NeuroanatomyA novel subpopulation of 5-HT type 3A receptor subunit immunoreactive interneurons in the rat basolateral amygdala
Section snippets
Tissue preparation
A total of 14 male Sprague–Dawley rats (250–350 g; Harlan, Indianapolis, IN, USA) were used in this study. All experiments were carried out in accordance with the National Institutes of Health Guide for the Care and Use of Laboratory Animals and were approved by the Institutional Animal Use and Care Committee (IACUC) of the University of South Carolina. All efforts were made to minimize animal suffering and to use the minimum number of animals necessary to produce reliable scientific data.
Rats
Results
5-HT3AR staining in the forebrain at the level of the amygdala appeared to be identical to that obtained in previous studies using this antibody (Morales et al., 1998). Thus, there was a moderate density of nonpyramidal 5-HT3AR+ neurons in the neocortex, hippocampus, and striatum, as well as in the BLC of the amygdala (Fig. 1). In addition, immunostained cells were also seen in anterior portions of the cortical and medial amygdalar nuclei. 5-HT3AR+ neurons were of similar morphology in all of
Discussion
This is the first study to use double-labeling immunohistochemistry to study the phenotypes of 5-HT3AR+ neurons in the BLC. As suggested in a previous immunohistochemical study (Morales et al., 1998), 5-HT3AR+ neurons in the amygdala were nonpyramidal interneurons that were mainly located in the BLC. The present investigation demonstrated that virtually all neurons in the BLC that expressed 5-HT3AR protein were GABA-immunoreactive, consistent with a previous report that 70–95% of telencephalic
Conclusion
In summary, some of the neurons expressing 5-HT3AR subunit protein in the BLC belong to a subpopulation of large GABAergic interneurons that are CCK+ (type L CCK+ interneurons), but most appear to constitute a previously unrecognized subpopulation of medium-sized GABAergic interneurons. In the awake state typical serotonergic neurons in the raphe fire in a clock-like manner at about 1–2 Hz, depending on the state of behavioral arousal (Jacobs and Azmitia, 1992). Thus, 5-HT3AR+ interneurons in
Acknowledgment
We are very grateful for the generous donation of the antibody to the 5-HT3AR subunit (antiserum 0165) by Dr. Marisela Morales (National Institute on Drug Abuse, Baltimore, MD, USA), the mouse anti-CCK antibody (antibody #9303) and mouse anti-VIP antibody (antibody CURE-V55) by Dr. John Walsh (CURE/Digestive Diseases Research Center, Antibody/RIA Core, NIH Grant #DK41301, Los Angeles, CA, USA), and the mouse anti-GABA antibody (antibody 115-AD5-A9) by Dr. Ismo Virtanen (University of Helsinki,
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