tissue architecture
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2022 ◽  
Author(s):  
Chih-Wei Hsu ◽  
Juan Cerda ◽  
Jason M Kirk ◽  
Williamson D. Turner ◽  
Tara L. Rasmussen ◽  
...  

Tissue clearing for whole organ cell profiling has revolutionized biology and imaging for exploration of organs in three-dimensional space without compromising tissue architecture. But complicated, laborious procedures, or expensive equipment, as well as the use of hazardous, organic solvents prevents the widespread adoption of these methods. Here we report a simple and rapid tissue clearing method, EZ Clear, that can clear whole adult mouse organs in 48 hours in just three simple steps. Samples stay at room temperature and remain hydrated throughout the clearing process, preserving endogenous and synthetic fluorescence, without altering sample size. After wholemount clearing and imaging, EZ Cleared samples can be further processed for downstream embedding and cryosectioning followed by standard histology or immunostaining, without loss of endogenous or synthetic fluorescence signal. Overall, the simplicity, speed, and flexibility of EZ Clear make it easy to adopt and apply to diverse approaches in biomedical research.


PLoS ONE ◽  
2021 ◽  
Vol 16 (12) ◽  
pp. e0261417
Author(s):  
R. C. M. Vulders ◽  
R. C. van Hoogenhuizen ◽  
E. van der Giessen ◽  
P. J. van der Zaag

The use of clearing agents has provided new insights in various fields of medical research (developmental biology, neurology) by enabling examination of tissue architecture in 3D. One of the challenges is that clearing agents induce tissue shrinkage and the shrinkage rates reported in the literature are incoherent. Here, we report that for a classical clearing agent, benzyl-alcohol benzyl-benzoate (BABB), the shrinkage decreases significantly with increasing sample size, and present an analytical formula describing this.


2021 ◽  
Author(s):  
Anadika R Prasad ◽  
Matthew P Bostock ◽  
Ines Lago-Baldaia ◽  
Zaynab Housseini ◽  
Vilaiwan M Fernandes

Precise neuronal numbers are required for circuit formation and function. Known strategies to control neuronal numbers involve regulating either cell proliferation or survival. In the developing Drosophila visual system photoreceptors from the eye-disc induce their target field, the lamina, one column at a time. Although each column initially contains ~6 precursors, only 5 differentiate into neurons of unique identities (L1-L5); the extra precursor undergoes apoptosis. We uncovered that Hedgehog signalling patterns columns, such that the 2 precursors experiencing the lowest signalling activity are specified as L5s; only one differentiates while the other extra precursor dies. We showed that a glial population called the outer chiasm giant glia (xgO), which reside below the lamina, relays differentiation signals from photoreceptors to induce L5 differentiation. The precursors nearest to xgO differentiate into L5s and antagonise inductive signalling to prevent the extra precursors from differentiating, resulting in their death. Thus, tissue architecture and feedback from young neurons fine-tune differentiation signals from glia to limit the number of neurons induced.


2021 ◽  
Author(s):  
Jingnan Liu ◽  
Yuanbing Zhang ◽  
Youfang Zhou ◽  
Qiao-Qi Wang ◽  
Kang Ding ◽  
...  

ABSTRACTTissue architecture determines its unique physiology and function. How these properties are intertwined has remained unclear. Here, we show that the metabolic enzyme CTP synthase (CTPS) form filamentous structures termed cytoophidia along the adipocyte cortex in Drosophila adipose tissue. Interestingly, loss of cytoophidia, whether due to reduced CTPS expression or a point mutation that specifically abrogates its polymerization ability, leads to downregulated Collagen-Integrin signaling, weakened adipocyte adhesion, and defective adipose architecture. Strikingly, CTPS specifically binds with Integrin subunit α2, which influences Integrin function and Collagen IV deposition. cytoophidia promote Collagen IV mRNA expression and thus its extracellular deposition to strengthen adipocyte adhesion. Remarkably, Collagen IV-Integrin signaling reciprocally regulates cytoophidium formation at a post-translational level. Together, we demonstrate that a positive feedback signaling loop containing both cytoophidia and Integrin adhesion complex couples tissue architecture and metabolism in the fly adipose.


Pharmaceutics ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 1994
Author(s):  
Josef Jampilek ◽  
Daniela Placha

Since the worldwide incidence of bone disorders and cartilage damage has been increasing and traditional therapy has reached its limits, nanomaterials can provide a new strategy in the regeneration of bones and cartilage. The nanoscale modifies the properties of materials, and many of the recently prepared nanocomposites can be used in tissue engineering as scaffolds for the development of biomimetic materials involved in the repair and healing of damaged tissues and organs. In addition, some nanomaterials represent a noteworthy alternative for treatment and alleviating inflammation or infections caused by microbial pathogens. On the other hand, some nanomaterials induce inflammation processes, especially by the generation of reactive oxygen species. Therefore, it is necessary to know and understand their effects in living systems and use surface modifications to prevent these negative effects. This contribution is focused on nanostructured scaffolds, providing a closer structural support approximation to native tissue architecture for cells and regulating cell proliferation, differentiation, and migration, which results in cartilage and bone healing and regeneration.


PLoS ONE ◽  
2021 ◽  
Vol 16 (11) ◽  
pp. e0259332
Author(s):  
Anna Fomitcheva-Khartchenko ◽  
Maria Anna Rapsomaniki ◽  
Bettina Sobottka ◽  
Peter Schraml ◽  
Govind V. Kaigala

A new workflow for protein-based tumor heterogeneity probing in tissues is here presented. Tumor heterogeneity is believed to be key for therapy failure and differences in prognosis in cancer patients. Comprehending tumor heterogeneity, especially at the protein level, is critical for tracking tumor evolution, and showing the presence of different phenotypical variants and their location with respect to tissue architecture. Although a variety of techniques is available for quantifying protein expression, the heterogeneity observed in the tissue is rarely addressed. The proposed method is validated in breast cancer fresh-frozen tissues derived from five patients. Protein expression is quantified on the tissue regions of interest (ROI) with a resolution of up to 100 μm in diameter. High heterogeneity values across the analyzed patients in proteins such as cytokeratin 7, β-actin and epidermal growth factor receptor (EGFR) using a Shannon entropy analysis are observed. Additionally, ROIs are clustered according to their expression levels, showing their location in the tissue section, and highlighting that similar phenotypical variants are not always located in neighboring regions. Interestingly, a patient with a phenotype related to increased aggressiveness of the tumor presents a unique protein expression pattern. In summary, a workflow for the localized extraction and protein analysis of regions of interest from frozen tissues, enabling the evaluation of tumor heterogeneity at the protein level is presented.


2021 ◽  
Author(s):  
Finosh Thankam ◽  
Victoria E. D. Wilson ◽  
Mohamed M Radwan ◽  
Aleem Siddique ◽  
Devendra K Agrawal

Abstract Aims: Expression status of pro-resolving lipid mediators (PLM) and receptors in the post-CABG coronary arteries are largely unknown. Here, we aim to investigate the expression of PLMs in the atherosclerotic post-CABG swine LAD compared to without CABG (LAD-AS), and in isolated coronary artery smooth muscle cells (CASMCs) cultured under ischemia. Methodology: The arteries of interest were harvested from post-CABG atherosclerotic swine and the histomorphology and the expression status of key PLM mediators were quantified using immunostaining. SMCs were cultured under ischemia and confirmed the expression on PLM mediators at transcript and protein level.Results: The histomorphometric analysis revealed considerable alterations in the tissue architecture in LAD-CABG and LAD-AS arteries compared to control. PLM synthetic enzyme 5LPOX was significantly upregulated in LAD-CABG and LAD-AS whereas the other mediators including 12LPOX, 15LPOX, COX2, ChemR23, GPCR18, GPCR120 were decreased in LAD-CABG than control. LPOX enzymes and PLM receptors were upregulated in ischemic CASMCs with respect to control. Western blot showed the upregulation of 5LPOX, and ChemR23. Additionally, higher level of RvE1 was observed in ischemic control CASMCs which was decreased following reperfusion. Conclusion: These findings suggest that the CASMCs withstand the ischemia-triggered proinflammatory episodes by increasing the secretion of RvE1 mediated through 5LPOX and ChemR23 signaling.


Author(s):  
Djallel Eddine Houari ADLI ◽  
Mostapha BRAHMI ◽  
Kaddour ZIANI ◽  
Wafaa ARABI ◽  
Khaled KAHLOULA ◽  
...  

The present study was carried out in order to evaluate, on the one hand, the modifications induced by manganese chloride according to a neurobehavioral, biochemical and histological approach in developing Wistar rats and, on the other hand, to test the effectiveness of Foeniculum vulgare (fennel) essential oil (FEO) in restoring or not the harmful effects of manganese chloride (MnCl2). The characterization of this essential oil by gas chromatography showed that the major component is E-anethole (34.7907%). The administration of FEO corrected the depressive state, anxiety and locomotor hypoactivity respectively observed in rats exposed to MnCl2, thus, the FEO restored the activity of the various antioxidant enzymes with a clear improvement of the cerebral tissue architecture in intoxicated rats treated with FEO. In conclusion, the FEO has a beneficial effect on the nervous system of rats intoxicated with MnCl2 which justifies the importance of this oil in traditional medicine.


2021 ◽  
Vol 8 ◽  
Author(s):  
Yu-Sheng Wang ◽  
Jia Guo

The ability to quantify a large number of varied transcripts in single cells in their native spatial context is crucial to accelerate our understanding of health and disease. Bulk cell RNA analysis masks the heterogeneity in the cell population, while the conventional RNA imaging approaches suffer from low multiplexing capacity. Recent advances in multiplexed fluorescence in situ hybridization (FISH) methods enable comprehensive RNA profiling in individual cells in situ. These technologies will have wide applications in many biological and biomedical fields, including cell type classification, signaling network analysis, tissue architecture, disease diagnosis and patient stratification, etc. In this minireview, we will present the recent technological advances of multiplexed single-cell in situ RNA profiling assays, discuss their advantages and limitations, describe their biological applications, highlight the current challenges, and propose potential solutions.


2021 ◽  
Author(s):  
Richard C Lavin ◽  
Shumin Tan

A hallmark of Mycobacterium tuberculosis (Mtb) infection with critical impact on disease development and outcome is the marked heterogeneity that exists, spanning differences in lesion types to changes in microenvironment as the infection progresses1-7. A mechanistic understanding of how this heterogeneity affects Mtb growth and treatment efficacy necessitates single bacterium-level studies in the context of intact host tissue architecture; however, such an evaluation has been technically challenging. Here, we exploit fluorescent reporter Mtb strains and the C3HeB/FeJ murine model in an integrated imaging approach to study microenvironment heterogeneity within a single lesion in situ, and analyze how these differences relate to non-uniformity in Mtb replication state, activity, and drug efficacy. We show that the pH and chloride environments differ spatially in a caseous necrotic lesion, with increased acidity and chloride levels in the lesion cuff versus the necrotic core. Conversely, a higher percentage of Mtb in the necrotic core versus the lesion cuff were in an actively replicating state, and correspondingly active in transcription and translation. Finally, examination of three first-line anti-tubercular drugs showed that efficacy of isoniazid was strikingly poor against bacteria in the lesion cuff. Our study reveals spatial relationships of intra-lesion heterogeneity, sheds light on important considerations in the development of anti-tubercular treatment strategies, and establishes a foundational framework for Mtb infection heterogeneity analysis at the single cell level in situ.


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