PMID- 23967349
OWN - NLM
STAT- MEDLINE
DCOM- 20140514
LR  - 20211021
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 8
IP  - 8
DP  - 2013
TI  - Characterization of GABAergic neurons in the mouse lateral septum: a double 
      fluorescence in situ hybridization and immunohistochemical study using tyramide 
      signal amplification.
PG  - e73750
LID - 10.1371/journal.pone.0073750 [doi]
LID - e73750
AB  - Gamma-aminobutyric acid (GABA) neurotransmission in the lateral septum (LS) is 
      implicated in modulating various behavioral processes, including emotional 
      reactivity and maternal behavior. However, identifying the phenotype of GABAergic 
      neurons in the CNS has been hampered by the longstanding inability to reliably 
      detect somal immunoreactivity for GABA or glutamic acid decarboxylase (GAD), the 
      enzyme that produces GABA. In this study, we designed unique probes for both 
      GAD65 (GAD2) and GAD67 (GAD1), and used fluorescence in Situ hybridization (FISH) 
      with tyramide signal amplification (TSA) to achieve unequivocal detection of cell 
      bodies of GABAergic neurons by GAD mRNAs. We quantitatively characterized the 
      expression and chemical phenotype of GABAergic neurons across each subdivision of 
      LS and in cingulate cortex (Cg) and medial preoptic area (MPOA) in female mice. 
      Across LS, almost all GAD65 mRNA-expressing neurons were found to contain GAD67 
      mRNA (approximately 95-98%), while a small proportion of GAD67 mRNA-containing 
      neurons did not express GAD65 mRNA (5-14%). Using the neuronal marker NeuN, 
      almost every neuron in LS (> 90%) was also found to be GABA-positive. Interneuron 
      markers using calcium-binding proteins showed that LS GABAergic neurons displayed 
      immunoreactivity for calbindin (CB) or calretinin (CR), but not parvalbumin (PV); 
      almost all CB- or CR-immunoreactive neurons (98-100%) were GABAergic. The 
      proportion of GABAergic neurons immunoreactive for CB or CR varied depending on 
      the subdivisions examined, with the highest percentage of colocalization in the 
      caudal intermediate LS (LSI) (approximately 58% for CB and 35% for CR). These 
      findings suggest that the vast majority of GABAergic neurons within the LS have 
      the potential for synthesizing GABA via the dual enzyme systems GAD65 and GAD67, 
      and each subtype of GABAergic neurons identified by distinct calcium-binding 
      proteins may exert unique roles in the physiological function and neuronal 
      circuitry of the LS.
FAU - Zhao, Changjiu
AU  - Zhao C
AD  - Department of Zoology, University of Wisconsin-Madison, Madison, Wisconsin, USA. 
      czhou23@wisc.edu
FAU - Eisinger, Brian
AU  - Eisinger B
FAU - Gammie, Stephen C
AU  - Gammie SC
LA  - eng
GR  - R01 MH085642/MH/NIMH NIH HHS/United States
GR  - R01 MH 085642/MH/NIMH NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20130813
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (RNA, Messenger)
RN  - 56-12-2 (gamma-Aminobutyric Acid)
RN  - EC 4.1.1.15 (Glutamate Decarboxylase)
RN  - EC 4.1.1.15 (glutamate decarboxylase 1)
RN  - EC 4.1.1.15 (glutamate decarboxylase 2)
SB  - IM
MH  - Animals
MH  - Female
MH  - GABAergic Neurons/*metabolism
MH  - Gene Expression
MH  - Glutamate Decarboxylase/genetics/metabolism
MH  - Gyrus Cinguli/metabolism
MH  - Immunohistochemistry
MH  - Mice
MH  - Microscopy, Fluorescence
MH  - Preoptic Area/metabolism
MH  - Protein Binding
MH  - Protein Transport
MH  - RNA, Messenger/genetics/metabolism
MH  - Septal Nuclei/*metabolism
MH  - gamma-Aminobutyric Acid/metabolism
PMC - PMC3742568
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2013/08/24 06:00
MHDA- 2014/05/16 06:00
PMCR- 2013/08/13
CRDT- 2013/08/23 06:00
PHST- 2013/04/24 00:00 [received]
PHST- 2013/07/26 00:00 [accepted]
PHST- 2013/08/23 06:00 [entrez]
PHST- 2013/08/24 06:00 [pubmed]
PHST- 2014/05/16 06:00 [medline]
PHST- 2013/08/13 00:00 [pmc-release]
AID - PONE-D-13-16892 [pii]
AID - 10.1371/journal.pone.0073750 [doi]
PST - epublish
SO  - PLoS One. 2013 Aug 13;8(8):e73750. doi: 10.1371/journal.pone.0073750. eCollection 
      2013.