Neurons Containing the Enzyme Nicotinamide Adenine Dinucleotide Phosphate (NADPH)-Diaphorase in Organotypic Static Slice Cultures of Rat Cerebral Cortices: Developmental Study.

Abstract

The present study concerns the first trial of cultivation of neonatal rat cerebral cortical neurons containing the enzyme nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase [NADPH-d (+) neurons] in the organotypic static slice cultures. NADPH-d (+) neurons were scattered throughout the cortical layers and were highly concentrated in layers VI and II/III at the initial period, i. e. day 0, in culture of 7-day-old cerebral cortex. This distribution pattern was essentially the same as those seen after 28days in culture and in 35-day old rats. There were no apparent changes in the number of NADPHd (+) cells in the cortex at each time period examined (0, 10, 19 and 28days in culture). The size of cortical NADPH-d (+) neurons increased significantly, particularly during the first 10days, in culture. The numbers of primary neurites and their branching points, and the total neuritic length all increased significantly up to 19 days, but no further increase was observed at 28 days in culture. Also, the morphological features of NADPH-d (+) neurons after 19 and 28days in culture were almost the same as those in 35-day-old rats. Thus, cortical NADPH-d (+) neurons developed and reached the maturate state after 19 days in the cultivation system used here. These findings suggest that the organotypic static slice culture technique provides successful cultivation of cortical NADPH-d (+) neurons with normal-like development in situ, and should therefore be useful for studying the biology of these cells.