scholarly journals Null cyp1b1 Activity in Zebrafish Leads to Variable Craniofacial Defects Associated with Altered Expression of Extracellular Matrix and Lipid Metabolism Genes

2021 ◽  
Author(s):  
Susana Alexandre-Moreno ◽  
Juan-Manuel Bonet-Fernández ◽  
Raquel Atienzar-Aroca ◽  
José-Daniel Aroca-Aguilar ◽  
Julio Escribano
Keyword(s):  
Extracellular Matrix ◽  
Lipid Metabolism ◽  
Molecular Mechanisms ◽  
Congenital Glaucoma ◽  
Incomplete Penetrance ◽  
Loss Of Function ◽  
Altered Expression ◽  
Matrix Gene ◽  
Lipid Metabolism Genes ◽  
Craniofacial Defects

CYP1B1 loss-of-function (LoF) is the main known genetic alteration present in recessive primary congenital glaucoma (PCG), an infrequent disease characterized by delayed embryonic development of the ocular iridocorneal angle and caused by poorly understood molecular mechanisms. To model CYP1B1 LoF underlying PCG, we developed a cyp1b1 knockout (KO) zebrafish line using CRISPR/Cas9 genome editing. This line carries the c.535_667del frameshift mutation that results in a 72% mRNA reduction with residual mRNA predicted to produce an inactive truncated protein (p.(His179Glyfs*6)). Craniofacial defects and jaw maldevelopment were observed in 23% of somatic mosaic F0 crispant larvae (144 hpf). These early phenotypes were not detected in KO F3 larvae (144 hpf) but 27% of adult fishes (4 months) showed uni or bilateral craniofacial alterations, indicating the existence of incomplete penetrance and variable expressivity. These phenotypes increased to 86% in the adult offspring of inbred progenitors with craniofacial defects. No glaucoma-related phenotypes were observed in the cyp1b1 mutants. Transcriptomic analyses of the offspring (7dpf) of KO cyp1b1 progenitors with adult-onset craniofacial defects revealed that differentially expressed genes were functionally enriched in groups related with extracellular matrix and cell adhesion, cell growth and proliferation, lipid metabolism (retinoids, steroids, and fatty acids, and oxidation-reduction processes which included several cytochrome P450 genes) and inflammation. In summary, this study shows the complexity of phenotypes and molecular pathways associated with cyp1b1 LoF, with species-dependency, and provides evidence for dysregulation of extracellular matrix gene expression as one of the mechanisms underlaying pathogenicity associated with cyp1b1 disruption.

scholarly journals Null cyp1b1 Activity in Zebrafish Leads to Variable Craniofacial Defects Associated with Altered Expression of Extracellular Matrix and Lipid Metabolism Genes

2021 ◽  
Vol 22 (12) ◽  
pp. 6430
Author(s):  
Susana Alexandre-Moreno ◽  
Juan-Manuel Bonet-Fernández ◽  
Raquel Atienzar-Aroca ◽  
José-Daniel Aroca-Aguilar ◽  
Julio Escribano
Keyword(s):  
Extracellular Matrix ◽  
Lipid Metabolism ◽  
Molecular Mechanisms ◽  
Congenital Glaucoma ◽  
Incomplete Penetrance ◽  
Loss Of Function ◽  
Altered Expression ◽  
Matrix Gene ◽  
Lipid Metabolism Genes ◽  
Craniofacial Defects

CYP1B1 loss of function (LoF) is the main known genetic alteration present in recessive primary congenital glaucoma (PCG), an infrequent disease characterized by delayed embryonic development of the ocular iridocorneal angle; however, the underlying molecular mechanisms are poorly understood. To model CYP1B1 LoF underlying PCG, we developed a cyp1b1 knockout (KO) zebrafish line using CRISPR/Cas9 genome editing. This line carries the c.535_667del frameshift mutation that results in the 72% mRNA reduction with the residual mRNA predicted to produce an inactive truncated protein (p.(His179Glyfs*6)). Microphthalmia and jaw maldevelopment were observed in 23% of F0 somatic mosaic mutant larvae (144 hpf). These early phenotypes were not detected in cyp1b1-KO F3 larvae (144 hpf), but 27% of adult (four months) zebrafish exhibited uni- or bilateral craniofacial alterations, indicating the existence of incomplete penetrance and variable expressivity. These phenotypes increased to 86% in the adult offspring of inbred progenitors with craniofacial defects. No glaucoma-related phenotypes were observed in cyp1b1 mutants. Transcriptomic analyses of the offspring (seven dpf) of cyp1b1-KO progenitors with adult-onset craniofacial defects revealed functionally enriched differentially expressed genes related to extracellular matrix and cell adhesion, cell growth and proliferation, lipid metabolism (retinoids, steroids and fatty acids and oxidation–reduction processes that include several cytochrome P450 genes) and inflammation. In summary, this study shows the complexity of the phenotypes and molecular pathways associated with cyp1b1 LoF, with species dependency, and provides evidence for the dysregulation of extracellular matrix gene expression as one of the mechanisms underlying the pathogenicity associated with cyp1b1 disruption.

scholarly journals Acupuncture on ST36, CV4 and KI1 Suppresses the Progression of Methionine- and Choline-Deficient Diet-Induced Nonalcoholic Fatty Liver Disease in Mice

Metabolites ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 299 ◽  
Author(s):  
Xiangjin Meng ◽  
Xin Guo ◽  
Jing Zhang ◽  
Junji Moriya ◽  
Junji Kobayashi ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Liver Disease ◽  
Fatty Liver ◽  
Nonalcoholic Fatty Liver Disease ◽  
Fatty Liver Disease ◽  
Molecular Mechanisms ◽  
Acupuncture Treatment ◽  
Lipid Analysis ◽  
Altered Expression ◽  
Nonalcoholic Fatty Liver

Nonalcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases worldwide, and its treatment remain a constant challenge. A number of clinical trials have shown that acupuncture treatment has beneficial effects for patients with NAFLD, but the molecular mechanisms underlying its action are still largely unknown. In this study, we established a mouse model of NAFLD by administering a methionine- and choline-deficient (MCD) diet and selected three acupoints (ST36, CV4, and KI1) or nonacupoints (sham) for needling. We then investigated the effects of acupuncture treatment on the progression of NAFLD and the underlying mechanisms. After two weeks of acupuncture treatment, the liver in the needling-nonapcupoint group (NG) mice appeared pale and yellowish in color, while that in the needling-acupoint group (AG) showed a bright red color. Histologically, fewer lipid droplets and inflammatory foci were observed in the AG liver than in the NG liver. Furthermore, the expression of proinflammatory signaling factors was significantly downregulated in the AG liver. A lipid analysis showed that the levels of triglyceride (TG) and free fatty acid (FFA) were lower in the AG liver than in the NG liver, with an altered expression of lipid metabolism-related factors as well. Moreover, the numbers of 8-hydroxy-2′-deoxyguanosine (8-OHdG)-positive hepatocytes and levels of hepatic thiobarbituric acid reactive substances (TBARS) were significantly lower in AG mice than in NG mice. In line with these results, a higher expressions of antioxidant factors was found in the AG liver than in the NG liver. Our results indicate that acupuncture repressed the progression of NAFLD by inhibiting inflammatory reactions, reducing oxidative stress, and promoting lipid metabolism of hepatocytes, suggesting that this approach might be an important complementary treatment for NAFLD.

scholarly journals Altered expression patterns of lipid metabolism genes in an animal model of HCV core-related, nonobese, modest hepatic steatosis

BMC Genomics ◽  
2008 ◽  
Vol 9 (1) ◽  
pp. 109 ◽  
Author(s):  
Ming-Ling Chang ◽  
Chau-Ting Yeh ◽  
Jeng-Chang Chen ◽  
Chau-Chun Huang ◽  
Shi-Ming Lin ◽  
...  
Keyword(s):  
Animal Model ◽  
Lipid Metabolism ◽  
Hepatic Steatosis ◽  
Expression Patterns ◽  
Altered Expression ◽  
Lipid Metabolism Genes

scholarly journals Bioreactor-Controlled Physoxia Regulates TGF-β Signaling to Alter Extracellular Matrix Synthesis by Human Chondrocytes

2019 ◽  
Vol 20 (7) ◽  
pp. 1715 ◽  
Author(s):  
Holger Jahr ◽  
Seval Gunes ◽  
Annika-Ricarda Kuhn ◽  
Sven Nebelung ◽  
Thomas Pufe
Keyword(s):  
Extracellular Matrix ◽  
Oxygen Tension ◽  
Molecular Mechanisms ◽  
Matrix Metalloproteases ◽  
Human Chondrocytes ◽  
Marker Genes ◽  
Matrix Synthesis ◽  
Altered Expression ◽  
Positive Effects ◽  
Marker Expression

Culturing articular chondrocytes under physiological oxygen tension exerts positive effects on their extracellular matrix synthesis. The underlying molecular mechanisms which enhance the chondrocytic phenotype are, however, still insufficiently elucidated. The TGF-β superfamily of growth factors, and the prototypic TGF-β isoforms in particular, are crucial in maintaining matrix homeostasis of these cells. We employed a feedback-controlled table-top bioreactor to investigate the role of TGF-β in microtissues of human chondrocytes over a wider range of physiological oxygen tensions (i.e., physoxia). We compared 1%, 2.5%, and 5% of partial oxygen pressure (pO2) to the ‘normoxic’ 20%. We confirmed physoxic conditions through the induction of marker genes (PHD3, VEGF) and oxygen tension-dependent chondrocytic markers (SOX9, COL2A1). We identified 2.5% pO2 as an oxygen tension optimally improving chondrocytic marker expression (ACAN, COL2A1), while suppressing de-differentiation markers (COL1A1, COL3A1). Expression of TGF-β isoform 2 (TGFB2) was, relatively, most responsive to 2.5% pO2, while all three isoforms were induced by physoxia. We found TGF-β receptors ALK1 and ALK5 to be regulated by oxygen tension on the mRNA and protein level. In addition, expression of type III co-receptors betaglycan and endoglin appeared to be regulated by oxygen tension as well. R-Smad signaling confirmed that physoxia divergently regulated phosphorylation of Smad1/5/8 and Smad2/3. Pharmacological inhibition of canonical ALK5-mediated signaling abrogated physoxia-induced COL2A1 and PAI-1 expression. Physoxia altered expression of hypertrophy markers and that of matrix metalloproteases and their activity, as well as expression ratios of specific proteins (Sp)/Krüppel-like transcription factor family members SP1 and SP3, proving a molecular concept of ECM marker regulation. Keeping oxygen levels tightly balanced within a physiological range is important for optimal chondrocytic marker expression. Our study provides novel insights into transcriptional regulations in chondrocytes under physoxic in vitro conditions and may contribute to improving future cell-based articular cartilage repair strategies.

scholarly journals Genome-wide expression and network analyses of mutants in key brassinosteroid signaling genes

BMC Genomics ◽  
2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Razgar Seyed Rahmani ◽  
Tao Shi ◽  
Dongzhi Zhang ◽  
Xiaoping Gou ◽  
Jing Yi ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Signaling Pathway ◽  
Molecular Mechanisms ◽  
Differential Expression Analysis ◽  
Transcriptional Level ◽  
Partial Recovery ◽  
Hormone Signaling ◽  
Loss Of Function ◽  
Altered Expression ◽  
Network Analyses

Abstract Background Brassinosteroid (BR) signaling regulates plant growth and development in concert with other signaling pathways. Although many genes have been identified that play a role in BR signaling, the biological and functional consequences of disrupting those key BR genes still require detailed investigation. Results Here we performed phenotypic and transcriptomic comparisons of A. thaliana lines carrying a loss-of-function mutation in BRI1 gene, bri1–5, that exhibits a dwarf phenotype and its three activation-tag suppressor lines that were able to partially revert the bri1–5 mutant phenotype to a WS2 phenotype, namely bri1–5/bri1–1D, bri1–5/brs1–1D, and bri1–5/bak1–1D. From the three investigated bri1–5 suppressors, bri1–5/bak1–1D was the most effective suppressor at the transcriptional level. All three bri1–5 suppressors showed altered expression of the genes in the abscisic acid (ABA signaling) pathway, indicating that ABA likely contributes to the partial recovery of the wild-type phenotype in these bri1–5 suppressors. Network analysis revealed crosstalk between BR and other phytohormone signaling pathways, suggesting that interference with one hormone signaling pathway affects other hormone signaling pathways. In addition, differential expression analysis suggested the existence of a strong negative feedback from BR signaling on BR biosynthesis and also predicted that BRS1, rather than being directly involved in signaling, might be responsible for providing an optimal environment for the interaction between BRI1 and its ligand. Conclusions Our study provides insights into the molecular mechanisms and functions of key brassinosteroid (BR) signaling genes, especially BRS1.

scholarly journals Genome-Wide Expression and Network Analyses of Mutants in Key Brassinosteroid Signaling Genes

2020 ◽  
Author(s):  
Razgar Seyed Rahmani ◽  
Tao Shi ◽  
Dongzhi Zhang ◽  
Xiaoping Gou ◽  
Jing Yi ◽  
...  
Keyword(s):  
Signaling Pathways ◽  
Signaling Pathway ◽  
Molecular Mechanisms ◽  
Differential Expression Analysis ◽  
Transcriptional Level ◽  
Partial Recovery ◽  
Hormone Signaling ◽  
Loss Of Function ◽  
Altered Expression ◽  
Network Analyses

Abstract BackgroundBrassinosteroid (BR) signaling regulates plant growth and development in concert with other signaling pathways. Although many genes have been identified that play a role in Brassinosteroid (BR) signaling, the biological and functional consequences of disrupting those key BR genes still requires detailed investigation.ResultsHere we performed phenotypic and transcriptomic comparisons of A. thaliana lines carrying a loss-of-function mutation in BRI1 gene, bri1-5, that exhibits a dwarf phenotype along with its three activation-tag suppressor lines that were able to partially revert the bri1-5 mutant phenotype to a WT phenotype, namely bri1-5/bri1-1D, bri1-5/brs1-1D, bri1-5/bak1-1D. From the three investigated bri1-5 suppressors, bri1-5/bak1-1D was the most effective suppressor at the transcriptional level. All three bri1-5 suppressors showed altered expression of the genes in the abscisic acid (ABA signaling) pathway, indicating that ABA likely contributes to the partial recovery of the wild type phenotype in these bri1-5 suppressors. Network analysis revealed crosstalk between BR and other phytohormone signaling pathways, suggesting that interference with one hormone signaling pathway affects other hormone signaling pathways. In addition, differential expression analysis suggested the existence of a strong negative feedback from BR signaling on BR biosynthesis and also predicted that BRS1, rather than being directly involved in signaling, is likely responsible for providing an optimal environment for the interaction between BRI1 and its ligand. ConclusionsOur study provides insights into the molecular mechanisms and functions of key brassinosteroid (BR) signaling genes, especially BRS1.

scholarly journals Expression of lipid metabolism genes provides new insights into intramuscular fat deposition in Laiwu pigs

2020 ◽  
Vol 33 (3) ◽  
pp. 390-397 ◽  
Author(s):  
Hui Wang ◽  
Jin Wang ◽  
Dan-dan Yang ◽  
Zong-li Liu ◽  
Yong-qing Zeng ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Lipid Metabolism ◽  
Fatty Acid Synthase ◽  
Molecular Mechanisms ◽  
Intramuscular Fat ◽  
Fat Deposition ◽  
Fatty Acid Binding ◽  
Fat Removal ◽  
Lipid Metabolism Genes ◽  
Imf Content

Objective: The objective of this study was to measure the special expression pattern of lipid metabolism genes and investigate the molecular mechanisms underlying intramuscular fat (IMF) deposition in Longissimus dorsi muscle of Laiwu pigs.Methods: Thirty-six pigs (Laiwu n = 18; Duroc×Landrace×Yorkshire n = 18) were used for the measurement of the backfat thickness, marbling score, IMF content, and expression of lipid metabolism genes.Results: Significant correlations were found between IMF content and the mRNA expression of lipid metabolism genes. Of the 14 fat deposition genes measured, fatty acid synthase (FASN) showed the strongest correlation (r = 0.75, p = 0.001) with IMF content, and of the 6 fat removal genes, carnitine palmitoyl transferase 1B (CPT1B) exhibited the greatest negative correlation (r = –0.66, p = 0.003) with IMF content in Laiwu pig. Multiple regression analysis showed that CPT1B, FASN, solute carrier family 27 member 1 (SLC27A1), and fatty acid binding protein 3 (FABP3) contributed 38% of the prediction value for IMF content in Laiwu pigs. Of these four variables, CPT1B had the greatest contribution to IMF content (14%) followed by FASN (11%), SLC27A1 (9%), and FABP3 (4%).Conclusion: Our results indicate that the combined effects of an upregulation in fat deposition genes and downregulation in fat removal genes promotes IMF deposition in Laiwu pigs.

scholarly journals An integrated omics analysis reveals the gene expression profiles of rapeseed, castor bean, and maize for seed oil biosynthesis

2021 ◽  
Author(s):  
Nian Liu ◽  
Jing Liu ◽  
Shihang Fan ◽  
Hongfang Liu ◽  
Xue-Rong Zhou ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Castor Bean ◽  
Oil Content ◽  
Molecular Mechanisms ◽  
Seed Oil ◽  
Lipid Biosynthesis ◽  
Biosynthetic Enzymes ◽  
Seed Oil Content ◽  
Oil Biosynthesis ◽  
Lipid Metabolism Genes

Abstract Background Seed storage lipids are valuable for human diet and for the sustainable development of mankind. In recent decades, many lipid metabolism genes and pathways have been identified, but the molecular mechanisms that underlie species differences in seed oil biosynthesis are not fully understood. Results To investigate the molecular mechanisms of seed oil accumulation in different species, we performed comparative genome and transcriptome analyses of rapeseed and castor bean, which have high seed oil contents, and maize, which has a low seed oil content. The results uncovered the molecular mechanism of the low and high seed oil content in maize and castor bean, respectively. Transcriptome analyses showed that more than 61% of the lipid- and carbohydrate-related genes were regulated in rapeseed and castor bean, but only 20.1% of the lipid-related genes and 22.5% of the carbohydrate-related genes were regulated in maize. Compared to rapeseed and castor bean, fewer lipid biosynthesis genes but more lipid metabolism genes were regulated in the maize embryo. More importantly, most maize genes encoding lipid-related transcription factors, triacylglycerol (TAG) biosynthetic enzymes, pentose phosphate pathway (PPP) and Calvin Cycle proteins were not regulated during seed oil synthesis, despite the presence of many homologs in the maize genome. These results revealed the molecular underpinnings of the low seed oil content in maize. In castor bean, we observed differential regulation of vital oil biosynthetic enzymes and extremely high expression levels of oil biosynthetic genes, which were consistent with the rapid accumulation of oil in castor bean developing seeds. Conclusions Compared to oil seed (rapeseed and castor bean), less oil biosynthetic genes were regulated during the seed development in non-oil seed (maize). These results shed light on molecular mechanisms of lipid biosynthesis in rapeseed, castor bean, and maize. They can provide information on key target genes that may be useful for future experimental manipulation of oil production in oilseed crops.

Distinct functional classes of PDGFRB pathogenic variants in primary familial brain calcification

2021 ◽  
Author(s):  
Sandrine Lenglez ◽  
Ariane Sablon ◽  
Gilles Fénelon ◽  
Anne Boland ◽  
Jean-François Deleuze ◽  
...  
Keyword(s):  
Growth Factor ◽  
Molecular Mechanisms ◽  
Residual Activity ◽  
Receptor Expression ◽  
Platelet Derived Growth Factor ◽  
Incomplete Penetrance ◽  
Loss Of Function ◽  
Primary Familial Brain Calcification ◽  
Brain Calcification ◽  
The Impact

Abstract Platelet-derived growth factor receptor beta (PDGFRB) is one of the genes associated with primary familial brain calcification (PFBC), an inherited neurological disease (OMIM:173410). Genetic analysis of patients and families revealed at least 13 PDGFRB heterozygous missense variants, including two novel ones described in the present report. Limited experimental data published on five of these variants had suggested that they decrease the receptor activity. No functional information was available on the impact of variants located within the receptor extracellular domains. Here, we performed a comprehensive molecular analysis of PDGFRB variants linked to PFBC. Mutated receptors were transfected in various cell lines to monitor receptor expression, signaling, mitogenic activity, and ligand binding. Four mutants caused a complete loss of tyrosine kinase activity in multiple assays. One of the novel variants, p.Pro154Ser, decreased the receptor expression and abolished binding of platelet-derived growth factor (PDGF-BB). Others showed a partial loss of function related to reduced expression or signaling. Combining clinical, genetic and molecular data, we consider nine variants as pathogenic or likely pathogenic, three as benign or likely benign and one as a variant of unknown significance. We discuss the possible relationship between the variant residual activity, incomplete penetrance, brain calcification and neurological symptoms. In conclusion, we identified distinct molecular mechanisms whereby PDGFRB variants may result in a receptor loss of function. This work will facilitate genetic counselling in PFBC.

scholarly journals CPAMD8 loss-of-function underlies non-dominant congenital glaucoma with variable anterior segment dysgenesis and abnormal extracellular matrix

2020 ◽  
Vol 139 (10) ◽  
pp. 1209-1231 ◽  
Author(s):  
Juan-Manuel Bonet-Fernández ◽  
José-Daniel Aroca-Aguilar ◽  
Marta Corton ◽  
Ana-Isabel Ramírez ◽  
Susana Alexandre-Moreno ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Anterior Segment ◽  
Congenital Glaucoma ◽  
Loss Of Function ◽  
Anterior Segment Dysgenesis
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