Postnatal developmental profile of urocortin 1 and cocaine- and amphetamine-regulated transcript in the perioculomotor region of C57BL/6J mice

• Published in: Brain research 2010-03, Vol.1319, p.33-43
• Main Authors: Cservenka, Anita, Spangler, Erika, Cote, Dawn M, Ryabinin, Andrey E
• Format: Article
• Language: English
• Subjects: Animals; Animals, Newborn; Biological and medical sciences; Brain - growth & development; Brain - metabolism; Corticotropin releasing factor; Development; Development. Senescence. Regeneration. Transplantation; Edinger-Westphal; Fluorescent Antibody Technique; Fundamental and applied biological sciences. Psychology; Gene Expression Regulation, Developmental; Immunohistochemistry; Male; Mice; Mice, Inbred C57BL; Nerve Tissue Proteins - genetics; Nerve Tissue Proteins - metabolism; Neurology; Neurons - metabolism; Neuropeptide; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - metabolism; Subgriseal; Urocortins - genetics; Urocortins - metabolism; Vertebrates: nervous system and sense organs
• ISSN: 0006-8993; 1872-6240
• DOI: 10.1016/j.brainres.2010.01.003

Abstract Urocortin 1 (Ucn 1) is an endogenous corticotropin releasing factor (CRF)-related peptide. Ucn 1 is most highly expressed in the perioculomotor urocortin containing neurons (pIIIu), previously known as the non-preganglionic Edinger-Westphal nucleus (npEW). Various studies indicate that these cells are involved in stress adaptation and the regulation of ethanol (EtOH) intake. However, the developmental trajectory of these neurons remained unexamined. Expression of the cocaine- and amphetamine-regulated transcript (CART), which co-localizes with Ucn 1 in the perioculomotor area (pIII) has been examined prenatally, but not postnatally. The goal of the current study was to characterize the ontogenetic profile of Ucn 1 and CART during postnatal development in C57BL/6J (B6) mice. B6 mice were bred, and brains were collected at postnatal days (PND) 1, 4, 8, 12, 16, 24 and 45. Brightfield immunohistochemical staining for Ucn 1 and CART showed that Ucn 1-immunoreactivity (ir) was absent at PND 1, while CART-ir was already apparent in pIIIu at birth, a finding indicating that although the pIIIu neurons have already migrated to their adult position, Ucn 1 expression is triggered in them at later postnatal stages. Ucn 1-ir gradually increased with age, approaching adult levels at PND 16. This developmental profile was confirmed by double-immunofluorescence, which showed that Ucn 1 was absent in CART-positive cells of pIII at PND 4 and that Ucn 1 and CART are strongly but not completely co-localized in pIII at PND 24. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analysis confirmed that Ucn 1 mRNA levels are significantly lower at PND 4 and PND 12 than in adult animals. The lack of brain Ucn 1 immunoreactivity at birth and the gradual postnatal increase in Ucn 1 in pIIIu suggests that this peptide plays a greater behavioral role in adulthood than during the early postnatal development of an organism.