Subcellular localization of endogenous IAA during poplar leaf rhizogenesis revealed by in situ immunocytochemistry

2014 ◽  
Vol 8 (5) ◽  
pp. 377-386 ◽  
Author(s):  
Ningguang Dong ◽  
Ying Gao ◽  
Yanbin Hao ◽  
Weilun Yin ◽  
Dong Pei
Keyword(s):  
Subcellular Localization ◽  
Endogenous Iaa ◽  
Poplar Leaf

Subcellular localization of phosducin in rod photoreceptors

2005 ◽  
Vol 22 (1) ◽  
pp. 19-25 ◽  
Author(s):  
JING CHEN ◽  
TATSURO YOSHIDA ◽  
KOICHI NAKANO ◽  
MARK W. BITENSKY
Keyword(s):  
Subcellular Localization ◽  
Dark Adaptation ◽  
Outer Segment ◽  
Photoreceptor Cells ◽  
Immunogold Labeling ◽  
Synaptic Function ◽  
Immunogold Electron Microscopy ◽  
Rod Photoreceptors ◽  
Relative Paucity

Phosducin (Pd) is a 28-kD phosphoprotein whose expression in retina appears limited to photoreceptor cells. Pd binds to the β,γ subunits of transducin (Gt). Their binding affinity is markedly diminished by Pd phosphorylation. While Pd has long been regarded as a candidate for the regulation of Gt, the molecular details of Pd function remain unclear. This gap in understanding is due in part to a lack of precise information concerning the total amount and subcellular localization of rod Pd. While earlier studies suggested that Pd was a rod outer segment (ROS) protein, recent findings have demonstrated that Pd is distributed throughout the rod. In this report, the subcellular distribution and amounts of rat Pd are quantified with immunogold electron microscopy. After light or dark adaptation, retinal tissues were fixedin situand prepared for ultrathin sectioning and immunogold labeling. Pd concentrations were analyzed over the entire length of the rod. The highest Pd labeling densities were found in the rod synapse. Less intense Pd staining was observed in the ellipsoid and myoid regions, while minimal labeling densities were found in the ROS and the rod nucleus. In contrast with rod Gt, no evidence was found for light-dependent movement of Pd between inner and outer segments. There is a relative paucity of Pd in the ROS as compared with the large amounts of Gtfound there. This does not support the earlier idea that Pd could modulate Gtactivity by controlling its concentration. On the other hand, the presence of Pd in the nucleus is consistent with its possible role as a regulator of transcription. The functions of Pd in the ellipsoid and myoid regions remain unclear. The highest concentration of Pd was found at the rod synapse, consistent with a suggested role for Pd in the regulation of synaptic function.

scholarly journals In situ subcellular localization of epitope-tagged human and rabbit carboxylesterases

Cytometry ◽  
1998 ◽  
Vol 32 (3) ◽  
pp. 223-232 ◽  
Author(s):  
Philip M. Potter ◽  
Judith S. Wolverton ◽  
Christopher L. Morton ◽  
David O. Whipple ◽  
Mary K. Danks
Keyword(s):  
Subcellular Localization

scholarly journals Visualizing adenosine to inosine RNA editing in single mammalian cells

2016 ◽  
Author(s):  
Ian A. Mellis ◽  
Rohit K. Gupte ◽  
Arjun Raj ◽  
Sara H. Rouhanifard
Keyword(s):  
Subcellular Localization ◽  
Rna Editing ◽  
Nuclear Localization ◽  
Mammalian Cells ◽  
Single Cells ◽  
Frequent Type ◽  
Rna Fish

AbstractConversion of adenosine bases to inosine in RNA is a frequent type of RNA editing, but important details about its biology, including subcellular localization, remain unknown due to a lack of imaging tools. We developed an RNA FISH strategy we called inoFISH that enables us to directly visualize and quantify adenosine-to-inosine edited transcripts in situ. Applying this tool to three edited transcripts (GRIA2, EIF2AK2 and NUP43), we found that editing of these transcripts is not correlated with nuclear localization nor paraspeckle association, and that NUP43 exhibits constant editing rates between single cells while the rates for GRIA2 vary.

Changes in mitochondrial surface charge mediate recruitment of signaling molecules during apoptosis

2011 ◽  
Vol 300 (1) ◽  
pp. C33-C41 ◽  
Author(s):  
Bryan Heit ◽  
Tony Yeung ◽  
Sergio Grinstein
Keyword(s):  
Cell Death ◽  
Programmed Cell Death ◽  
Subcellular Localization ◽  
Surface Charge ◽  
Electrostatic Interactions ◽  
Negative Charge ◽  
Cytochrome C Release ◽  
Cationic Proteins ◽  
Over Expression

Electrostatic interactions with negative lipids contribute to the subcellular localization of polycationic proteins. In situ measurements using cytosolic probes of surface charge indicate that normal mitochondria are not noticeably electronegative. However, during apoptosis mitochondria accrue negative charge and acquire the ability to attract cationic proteins, including K-Ras. The marked increase in the surface charge of mitochondria occurs early in apoptosis, preceding depolarization of their inner membrane, cytochrome c release, and flipping of phosphatidylserine across the plasmalemma. Using novel biosensors, we determined that the increased electronegativity of the mitochondria coincided with and was likely attributable to increased exposure of cardiolipin, which is dianionic. Ectopic (over)expression of cardiolipin-binding proteins precluded the increase in surface charge and inhibited apoptosis, implying that mitochondrial exposure of negatively charged lipids is required for progression of programmed cell death.

In situ localization of mRNAs and proteins with both light and Electron Microscopy

1991 ◽  
Vol 49 ◽  
pp. 50-51
Author(s):  
J. A. Pollock ◽  
Seymour Benzer ◽  
T. Deerinck ◽  
M. Martone ◽  
M. H. Ellisman
Keyword(s):  
Subcellular Localization ◽  
Regional Differences ◽  
Light And Electron Microscopy ◽  
Molecular Probes ◽  
Tremendous Amount ◽  
Electron Microscopes ◽  
And Function ◽  
Microscopic Techniques ◽  
Direct Microscopic Observation

Cell biological analysis through direct microscopic observation has adopted molecular biological techniques to study the expression of particular genes. With antibodies and other molecular probes, the specific subcellular localization of a given type of protein can readily be visualized using conventional light microscopes, Confocal microscopes and electron microscopes. A tremendous amount of information has been reported on the synthesis, processing and transport of many classes of proteins resulting from the direct visualization of protein expression in the tissue. Recently, the specific subcellular localization of mRNA transcripts has been reported in several systems relating the localization of the RNA to the localization and function of the proteins they encode (Garner, et al., 1988; Macdonald and Struhl, 1988; Singer, et al., 1989; Yisraeli and Melton, 1988, 1990; Pollock et al., 1990). The success of these studies have relied on the large physical dimensions of the cells used so that the non-isotopic, light microscopic techniques employed could resolve distinct regional differences in the RNA signal.

scholarly journals Slc4a8 in the Kidney: Expression, Subcellular Localization and Role in Salt Reabsorption

2018 ◽  
Vol 50 (4) ◽  
pp. 1361-1375 ◽  
Author(s):  
Jie Xu ◽  
Sharon Barone ◽  
Kamyar Zahedi ◽  
Marybeth Brooks ◽  
Manoocher Soleimani
Keyword(s):  
In Situ Hybridization ◽  
Subcellular Localization ◽  
Collecting Duct ◽  
Basolateral Membrane ◽  
Acid Base ◽  
Duct Cells ◽  
Collecting Ducts ◽  
Collecting Duct Cells ◽  
Medullary Collecting Duct

Background/Aims: The sodium-dependent bicarbonate transporter Slc4a8 (a.k.a NDCBE) mediates the co-transport of sodium and bicarbonate in exchange for chloride. It is abundantly detected in the brain, with low expression levels in the kidney. The cell distribution and subcellular localization of Slc4a8 in the kidney and its role in acid/base and electrolyte homeostasis has been the subject of conflicting reports. There are no conclusive localization or functional studies to pinpoint the location and demonstrate the function of Slc4a8 in the kidney. Methods: Molecular techniques, including RT-PCR and in situ hybridization, were performed on kidney sections and tagged epitopes were used to examine the membrane targeting of Slc4a8 in polarized kidney cells. Crispr/Cas9 was used to generate and examine Slc4a8 KO mice. Results: Zonal distribution and in situ hybridization studies showed very little expression for Slc4a8 (NDCBE) in the cortex or in cortical collecting ducts (CCD). Slc4a8 was predominantly detected in the outer and inner medullary collecting ducts (OMCD and IMCD), and was targeted to the basolateral membrane of osmotically tolerant MDCK cells. Slc4a8 KO mice did not show any abnormal salt or bicarbonate wasting under baseline conditions or in response to bicarbonate loading, salt restriction or furosemide-induced diuresis. Conclusion: Slc4a8 (NDCBE) is absent in the CCD and is predominantly localized on the basolateral membrane of medullary collecting duct cells. Further, Slc4a8 deletion does not cause significant acid base or electrolyte abnormalities in pathophysiologic states. Additional studies are needed to examine the role of Slc4a8 (NDCBE) in intracellular pH and volume regulation in medullary collecting duct cells.

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