A Combined Neuroanatomy, Ex Vivo Imaging, and Immunohistochemistry Defined MRI Mask for the Human Paraventricular Nucleus of the Thalamus

The paraventricular nucleus of the thalamus (PVT) is an evolutionarily conserved midline thalamic structure known to contribute to arousal, interoceptive states, and motivated behaviors. Yet, a consensus anatomical definition of the human PVT across tissue-based and MRI-based approaches remains elusive, thereby limiting reliable translation between its cellular characteristics and in vivo functional connectivity. To address this challenge, we describe a histologically informed PVT segmentation compatible with standard 3 T MR imaging pipelines. We performed postmortem anatomical MRI scans on an intact whole brain and an excised thalamic block, manually segmented the PVT at high resolution using ex vivo calretinin staining and neuroanatomical landmarks, registered the resulting image-label pair to a commonly used MRI template space (Montreal Neurological Institute's MNI152), and performed a comparative reanalysis using this newly defined mask. This tissue-grounded PVT mask largely overlaps spatially with existing MRI-based PVT masks, with the exception of additional voxels posteriorly. Importantly, the functional connectivity patterns of this tissue-grounded mask are highly consistent with those previously reported. Collectively, this multimodal definition of the human PVT balances tissue-based ground truth with in vivo MRI features, providing a valuable resource for advancing translation between cellular level features identified by histology and in vivo functional connectivity at the meso/macro scale in the understudied human PVT.