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Tissue Differentiation (tissue + differentiation)
Selected AbstractsGene expression in skeletal tissues: application of laser capture microdissectionJOURNAL OF MICROSCOPY, Issue 1 2005D. Benoyahu Summary Tissue differentiation is based on the expression of transcription factors, receptors for cytokines, and nuclear receptors that regulate a specific phenotype. The purpose of this study was to select cells from various skeletal tissues in order to analyse differential gene expression of cells in the native environment in vivo. It is a difficult task to obtain cells from skeletal tissues, such as cartilage, periost, bone and muscle, that are structured together and do not exist as individual organs. We used laser capture microdissection which permits the selection and isolation of individual cells from tissue sections. The RNA isolated from these tissues was used for reverse transcriptase-polymerase chain reactions for molecular analysis. We analysed the expression of transcription factors (cFOS, cbfa1, MyoD), receptors for cytokines, nuclear receptors, alkaline phosphatase and the structural proteins osteocalcin and collagen II. The results obtained demonstrate differential patterns of gene expression according to the tissue arrangement in their native in vivo environment, with reliable interpretation of the functions of the analysed genes in the context of intact skeletal tissue physiology. [source] Involvement of BMP-4/msx-1 and FGF pathways in neural induction in the Xenopus embryoDEVELOPMENT GROWTH & DIFFERENTIATION, Issue 4 2000Akihiko Ishimura The msx homeodomain protein is a downstream transcription factor of the bone morphogenetic protein (BMP)-4 signal and a key regulator for neural tissue differentiation. Xmsx-1 antagonizes the dorsal expression of noggin and cerberus, as revealed by in situ hybridization and reverse transcription,polymerase chain reaction assays. In animal cap explants, Xmsx-1 and BMP-4 inhibit the neural tissue differentiation induced by noggin or cerberus. A loss-of-function study using the Xmsx-1/VP-16 fusion construct indicated that neural tissue formation was directly induced by the injection of fusion ribonucleic acid, although the expression of neural cell adhesion molecule (N-CAM) in the cap was less than that in the cap injected with tBR or noggin. In contrast to the single cap assay, unexpectedly, both BMP-4 and Xmsx-1 failed to inhibit neurulation in the ectodermal explants to which the organizer mesoderm was attached. The results of cell-lineage tracing experiments indicated that the neural cells were differentiated from the animal pole tissue where the excess RNA of either BMP-4 or Xmsx-1 was injected, whereas notochord was differentiated from the organizer mesoderm. Neural tissue differentiated from BMP-4 -injected ectodermal cells strongly expressed posterior neural markers, such as hoxB9 and krox20, suggesting that the posterior neural cells differentiated regardless of the existence of the BMP signal. The introduction of a dominant-negative form of the fibroblast growth factor (FGF) receptor (XFD) into the ectodermal cells drastically reduced the expression of pan and posterior neural markers (N-CAM and hoxB-9) if co-injected with BMP-4 RNA, although XFD alone at the same dose did not shut down the expression of N-CAM in the combination explants. Therefore, it is proposed that an FGF-related molecule was involved in the direct induction of posterior neural tissue in the inducing signals from the organizer mesoderm in vivo. [source] Neuro-mesodermal patterns in artificially deformed embryonic explants: A role for mechano-geometry in tissue differentiationDEVELOPMENTAL DYNAMICS, Issue 3 2010E.S. Kornikova Abstract The mutual arrangement of neural and mesodermal rudiments in artificially bent double explants of Xenopus laevis suprablastoporal areas was compared with that of intact explants. While some of the bent explants straightened or became spherical, most retained and actively reinforced the imposed curvature, creating folds on their concave sides and expanding convex surfaces. In the intact explants, the arrangement of neural and mesodermal rudiments exhibited a distinct antero-posterior polarity, with some variability. In the bent explants, this polarity was lost: the neural rudiments were shifted towards concave while the mesodermal tissues moved towards the convex side, embracing the neural rudiments in a horseshoe-shaped manner. We associate these drastic changes in neuro-mesodermal patterning with the active extension and contraction of the convex and concave sides, respectively, triggered by the imposed deformations. We speculate that similar events are responsible for the establishment of neuro-mesodermal patterns during normal development. Developmental Dynamics 239:885,896, 2010. © 2010 Wiley-Liss, Inc. [source] PPAR: a therapeutic target in Parkinson's diseaseJOURNAL OF NEUROCHEMISTRY, Issue 2 2008Rajnish K. Chaturvedi Abstract Parkinson's disease (PD) is a progressive and chronic neurodegenerative disorder, characterized by progressive loss of dopaminergic neurons in substantia nigra. The etiology and pathogenesis of PD is still elusive, however, a large body of evidence suggests a prominent role of oxidative stress, inflammation, apoptosis, mitochondrial dysfunction and proteosomal dysfunction in the pathogenesis of PD. Due to multifactorial nature of the disease, currently available drug therapy cannot halt / slow down the disease progression, and only provides symptomatic relief. Peroxisome proliferator-activated receptor (PPAR), a member of nuclear receptor superfamily, regulates development, tissue differentiation, inflammation, mitochondrial function, wound healing, lipid metabolism and glucose metabolism. Recently, several PPAR agonists were shown to exert neuroprotective activity against oxidative damage, inflammation and apoptosis in several neurodegenerative disorders including Alzheimer's, Parkinson's, Huntington's, amyotrophic lateral sclerosis and multiple sclerosis. Similarly, regular intake of PPAR activating non-steroidal anti-inflammatory drugs such as indomethacin and ibuprofen was associated with reduced incidence and progression of neurodegenerative disorders in several epidemiological studies. In this article, we review studies relating to the neuroprotective effect of PPAR agonists in in vitro and in vivo models of PD. Similarly, the pharmacological mechanism in neuroprotective actions of PPAR agonists is also reviewed. In conclusion, PPAR agonists exert neuroprotective actions by regulating the expression of a set of genes involved in cell survival processes, and could be a therapeutic target in debilitating neurodegenerative illnesses such as PD. [source] CYR61 (CCN1) Protein Expression during Fracture Healing in an Ovine Tibial Model and Its Relation to the Mechanical Fixation StabilityJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 2 2006Jasmin Lienau Abstract The formation of new blood vessels is a prerequisite for bone healing. CYR61 (CCN1), an extracellular matrix-associated signaling protein, is a potent stimulator of angiogenesis and mesenchymal stem cell expansion and differentiation. A recent study showed that CYR61 is expressed during fracture healing and suggested that CYR61 plays a significant role in cartilage and bone formation. The hypothesis of the present study was that decreased fixation stability, which leads to a delay in healing, would lead to reduced CYR61 protein expression in fracture callus. The aim of the study was to quantitatively analyze CYR61 protein expression, vascularization, and tissue differentiation in the osteotomy gap and relate to the mechanical fixation stability during the course of healing. A mid-shaft osteotomy of the tibia was performed in two groups of sheep and stabilized with either a rigid or semirigid external fixator, each allowing different amounts of interfragmentary movement. The sheep were sacrificed at 2, 3, 6, and 9 weeks postoperatively. The tibiae were tested biomechanically and histological sections from the callus were analyzed immunohistochemically with regard to CYR61 protein expression and vascularization. Expression of CYR61 protein was upregulated at the early phase of fracture healing (2 weeks), decreasing over the healing time. Decreased fixation stability was associated with a reduced upregulation of the CYR61 protein expression and a reduced vascularization at 2 weeks leading to a slower healing. The maximum cartilage callus fraction in both groups was reached at 3 weeks. However, the semirigid fixator group showed a significantly lower CYR61 immunoreactivity in cartilage than the rigid fixator group at this time point. The fraction of cartilage in the semirigid fixator group was not replaced by bone as quickly as in the rigid fixator group leading to an inferior histological and mechanical callus quality at 6 weeks and therefore to a slower healing. The results supply further evidence that CYR61 may serve as an important regulator of bone healing. © 2005 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res [source] Requirement of spermidine for developmental transitions in Aspergillus nidulansMOLECULAR MICROBIOLOGY, Issue 3 2002Yuan Jin Summary Deletion of the spermidine synthase gene in the fungus Aspergillus nidulans results in a strain, ,spdA, which requires spermidine for growth and accumulates putrescine as the sole polyamine. Vegetative growth but not sporulation or sterigmatocystin production is observed when ,spdA is grown on media supplemented with 0.05,0.10 mM exogenous spermidine. Supplementation of ,spdA with , 0.10 mM spermidine restores sterigmatocystin production and , 0.50 mM spermidine produces a phenotype with denser asexual spore production and decreased radial hyphal growth compared with the wild type. ,spdA spores germinate in unsupplemented media but germ tube growth ceases after 8 h upon which time the spores swell to approximately three times their normal diameter. Hyphal growth is resumed upon addition of 1.0 mM spermidine. Suppression of a G protein signalling pathway could not force asexual sporulation and sterigmatocystin production in ,spdA strains grown in media lacking spermidine but could force both processes in ,spdA strains supplemented with 0.05 mM spermidine. These results show that increasing levels of spermidine are required for the transitions from (i) germ tube to hyphal growth and (ii) hyphal growth to tissue differentiation and secondary metabolism. Suppression of G protein signalling can over-ride the spermidine requirement for the latter but not the former transition. [source] Time-course of lipoxygenase, antioxidant enzyme activities and H2O2 accumulation during the early stages of Rhizobium,legume symbiosisNEW PHYTOLOGIST, Issue 1 2001Pablo Bueno Summary ,,The involvement of lipoxygenase and antioxidant enzyme activities as well as hydrogen peroxide (H2O2) accumulation are reported during early infection steps in alfalfa (Medicago sativa) roots inoculated either with a wild type Sinorhizobium meliloti or with a mutant defective in Nod-factor synthesis (Nod C,). ,,Compatibility between M. sativa and Rhizobium correlates, at least in part, with an increase in the activities of these enzymes, particularly catalase and lipoxygenase, during the preinfection period (up to 12 h). The mutant strain, defective in Nod-factor biosynthesis, showed a decrease in all enzyme activities assayed, and an increase in H2O2 accumulation. ,,Enhancement of scavenging activities for several reactive oxygen species correlated with compatibility of the S. meliloti,alfalfa symbiosis, whereas the Nod C, strain triggered a defence response. Nod factors were essential to suppress this response. ,,Increase in lipoxygenase and lipid hydroperoxide decomposing activities, observed during the first hours after inoculation with a compatible strain, could be related to tissue differentiation and/or the production of signal molecules involved in autoregulation of nodulation by the plant. [source] Technical note: Terahertz imaging of ancient mummies and boneAMERICAN JOURNAL OF PHYSICAL ANTHROPOLOGY, Issue 3 2010Lena Öhrström Abstract Ancient mummified soft-tissues are a unique source to study the evolution of disease. Diagnostic imaging of such historic tissues is of foremost interest in paleoanthropology or paleopathology, with conventional x-ray and computed tomography (CT) being the gold-standard. Longer wavelength radiation in the far-infrared or Terahertz region allows diagnostic close-to-surface tissue differentiation of bone morphology while being harmless to human cells. The aim of this study is to show the feasibility and the morpho-diagnostic impact of THz imaging of historic remains. Images of an artificially embalmed ancient Egyptian human mummy hand, an artificially embalmed ancient Egyptian mummified fish and a macerated human lumbar vertebra were obtained by THz-pulse imaging and compared with conventional X-ray and CT images. Although conventional x-ray imaging provides higher spatial resolution, we found that THz-imaging is well-suited for the investigation of ancient mummified soft tissue and embalming-related substances / wrappings. In particular, bone and cartilaginous structures can be well differentiated from surrounding soft-tissues and bandage-wrappings by THz imaging. Furthermore, THz-pulse imaging also measures the time-delay of the pulsed signal when passing through the sample, which provides supplementary information on the optical density of the sample that is not obtained by X-ray and CT. Terahertz radiation provides a completely non-invasive diagnostic imaging modality for historic dry specimens. We anticipate this modality also to be used for detection of hidden objects in historic samples such as funerary amulets still in situ in wrapped mummies, as well as potentially for the identification of spectral signatures from chemical substances, e.g., in embalming essences.. Am J Phys Anthropol, 2010. © 2010 Wiley-Liss, Inc. [source] Transcriptional profiling and biochemical analysis of mechanically induced cartilaginous tissues in a rat modelARTHRITIS & RHEUMATISM, Issue 4 2010Kristy T. Salisbury Palomares Objective To characterize patterns of molecular expression that lead to cartilage formation in vivo in a postnatal setting, by profiling messenger RNA expression across the time course of mechanically induced chondrogenesis. Methods Retired breeder Sprague-Dawley rats underwent a noncritical-sized transverse femoral osteotomy. Experimental animals (n = 45) were subjected to bending stimulation (60° cyclic motion in the sagittal plane for 15 minutes/day) of the osteotomy gap beginning on day 10 after the operation. Control animals (n = 32) experienced continuous rigid fixation. Messenger RNA isolated on days 10, 17, 24, and 38 after surgery was analyzed using a microarray containing 608 genes involved in skeletal development, tissue differentiation, fracture healing, and mechanotransduction. The glycosaminoglycan (GAG) content in the stimulated tissues was compared with that in native articular cartilage as a means of assessing the progression of chondrogenic development of the tissues. Results The majority of the 100 genes that were differentially expressed were up-regulated in response to mechanical stimulation. Many of these genes are associated with articular cartilage development and maintenance, diarthrodial joint development, cell adhesion, extracellular matrix synthesis, signal transduction, and skeletal development. Quantitative real-time polymerase chain reaction results were consistent with the microarray findings. The GAG content of the stimulated tissues increased over time and was no different from that of articular cartilage on day 38 after surgery. Conclusion Our findings indicate that mechanical stimulation causes up-regulation of genes that are principally involved in joint cavity morphogenesis and critical to articular cartilage function. Further study of this type of stimulation may identify key signaling events required for postnatal hyaline cartilage formation. [source] Developmental microRNA expression profiling of murine embryonic orofacial tissueBIRTH DEFECTS RESEARCH, Issue 7 2010Partha Mukhopadhyay Abstract BACKGROUND: Orofacial development is a multifaceted process involving precise, spatio-temporal expression of a panoply of genes. MicroRNAs (miRNAs), the largest family of noncoding RNAs involved in gene silencing, represent critical regulators of cell and tissue differentiation. MicroRNA gene expression profiling is an effective means of acquiring novel and valuable information regarding the expression and regulation of genes, under the control of miRNA, involved in mammalian orofacial development. METHODS: To identify differentially expressed miRNAs during mammalian orofacial ontogenesis, miRNA expression profiles from gestation day (GD) -12, -13 and -14 murine orofacial tissue were compared utilizing miRXplore microarrays from Miltenyi Biotech. Quantitative real-time PCR was utilized for validation of gene expression changes. Cluster analysis of the microarray data was conducted with the clValid R package and the UPGMA clustering method. Functional relationships between selected miRNAs were investigated using Ingenuity Pathway Analysis. RESULTS: Expression of over 26% of the 588 murine miRNA genes examined was detected in murine orofacial tissues from GD-12,GD-14. Among these expressed genes, several clusters were seen to be developmentally regulated. Differential expression of miRNAs within such clusters wereshown to target genes encoding proteins involved in cell proliferation, cell adhesion, differentiation, apoptosis and epithelial-mesenchymal transformation, all processes critical for normal orofacial development. CONCLUSIONS: Using miRNA microarray technology, unique gene expression signatures of hundreds of miRNAs in embryonic orofacial tissue were defined. Gene targeting and functional analysis revealed that the expression of numerous protein-encoding genes, crucial to normal orofacial ontogeny, may be regulated by specific miRNAs. Birth Defects Research (Part A), 2010. © 2010 Wiley-Liss, Inc. [source] Imprinting Status of G,S, NESP55, and XL,s in Cell Cultures Derived from Human Embryonic Germ Cells: GNAS Imprinting in Human Embryonic Germ CellsCLINICAL AND TRANSLATIONAL SCIENCE, Issue 5 2009Janet L. Crane M.D. Abstract GNAS is a complex gene that through use of alternative first exons encodes signaling proteins G,s and XL,s plus neurosecretory protein NESP55. Tissue-specific expression of these proteins is regulated through reciprocal genomic imprinting in fully differentiated and developed tissue. Mutations in GNAS account for several human disorders, including McCune-Albright syndrome and Albright hereditary osteodystrophy, and further knowledge of GNAS imprinting may provide insights into variable phenotypes of these disorders. We therefore analyzed expression of G,s, NESP55, and XL,s prior to tissue differentiation in cell cultures derived from human primordia germ cells. We found that the expression of G,s was biallelic (maternal allele: 52.6%± 2.5%; paternal allele: 47.2%± 2.5%; p= 0.07), whereas NESP55 was expressed preferentially from the maternal allele (maternal allele: 81.9%± 10%; paternal allele: 18.1%± 10%; p= 0.002) and XL,s was preferentially expressed from the paternal allele (maternal allele: 2.7%± 0.3%; paternal allele: 97.3%± 0.3%; p= 0.007). These results demonstrate that imprinting of NESP55 occurs very early in development, although complete imprinting appears to take place later than 5,11 weeks postfertilization, and that imprinting of XL,s occurs very early postfertilization. By contrast, mprinting of G,s most likely occurs after 11 weeks postfertilization and after tissue differentiation. [source] | |||||||||||||