Molecular phenotyping of transient postnatal tyrosine hydroxylase neurons in the rat bed nucleus of the stria terminalis

• Published in: Journal of chemical neuroanatomy 2017-07, Vol.82, p.29-38
• Main Author: Carter, David A.
• Format: Article
• Language: English
• Subjects: Animals; Animals, Newborn; Bed nucleus of the stria terminalis transcription factor; Egr1; Female; Male; Neurons - chemistry; Neurons - enzymology; NR4A2; Phenotype; Rats; Rats, Transgenic; Septal Nuclei - chemistry; Septal Nuclei - enzymology; Somatostatin; Tyrosine 3-Monooxygenase - analysis; Tyrosine 3-Monooxygenase - genetics; Tyrosine 3-Monooxygenase - metabolism; Tyrosine hydroxylase
• ISSN: 0891-0618; 1873-6300
• DOI: 10.1016/j.jchemneu.2017.04.002

•Transient BNSTLI TH+ve/EGR1+ve neurons are present in both male and female rats.•The molecular phenotype of BNSTLI neurons is defined using 13 protein markers.•The immature BNSTLI phenotype is independent of SOX2, REST and RBFOX2.•NR4A2/NURR1 is primarily cytoplasmic in BNSTLI TH+ve neurons.•TH+ve neurons in postnatal BNSTLI lack FOXA1, FOXA2 and FOXO1. The bed nucleus of the stria terminalis (BNST) is a complex integrative centre in the forebrain, composed of multiple sub-nuclei, each with discrete populations of neurons. Progress in understanding BNST function, both in the adult and during postnatal maturation, is dependent upon a more complete characterization of neuronal phenotypes in the BNST. The aim of the current study was to define the molecular phenotype of one postnatal BNST neuronal population, in order to identify molecular factors that may underlie both (protein marker-related) immaturity, and secondly, the transience of this phenotype. This BNST population was originally identified by high, but transient expression of the EGR1 transcription factor (TF) in postnatal rat lateral intermediate BNST (BNSTLI). The current results confirm a high level of Egr1 activation in postnatal day 10 (PN10) male BNSTLI that is lost at PN40, and now demonstrate a similar pattern of transient activation in female brains. Apparent cellular immaturity in this population, as indicated by low levels of the adult neuronal marker NeuN/RBFOX3, was found to be uncorrelated with both key neuronal regulator protein expression (SOX2 and REST), and also RBFOX2 protein levels. The BNSTLI neurons have a partial catecholaminergic phenotype (tyrosine hydroxylase-positive/dopa decarboxylase-negative; TH+ve/DDC−ve) that is lost at PN40. In contrast, the co-expressed neuropeptide, somatostatin, is maintained, albeit at lower levels, at PN40. The transcriptional basis of the transient and partial catecholaminergic phenotype was investigated by analysing TFs known to maintain adult dopaminergic (TH+ve/DDC+ve) neuronal phenotypes. The BNSTLI neurons were shown to lack forkhead TFs including FOXA1, FOXA2 and FOXO1. In addition, the BNSTLI neurons had low, primarily cytoplasmic, expression of NR4A2/NURR1, an orphan nuclear receptor that is critical for adult maintenance of midbrain dopamine neurons. These results detail the molecular features of an immature neuronal phenotype, and reveal TF deficiencies that may underlie postnatal transience of the phenotype.