Calretinin in the entorhinal cortex of the rat: Distribution, morphology, ultrastructure of neurons, and co-localization with γ-aminobutyric acid and parvalbumin

• Published in: Journal of comparative neurology (1911) 2000-09, Vol.425 (2), p.177-192
• Main Authors: Wouterlood, Floris G., van Denderen, Jacqueline C.M., van Haeften, Theo, Witter, Menno P.
• Format: Article
• Language: English
• Subjects: Animals; Axons - metabolism; Axons - ultrastructure; Calbindin 2; calcium-binding proteins; calretinin; Cell Size - physiology; co-localization; Dendrites - metabolism; Dendrites - ultrastructure; double-label confocal laser scanning microscopy; entorhinal cortex; Entorhinal Cortex - metabolism; Entorhinal Cortex - ultrastructure; Female; GABA; gamma-Aminobutyric Acid - metabolism; immunofluorescence; Nerve Fibers - metabolism; Nerve Fibers - ultrastructure; Neurons - classification; Neurons - metabolism; Neurons - ultrastructure; parvalbumin; Parvalbumins - metabolism; Presynaptic Terminals - metabolism; Presynaptic Terminals - ultrastructure; Rats; Rats, Wistar - anatomy & histology; Rats, Wistar - metabolism; S100 Calcium Binding Protein G - metabolism; ultrastructure
• ISSN: 0021-9967; 1096-9861
• DOI: 10.1002/1096-9861(20000918)425:2<177::AID-CNE2>3.0.CO;2-G

Calretinin is a marker that differentially labels neurons in the central nervous system. We used this marker to distinguish subtypes of neurons within the general population of neurons in the entorhinal cortex of the rat. The distribution, morphology, and ultrastructure of calretinin‐immunopositive neurons in this cortical area were documented. We further analyzed the co‐localization of the marker with γ‐aminobutyric acid (GABA) and studied whether calretinin‐positive neurons project to the hippocampal formation. Methods used included single‐label immunocytochemistry at the light and electron microscopic level, retrograde tracing combined with immunocytochemistry, and double‐label confocal laser scanning microscopy (CLSM). The entorhinal cortex contained calretinin‐positive cells in a scattered fashion, in all layers except layer IV (lamina dissecans). Bipolar and multipolar dendritic configurations were present, displaying smooth dendrites. Bipolar cells had a uniform morphology whereas the multipolar calretinin cell population consisted of large neurons, cells with long ascending dendrites, horizontally oriented neurons, and small spherical cells. Retrograde tracing combined with immunocytochemistry showed that calretinin is not present in cells projecting to the hippocampus. Few synapic contacts between calretinin‐positive axon terminals and immunopositive cell bodies and dendrites were seen. Most axon terminals of calretinin fibers formed asymmetrical synapses, and immunopositive axons were always unmyelinated. Results obtained in the CLSM indicate that calretinin co‐exists in only 18–20% of the GABAergic cell population (mostly small spherical and bipolar cells). Thus, the entorhinal cortex contains two classes of calretinin interneurons: GABA positive and GABA negative. The first class is presumably a classical, GABAergic inhibitory interneuron. The finding of calretinin‐immunoreactive axon terminals with asymmetrical synapses suggests that the second class of calretinin neuron is a novel type of a (presumably excitatory) interneuron. J. Comp. Neurol. 425:177–192, 2000. © 2000 Wiley‐Liss, Inc.