GABA immunoreactivity in hypothalamic neurons and growth cones in early development in vitro before synapse formation.
GABA (gamma-aminobutyrate) is the most prevalent inhibitory transmitter in the mature hypothalamus. In contrast, in the developing hypothalamus, GABA may exert depolarizing actions leading to neuronal excitation. To determine whether GABA is present in hypothalamic neurons early in development, and whether there is a preferential expression in axonal growth cones, immunogold and peroxidase studies were used with light and whole mount transmission electron microscopy. At embryonic day 15, a stage of development at the beginning of hypothalamic neurogenesis, histological sections showed GABA immunoreactivity in fibers and weakly stained perikarya. Hypothalamic neurons (13%) cultured at embryonic day 15 were immunoreactive after 1 day in vitro. The percentage of neurons stained, and the intensity of staining increased during the next few days to 39% at 4 days in vitro. Neuritic growth cones, including lamellipodia and long filopodia, showed strong immunoreactivity before synaptogenesis. By using neuronal whole mounts studied with transmission electron microscopy and GABA silver-enhanced immunogold staining, a quantitative comparison of growth cones after a day and a half in culture revealed that the growth cone of the longest process, the putative axon, had a greater level of immunogold labeling than that of the shorter processes, the putative dendrites. This finding is one of the earliest biochemical differences between putative axons and dendrites. Astrocytes in the same cultures showed no immunolabeling. These results indicate that GABA is present very early in the development of hypothalamic neurons and is in a position to be released.
van den Pol
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AN
Section of Neurosurgery, Yale University School of Medicine, New Haven, Connecticut 06520, USA. anthony.vandenpol@Yale.edu
D000818:Animals; D001369:Axons / Q000502:physiology / Q000648:ultrastructure; D002454:Cell Differentiation; D003712:Dendrites / Q000502:physiology / Q000648:ultrastructure; D005260:Female; D007031:Hypothalamus / Q000196:embryology / Q000378:metabolism* / Q000648:ultrastructure; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D009474:Neurons / Q000378:metabolism* / Q000648:ultrastructure; D011247:Pregnancy; D051381:Rats; D017207:Rats, Sprague-Dawley; D013569:Synapses / Q000502:physiology* / Q000648:ultrastructure; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
D005680:gamma-Aminobutyric Acid