamine was sufficient to essentially overcome this chondrogenic defect in both micromass and ex vivo explant assays of Prx1-Cre:Ptc1(c/c) limbs. These data demonstrate for the first time the inhibitory effect of cell autonomously activated hedgehog signaling on chondrogenesis, and stress the importance of PTC1 in maintaining strict control of signaling levels during this phase of skeletal development.
- Published
- 2010
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21. The molecular regulation of vertebrate limb patterning.
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Butterfield NC, McGlinn E, and Wicking C
- Subjects
- Animals, Embryonic Induction, Extremities anatomy & histology, Extremities embryology, Extremities physiology, Gene Expression Regulation, Developmental, Hedgehog Proteins genetics, Hedgehog Proteins metabolism, Intercellular Signaling Peptides and Proteins metabolism, Kruppel-Like Transcription Factors genetics, Kruppel-Like Transcription Factors metabolism, Models, Biological, Nerve Tissue Proteins genetics, Nerve Tissue Proteins metabolism, Signal Transduction, Vertebrates anatomy & histology, Vertebrates embryology, Vertebrates physiology, Body Patterning physiology, Extremities growth & development, Vertebrates growth & development
- Abstract
The limb has long been considered a paradigm for organogenesis because of its simplicity and ease of manipulation. However, it has become increasingly clear that the processes required to produce a perfectly formed limb involve complex molecular interactions across all three axes of limb development. Old models have evolved with acquisition of molecular knowledge, and in more recent times mathematical modeling approaches have been invoked to explain the precise spatio-temporal regulation of gene networks that coordinate limb patterning and outgrowth. This review focuses on recent advances in our understanding of vertebrate limb development, highlighting the signaling interactions required to lay down the pattern on which the processes of differentiation will act to ultimately produce the final limb., (Copyright 2010 Elsevier Inc. All rights reserved.)
- Published
- 2010
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22. The metalloendopeptidase gene Pitrm1 is regulated by hedgehog signaling in the developing mouse limb and is expressed in muscle progenitors.
- Author
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Town L, McGlinn E, Fiorenza S, Metzis V, Butterfield NC, Richman JM, and Wicking C
- Subjects
- Animals, Embryo, Mammalian, Gene Expression Regulation, Developmental, Gene Expression Regulation, Enzymologic, Hedgehog Proteins genetics, Hedgehog Proteins metabolism, Metalloendopeptidases metabolism, Mice, Mice, Transgenic, Muscle, Skeletal metabolism, Myoblasts, Skeletal metabolism, PAX3 Transcription Factor, Paired Box Transcription Factors metabolism, Signal Transduction genetics, Tissue Distribution, Extremities embryology, Hedgehog Proteins physiology, Metalloendopeptidases genetics, Muscle, Skeletal embryology, Stem Cells metabolism
- Abstract
Pitrm1 is a zinc metalloendopeptidase that has been implicated in Alzheimer's disease and mitochondrial peptide degradation, but to date no major role in embryonic development has been documented. In a screen for genes regulated by hedgehog signaling in the mouse limb, we showed that expression of Pitrm1 is upregulated in response to loss of the Gli3 transcription factor. Here we confirm spatial changes in Pitrm1 expression in the Gli3 mutant mouse limb and examine Pitrm1 expression in Shh null and Ptch1 conditional deletion mouse mutants. In wild-type mice, Pitrm1 is expressed in a number of developing tissues known to be patterned by Sonic hedgehog, including the limbs, face, cortex, hippocampus, cerebellum, tectum, sub-mandibular gland, lung, genital tubercle, hair follicles, and the enamel knot of the teeth. Additionally, Pitrm1 is expressed in Pax3-expressing myoblast progenitors in the limb, the dermomyotome, and developing muscles of the face and torso., ((c) 2009 Wiley-Liss, Inc.)
- Published
- 2009
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23. Patched 1 is a crucial determinant of asymmetry and digit number in the vertebrate limb.
- Author
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Butterfield NC, Metzis V, McGlinn E, Bruce SJ, Wainwright BJ, and Wicking C
- Subjects
- Animals, Apoptosis, Embryo, Mammalian cytology, Embryo, Mammalian metabolism, Female, Hedgehog Proteins genetics, Kruppel-Like Transcription Factors genetics, Male, Mice, Mice, Inbred C57BL, Nerve Tissue Proteins genetics, Patched Receptors, Patched-1 Receptor, Receptors, Cell Surface genetics, Signal Transduction, Up-Regulation, Zinc Finger Protein Gli3, Body Patterning, Extremities embryology, Gene Expression Regulation, Developmental, Receptors, Cell Surface metabolism
- Abstract
The vertebrate hedgehog receptor patched 1 (Ptc1) is crucial for negative regulation of the sonic hedgehog (Shh) pathway during anterior-posterior patterning of the limb. We have conditionally inactivated Ptc1 in the mesenchyme of the mouse limb using Prx1-Cre. This results in constitutive activation of hedgehog (Hh) signalling during the early stages of limb budding. Our data suggest that variations in the timing and efficiency of Cre-mediated excision result in differential forelimb and hindlimb phenotypes. Hindlimbs display polydactyly (gain of digits) and a molecular profile similar to the Gli3 mutant extra-toes. Strikingly, forelimbs are predominantly oligodactylous (displaying a loss of digits), with a symmetrical, mirror-image molecular profile that is consistent with re-specification of the anterior forelimb to a posterior identity. Our data suggest that this is related to very early inactivation of Ptc1 in the forelimb perturbing the gene regulatory networks responsible for both the pre-patterning and the subsequent patterning stages of limb development. These results establish the importance of the downstream consequences of Hh pathway repression, and identify Ptc1 as a key player in limb patterning even prior to the onset of Shh expression.
- Published
- 2009
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24. Expression of the NET family member Zfp503 is regulated by hedgehog and BMP signaling in the limb.
- Author
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McGlinn E, Richman JM, Metzis V, Town L, Butterfield NC, Wainwright BJ, and Wicking C
- Subjects
- Animals, Bone Morphogenetic Proteins genetics, Carrier Proteins genetics, Chick Embryo, Embryo, Mammalian anatomy & histology, Embryo, Mammalian physiology, Embryo, Nonmammalian anatomy & histology, Embryo, Nonmammalian physiology, Extremities physiology, Female, Hedgehog Proteins genetics, In Situ Hybridization, Intracellular Signaling Peptides and Proteins, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, Nerve Tissue Proteins genetics, Nuclear Proteins genetics, Bone Morphogenetic Proteins metabolism, Carrier Proteins metabolism, Extremities anatomy & histology, Extremities embryology, Gene Expression Regulation, Developmental, Hedgehog Proteins metabolism, Nerve Tissue Proteins metabolism, Nuclear Proteins metabolism, Signal Transduction physiology
- Abstract
The NET/Nlz family of zinc finger transcription factors contribute to aspects of developmental growth and patterning across evolutionarily diverse species. To date, however, these molecules remain largely uncharacterized in mouse and chick. We previously reported that limb bud expression of Zfp503, the mouse orthologue of zebrafish nlz2/znf503, is dependent on Gli3. Here, we show that Zfp503/Znf503 is expressed in a restricted pattern during mouse and chick embryogenesis, with particularly dynamic expression in the developing limbs, face, somites, and brain. We also add to our previous data on Gli3 regulation by showing that the anterior domain of Zfp503 expression in the mouse limb is responsive to genetic and nongenetic manipulation of hedgehog signaling. Finally, we demonstrate that posterior expression of Znf503 in the chick limb is responsive to bone morphogenetic protein (BMP) signaling, indicating that Zfp503/Znf503 may act at the nexus of multiple signaling pathways in development., ((c) 2008 Wiley-Liss, Inc.)
- Published
- 2008
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25. A genome-wide screen for modifiers of transgene variegation identifies genes with critical roles in development.
- Author
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Ashe A, Morgan DK, Whitelaw NC, Bruxner TJ, Vickaryous NK, Cox LL, Butterfield NC, Wicking C, Blewitt ME, Wilkins SJ, Anderson GJ, Cox TC, and Whitelaw E
- Subjects
- Animals, Female, Genes, Lethal, Genome, Heterozygote, Histone Deacetylase 1, Histone Deacetylases metabolism, Male, Mice, Mice, Inbred C57BL, Mice, Inbred Strains, Mice, Transgenic, Transcription Factors metabolism, Williams Syndrome physiopathology, Embryonic Development, Epigenesis, Genetic
- Abstract
Background: Some years ago we established an N-ethyl-N-nitrosourea screen for modifiers of transgene variegation in the mouse and a preliminary description of the first six mutant lines, named MommeD1-D6, has been published. We have reported the underlying genes in three cases: MommeD1 is a mutation in SMC hinge domain containing 1 (Smchd1), a novel modifier of epigenetic gene silencing; MommeD2 is a mutation in DNA methyltransferase 1 (Dnmt1); and MommeD4 is a mutation in Smarca 5 (Snf2h), a known chromatin remodeler. The identification of Dnmt1 and Smarca5 attest to the effectiveness of the screen design., Results: We have now extended the screen and have identified four new modifiers, MommeD7-D10. Here we show that all ten MommeDs link to unique sites in the genome, that homozygosity for the mutations is associated with severe developmental abnormalities and that heterozygosity results in phenotypic abnormalities and reduced reproductive fitness in some cases. In addition, we have now identified the underlying genes for MommeD5 and MommeD10. MommeD5 is a mutation in Hdac1, which encodes histone deacetylase 1, and MommeD10 is a mutation in Baz1b (also known as Williams syndrome transcription factor), which encodes a transcription factor containing a PHD-type zinc finger and a bromodomain. We show that reduction in the level of Baz1b in the mouse results in craniofacial features reminiscent of Williams syndrome., Conclusions: These results demonstrate the importance of dosage-dependent epigenetic reprogramming in the development of the embryo and the power of the screen to provide mouse models to study this process.
- Published
- 2008
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26. Identification and analysis of novel genes expressed in the mouse embryonic facial primordia.
- Author
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Bennetts JS, Fowles LF, Butterfield NC, Berkman JL, Teasdale RD, Simpson F, and Wicking C
- Subjects
- Animals, Computational Biology, Gene Expression Profiling, Genetic Vectors, HeLa Cells, Humans, In Situ Hybridization, Mice, Sequence Analysis, DNA, Transfection, Craniofacial Abnormalities genetics, Face embryology, Gene Expression Regulation, Developmental, Gene Library
- Abstract
Craniofacial anomalies are a common feature of human congenital dysmorphology syndromes, suggesting that genes expressed in the developing face are likely to play a wider role in embryonic development. To facilitate the identification of genes involved in embryogenesis, we previously constructed an enriched cDNA library by subtracting adult mouse liver cDNA from that of embryonic day (E)10.5 mouse pharyngeal arch cDNA. From this library, 273 unique clones were sequenced and known proteins binned into functional categories in order to assess enrichment of the library (1). We have now selected 31 novel and poorly characterised genes from this library and present bioinformatic analysis to predict proteins encoded by these genes, and to detect evolutionary conservation. Of these genes 61% (19/31) showed restricted expression in the developing embryo, and a subset of these was chosen for further in silico characterisation as well as experimental determination of subcellular localisation based on transient transfection of predicted full-length coding sequences into mammalian cell lines. Where a human orthologue of these genes was detected, chromosomal localisation was determined relative to known loci for human congenital disease.
- Published
- 2006
- Full Text
- View/download PDF
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