Single-Cell RT-PCR Identification of Genes Expressed by Human Islet Endocrine Cells

2009 ◽  
pp. 73-86 ◽  
Author(s):  
Dany Muller ◽  
Peter M. Jones ◽  
Shanta J. Persaud
Keyword(s):  
Single Cell ◽  
Endocrine Cells ◽  
Human Islet ◽  
Rt Pcr ◽  
Pcr Identification

scholarly journals P2X2 receptors differentiate placodal vs. neural crest C-fiber phenotypes innervating guinea pig lungs and esophagus

2008 ◽  
Vol 295 (5) ◽  
pp. L858-L865 ◽  
Author(s):  
Kevin Kwong ◽  
Marian Kollarik ◽  
Christina Nassenstein ◽  
Fei Ru ◽  
Bradley J. Undem
Keyword(s):  
Guinea Pig ◽  
Neural Crest ◽  
Single Cell ◽  
Dorsal Root Ganglia ◽  
Dorsal Root ◽  
Receptor Activation ◽  
Rt Pcr ◽  
C Fibers ◽  
Embryonic Origin ◽  
Ganglion Neurons

The lungs and esophagus are innervated by sensory neurons with somata in the nodose, jugular, and dorsal root ganglion. These sensory ganglia are derived from embryonic placode (nodose) and neural crest tissues (jugular and dorsal root ganglia; DRG). We addressed the hypothesis that the neuron's embryonic origin (e.g., placode vs. neural crest) plays a greater role in determining particular aspects of its phenotype than the environment in which it innervates (e.g., lungs vs. esophagus). This hypothesis was tested using a combination of extracellular and patch-clamp electrophysiology and single-cell RT-PCR from guinea pig neurons. Nodose, but not jugular C-fibers innervating the lungs and esophagus, responded to α,β-methylene ATP with action potential discharge that was sensitive to the P2X3 (P2X2/3) selective receptor antagonist A-317491. The somata of lung- and esophagus-specific sensory fibers were identified using retrograde tracing with a fluorescent dye. Esophageal- and lung-traced neurons from placodal tissue (nodose neurons) responded similarly to α,β-methylene ATP (30 μM) with a large sustained inward current, whereas in neurons derived from neural crest tissue (jugular and DRG neurons), the same dose of α,β-methylene ATP resulted in only a transient rapidly inactivating current or no detectable current. It has been shown previously that only activation of P2X2/3 heteromeric receptors produce sustained currents, whereas homomeric P2X3 receptor activation produces a rapidly inactivating current. Consistent with this, single-cell RT-PCR analysis revealed that the nodose ganglion neurons innervating the lungs and esophagus expressed mRNA for P2X2 and P2X3 subunits, whereas the vast majority of jugular and dorsal root ganglia innervating these tissues expressed only P2X3 mRNA with little to no P2X2 mRNA expression. We conclude that the responsiveness of C-fibers innervating the lungs and esophagus to ATP and other purinergic agonists is determined more by their embryonic origin than by the environment of the tissue they ultimately innervate.

Single-Cell RT-PCR cDNA Subtraction

2003 ◽  
pp. 601-609 ◽  
Author(s):  
Damian L. Weaver ◽  
César Núñez ◽  
Clare Brunet ◽  
Victoria Bostock ◽  
Gerard Brady
Keyword(s):  
Single Cell ◽  
Rt Pcr ◽  
Cdna Subtraction

Single-cell RT-PCR analysis of GIRK channels expressed in rat locus coeruleus and nucleus basalis neurons

2004 ◽  
Vol 358 (1) ◽  
pp. 63-67 ◽  
Author(s):  
Takeharu Kawano ◽  
Peng Zhao ◽  
Shigehiro Nakajima ◽  
Yasuko Nakajima
Keyword(s):  
Locus Coeruleus ◽  
Single Cell ◽  
Nucleus Basalis ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Girk Channels

scholarly journals Gene Expression Analysis by Multiplex Single-Cell RT-PCR

2019 ◽  
pp. 139-154 ◽  
Author(s):  
Ludovic Tricoire ◽  
Bruno Cauli ◽  
Bertrand Lambolez
Keyword(s):  
Gene Expression ◽  
Single Cell ◽  
Expression Analysis ◽  
Gene Expression Analysis ◽  
Rt Pcr

scholarly journals Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning

2008 ◽  
Vol 329 (1-2) ◽  
pp. 112-124 ◽  
Author(s):  
Thomas Tiller ◽  
Eric Meffre ◽  
Sergey Yurasov ◽  
Makoto Tsuiji ◽  
Michel C. Nussenzweig ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
B Cells ◽  
Single Cell ◽  
Expression Vector ◽  
Rt Pcr ◽  
Efficient Generation ◽  
Human B Cells

Composition and function of macroencapsulated human embryonic stem cell-derived implants: comparison with clinical human islet cell grafts

2014 ◽  
Vol 307 (9) ◽  
pp. E838-E846 ◽  
Author(s):  
Evi Motté ◽  
Edit Szepessy ◽  
Krista Suenens ◽  
Geert Stangé ◽  
Myriam Bomans ◽  
...  
Keyword(s):  
Islet Cell ◽  
Endocrine Cells ◽  
Large Scale ◽  
Therapeutic Potential ◽  
Ex Vivo ◽  
Embryonic Stem ◽  
Human Islet ◽  
Insulin Biosynthesis ◽  
Β Cells ◽  
C Peptide

β-Cells generated from large-scale sources can overcome current shortages in clinical islet cell grafts provided that they adequately respond to metabolic variations. Pancreatic (non)endocrine cells can develop from human embryonic stem (huES) cells following in vitro derivation to pancreatic endoderm (PE) that is subsequently implanted in immune-incompetent mice for further differentiation. Encapsulation of PE increases the proportion of endocrine cells in subcutaneous implants, with enrichment in β-cells when they are placed in TheraCyte-macrodevices and predominantly α-cells when they are alginate-microencapsulated. At posttransplant (PT) weeks 20–30, macroencapsulated huES implants presented higher glucose-responsive plasma C-peptide levels and a lower proinsulin-over-C-peptide ratio than human islet cell implants under the kidney capsule. Their ex vivo analysis showed the presence of single-hormone-positive α- and β-cells that exhibited rapid secretory responses to increasing and decreasing glucose concentrations, similar to isolated human islet cells. However, their insulin secretory amplitude was lower, which was attributed in part to a lower cellular hormone content; it was associated with a lower glucose-induced insulin biosynthesis, but not with lower glucagon-induced stimulation, which together is compatible with an immature functional state of the huES-derived β-cells at PT weeks 20–30. These data support the therapeutic potential of macroencapsulated huES implants but indicate the need for further functional analysis. Their comparison with clinical-grade human islet cell grafts sets references for future development and clinical translation.

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