The Bcl-2 and NeuN Expressions and Morphological Changes in Organotypic Explant Culture of Rat Hippocampus

J Korean Neurol Assoc > Volume 22(4); 2004 > Article

Journal of the Korean Neurological Association 2004;22(4): 368-374.

쥐 해마의 기관형적 절편배양 과정에서 Bcl-2와 NeuN 단백의 발현 양상 및 형태학적 관찰

윤영철, 권오상 채수안

중앙대학교 의과대학 신경과학교실, 소아과학교실

The Bcl-2 and NeuN Expressions and Morphological Changes in Organotypic Explant Culture of Rat Hippocampus

Young-Chul Youn

Departments of Neurology and Pediatrics*, Chung-Ang University College of Medicine, Seoul, Korea

Abstract

Background: Bcl-2 protein is essential for neurodevelopment and it is implicated in various neurodegenerative diseases. However, little is known about the normal developmental expression of Bcl-2 and NeuN protein in hippocampus slice cultures. We performed this study to assess their morphological changes and developmental expressions of Bcl-2 and NeuN protein.
Methods: We cultured the hippocampus of an eight days postnatal Sprague-Dawley (SD) rat in slices of 450 m. We observed the morphological development and differentiation of the hippocampus slices. Additionally, we measured the developmental expression of Bcl-2 and NeuN, -actin protein by western blotting.
Results: The hippocampal slice cultures revealed matured structures as early as 6 days in vitro (DIV) and a relatively high degree of tissue organization until 18 DIV. After 21 DIV, there were migration of the cells away from the margins of the slices. NeuN expressions at 7, 13, and 19 DIV were 1.42±0.38 (meanSD, NeuN/ beta-actin OD ratio, insert number=4, each insert had 5 culture slices), 2.95±0.53, 1.58±0.42 respectively. Bcl-2 expression at 6, 12, and 18 DIV were 0.56±0.18 (mean±SD, Bcl-2/beta-actin OD ratio, insert number=4, each insert had 5 culture slices), 1.22±0.28, 1.03±0.25.
Conclusions: Organotypic slice cultures of a rat's hippocampus shows morphological maturation within 14 DIV and the highest NeuN and Bcl-2 level at about 13 DIV. It is expected that these findings would be useful as baseline data in the field of research on neuronal development and aging processes.Key Words: Organotypic culture, Hippocampus, Bcl-2, NeuN