Hippocampal corticotropin releasing hormone: pre- and postsynaptic location and release by stress
Section snippets
Animals and tissue handling
Brains of immature Sprague–Dawley-derived rats (approximately P18, when density of CRH-expressing neurons in hippocampal pyramidal cell layers is maximal; Chen et al., 2001a) and young adult mice (C57/BL6) were used. Animals were born and maintained in a quiet, uncrowded, NIH-approved facility on a 12-h light/dark cycle, and brains were harvested under relatively stress-free conditions (Yan et al., 1998). Animals were undisturbed for 24 h prior to experiments, and were then deeply anesthetized
Presumed releasable pools of hippocampal CRH are located in axon terminals of GABAergic interneurons
Consistent with earlier studies (Sakanaka et al., 1987, Yan et al., 1998, Chen et al., 2001a), CRH was expressed in basket-type interneurons within the hippocampal pyramidal cell layer (Fig. 1A). These CRH-immunoreactive somata co-expressed mRNA for the GABA synthesizing enzyme glutamic acid decarboxylase (GAD67; Fig. 1B–D). Robust CRH-immunoreactivity was apparent within the perisomatic axonal ‘baskets’ surrounding principal cells (Fig. 1A, arrows), and double-labeling confirmed that
Discussion
The principal findings of these studies are: (1) The hippocampal CRH ‘system’ is organized with CRH expressed in GABAergic interneurons and the mediating receptor on dendritic spines of principal cells, (2) stress induces molecular/ transcriptional activation of hippocampal pyramidal cells, (3) stress-induced activation of a subset of hippocampal neurons requires activation of CRH receptors expressed by these cells, and (4) blocking central CRH receptors without interfering with the peripheral
Acknowledgements
The authors thank Karin Mews and Aniko Schneider for help with the EM studies, and M. Hinojosa for excellent editorial assistance. Grant number: NS28912, NS39307 (TZB), SFB 505 (MF). Grant sponsor: NIH and Deutsche Forschungsgemeinschaft.
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