TY - JOUR
T1 - Characterization of genetic differences within the centrallyprojecting Edinger-Westphal nucleus of C57BL/6J and DBA/2J mice by expression profiling
AU - Giardino, William J.
AU - Cote, Dawn M.
AU - Li, Ju
AU - Ryabinin, Andrey E.
PY - 2012
Y1 - 2012
N2 - 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.
AB - 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.
KW - Alcohol
KW - Edinger-westphal
KW - Immediate early gene
KW - Inducible transcription factor
KW - Midbrain
KW - Neuropeptide
KW - Oculomotor
KW - Urocortin
UR - http://www.scopus.com/inward/record.url?scp=84856774930&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84856774930&partnerID=8YFLogxK
U2 - 10.3389/fnana.2012.00005
DO - 10.3389/fnana.2012.00005
M3 - Article
C2 - 22745629
AN - SCOPUS:84856774930
SN - 1662-5129
JO - Frontiers in Neuroanatomy
JF - Frontiers in Neuroanatomy
IS - JANUARY 2012
M1 - 5
ER -