Osteogenic Mechanisms of Basal Ganglia Calcification and its ex vivo Model in the Hypoparathyroid Milieu

Endocrinology ◽  
2021 ◽  
Vol 162 (4) ◽  
Author(s):  
Parmita Kar ◽  
Tabin Millo ◽  
Soma Saha ◽  
Samrina Mahtab ◽  
Shipra Agarwal ◽  
...  
Keyword(s):  
Basal Ganglia ◽  
Gray Matter ◽  
Molecular Mechanisms ◽  
Ex Vivo ◽  
Ex Vivo Model ◽  
Caudate Nuclei ◽  
Basal Ganglia Calcification ◽  
Cortical Gray Matter ◽  
Autopsy Tissues ◽  
High Phosphate

Abstract Context Basal-ganglia calcification (BGC) is common (70%) in patients with chronic hypoparathyroidism. Interestingly, cortical gray matter is spared from calcification. The mechanism of BGC, role of hyperphosphatemia, and modulation of osteogenic molecules by parathyroid hormone (PTH) in its pathogenesis is not clear. Objective We assessed the expression of a large repertoire of molecules with proosteogenic or antiosteogenic effects, including neuroprogenitor cells in caudate, dentate, and cortical gray matter from normal autopsy tissues. The effect of high phosphate and PTH was assessed in an ex vivo model of BGC using striatum tissue culture of the Sprague-Dawley rat. Methods The messenger RNA and protein expression of 39 molecules involved in multiple osteogenic pathways were assessed in 25 autopsy tissues using reverse-transcriptase polymerase chain reaction, Western blot, and immunofluorescence. The striatal culture was maintained in a hypoparathyroid milieu for 24 days with and without (a) high phosphate (10-mm β-glycerophosphate) and (b) PTH(1-34) (50 ng/mL Dulbecco’s modified Eagle’s medium–F12 media) for their effect on striatal calcification and osteogenic molecules. Results Procalcification molecules (osteonectin, β-catenin, klotho, FZD4, NT5E, LRP5, WNT3A, collagen-1α, and SOX2-positive neuroprogenitor stem cells) had significantly higher expression in the caudate than gray matter. Caudate nuclei also had higher expression of antiosteogenic molecules (osteopontin, carbonic anhydrase-II [CA-II], MGP, sclerostin, ISG15, ENPP1, and USP18). In an ex vivo model, striatum culture showed an increased propensity for calcified nodules with mineral deposition similar to that of bone tissue on Fourier-transformed infrared spectroscopy, alizarin, and von Kossa stain. Mineralization in striatal culture was enhanced by high phosphate and decreased by exogenous PTH through increased expression of CA-II. Conclusion This study provides a conceptual advance on the molecular mechanisms of BGC and the possibility of PTH therapy to prevent this complication in a hypoparathyroid milieu.

scholarly journals Earliest Mechanisms of Dopaminergic Neurons Sufferance in a Novel Slow Progressing Ex Vivo Model of Parkinson Disease in Rat Organotypic Cultures of Substantia Nigra

2019 ◽  
Vol 20 (9) ◽  
pp. 2224 ◽  
Author(s):  
Matteo Dal Ben ◽  
Rosario Bongiovanni ◽  
Simone Tuniz ◽  
Emanuela Fioriti ◽  
Claudio Tiribelli ◽  
...  
Keyword(s):  
Substantia Nigra ◽  
Parkinson Disease ◽  
Dopaminergic Neurons ◽  
Molecular Mechanisms ◽  
Ex Vivo ◽  
Ex Vivo Model ◽  
Redox Imbalance ◽  
Clinical Phase ◽  
Protein Clearance ◽  
Persistent Inflammation

The current treatments of Parkinson disease (PD) are ineffective mainly due to the poor understanding of the early events causing the decline of dopaminergic neurons (DOPAn). To overcome this problem, slow progressively degenerating models of PD allowing the study of the pre-clinical phase are crucial. We recreated in a short ex vivo time scale (96 h) all the features of human PD (needing dozens of years) by challenging organotypic culture of rat substantia nigra with low doses of rotenone. Thus, taking advantage of the existent knowledge, the model was used to perform a time-dependent comparative study of the principal possible causative molecular mechanisms undergoing DOPAn demise. Alteration in the redox state and inflammation started at 3 h, preceding the reduction in DOPAn number (pre-diagnosis phase). The number of DOPAn declined to levels compatible with diagnosis only at 12 h. The decline was accompanied by a persistent inflammation and redox imbalance. Significant microglia activation, apoptosis, a reduction in dopamine vesicle transporters, and the ubiquitination of misfolded protein clearance pathways were late (96 h, consequential) events. The work suggests inflammation and redox imbalance as simultaneous early mechanisms undergoing DOPAn sufferance, to be targeted for a causative treatment aimed to stop/delay PD.

scholarly journals Expression of Osteogenic Molecules in the Caudate Nucleus and Gray Matter and Their Potential Relevance for Basal Ganglia Calcification in Hypoparathyroidism

2014 ◽  
Vol 99 (5) ◽  
pp. 1741-1748 ◽  
Author(s):  
Ravinder Goswami ◽  
Tabin Millo ◽  
Shruti Mishra ◽  
Madhuchhanda Das ◽  
Mansi Kapoor ◽  
...  
Keyword(s):  
Basal Ganglia ◽  
Caudate Nucleus ◽  
Gray Matter ◽  
Basal Ganglia Calcification

Quantitative high-field imaging of sub-cortical gray matter in multiple sclerosis

2011 ◽  
Vol 18 (4) ◽  
pp. 433-441 ◽  
Author(s):  
R Marc Lebel ◽  
Amir Eissa ◽  
Peter Seres ◽  
Gregg Blevins ◽  
Alan H Wilman
Keyword(s):  
Multiple Sclerosis ◽  
Basal Ganglia ◽  
Gray Matter ◽  
High Field ◽  
Red Nucleus ◽  
Quantitative Mri ◽  
Relaxation Rates ◽  
Relapsing Remitting ◽  
Cortical Gray Matter ◽  
Very High

Background: In addition to neuronal injury, inflammatory, and demyelinating processes, evidence suggests multiple sclerosis (MS) is also associated with increased iron deposition in the basal ganglia. Magnetic resonance imaging (MRI), particularly at very high field strengths, is sensitive to iron accumulation and may enable visualization and quantification of iron associated with MS. Objectives: To investigate the sub-cortical gray matter in patients with early-stage relapsing–remitting MS using multiple, and novel, quantitative MRI measures at very high field. Methods: In total, 22 patients with relapsing–remitting MS and 22 control subjects were imaged at 4.7 Tesla. Transverse relaxation rates (R2 and R2*) and susceptibility phase were quantified in four basal ganglia nuclei, the thalamus, and the red nuclei. Parameters in patients with MS were compared with those in healthy subjects and correlated with clinical scores. Results: Significant abnormalities were observed in most structures, most notably in the pulvinar sub-nucleus. Significant correlations with disability were observed in the pulvinar; marginally significant correlations were also observed in the thalamus and red nucleus. No significant correlations were observed with duration since index relapse. Conclusions: Widespread abnormalities are present in the deep gray matter nuclei of patients recently diagnosed with MS; these abnormalities can be detected via multi-modal high-field MRI. Imaging metrics, particularly R2*, relate to disease severity in the pulvinar and other gray matter regions.

scholarly journals Stretch increases alveolar type 1 cell number in fetal lungs through ROCK-Yap/Taz pathway

2021 ◽  
Author(s):  
Tram Mai Nguyen ◽  
Johannes van der Merwe ◽  
Linda Elowsson Rendin ◽  
Anna-Karin Larsson-Callerfelt ◽  
Jan Deprest ◽  
...  
Keyword(s):  
Lung Development ◽  
Molecular Mechanisms ◽  
Ex Vivo ◽  
Fluid Pressure ◽  
Fetal Lung ◽  
In Vivo Model ◽  
Alveolar Type ◽  
Ex Vivo Model

Accurate fluid pressure in the fetal lung is critical for its development, especially at the beginning of the saccular stage when alveolar epithelial type 1 (AT1) and type 2 (AT2) cells differentiate from the epithelial progenitors. Despite our growing understanding of the role of physical forces in lung development, the molecular mechanisms that regulate the transduction of mechanical stretch to alveolar differentiation remain elusive. To simulate lung distension, we optimized both an ex vivo model with precision cut lung slices and an in vivo model of fetal tracheal occlusion. Increased mechanical tension showed to improve alveolar maturation and differentiation towards AT1. By manipulating ROCK pathway, we demonstrate that stretch-induced Yap/Taz activation promotes alveolar differentiation towards AT1 phenotype via ROCK activity. Our findings show that balanced ROCK-Yap/Taz signaling is essential to regulate AT1 differentiation in response to mechanical stretching of the fetal lung, which might be helpful in improving lung development and regeneration.

A Novel Porcine Ex Vivo Retina Culture Model for Oxidative Stress Induced by H2O2

2017 ◽  
Vol 45 (1) ◽  
pp. 11-25 ◽  
Author(s):  
José Hurst ◽  
Sandra Kuehn ◽  
Adelina Jashari ◽  
Teresa Tsai ◽  
Karl Ulrich Bartz-Schmidt ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Molecular Mechanisms ◽  
Ganglion Cells ◽  
Ex Vivo ◽  
Laboratory Animals ◽  
Ex Vivo Model ◽  
Cd11b Expression ◽  
Hsp70 Mrna ◽  
New Therapeutic Approaches ◽  
Culture Models

Oxidative stress is a key player in many ophthalmic diseases. However, the role of oxidative stress in most degenerative processes is not yet known. Therefore, accurate and practical models are required to efficiently screen for therapeutics. Porcine eyes are closely related to the human eye, and can be obtained from the abattoir as a by-product of the food industry. Therefore, they offer excellent opportunities for the development of culture models with which to pre-screen potential therapies, while reducing the use of laboratory animals. To induce oxidative stress, organotypic cultures of porcine retina were treated with different doses of hydrogen peroxide (H2O2; 100, 300 and 500μM) for three hours. On days 3 and 8, the retinas were conserved for histological and Western blotting analyses and for evaluation of gene expression, which determined the number of retinal ganglion cells (RGCs), the activation state of glial cells, and the expression levels of several oxidative stress markers. H2O2 treatment led to a reduction in the number of RGCs and to an increase in apoptotic RGCs. In addition, a dose-dependent increase of microglia and an elevation of CD11b expression was observed. On day 3, a reduction of IL-1β, and an increase of iNOS, as well as of HSP70 mRNA were found. On day 8, an increase in TNF-α and IL-1β mRNA expression was detected. In conclusion, this ex vivo model offers an opportunity to study the molecular mechanisms underlying certain eye disorders and to test new therapeutic approaches to diminish the effects of oxidative stress.

Tissue Factor Localization in Non-Human Primate Cerebral Tissue

1992 ◽  
Vol 68 (06) ◽  
pp. 642-647 ◽  
Author(s):  
Gregory J del Zoppo ◽  
Jian-Qing Yu ◽  
Brian R Copeland ◽  
Winston S Thomas ◽  
Jacob Schneiderman ◽  
...  
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
White Matter ◽  
Basal Ganglia ◽  
Tissue Factor ◽  
Gray Matter ◽  
Sandwich Elisa ◽  
Procoagulant Activity ◽  
Cortical Gray Matter ◽  
Human Primate

SummaryTissue factor (TF), the principal procoagulant of human brain, resides in specific regions of the non-human primate central nervous system. Immunohistochemical studies employing murine anti-human TF monoclonal antibodies (MoAbs) detected TF antigen in the cortex, basal ganglia, cerebellum, and cervical spinal cord in three normal baboon subjects. Although significantly less prominent than human cortical gray matter, a distinct partition of TF in gray matter >white matter was noted. The gray matter predilection of TF was confirmed in primate temporal and parietal lobe cortex by both sandwich ELISA and one-stage coagulation assay. Variation in the relative quantity of TF antigen was observed by ELISA among the three subjects studied. Procoagulant activity followed the pattern of TF antigen (cortical gray matter >basal ganglia ≥cerebellum >cortical white matter), and was 96.5–98.5% inhibitable by a function inhibiting anti-human TF MoAb combination. TF antigen was associated with the microvasculature of all cerebral tissues studied, and spared capillaries most selectively in the cerebral cortex, basal ganglia, and cerebellum. These findings suggest a highly specific ordering of TF antigen and related procoagulant activity in the central nervous system of the baboon, confined primarily to gray matter parenchyma, and to the non-capillary microvasculature.

scholarly journals An ex vivo model to induce early fibrosis-like changes in human precision-cut lung slices

2017 ◽  
Vol 312 (6) ◽  
pp. L896-L902 ◽  
Author(s):  
Hani N. Alsafadi ◽  
Claudia A. Staab-Weijnitz ◽  
Mareike Lehmann ◽  
Michael Lindner ◽  
Britta Peschel ◽  
...  
Keyword(s):  
Growth Factor ◽  
Molecular Mechanisms ◽  
Transforming Growth Factor ◽  
Ex Vivo ◽  
Disease Onset ◽  
Water Soluble ◽  
High Morbidity ◽  
Ex Vivo Model ◽  
Early Fibrosis ◽  
Precision Cut Lung Slices

Idiopathic pulmonary fibrosis (IPF) is a devastating chronic interstitial lung disease (ILD) characterized by lung tissue scarring and high morbidity. Lung epithelial injury, myofibroblast activation, and deranged repair are believed to be key processes involved in disease onset and progression, but the exact molecular mechanisms behind IPF remain unclear. Several drugs have been shown to slow disease progression, but treatments that halt or reverse IPF progression have not been identified. Ex vivo models of human lung have been proposed for drug discovery, one of which is precision-cut lung slices (PCLS). Although PCLS production from IPF explants is possible, IPF explants are rare and typically represent end-stage disease. Here we present a novel model of early fibrosis-like changes in human PCLS derived from patients without ILD/IPF using a combination of profibrotic growth factors and signaling molecules (transforming growth factor-β, tumor necrosis factor-α, platelet-derived growth factor-AB, and lysophosphatidic acid). Fibrotic-like changes of PCLS were qualitatively analyzed by histology and immunofluorescence and quantitatively by water-soluble tetrazolium-1, RT-qPCR, Western blot analysis, and ELISA. PCLS remained viable after 5 days of treatment, and fibrotic gene expression ( FN1, SERPINE1, COL1A1, CTGF, MMP7, and ACTA2) increased as early as 24 h of treatment, with increases in protein levels at 48 h and increased deposition of extracellular matrix. Alveolar epithelium reprogramming was evident by decreases in surfactant protein C and loss of HOPX. In summary, using human-derived PCLS, we established a novel ex vivo model that displays characteristics of early fibrosis and could be used to evaluate novel therapies and study early-stage IPF pathomechanisms.

scholarly journals Increased HLA-DR expression and cortical demyelination in MS links with HLA-DR15

2019 ◽  
Vol 7 (2) ◽  
pp. e656 ◽  
Author(s):  
Lukas Simon Enz ◽  
Thomas Zeis ◽  
Daniela Schmid ◽  
Florian Geier ◽  
Franziska van der Meer ◽  
...  
Keyword(s):  
Human Brain ◽  
Gray Matter ◽  
Immunohistochemical Analysis ◽  
Bimodal Distribution ◽  
Lesion Size ◽  
Cortical Lesion ◽  
Hla Dr ◽  
Cortical Gray Matter ◽  
Autopsy Tissues ◽  
The Brain

ObjectiveTo investigate molecular changes in multiple sclerosis (MS) normal-appearing cortical gray matter (NAGM).MethodsWe performed a whole-genome gene expression microarray analysis of human brain autopsy tissues from 64 MS NAGM samples and 42 control gray matter samples. We further examined our cases by HLA genotyping and performed immunohistochemical and immunofluorescent analysis of all human brain tissues.ResultsHLA-DRB1 is the transcript with highest expression in MS NAGM with a bimodal distribution among the examined cases. Genotyping revealed that every case with the MS-associated HLA-DR15 haplotype also shows high HLA-DRB1 expression and also of the tightly linked HLA-DRB5 allele. Quantitative immunohistochemical analysis confirmed the higher expression of HLA-DRB1 in HLA-DRB1*15:01 cases at the protein level. Analysis of gray matter lesion size revealed a significant increase of cortical lesion size in cases with high HLA-DRB1 expression.ConclusionsOur data indicate that increased HLA-DRB1 and -DRB5 expression in the brain of patients with MS may be an important factor in how the HLA-DR15 haplotype contributes to MS pathomechanisms in the target organ.

scholarly journals Cooccurrence of Multiple Sclerosis and Idiopathic Basal Ganglia Calcification

2015 ◽  
Vol 2015 ◽  
pp. 1-3
Author(s):  
M. Abedini ◽  
N. Karimi ◽  
N. Tabrizi
Keyword(s):  
Multiple Sclerosis ◽  
Central Nervous System ◽  
Nervous System ◽  
Basal Ganglia ◽  
Gray Matter ◽  
Neurodegenerative Disorder ◽  
Rare Event ◽  
Basal Ganglia Calcification ◽  
Brain Calcification ◽  
Idiopathic Basal Ganglia Calcification

Multiple sclerosis (MS) is a chronic inflammatory demyelinating and neurodegenerative disease of central nervous system that affects both white and gray matter. Idiopathic calcification of the basal ganglia is a rare neurodegenerative disorder of unknown cause that is characterized by sporadic or familial brain calcification. Concurrence of multiple sclerosis (MS) and idiopathic basal ganglia calcification (Fahr’s disease) is very rare event. In this study, we describe a cooccurrence of idiopathic basal ganglia calcification with multiple sclerosis. The association between this disease and MS is unclear and also maybe probably coincidental.

scholarly journals Mapping the subcortical connectivity of the human default mode network

2021 ◽  
Author(s):  
Jian Li ◽  
William H. Curley ◽  
Bastien Guerin ◽  
Darin D. Dougherty ◽  
Adrian V. Dalca ◽  
...  
Keyword(s):  
Basal Ganglia ◽  
Default Mode Network ◽  
Basal Forebrain ◽  
Ex Vivo ◽  
7 Tesla ◽  
Human Consciousness ◽  
Connectivity Map ◽  
Caudate Nuclei ◽  
Default Mode ◽  
Human Connectome Project

The default mode network (DMN) mediates self-awareness and introspection, core components of human consciousness. Therapies to restore consciousness in patients with severe brain injuries have historically targeted subcortical sites in the brainstem, thalamus, hypothalamus, basal forebrain, and basal ganglia, with the goal of reactivating cortical DMN nodes. However, the subcortical connectivity of the DMN has not been fully mapped and optimal subcortical targets for therapeutic neuromodulation of consciousness have not been identified. In this work, we created a comprehensive map of DMN subcortical connectivity by combining high-resolution functional and structural datasets with advanced signal processing methods. We analyzed 7 Tesla resting-state functional MRI (rs-fMRI) data from 168 healthy volunteers acquired in the Human Connectome Project. The rs-fMRI blood-oxygen-level-dependent (BOLD) data were temporally synchronized across subjects using the BrainSync algorithm. Cortical and subcortical DMN nodes were jointly analyzed and identified at the group level by applying a novel Nadam-Accelerated SCAlable and Robust (NASCAR) tensor decomposition method to the synchronized dataset. The subcortical connectivity map was then overlaid on a 7 Tesla 100 micron ex vivo MRI dataset for neuroanatomic analysis using automated segmentation of nuclei within the brainstem, thalamus, hypothalamus, basal forebrain, and basal ganglia. We further compared the NASCAR subcortical connectivity map with its counterpart generated from canonical seed-based correlation analyses. The NASCAR method revealed that BOLD signal in the central lateral nucleus of the thalamus and ventral tegmental area of the midbrain is strongly correlated with that of the DMN. In an exploratory analysis, additional subcortical sites in the median and dorsal raphe, lateral hypothalamus, and caudate nuclei were correlated with the cortical DMN. We also found that the putamen and globus pallidus are negatively correlated (i.e., anti-correlated) with the DMN, providing rs-fMRI evidence for the mesocircuit hypothesis of human consciousness, whereby a striatopallidal feedback system modulates anterior forebrain function via disinhibition of the central thalamus. Seed-based analyses yielded similar subcortical DMN connectivity, but the NASCAR result showed stronger contrast and better spatial alignment with dopamine immunostaining data. The DMN subcortical connectivity map identified here advances understanding of the subcortical regions that contribute to human consciousness and can be used to inform the selection of therapeutic targets in clinical trials for patients with disorders of consciousness.

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