Organization and ultrastructure of 5-bromo-2'-deoxiuridine (BrdU)-immunoreactive cells in neurogenic niche of lateral ventricle in Long-Evans rats (Rattus norvegicus)

• Grant number: 11/19747-3
• Support Opportunities: Regular Research Grants
• Start date: March 01, 2012
• End date: February 28, 2014
• Field of knowledge: Biological Sciences - Morphology - Anatomy
• Principal Investigator: Maria Inês Nogueira
• Grantee: Maria Inês Nogueira
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Ii-Sei Watanabe

Abstract

Neural cells retain their division process during the adult phase of development. Such activity is restricted to two proliferative niches in brain, among which the ventricular zone. This region surrounds the ependyma and subependymal zone (SEZ) of lateral ventricle, and even adjacent, SEZ cells differ in ultrastructural and neurochemical aspects. Initial studies observed cell proliferation exclusively in SEZ, but increasing studies suggest that ependymal cells trigger cell proliferation in the brain. Currently, arguments are based mainly on identification of co-localization of cell division cycle markers with neurochemical characteristics of ependymal cells, observed by confocal laser microscopy. To adequately explore and contribute to establishment of a putative new role to ependymal cells, it becomes necessary to characterize the ultrastructural level of the neural elements dividing the ependyma, which is the goal of this project. In this way, we propose to study the ultrastructural details of 5-bromo-2'-deoxyuridine (BrdU)-immunoreactive cells at ependyma, as well as their probable derivations in SEZ, and contribute to knowledgement of in situ three-dimensional organization of ependymal and SEZ cells. We use adult Long-Evans rats. For chemical induction of cell proliferation (icv administration of 1 mL Ara-C, 2% in sterile saline), animals receive the mitotic cycle marker BrdU (ip administration of 200 mg / kg) in a single doses. Subgroups of animals will be analyzed 24 h, 72 h or 7 days after administration of BrdU. Anti-BrdU immunohistochemistry findings will be examined by transmission electron microscopy. By high-resolution scanning electron microscopy, we plan to demonstrate the in situ tridimensional aspects of SEZ cells. Our preliminary results demonstrated newest and detailed citoarchitetonic aspects of ependymal layer and SEZ. Moreover, high-resolution scanning electron microscopy method allowed us to visualize the organization of ependymal surface and its constituents. Further studies, following the methodology implemented by our research group, will establish an appropriate treatment of samples, so permitting the comprehension of SEZ organization. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

DOS SANTOS HAEMMERLE, CARLOS ALEXANDRE; NOGUEIRA, MARIA INES; WATANABE, II-SEI. The neural elements in the lining of the ventricular-subventricular zone: making an old story new by high-resolution scanning electron microscopy. FRONTIERS IN NEUROANATOMY, v. 9, OCT 28 2015. (11/19747-3, 10/17519-0)

TAKADA, S. H.; DOS SANTOS HAEMMERLE, C. A.; MOTTA-TEIXEIRA, L. C.; MACHADO-NILS, A. V.; LEE, V. Y.; TAKASE, L. F.; CRUZ-RIZZOLO, R. J.; KIHARA, A. H.; XAVIER, G. F.; WATANABE, I. -S.; et al. NEONATAL ANOXIA IN RATS: HIPPOCAMPAL CELLULAR AND SUBCELLULAR CHANGES RELATED TO CELL DEATH AND SPATIAL MEMORY. Neuroscience, v. 284, p. 247-259, JAN 22 2015. (11/19747-3, 10/01488-9)