Refined anatomical isolation of functional sleep circuits exhibits distinctive regional and circadian gene transcriptional profiles.
Powerful new approaches to study molecular variation in distinct neuronal populations have recently been developed enabling a more precise investigation of the control of neural circuits involved in complex behaviors such as wake and sleep. We applied laser capture microdissection (LCM) to isolate precise brain nuclei from rat CNS at opposing circadian time points associated with wake and sleep. Discrete anatomical and temporal analysis was performed to examine the extent of variation in the transcriptional control associated with both identifiable anatomical nuclei and with light/dark cycle. Precise isolation of specific brain nuclei regulating sleep and arousal, including the LC, SCN, TMN, VTA, and VLPO, demonstrated robust changes in gene expression. Many of these differences were not observed in previous studies where whole brain lysates or gross dissections were used to probe for changes in gene expression. The robust and differential profiles of genomic data obtained from the approaches used herein underscore the requirement for careful anatomical refinement in CNS gene expression studies designed to understand genomic control within behaviorally-linked, but functionally isolated brain nuclei.
Winrow
Christopher J
CJ
Depression and Circadian Disorders Department, Merck Research Laboratories, West Point, PA 19486, USA.
Tanis
Keith Q
KQ
Rigby
Alison M
AM
Taylor
Rhonda R
RR
Serikawa
Kyle
K
McWhorter
Mollie
M
Tokiwa
George Y
GY
Marton
Matthew J
MJ
Stone
David J
DJ
Koblan
Kenneth S
KS
Renger
John J
JJ
D000818:Animals; D001143:Arousal / Q000235:genetics; D001921:Brain / Q000033:anatomy & histology / Q000378:metabolism*; D001931:Brain Mapping / Q000379:methods; D002940:Circadian Rhythm / Q000235:genetics*; D003624:Darkness; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000235:genetics*; D007834:Lasers; D008027:Light; D008297:Male; D042282:Microdissection / Q000379:methods; D009415:Nerve Net / Q000033:anatomy & histology / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000378:metabolism; D009434:Neural Pathways / Q000033:anatomy & histology / Q000378:metabolism; D010775:Photic Stimulation; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012890:Sleep / Q000235:genetics*; D015533:Transcriptional Activation / Q000235:genetics
D009419:Nerve Tissue Proteins; D012333:RNA, Messenger