Study of stimulus-secretion coupling in single cells using antisense oligodeoxynucleotides and patch-clamp techniques to inhibit specific protein expression

1994 ◽  
Vol 14 (5) ◽  
pp. 539-556 ◽  
Author(s):  
Pierre-Marie Lledo ◽  
William T. Mason ◽  
Robert Zorec
Keyword(s):  
Patch Clamp ◽  
Protein Expression ◽  
Single Cells ◽  
Specific Protein ◽  
Antisense Oligodeoxynucleotides ◽  
Stimulus Secretion Coupling

scholarly journals Parental Allele-Specific Protein Expression in Single Cells In Vivo

2018 ◽  
Author(s):  
Chiu-An Lo ◽  
Brian E. Chen
Keyword(s):  
Protein Expression ◽  
Single Cells ◽  
Specific Protein ◽  
The Other ◽  
Parental Allele ◽  
Allele Specific ◽  
Parent Of Origin ◽  
Ribosomal Protein L13a ◽  
Over Time

AbstractAllelic expression from each parent-of-origin is important as a backup and to ensure that enough protein products of a gene are produced. Thus far, it is not known how each cell throughout a tissue differs in parental allele expression at the level of protein synthesis. Here, we measure the expression of the Ribosomal protein L13a (Rpl13a) from both parental alleles simultaneously in single cells in the living animal. We use genome-edited Drosophila that have a quantitative reporter of protein synthesis inserted into the endogenous Rpl13a locus. We find that individual cells can have large (>10-fold) differences in protein expression between the two parental alleles. Cells can produce protein from only one allele oftentimes, and time-lapse imaging of protein production from each parental allele in each cell showed that the imbalance in expression from one parental allele over the other can invert over time.One sentence summaryParental allele-specific protein expression varies widely across cells and over time.HighlightsWe used genome editing to insert a quantifiable protein translation reporter into the endogenous Ribosomal protein L13a gene and thus track the protein expression of both parental alleles simultaneously in every single cell in the awake animal.Cells can have a large difference in protein expression for one parental allele over the other, and this can invert over time, and can occur in clusters of cells within a tissue.We demonstrate the highly variable nature of heterozygous and homozygous definitions across single cells, and over time.Our study demonstrates a new paradigm that can be used to examine inherited epigenetic control of expression from a specific parent-of-origin allele across single clonal cells from a common progenitor in vivo.

Tissue Distribution and Gender-Specific Protein Expression of Cytochrome P450 in five Mouse Genotypes with a Background of FVB

2018 ◽  
Vol 35 (6) ◽  
Author(s):  
Jiamei M. Chen ◽  
Qisong S. Zhang ◽  
Xiaoyan Y. Li ◽  
Xia Gong ◽  
Yanjiao J. Ruan ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Protein Expression ◽  
Tissue Distribution ◽  
Specific Protein ◽  
And Gender ◽  
Gender Specific

Fluoride exposure inhibits protein expression and enzyme activity in the lung-stage larvae ofAscaris suum

Parasitology ◽  
2006 ◽  
Vol 133 (4) ◽  
pp. 497-508 ◽  
Author(s):  
M. K. ISLAM ◽  
T. MIYOSHI ◽  
M. YAMADA ◽  
M. A. ALIM ◽  
X. HUANG ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression Profiles ◽  
Immunoblot Analysis ◽  
Specific Protein ◽  
Inorganic Pyrophosphatase ◽  
2D Electrophoresis ◽  
Peptide Mass ◽  
Enzyme Families ◽  
Protein Expression Profiles ◽  
Peptide Mass Spectrometry

Sodium fluoride (NaF) is an anion that has been previously shown to block the moulting process ofAscaris suumlarvae. This study describes moulting and development-specific protein expression profiles ofA. suumlung-stage L3 (AsLL3) following NaF exposure. AsLL3s cultured in the presence or absence of NaF were prepared for protein analysis using two-dimensional (2D) electrophoresis. NaF exposure inhibited at least 22 proteins in AsLL3 compared with moulted larvae (i.e. AsLL4). A further comparison of AsLL4 with those of pre-cultured AsLL3 and NaF-exposed AsLL3 revealed 8 stage-specifically and 4 over-expressed proteins. Immunoblot analysis revealed an inhibition by NaF of 19 immunoreactive proteins. Enzyme assay and immunochemical data showed an inhibition of the moulting-specific inorganic pyrophosphatase activity by 41% and a decreased expression in NaF-treated larvae, indicating its significance in the moulting process. A protein spot associated with NaF inhibition was isolated and identified by peptide mass spectrometry and bioinformatics approaches to be a member of 3–hydroxyacyl–CoA dehydrogenase/short-chain dehydrogenase enzyme families. These results have implications for the identification of proteins specific to the moulting process as potential chemotherapeutic targets.

Patch-Clamp Recording and RT-PCR on Single Cells

2003 ◽  
pp. 193-232 ◽  
Author(s):  
Bertrand Lambolez ◽  
Etienne Audinat ◽  
Pascal Bochet ◽  
Jean Rossier
Keyword(s):  
Patch Clamp ◽  
Single Cells ◽  
Rt Pcr ◽  
Patch Clamp Recording

Understanding Control of Exocytotic Secretion in Single Adenohypophysial Cells

Physiology ◽  
1994 ◽  
Vol 9 (5) ◽  
pp. 219-223
Author(s):  
Robert Zorec ◽  
G. Zupančič ◽  
M. Rupnik ◽  
L. Kocmur ◽  
S. Grilc ◽  
...  
Keyword(s):  
Patch Clamp ◽  
Cellular Level ◽  
Membrane Capacitance ◽  
Secretory Cells ◽  
Patch Clamp Technique ◽  
Clamp Technique ◽  
Stimulus Secretion Coupling ◽  
Insight Into

Stimulus-secretion coupling at the cellular level is studied by measuring changes in membrane capacitance in a variety of secretory cells. Attempts to gain new insight into the control of exocytosis in adenohypophysial cells by the patch-clamp technique are briefly outlined.

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