Region and cell selective induction of Zbtb16/Plzf by multiple stressors in the adult murine hypothalamus

Stress activates the hypothalamic–pituitary–adrenal (HPA) axis, a neuroendocrine system that regulates feeding, reproduction, aggression, and other homeostatic functions. Stressors markedly alter gene expression along the HPA axis and within hypothalamic nuclei that drive it, including the paraventricular nucleus (PVN). To identify genetic regulators of PVN stress responses, adult mice were exposed to two hours of multimodal stress, and gene expression profiles were assessed using bulk RNA sequencing. We identified the transcription factor zinc finger and BTB domain–containing 16 (Zbtb16), also known as PLZF, as a stress-responsive glucocorticoid receptor (GR) target in the PVN. Zbtb16 mRNA increased twofold in male and female mice within two hours of restraint stress or injection of the synthetic glucocorticoid dexamethasone (DEX). Immunohistochemistry confirmed Zbtb16 protein expression and localization in the PVN after 20 minutes of restraint stress followed by 4 hours of recovery. In situ hybridization further confirmed Zbtb16 mRNA colocalization in the PVN where antibody immunoreactivity was observed. Cellular analyses revealed robust Zbtb16 expression in CRH neurons, cells typically activated after stress, as indicated by colocalization with c-FOS. To assess regulation by immune stress, adult mice were injected with tumor necrosis factor-α (TNFα). Additional spatial immunohistochemical analyses demonstrated region-specific and cell type–specific expression patterns: Zbtb16 colocalized with CRH neurons in the mid-PVN but was more prominent in astrocytes surrounding the PVN. Together, these findings identify Zbtb16 as a glucocorticoid- and cytokine-inducible transcriptional regulator with region- and cell type–specific roles in PVN stress circuitry. Other genes from the original screen are now under investigation. Together with stereotaxic injections of glial-selective toxins, an emerging goal is to define cytokine contributions to PVN function. Supported by ORWH-U54-MH118919. This abstract was presented at the American Physiology Summit 2026 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.