Characterization of genetic differences within the centrallyprojecting Edinger-Westphal nucleus of C57BL/6J and DBA/2J mice by expression profiling

William J. Giardino, Dawn M. Cote, Ju Li, Andrey Ryabinin

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Detailed examination of the midbrain Edinger-Westphal nucleus (EW) revealed the existence of two distinct nuclei. One population of preganglionic neurons was found to control oculomotor functions (EWpg), and a separate population of centrally-projecting neurons (EWcp) was found to contain stress- and feeding-related neuropeptides. Although it has been shown that EWcp neurons are highly responsive to drugs of abuse and behavioral stress, a genetic characterization of the EWcp was needed. To identify genetic differences in the EWcp of inbred mouse strains that differ in behaviors relevant to EWcp function, we used publicly-available tools from the Allen Brain Atlas to identify 68 transcripts that were selectively-expressed in the EWcp, and examined their expression within tissue punch microdissection samples containing the EWcp of adult male C57BL/6J (B6) and DBA/2J (D2) mice. Using 96-well quantitative real-time PCR (qPCR) arrays that included the EWcp-specific genes, several other genes of interest, and five housekeeping genes, we identified strain differences in expression of 11 EWcpspecific genes (BC023892, Btg3, Bves, Cart, Cck, Ghsr, Neto1, Postn, Ptprn, Rcn1, and Ucn), two immediate early genes (Egr1 and Fos), and one dopamine-related gene (Drd5). All significant expression differences were greater in B6 vs. D2 mice, and several of these were verified either at the protein level using immunohistochemistry (IHC) or in silico using microarray data sets from whole brain and other brain areas. These results demonstrate a significant advance in our understanding of the EWcp on three levels. First, we generated a list of EWcp-specific genes (most of which had not yet been reported within the EWcp in the literature) that will be informative for future studies of EWcp function. Second, due to similarity in results from qPCR and IHC, we revealed that strain differences in basal EWcp neuropeptide content are accounted for by differential transcription and number of peptidergic neurons, rather than by differential rates of peptide release. And third, our identification of differentially-expressed EWcpspecific genes between B6 and D2 mice may hold powerful insight into the neurogenetic contributions of the EWcp to stress- and addiction-related behaviors.

Original languageEnglish (US)
Article number5
JournalFrontiers in Neuroanatomy
Issue numberJANUARY 2012
DOIs
StatePublished - 2012

Fingerprint

Inbred DBA Mouse
Genes
Neurons
Neuropeptides
Brain
Immunohistochemistry
Microdissection
Inbred Strains Mice
Immediate-Early Genes
Atlases
Essential Genes
Street Drugs
Mesencephalon
Computer Simulation
Population
Edinger-Westphal Nucleus
Real-Time Polymerase Chain Reaction
Dopamine
Peptides
Proteins

Keywords

  • Alcohol
  • Edinger-westphal
  • Immediate early gene
  • Inducible transcription factor
  • Midbrain
  • Neuropeptide
  • Oculomotor
  • Urocortin

ASJC Scopus subject areas

  • Anatomy
  • Neuroscience (miscellaneous)
  • Cellular and Molecular Neuroscience

Cite this

Characterization of genetic differences within the centrallyprojecting Edinger-Westphal nucleus of C57BL/6J and DBA/2J mice by expression profiling. / Giardino, William J.; Cote, Dawn M.; Li, Ju; Ryabinin, Andrey.

In: Frontiers in Neuroanatomy, No. JANUARY 2012, 5, 2012.

Research output: Contribution to journalArticle

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