scholarly journals The Drosophila tango gene encodes a bHLH-PAS protein that is orthologous to mammalian Arnt and controls CNS midline and tracheal development

Development ◽  
1997 ◽  
Vol 124 (22) ◽  
pp. 4571-4582 ◽  
Author(s):  
M. Sonnenfeld ◽  
M. Ward ◽  
G. Nystrom ◽  
J. Mosher ◽  
S. Stahl ◽  
...  
Keyword(s):  
Cell Culture ◽  
Gene Dosage ◽  
Cell Lineage ◽  
Regulatory System ◽  
In Vivo Studies ◽  
Enhancer Element ◽  
Evolutionarily Conserved ◽  
Cns Midline ◽  
Functionally Diverse

The Drosophila single-minded and trachealess bHLH-PAS genes control transcription and development of the CNS midline cell lineage and tracheal tubules, respectively. We show that Single-minded and Trachealess activate transcription by forming dimers with the Drosophila Tango protein that is an orthologue of the mammalian Arnt protein. Both cell culture and in vivo studies show that a DNA enhancer element acts as a binding site for both Single-minded::Tango and Trachealess::Tango heterodimers and functions in controlling CNS midline and tracheal transcription. Isolation and analysis of tango mutants reveal CNS midline and tracheal defects, and gene dosage studies demonstrate in vivo interactions between single-minded::tango and trachealess::tango. These experiments support the existence of an evolutionarily conserved, functionally diverse bHLH-PAS protein regulatory system.

scholarly journals Which cytochrome P450 metabolizes phenazepam? Step by step in silico, in vitro, and in vivo studies

2018 ◽  
Vol 33 (2) ◽  
pp. 65-73 ◽  
Author(s):  
Dmitriy V. Ivashchenko ◽  
Anastasia V. Rudik ◽  
Andrey A. Poloznikov ◽  
Sergey V. Nikulin ◽  
Valeriy V. Smirnov ◽  
...  
Keyword(s):  
Cell Culture ◽  
Cytochrome P450 ◽  
In Silico ◽  
In Vivo Studies ◽  
Cyp3a Activity ◽  
Cell Culture Study ◽  
Study Results ◽  
Three Stages

Abstract Background: Phenazepam (bromdihydrochlorphenylbenzodiazepine) is the original Russian benzodiazepine tranquilizer belonging to 1,4-benzodiazepines. There is still limited knowledge about phenazepam’s metabolic liver pathways and other pharmacokinetic features. Methods: To determine phenazepam’s metabolic pathways, the study was divided into three stages: in silico modeling, in vitro experiment (cell culture study), and in vivo confirmation. In silico modeling was performed on the specialized software PASS and GUSAR to evaluate phenazepam molecule affinity to different cytochromes. The in vitro study was performed using a hepatocytes’ cell culture, cultivated in a microbioreactor to produce cytochrome P450 isoenzymes. The culture medium contained specific cytochrome P450 isoforms inhibitors and substrates (for CYP2C9, CYP3A4, CYP2C19, and CYP2B6) to determine the cytochrome that was responsible for phenazepam’s metabolism. We also measured CYP3A activity using the 6-betahydroxycortisol/cortisol ratio in patients. Results: According to in silico and in vitro analysis results, the most probable metabolizer of phenazepam is CYP3A4. By the in vivo study results, CYP3A activity decreased sufficiently (from 3.8 [95% CI: 2.94–4.65] to 2.79 [95% CI: 2.02–3.55], p=0.017) between the start and finish of treatment in patients who were prescribed just phenazepam. Conclusions: Experimental in silico and in vivo studies confirmed that the original Russian benzodiazepine phenazepam was the substrate of CYP3A4 isoenzyme.

scholarly journals Long Term Pharmacological Perturbation of Autophagy in Mice: Are HCQ Injections a Relevant Choice?

Biomedicines ◽  
2020 ◽  
Vol 8 (3) ◽  
pp. 47 ◽  
Author(s):  
Jean-Daniel Masson ◽  
Benoit Blanchet ◽  
Baptiste Periou ◽  
François-Jérôme Authier ◽  
Baharia Mograbi ◽  
...  
Keyword(s):  
Mouse Models ◽  
In Vivo Studies ◽  
Loss Of Function ◽  
Evolutionarily Conserved ◽  
Atg Genes ◽  
Conditional Inhibition ◽  
The Impact ◽  
Catabolic Process

Macroautophagy (hereafter referred to as autophagy) is an evolutionarily conserved catabolic process whose loss-of-function has been linked to a growing list of pathologies. Knockout mouse models of key autophagy genes have been instrumental in the demonstration of the critical functions of autophagy, but they display early lethality, neurotoxicity and unwanted autophagy-independent phenotypes, limiting their applications for in vivo studies. To avoid problems encountered with autophagy-null transgenic mice, we investigated the possibility of disturbing autophagy pharmacologically in the long term. Hydroxychloroquine (HCQ) ip injections were done in juvenile and adult C57bl/6j mice, at range doses adapted from the human malaria prophylactic treatment. The impact on autophagy was assessed by western-blotting, and juvenile neurodevelopment and adult behaviours were evaluated for four months. Quite surprisingly, our results showed that HCQ treatment in conditions used in this study neither impacted autophagy in the long term in several tissues and organs nor altered neurodevelopment, adult behaviour and motor capabilities. Therefore, we recommend for future long-term in vivo studies of autophagy, to use genetic mouse models allowing conditional inhibition of selected Atg genes in appropriate lineage cells instead of HCQ treatment, until it could be successfully revisited using higher HCQ doses and/or frequencies with acceptable toxicity.

scholarly journals Loss of the histone chaperone ASF1B reduces female reproductive capacity in mice

Reproduction ◽  
2016 ◽  
Vol 151 (5) ◽  
pp. 477-489 ◽  
Author(s):  
S Messiaen ◽  
J Guiard ◽  
C Aigueperse ◽  
I Fliniaux ◽  
S Tourpin ◽  
...  
Keyword(s):  
Gonad Development ◽  
Cell Lineage ◽  
Reproductive Capacity ◽  
Chromatin Modifications ◽  
Mouse Development ◽  
Differentiation Potential ◽  
Female Mice ◽  
Evolutionarily Conserved ◽  
Meiotic Entry

Abstract Anti-silencing function 1 (ASF1) is an evolutionarily conserved histone H3–H4 chaperone involved in the assembly/disassembly of nucleosome and histone modification. Two paralogous genes, Asf1a and Asf1b, exist in the mouse genome. Asf1a is ubiquitously expressed and its loss causes embryonic lethality. Conversely, Asf1b expression is more restricted and has been less studied. To determine the in vivo function of Asf1b, we generated a Asf1b-deficient mouse line (Asf1bGT(ROSA-βgeo)437) in which expression of the lacZ reporter gene is driven by the Asf1b promoter. Analysis of β-galactosidase activity at early embryonic stages indicated a correlation between Asf1b expression and cell differentiation potential. In the gonads of both male and female, Asf1b expression was specifically detected in the germ cell lineage with a peak expression correlated with meiosis. The viability of Asf1b-null mice suggests that Asf1b is dispensable for mouse development. However, these mice showed reduced reproductive capacity compared with wild-type controls. We present evidence that the timing of meiotic entry and the subsequent gonad development are affected more severely in Asf1b-null female mice than in male mice. In female mice, in addition to subfertility related to altered gamete formation, variable defects compromising the development and/or survival of their offspring were also observed. Altogether, our data indicate the importance of Asf1b expression at the time of meiotic entry, suggesting that chromatin modifications may play a central role in this process.

scholarly journals Tumor Gene Targeting Using Microspheres: Cell Culture and in vivo Studies

Drug Delivery ◽  
1997 ◽  
Vol 4 (4) ◽  
pp. 263-267 ◽  
Author(s):  
Crispin R. Dass ◽  
E. E. Decruz ◽  
T. L. Walker ◽  
M. A. Burton
Keyword(s):  
Cell Culture ◽  
Gene Targeting ◽  
In Vivo Studies ◽  
Tumor Gene

Tissue engineering: a new frontier in physiological genomics

2007 ◽  
Vol 32 (1) ◽  
pp. 28-32 ◽  
Author(s):  
Matthew C. Petersen ◽  
Jozef Lazar ◽  
Howard J. Jacob ◽  
Tetsuro Wakatsuki
Keyword(s):  
Cell Culture ◽  
High Throughput Screening ◽  
3D Culture ◽  
3D Cell Culture ◽  
Cardiac Cells ◽  
In Vivo Studies ◽  
Specific Cell ◽  
Organ Systems ◽  
Engineered Tissue

Considerable progress has been made in the last decade in the engineering and construction of a number of artificial tissue types. These constructs are typically viewed from the perspective of possible sources for implant and transplant materials in the clinical arena. However, incorporation of engineered tissues, often referred to as three-dimensional (3D) cell culture, also offers the possibility for significant advancements in research for physiological genomics. These 3D systems more readily mimic the in vivo setting than traditional 2D cell culture, and offer distinct advantages over the in vivo setting for some organ systems. As an example, cardiac cells in 3D culture 1) are more accessible for siRNA studies, 2) can be engineered with specific cell types, and 3) offer the potential for high-throughput screening of gene function. Here the state-of-the-art is reviewed and the applications for engineered tissue in genomics research are proposed. The ability to use engineered tissue in combination with genomics creates a bridge between traditional cellular and in vivo studies that is critical to enabling the transition of genetic information into mechanistic understanding of disease processes.

scholarly journals Biomaterial-Assisted Regenerative Medicine

2021 ◽  
Vol 22 (16) ◽  
pp. 8657
Author(s):  
Teruki Nii ◽  
Yoshiki Katayama
Keyword(s):  
Cell Culture ◽  
Regenerative Medicine ◽  
Drug Screening ◽  
Drug Research ◽  
Drug Evaluation ◽  
Cell Activity ◽  
The Body ◽  
In Vivo Studies

This review aims to show case recent regenerative medicine based on biomaterial technologies. Regenerative medicine has arousing substantial interest throughout the world, with “The enhancement of cell activity” one of the essential concepts for the development of regenerative medicine. For example, drug research on drug screening is an important field of regenerative medicine, with the purpose of efficient evaluation of drug effects. It is crucial to enhance cell activity in the body for drug research because the difference in cell condition between in vitro and in vivo leads to a gap in drug evaluation. Biomaterial technology is essential for the further development of regenerative medicine because biomaterials effectively support cell culture or cell transplantation with high cell viability or activity. For example, biomaterial-based cell culture and drug screening could obtain information similar to preclinical or clinical studies. In the case of in vivo studies, biomaterials can assist cell activity, such as natural healing potential, leading to efficient tissue repair of damaged tissue. Therefore, regenerative medicine combined with biomaterials has been noted. For the research of biomaterial-based regenerative medicine, the research objective of regenerative medicine should link to the properties of the biomaterial used in the study. This review introduces regenerative medicine with biomaterial.

scholarly journals Cell lineage and cell cycling analyses of the 4d micromere using live imaging in the marine annelid Platynereis dumerilii

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
B Duygu Özpolat ◽  
Mette Handberg-Thorsager ◽  
Michel Vervoort ◽  
Guillaume Balavoine
Keyword(s):  
Cell Cycle ◽  
Single Cell ◽  
Cell Fate ◽  
Cell Lineage ◽  
Growth Zone ◽  
Live Imaging ◽  
In Vivo Studies ◽  
Platynereis Dumerilii ◽  
Cell Cycling

Cell lineage, cell cycle, and cell fate are tightly associated in developmental processes, but in vivo studies at single-cell resolution showing the intricacies of these associations are rare due to technical limitations. In this study on the marine annelid Platynereis dumerilii, we investigated the lineage of the 4d micromere, using high-resolution long-term live imaging complemented with a live-cell cycle reporter. 4d is the origin of mesodermal lineages and the germline in many spiralians. We traced lineages at single-cell resolution within 4d and demonstrate that embryonic segmental mesoderm forms via teloblastic divisions, as in clitellate annelids. We also identified the precise cellular origins of the larval mesodermal posterior growth zone. We found that differentially-fated progeny of 4d (germline, segmental mesoderm, growth zone) display significantly different cell cycling. This work has evolutionary implications, sets up the foundation for functional studies in annelid stem cells, and presents newly established techniques for live imaging marine embryos.

Impairment of autophagy by TTR V30M aggregates: in vivo reversal by TUDCA and curcumin

2016 ◽  
Vol 130 (18) ◽  
pp. 1665-1675 ◽  
Author(s):  
Cristina A. Teixeira ◽  
Maria do Rosário Almeida ◽  
Maria João Saraiva
Keyword(s):  
Cell Culture ◽  
Amyloid Fibrils ◽  
Transgenic Animals ◽  
Cellular Mechanism ◽  
In Vivo Studies ◽  
Autophagic Flux ◽  
Gi Tract ◽  
Alternative Therapeutic Approach

Transthyretin (TTR)-related amyloidoses are diseases characterized by extracellular deposition of amyloid fibrils and aggregates in tissues composed of insoluble misfolded TTR that becomes toxic. Previous studies have demonstrated the ability of small compounds in preventing and reversing TTR V30M deposition in transgenic mice gastrointestinal (GI) tract as well as lowering biomarkers associated with cellular stress and apoptotic mechanisms. In the present study we aimed to study TTR V30M aggregates effect in autophagy, a cellular mechanism crucial for cell survival that has been implicated in the development of several neurodegenerative diseases. We were able to demonstrate in cell culture that TTR V30M aggregates cause a partial impairment of the autophagic machinery as shown by p62 accumulation, whereas early steps of the autophagic flux remain unaffected as shown by autophagosome number evaluation and LC3 turnover assay. Our studies performed in TTR V30M transgenic animals demonstrated that tauroursodeoxycholic acid (TUDCA) and curcumin effectively reverse p62 accumulation in the GI tract pointing to the ability of both compounds to modulate autophagy additionally to mitigate apoptosis. Overall, our in vitro and in vivo studies establish an association between TTR V30M aggregates and autophagy impairment and suggest the use of autophagy modulators as an additional and alternative therapeutic approach for the treatment of TTR V30M-related amyloidosis.

Resistance to Fas-induced apoptosis in hepatocytes: role of GSH depletion by cell isolation and culture

2002 ◽  
Vol 283 (3) ◽  
pp. G709-G718 ◽  
Author(s):  
Lina Musallam ◽  
Chantal Éthier ◽  
Pierre Selim Haddad ◽  
Francine Denizeau ◽  
Marc Bilodeau
Keyword(s):  
Cell Culture ◽  
Cell Isolation ◽  
In Vivo Studies ◽  
Intracellular Glutathione ◽  
Mouse Hepatocytes ◽  
Induced Apoptosis ◽  
The Relationship ◽  
Apoptotic Induction

The involvement of reduction/oxidation (redox) state in cell sensitivity to apoptosis has been suggested by several studies in which induction of apoptosis was shown to require oxidative stress or GSH extrusion. On the other hand, biochemical studies of caspases revealed that their activation necessitates a reduced cysteine in their active site. This is ensured by maintaining intact intracellular glutathione status during apoptotic induction as reported by in vivo studies. Therefore, we investigated the relationship between intracellular glutathione levels and the sensitivity of mouse hepatocytes in culture to Fas-induced apoptosis as well as potential mechanisms responsible for this sensitivity. We found that total and reduced glutathione levels are decreased by one-half after cell isolation procedure and further decline by 25% during cell culture for 2 h in normal Williams' E medium. Cell culture in medium supplemented with cysteine and methionine maintains glutathione at a level similar to that measured just after cell isolation. Results show that the capacity of Fas to activate caspase-8 and to induce apoptosis requires important intracellular glutathione levels and high GSH/total glutathione ratio. In conclusion, the present study shows that intracellular glutathione plays an important role in maintaining the apoptotic machinery functional and is thus capable of transmitting the apoptotic signal.

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