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

Youn, Young Chul; Kwon, Oh Sang; Chae, Soo Ahn

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.