Quantitative ultrastructural differences between local and medial septal GABAergic axon terminals in the rat hippocampus.
Functionally distinct subsets of hippocampal inhibitory neurons exhibit large differences in the frequency, pattern and short-term plasticity of GABA release from their terminals. Heterogeneity is also evident in the ultrastructural features of GABAergic axon terminals examined in the electron microscope, but it is not known if or how this corresponds to interneuron subtypes. We investigated the feasibility of separating morphologically distinct clusters of terminal types, using the approach of measuring several ultrastructural parameters of GABAergic terminals in the CA1 area of the rat hippocampus. Septo-hippocampal axon terminals were anterogradely labeled by biotinylated dextran amine and visualized by pre-embedding immunogold staining to delineate one homogeneous terminal population. Long series (100-150) of ultrathin sections were cut from stratum oriens and stratum radiatum of the CA1 area, and GABAergic terminals were identified by post-embedding immunogold staining. Stereologically unbiased samples of the total GABAergic axon terminal population and a random sample of the septal axon terminals were reconstructed in 3D, and several of their parameters were measured (e.g. bouton volume, synapse surface, volume occupied by vesicles, mitochondria volume). Septal terminals demonstrated significantly larger mean values for most parameters than the total population of local GABAergic terminals. There was no significant difference between terminals reconstructed in the basal and apical dendritic regions of pyramidal cells, neither for the septal nor for the local population. Importantly, almost all parameters were highly correlated, precluding the possibility of clustering the local terminals into non-overlapping subsets. Factor and cluster analysis confirmed these findings. Our results suggest that similarly to excitatory terminals, inhibitory terminals follow an "ultrastructural size principle," and that the terminals of different interneuron subtypes cannot be distinguished by ultrastructure alone.
Eyre
M D
MD
Institute of Experimental Medicine, Hungarian Academy of Sciences, P.O. Box 67, H-1450, Budapest, Hungary. eyre@koki.hu
Freund
T F
TF
Gulyas
A I
AI
D000818:Animals; D001710:Biotin / Q000031:analogs & derivatives; D003911:Dextrans; D005456:Fluorescent Dyes; D006624:Hippocampus / Q000502:physiology* / Q000648:ultrastructure*; D007091:Image Processing, Computer-Assisted; D007150:Immunohistochemistry; D008297:Male; D009412:Nerve Fibers / Q000502:physiology / Q000648:ultrastructure; D017729:Presynaptic Terminals / Q000502:physiology* / Q000648:ultrastructure*; D051381:Rats; D017208:Rats, Wistar; D005680:gamma-Aminobutyric Acid / Q000502:physiology*
D003911:Dextrans; D005456:Fluorescent Dyes; C076397:biotinylated dextran amine; D005680:gamma-Aminobutyric Acid; D001710:Biotin