ALERT: This system is being upgraded on Tuesday December 12. It will not be available
for use for several hours that day while the upgrade is in progress. Deposits to DukeSpace
will be disabled on Monday December 11, so no new items are to be added to the repository
while the upgrade is in progress. Everything should be back to normal by the end of
day, December 12.
Sensitive and precise quantification of insulin-like mRNA expression in Caenorhabditis elegans.
Abstract
Insulin-like signaling regulates developmental arrest, stress resistance and lifespan
in the nematode Caenorhabditis elegans. However, the genome encodes 40 insulin-like
peptides, and the regulation and function of individual peptides is largely uncharacterized.
We used the nCounter platform to measure mRNA expression of all 40 insulin-like peptides
as well as the insulin-like receptor daf-2, its transcriptional effector daf-16, and
the daf-16 target gene sod-3. We validated the platform using 53 RNA samples previously
characterized by high density oligonucleotide microarray analysis. For this set of
genes and the standard nCounter protocol, sensitivity and precision were comparable
between the two platforms. We optimized conditions of the nCounter assay by varying
the mass of total RNA used for hybridization, thereby increasing sensitivity up to
50-fold and reducing the median coefficient of variation as much as 4-fold. We used
deletion mutants to demonstrate specificity of the assay, and we used optimized conditions
to assay insulin-like gene expression throughout the C. elegans life cycle. We detected
expression for nearly all insulin-like genes and find that they are expressed in a
variety of distinct patterns suggesting complexity of regulation and specificity of
function. We identified insulin-like genes that are specifically expressed during
developmental arrest, larval development, adulthood and embryogenesis. These results
demonstrate that the nCounter platform provides a powerful approach to analyzing insulin-like
gene expression dynamics, and they suggest hypotheses about the function of individual
insulin-like genes.
Type
Journal articleSubject
AnimalsCaenorhabditis elegans
Insulin
Nucleic Acid Conformation
Oligonucleotide Array Sequence Analysis
RNA, Messenger
Reproducibility of Results
Permalink
https://hdl.handle.net/10161/10404Published Version (Please cite this version)
10.1371/journal.pone.0018086Publication Info
Baugh, L Ryan; Kurhanewicz, Nicole; & Sternberg, Paul W (2011). Sensitive and precise quantification of insulin-like mRNA expression in Caenorhabditis
elegans. PLoS One, 6(3). pp. e18086. 10.1371/journal.pone.0018086. Retrieved from https://hdl.handle.net/10161/10404.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
Collections
More Info
Show full item recordScholars@Duke
L. Ryan Baugh
Professor of Biology
The Baugh Lab is interested in phenotypic plasticity and physiological adaptation
to variable environmental conditions. We are using the roundworm C. elegans to understand
how animals adapt to starvation using primarily genetic and genomic approaches. We
are studying how development is governed by nutrient availability, how animals survive
starvation, and the long-term consequences of starvation including adult disease and
transgenerational epigenetic inheritance.

Articles written by Duke faculty are made available through the campus open access policy. For more information see: Duke Open Access Policy
Rights for Collection: Scholarly Articles
Works are deposited here by their authors, and represent their research and opinions, not that of Duke University. Some materials and descriptions may include offensive content. More info