scholarly journals Vascular Tissue-Specific Expression of Bnac4.BOR1;1c, an Efflux Boron Transporter Gene, is Regulated in Response to Boron Availability for Efficient Boron Acquisition in Brassica Napus

2021 ◽  
Author(s):  
Sheliang Wang ◽  
Ling Liu ◽  
Dan Zou ◽  
Yupu Huang ◽  
Zhe Zhao ◽  
...  
Keyword(s):  
Brassica Napus ◽  
Vascular Bundle ◽  
Vascular Tissue ◽  
Physiological Role ◽  
Mrna Abundance ◽  
Translation Efficiency ◽  
Specific Expression ◽  
Polar Localization ◽  
B Uptake

Abstract AimsBnaC4.BOR1;1c is required for B acquisition in Brassica napus (B. napus) under low B stress. This study aimed to reveal the B regulatory mechanism of BnaC4.BOR1;1c and its physiological roles in B translocation from roots to shoots and B distribution in shoots. MethodsTransgenic Arabidopsis plants expressing GUS (β-glucuronidase) under different promoters were generated and the mRNA, and GUS activity was quantitatively measured. The in-situ PCR and immunohistochemistry in B. napus were performed to investigate BnaC4.BOR1;1c expression pattern and localization. Furthermore, assays of B transport and distribution in wild type B. napus and BnaC4.BOR1;1c RNAi lines were carried out to elucidate its physiological roles. ResultsResults showed that BnaC4.BOR1;1c mRNA abundance is negatively correlated with B availability, which was mediated by the 29 nt sequence in the 5’ terminal region of 5’-UTR. Besides, the 5’-UTR simultaneously regulates protein expression level, most probably depending on the translation efficiency. BnaC4.BOR1;1c mainly localizes on the plasma membrane of vascular bundle cells in roots and shoots with a polar localization manner that is probably beneficial to B xylem loading in roots and B unloading from xylem to phloem in vascular bundle of shoots. Short-term 10B uptake analysis demonstrates that BnaC4.BOR1;1c preferentially distributes B to developing leaves and flowers under B deficiency. ConclusionThis study reveals combined regulatory action of mRNA abundance and translation efficiency mediated by the 5’-UTR in BnaC4.BOR1;1c in response to B availability and its physiological role in preferential B acquisition in developing tissues of B. napus.

In situ localization of storage protein mRNAs in developing meristems of Brassica napus embryos

Development ◽  
1991 ◽  
Vol 111 (2) ◽  
pp. 299-313 ◽  
Author(s):  
D.E. Fernandez ◽  
F.R. Turner ◽  
M.L. Crouch
Keyword(s):  
Brassica Napus ◽  
Storage Protein ◽  
Shoot Apical Meristem ◽  
Apical Meristem ◽  
Vascular Tissue ◽  
Leaf Primordia ◽  
Root Apical Meristem ◽  
Cdna Clones ◽  
Final Maturation

Probes derived from cDNA clones of napin and cruciferin, the major storage proteins of Brassica napus, and in situ hybridization techniques were used to examine changes in the spatial and temporal distribution of storage protein messages during the course of embryogeny, with a special emphasis on the developing apical meristems. Napin mRNAs begin to accumulate in the cortex of the axis during late heart stage, in the outer faces of the cotyledons during torpedo stage and in the inner faces of the cotyledons during cotyledon stage. Cruciferin mRNAs accumulate in a similar pattern but approximately 5 days later. Cells in the apical regions where root and shoot meristems develop do not accumulate storage protein messages during early stages of embryogeny. In the upper axis, the boundary between these apical cells and immediately adjacent cells that accumulate napin and cruciferin mRNAs is particularly distinct. Our analysis indicates that this boundary is not related to differences in tissue or cell type, but appears instead to be coincident with the site of a particular set of early cell divisions. A major change in the mRNA accumulation patterns occurs halfway through embryogeny, as the embryos enter maturation stage and start drying down. Final maturation of the shoot apical meristem is associated with the development of leaf primordia and the accumulation of napin mRNAs in the meristem, associated leaf primordia and vascular tissue. Cruciferin mRNAs accumulate only in certain zones of the shoot apical meristem and on the flanks of leaf primordia. Neither type of mRNA accumulates in the root apical meristem at any stage.

Detection and mapping of interactions between chromatin and the nuclear envelope in drosophila embryos

1995 ◽  
Vol 53 ◽  
pp. 764-765
Author(s):  
W.F. Marshall ◽  
A.F. Dernburg ◽  
B. Harmon ◽  
J.W. Sedat
Keyword(s):  
Nuclear Envelope ◽  
Chromatin Condensation ◽  
Three Dimensional ◽  
Physiological Role ◽  
Nuclear Surface ◽  
Intact Cells ◽  
Molecular Determinants ◽  
Surface Harmonic ◽  
Drosophila Embryos

Interactions between chromatin and nuclear envelope (NE) have been implicated in chromatin condensation, gene regulation, nuclear reassembly, and organization of chromosomes within the nucleus. To further investigate the physiological role played by such interactions, it will be necessary to determine which loci specifically interact with the nuclear envelope. This will not only facilitate identification of the molecular determinants of this interaction, but will also allow manipulation of the pattern of chromatin-NE interactions to probe possible functions. We have developed a microscopic approach to detect and map chromatin-NE interactions inside intact cells.Fluorescence in situ hybridization (FISH) is used to localize specific chromosomal regions within the nucleus of Drosophila embryos and anti-lamin immunofluorescence is used to detect the nuclear envelope. Widefield deconvolution microscopy is then used to obtain a three-dimensional image of the sample (Fig. 1). The nuclear surface is represented by a surface-harmonic expansion (Fig 2). A statistical test for association of the FISH spot with the surface is then performed.

A γGT-AT1A receptor transgene protects renal cortical structure in AT1 receptor-deficient mice

2004 ◽  
Vol 18 (3) ◽  
pp. 290-298 ◽  
Author(s):  
Thu H. Le ◽  
Michael I. Oliverio ◽  
Hyung-Suk Kim ◽  
Harmony Salzler ◽  
Rajesh C. Dash ◽  
...  
Keyword(s):  
Blood Pressure ◽  
Transgenic Mice ◽  
Proximal Tubule ◽  
Physiological Role ◽  
Renal Proximal Tubule ◽  
Hypoxanthine Phosphoribosyl Transferase ◽  
Wild Type ◽  
Specific Expression ◽  
Low Levels ◽  
Deficient Mice

To understand the physiological role of angiotensin type 1 (AT1) receptors in the proximal tubule of the kidney, we generated a transgenic mouse line in which the major murine AT1 receptor isoform, AT1A, was expressed under the control of the P1 portion of the γ-glutamyl transpeptidase (γGT) promoter. In transgenic mice, this promoter has been shown to confer cell-specific expression in epithelial cells of the renal proximal tubule. To avoid random integration of multiple copies of the transgene, we used gene targeting to produce mice with a single-copy transgene insertion at the hypoxanthine phosphoribosyl transferase ( Hprt) locus on the X chromosome. The physiological effects of the γGT-AT1A transgene were examined on a wild-type background and in mice with targeted disruption of one or both of the murine AT1 receptor genes ( Agtr1a and Agtr1b). On all three backgrounds, γGT-AT1A transgenic mice were healthy and viable. On the wild-type background, the presence of the transgene did not affect development, blood pressure, or kidney structure. Despite relatively low levels of expression in the proximal tubule, the transgene blunted the increase in renin expression typically seen in AT1-deficient mice and partially rescued the kidney phenotype associated with Agtr1a−/− Agtr1b−/− mice, significantly reducing cortical cyst formation by more than threefold. However, these low levels of cell-specific expression of AT1 receptors in the renal proximal tubule did not increase the low blood pressures or abolish sodium sensitivity, which are characteristic of AT1 receptor-deficient mice. Although our studies do not clearly identify a role for AT1 receptors in the proximal tubules of the kidney in blood pressure homeostasis, they support a major role for these receptors in modulating renin expression and in maintaining structural integrity of the renal cortex.

scholarly journals Engineering the mechanical and biological properties of nanofibrous vascular grafts for in situ vascular tissue engineering

2017 ◽  
Vol 9 (3) ◽  
pp. 035007 ◽  
Author(s):  
Jeffrey J D Henry ◽  
Jian Yu ◽  
Aijun Wang ◽  
Randall Lee ◽  
Jun Fang ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Vascular Tissue ◽  
Vascular Grafts ◽  
Biological Properties ◽  
Vascular Tissue Engineering

scholarly journals Localization of type 1 17beta-hydroxysteroid dehydrogenase mRNA and protein in syncytiotrophoblasts and invasive cytotrophoblasts in the human term villi

2000 ◽  
Vol 165 (2) ◽  
pp. 217-222 ◽  
Author(s):  
M Bonenfant ◽  
PR Provost ◽  
R Drolet ◽  
Y Tremblay
Keyword(s):  
In Situ Hybridization ◽  
Hydroxysteroid Dehydrogenase ◽  
Binding Activity ◽  
Placental Lactogen ◽  
Specific Expression ◽  
P450 Aromatase ◽  
Chain Cleavage ◽  
Extravillous Cytotrophoblasts

The 17beta-hydroxysteroid dehydrogenases (17beta-HSDs) play a key role in the synthesis of sex steroids. The hallmark of this family of enzymes is the interconversion, through their oxydoreductive reactivity at position C17, of 17-keto- and 17beta-hydroxy-steroids. Because this reaction essentially transforms steroids having low binding activity for the steroid receptor to their more potent 17beta-hydroxysteroids isoforms, it is crucial to the control of the physiological activities of both estrogens and androgens. The human placenta produces large amounts of progesterone and estrogens throughout pregnancy. The placental type 1 17beta-HSD enzyme (E17beta-HSD) catalyzes the reduction of the low activity estrogen, estrone, into the potent estrogen, estradiol. We studied the cell-specific expression of type 1 17beta-HSD in human term placental villous tissue by combining in situ hybridization to localize type 1 17beta-HSD mRNA with immunohistochemistry using an antibody against human placental lactogen, a trophoblast marker. Immunolocalization of E17beta-HSD was also performed. To ascertain whether other steroidogenic enzymes are present in the same cell type, cytochrome P450 cholesterol side-chain cleavage (P450scc), P450 aromatase, and type 1 3beta-hydroxysteroid dehydrogenase (3beta-HSD) were also localized by immunostaining. Our results showed that the syncytium is the major steroidogenic unit of the fetal term villi. In fact, type 1 17beta-HSD mRNA and protein, as well as P450scc, P450 aromatase, and 3beta-HSD immunoreactivities were found in these cells. In addition, our results revealed undoubtedly that extravillous cytotrophoblasts (CTBs), e.g. those from which cell columns of anchoring villous originate, also express the type 1 17beta-HSD gene. However, CTBs lying beneath the syncytial layer, e.g. those from which syncytiotrophoblasts develop, contained barely detectable amounts of type 1 17beta-HSD mRNA as determined by in situ hybridization. These findings, along with those from other laboratories confirm the primordial role of the syncytium in the synthesis of steroids during pregnancy. In addition, our results indicate for the first time that CTBs differentiating along the invasive pathway contain type 1 17beta-HSD mRNA.

scholarly journals Experimental approach to overexpress pituitary adenylate cyclase activating polypeptide (PACAP) specifically in the hypothalamus

2021 ◽  
Author(s):  
◽  
Yamna Rahim
Keyword(s):  
Energy Expenditure ◽  
Experimental Approach ◽  
Health Condition ◽  
Specific Expression ◽  
Steroidogenic Factor 1 ◽  
Adaptive Thermogenesis ◽  
Brown Adipose ◽  
Pituitary Adenylate Cyclase ◽  
The Brain

Obesity is adetrimental health condition that occurs when energy intake, exceeds energy expenditure. Pituitary adenylatecyclase-activating polypeptide (PACAP) regulates energy expenditure, including adaptive thermogenesis, through the hypothalamic-sympatheticnervous system-brown adipose tissue axis. We hypothesize that PACAP expression in the ventromedial nucleus (VMN) is required for adaptive thermogenesis. To assess this, our goal is to develop an animal model that expresses PACAP specifically in the VMN of the hypothalamus. As a first step to achieving this goal, we established a protocol to deliver an adeno-associatedvirus (AAV) expressing the visible protein eGFP under the control of a VMN-specific promoter, steroidogenic factor 1 (SF1) using stereotaxic surgery. A second step was to develop a protocol to detect PACAP mRNA in the brain using in situ hybridization. Our results showed that the stereotaxic protocol was successful and provides significant progress towards achieving PACAP-specific expression in the VMN.

scholarly journals Vasodilator-stimulated phosphoprotein protects against vascular inflammation and insulin resistance

2014 ◽  
Vol 307 (7) ◽  
pp. E571-E579 ◽  
Author(s):  
Andrew M. Cheng ◽  
Norma Rizzo-DeLeon ◽  
Carole L. Wilson ◽  
Woo Je Lee ◽  
Sanshiro Tateya ◽  
...  
Keyword(s):  
Insulin Resistance ◽  
Bone Marrow ◽  
Insulin Sensitivity ◽  
Vascular Tissue ◽  
Vascular Inflammation ◽  
Physiological Role ◽  
Whole Body ◽  
Aortic Tissue ◽  
Protective Effects ◽  
Downstream Mediator

Among the pleotropic effects of endothelial nitric oxide (NO) is protection against vascular inflammation during high-fat diet (HFD) feeding. The current work investigated the role of the enzyme vasodilatory-stimulated phosphoprotein (VASP) as a downstream mediator of the anti-inflammatory effect of NO signaling in vascular tissue. Relative to mice fed a low-fat diet (LFD), levels of VASP Ser239 phosphorylation, a marker of VASP activation, were dramatically reduced in aortic tissue of mice with obesity induced by consuming a HFD. As reported previously, the effect of the HFD was associated with increased aortic inflammation, as measured by increased NF-κB-dependent gene expression, and reduced vascular insulin sensitivity (including insulin-stimulated phosphorylation of eNOS and Akt). These effects of the HFD were recapitulated by VASP knockout, implying a physiological role for VASP to constrain inflammatory signaling and thereby maintain vascular insulin sensitivity. Conversely, overexpression of VASP in endothelial cells blocked inflammation and insulin resistance induced by palmitate. The finding that transplantation of bone marrow from VASP-deficient donors into normal recipients does not recapitulate the vascular effects of whole body VASP deficiency suggests that the protective effects of this enzyme are not mediated in immune or other bone marrow-derived cells. These studies implicate VASP as a downstream mediator of the NO/cGMP pathway that is both necessary and sufficient to protect against vascular inflammation and insulin resistance. As such, this work identifies VASP as a potential therapeutic target in the treatment of obesity-related vascular dysfunction.

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