scholarly journals Versican gene expression in human articular cartilage and comparison of mRNA splicing variation with aggrecan

1993 ◽  
Vol 291 (2) ◽  
pp. 361-367 ◽  
Author(s):  
J Grover ◽  
P J Roughley
Keyword(s):  
Alternative Splicing ◽  
Articular Cartilage ◽  
Regulatory Protein ◽  
Mrna Splicing ◽  
Human Articular Cartilage ◽  
Complement Regulatory Protein ◽  
Mature Adult ◽  
Epidermal Growth ◽  
Egf Domain

The chondrocytes in human articular cartilage from subjects of all ages express mRNAs for both of the aggregating proteoglycans aggrecan and versican, although the level of expression of versican mRNA is much lower than that of aggrecan mRNA. Aggrecan shows alternative splicing of the epidermal growth factor (EGF)-like domain within its C-terminal globular region, but there is no evidence for a major difference in situ in the relative expression of this domain with age. At all ages studied from birth to the mature adult, a greater proportion of transcripts lacked the EGF domain. The relative proportions of the two transcripts did not change upon culture and passage of isolated chondrocytes. In contrast, the neighbouring complement regulatory protein (CRP)-like domain was predominantly expressed irrespective of age, but cell culture did result in variation of the splicing of this domain. Versican possesses two EGF-like domains and one CRP-like domain, but at all ages the three domains were predominantly present in all transcripts. This situation persisted upon culture and passage of the chondrocytes. Thus, unlike aggrecan, the versican expressed by human articular cartilage does not appear to undergo alternative splicing of its C-terminal globular region, either in cartilage in situ or in chondrocytes in culture.

scholarly journals Human iPSC-derived RPE and retinal organoids reveal impaired alternative splicing of genes involved in pre-mRNA splicing in PRPF31 autosomal dominant retinitis pigmentosa

2017 ◽  
Author(s):  
Adriana Buskin ◽  
Lili Zhu ◽  
Valeria Chichagova ◽  
Basudha Basu ◽  
Sina Mozaffari-Jovin ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Retinitis Pigmentosa ◽  
Gene Editing ◽  
Autosomal Dominant ◽  
Mrna Processing ◽  
Mrna Splicing ◽  
Patient Specific ◽  
Rpe Cells ◽  
Autosomal Dominant Retinitis Pigmentosa

SummaryMutations in pre-mRNA processing factors (PRPFs) cause 40% of autosomal dominant retinitis pigmentosa (RP), but it is unclear why mutations in ubiquitously expressed PRPFs cause retinal disease. To understand the molecular basis of this phenotype, we have generated RP type 11 (PRPF31-mutated) patient-specific retinal organoids and retinal pigment epithelium (RPE) from induced pluripotent stem cells (iPSC). Impaired alternative splicing of genes encoding pre-mRNA splicing proteins occurred in patient-specific retinal cells and Prpf31+/− mouse retinae, but not fibroblasts and iPSCs, providing mechanistic insights into retinal-specific phenotypes of PRPFs. RPE was the most affected, characterised by loss of apical-basal polarity, reduced trans-epithelial resistance, phagocytic capacity, microvilli, and cilia length and incidence. Disrupted cilia morphology was observed in patient-derived-photoreceptors that displayed progressive features associated with degeneration and cell stress. In situ gene-editing of a pathogenic mutation rescued key structural and functional phenotypes in RPE and photoreceptors, providing proof-of-concept for future therapeutic strategies.eTOCPRPF31 is a ubiquitously expressed pre-mRNA processing factor that when mutated causes autosomal dominant RP. Using a patient-specific iPSC approach, Buskin and Zhu et al. show that retinal-specific defects result from altered splicing of genes involved in the splicing process itself, leading to impaired splicing, loss of RPE polarity and diminished phagocytic ability as well as reduced cilia incidence and length in both photoreceptors and RPE.HighlightsSuccessful generation of iPSC-derived RPE and photoreceptors from four RP type 11 patientsRPE cells express the mutant PRPF31 protein and show the lowest expression of wildtype proteinPRPF31 mutations result in altered splicing of genes involved in pre-mRNA splicing in RPE and retinal organoidsPrpf31 haploinsufficiency results in altered splicing of genes involved in pre-mRNA splicing in mouse retinaRPE cells display loss of polarity, reduced barrier function and phagocytosisPhotoreceptors display shorter and fewer cilia and degenerative featuresRPE cells display most abnormalities suggesting they might be the primary site of pathogenesisIn situ gene editing corrects the mutation and rescues key phenotypes

Functional in situ assessment of human articular cartilage using MRI: a whole-knee joint loading device

2017 ◽  
Vol 16 (6) ◽  
pp. 1971-1986 ◽  
Author(s):  
Sven Nebelung ◽  
Manuel Post ◽  
Stefan Raith ◽  
Horst Fischer ◽  
Matthias Knobe ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Knee Joint ◽  
Human Articular Cartilage ◽  
Joint Loading ◽  
Loading Device ◽  
In Situ Assessment ◽  
Knee Joint Loading

scholarly journals The action of human articular-cartilage metalloproteinase on proteoglycan and link protein. Similarities between products of degradation in situ and in vitro

1986 ◽  
Vol 237 (1) ◽  
pp. 117-122 ◽  
Author(s):  
I K Campbell ◽  
P J Roughley ◽  
J S Mort
Keyword(s):  
Hyaluronic Acid ◽  
Articular Cartilage ◽  
Interleukin 1 ◽  
Human Articular Cartilage ◽  
Link Protein ◽  
Protein Components ◽  
Stimulation Of ◽  
Hyaluronic Acid Binding

Interleukin 1 stimulation of human articular cartilage in organ culture produced the concomitant release of proteoglycan fragments and latent metalloproteinase. The released fragments ranged in size from that of almost intact proteoglycan subunits to the product of limiting digestion generated by the activated metalloproteinase. None of the fragments possessed the ability to interact with hyaluronic acid. Analysis of proteoglycan aggregate digested with the activated metalloproteinase showed that isolated hyaluronic acid-binding regions were produced from the proteoglycan subunits, and that the two higher-Mr link-protein components (Mr 48,000 and 44,000) were converted into the lowest-Mr component (Mr 41,000). Link protein extracted from cartilage under stimulation with interleukin 1 showed a similar conversion. These results suggest that interleukin 1 stimulates the release of latent metalloproteinase from chondrocytes and that a proportion of the enzyme is activated in situ in the cartilage matrix. The mode of action of the activated enzyme is compatible with a role in the changes in proteoglycan structure seen in aging.

The Inhomogeneous Mechanical Properties of the Pericellular Matrix of Articular Cartilage Measured In Situ by Atomic Force Microscopy

2009 ◽  
Author(s):  
Rebecca E. Wilusz ◽  
Eric M. Darling ◽  
Michael P. Bolognesi ◽  
Stefan Zauscher ◽  
Farshid Guilak
Keyword(s):  
Mechanical Properties ◽  
Atomic Force Microscopy ◽  
Articular Cartilage ◽  
Biomechanical Properties ◽  
Human Articular Cartilage ◽  
Pericellular Matrix ◽  
Force Microscopy ◽  
Narrow Region ◽  
Atomic Force

Articular cartilage is the connective tissue that lines the articulating surfaces of diarthrodial joints, providing a low-friction, load-bearing surface during joint motion. Articular cartilage comprises of a single cell type, the chondrocyte, embedded within an extensive extracellular matrix (ECM). Each chondrocyte is surrounded by a narrow region called the pericellular matrix (PCM) that is distinct from the ECM in both its biochemical composition [1] and biomechanical properties [2]. While multiple techniques have been used to measure the mechanical properties of the PCM, including micropipette aspiration of isolated chondrons [2], these studies required mechanical or enzymatic extraction of the chondrocyte and surrounding PCM (i.e., the “chondron” [1]) from the cartilage, and the influence of this isolation process on PCM properties is unknown. Atomic force microscopy (AFM) provides a high resolution form of nano- and microindentation approaches that can be used to measure local mechanical properties in situ [3,4]. The objective of this study was to use AFM to quantify the biomechanical properties of the ECM and PCM of human articular cartilage in situ.

scholarly journals A homing receptor-IgG chimera as a probe for adhesive ligands of lymph node high endothelial venules.

1990 ◽  
Vol 110 (6) ◽  
pp. 2221-2229 ◽  
Author(s):  
S R Watson ◽  
Y Imai ◽  
C Fennie ◽  
J S Geoffroy ◽  
S D Rosen ◽  
...  
Keyword(s):  
Regulatory Protein ◽  
Complement Regulatory Protein ◽  
Cloning And Sequencing ◽  
High Endothelial Venules ◽  
Homing Receptor ◽  
Protein Motifs ◽  
Calcium Dependent ◽  
Extracellular Region ◽  
Peripheral Lymph ◽  
Epidermal Growth

The binding of lymphocytes to high endothelial venules (HEV) within peripheral lymph nodes (pln) is thought to be mediated by a lectinlike adhesion molecule termed the pln homing receptor (pln HR). The cloning and sequencing of cDNAs encoding both murine and human pln HR revealed that these adhesion molecules contain protein motifs that are homologous to C-type or calcium dependent lectin domains as well as to epidermal growth factor (egf) and complement-regulatory protein domains. We have produced a novel, antibody-like form of the murine HR by joining the extracellular region of the receptor to a human IgG heavy chain. This antibody-like molecule is capable of recognizing carbohydrates, blocking the binding of lymphocytes to pln HEV, and serving as a histochemical reagent for the staining of pln HEV. This murine HR-IgG chimera should prove useful in analyzing the distribution of the HR ligand(s) in normal as well as in inflammatory states.

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