Calretinin and calbindin architecture of the midline thalamus associated with prefrontal-hippocampal circuitry

Abstract

The midline thalamus bi-directionally connects the medial prefrontal cortex (mPFC) and hippocampus (HC) creating a unique cortico-thalamo-cortico circuit fundamental to memory and executive function. While the anatomical connectivity of midline thalamus has been thoroughly investigated, little is known about its cellular organization within each nucleus. Here we used immunohistological techniques to examine cellular distributions in the midline thalamus based on the calcium binding proteins parvalbumin (PV), calretinin (CR), and calbindin (CB). We also examined these calcium binding proteins in a population of reuniens cells known to project to both mPFC and HC using a dual fluorescence retrograde adenoassociated virus (AAV) based tracing approach. These dual reuniens mPFC-HC projecting cells, in particular, are thought to be important for synchronizing mPFC and HC activity. First, we confirmed the absence of PV+ neurons in the midline thalamus. Second, we found a common pattern of CR+ and CB+ cells throughout midline thalamus with CR+ cells running along the nearby third ventricle (3V) and penetrating the midline. CB+ cells were consistently more lateral and toward the middle of the dorsal-ventral extent of the midline thalamus. Notably, single-labeled CR+ and CB+ zones were partially overlapping and included dual-labeled CR+/CB+ cells. Within RE, we also observed a CR and CB subzone specific diversity. Interestingly, dual mPFC-HC projecting neurons in RE expressed none of the calcium binding proteins examined, but were contained in nests of CR+ and CB+ cells. Overall, the midline thalamus was well organized into CR+ and CB+ rich zones distributed throughout the region, with dual mPFC-HC projecting cells in reuniens representing a unique cell population. These results provide a cytoarchitectural organization in the midline thalamus based on calcium binding protein expression, and sets the stage for future cell-type specific interrogations of the functional role of these different cell populations in mPFC-HC interactions.

Identifying the midline thalamus in humans in vivo

“…Whereas, RE and RH have been mostly studied in regards to their connections to regions implicated in various learning and memory processes, such as memory consolidation (3), spatial working memory (30,31), goal directed spatial navigation (32), temporal organization of memory (33), contextual fear memory (34) and behavioral flexibility (31). Interestingly, both PV and RE have neurons that project to both agranular medial prefrontal cortex and the hippocampus, and likely both are related to explicit memory processes under different conditions (35)(36)(37)(38).…”

Pathways for Memory, Cognition and Emotional Context: Hippocampal, Subgenual Area 25, and Amygdalar Axons Show Unique Interactions in the Primate Thalamic Reuniens Nucleus

“…In rodents, RE is characterized by its composition of projection neurons labeled with the calcium-binding proteins calbindin (CB) and calretinin (CR), but it lacks parvalbumin (PV) neurons (Arai et al, 1994;Bokor et al, 2002;Viena et al, 2021a). Thalamic CB and CR neurons give rise to the excitatory matrix thalamocortical system, which mostly innervates layers I-IIIa in wide axonal arbors and is thought to have a modulatory feedback-like role (Piantoni et al, 2016).…”