Populations of hindbrain glucagon-like peptide 1 (GLP1) neurons that innervate the hypothalamic PVH, thalamic PVT, or limbic forebrain BST have axon collaterals that reach all central regions innervated by GLP1 neurons.

Abstract

Neurons in the caudal nucleus of the solitary tract (cNTS) and intermediate reticular nucleus (IRt) that express the glucagon gene (Gcg) give rise to GLP1-immunopositive axons in the spinal cord and many subcortical brain regions. Central GLP1 receptor signaling contributes to motivated behavior and stress responses in rats and mice, in which hindbrain GLP1 neurons are activated to express cFos in a metabolic state-dependent manner. The present study examined whether GLP1 inputs to distinct brain regions arise from distinct subsets of Gcg-expressing neurons, and mapped the collective distribution of axon collaterals arising from projection-defined GLP1 neural populations. Using our Gcg-Cre knock-in rat model, Cre-dependent adeno-associated virus (AAV1) tracing was conducted in adult male and female rats to compare axonal projections of IRt vs. cNTS GLP1 neurons. Overlapping axonal projections were observed in all brain regions that receive GLP1 input, with the caveat that cNTS injections produced Cre-dependent labeling of some IRt neurons, and vice-versa. In additional experiments, specific diencephalic or limbic forebrain nuclei were microinjected with Cre-dependent retrograde AAVs (AAVrg) expressing reporters that fully labeled the axon collaterals of transduced GLP1 neural populations. AAVrg injected into each forebrain site labeled Gcg-expressing neurons in both the cNTS and IRt. The collective axon collaterals of these labeled neurons entered the spinal cord and every brain region previously reported to contain GLP1-positive axons. These results indicate that axons arising from populations of GLP1 neurons that innervate the thalamic PVT, hypothalamic PVH, and/or limbic forebrain BST collectively innervate all central regions that receive GLP1 axonal input.Significance statement Our novel anatomical findings indicate that target-defined populations of forebrain-projecting GLP1 neurons collectively project to downstream target regions in a widespread sprinkler-type manner, although collateralized axons arising from individual GLP1 projection neurons remain to be defined. Considered together with results from studies investigating the role of central GLP1 receptor signaling pathways in physiology and behavior, these findings support our emerging view that hindbrain Gcg-expressing neurons are positioned to simultaneously modulate synaptic transmission in widespread spinal cord, brainstem, hypothalamic, and limbic forebrain circuits in a metabolic state-dependent manner.