Search bioRxiv⌕ Search

Biology subjects

Milstead, R.

Publications and source records attributed to Milstead, R..

2 recordsLinked to original sources

Acute nicotine exposure produces a spatially organized transcriptional response in mouse olfactory cortex

Nicotine acts on nicotinic receptors throughout the forebrain. Most molecular studies of its effects, however, have focused on reward circuitry and have used bulk tissue, which cannot show which cells respond or how responding cells are arranged relative to one another. We previously found that a single nicotine exposure activates hippocampal astrocytes through the astrocyte-expressed kinase AKT2. Here we asked which cells respond to nicotine in the anterior olfactory cortex, a cholinergic-modulated region that encodes odor associations, and whether their astrocytic surroundings change. Male mice received a single intraperitoneal injection of nicotine or saline, and coronal sections spanning the anterior olfactory nucleus, piriform cortex and adjacent isocortex were sequencing 24 h later using Visium HD. Differential expression on nucleus-based segmentation was concentrated in olfactory excitatory neurons. Pathways centered on calcium-handling, synapse-assembly and extracellular-matrix programs, together with a lasting reduction of the immediate-early gene Egr1. Within the piriform region, the response localized to a continuous population of superficial pyramidal neurons expressing Cartpt and Penk. These neurons upregulated synapse-assembly, receptor-regulation and proteoglycan programs without changes to cell-type proportions. Nicotine also reduced expression of Sparcl1, which encodes astrocyte-secreted synaptogenic protein hevin, within one cell diameter of these neurons. Our study shows that a single nicotine exposure leaves a spatially organized transcriptional program on olfactory circuitry, centered on one neuronal population and the tissue at its border. These results identify candidate cells, genes and tissue locations for testing how nicotine alters sensory cue circuits.

neuroscience↗

AKT2 modulates astrocytic nicotine responses in vivo.

A greater understanding of the neurobiology of nicotine is needed to reduce or prevent chronic addiction, ameliorate detrimental nicotine withdrawal effects, and improve cessation rates. Nicotine binds and activates two astrocyte-expressed nicotinic acetylcholine receptors (nAChRs), 4{beta}2 and 7. Protein kinase B-{beta} (Pkb-{beta} or Akt2) expression is restricted to astrocytes in mice and humans and is activated by nicotine. To determine if AKT2 plays a role in astrocytic nicotinic responses, we generated astrocyte-specific Akt2 conditional knockout (cKO) and full Akt2 KO mice. For in/ex vivo studies, we examined mice exposed to chronic nicotine for two weeks in drinking water (200 g/mL) or following acute nicotine challenge (0.09, 0.2 mg/kg) after 24 hrs. Our in vitro studies used cultured mouse astrocytes to measure nicotine-dependent astrocytic responses. Sholl analysis was used to measure glial fibrillary acidic protein responses in astrocytes. Our data show wild-type (WT) mice exhibit increased astrocyte morphological complexity during acute nicotine exposure, with decreasing complexity during chronic nicotine use, whereas Akt2 cKO mice showed enhanced acute responses and reduced area following chronic exposure. In culture, we found 100 M nicotine sufficient for morphological changes and blocking 7 or 4{beta}2 nAChRs prevented observed morphologic changes. We performed conditioned place preference (CPP) in Akt2 cKO mice, which revealed reduced nicotine preference in cKO mice compared to controls. Finally, we performed RNASeq comparing nicotine- and LPS-mediated gene expression, identifying robust differences between these two astrocytic stimuli. These findings show the importance of nAChRs and AKT2 signaling in the astrocytic response to nicotine. Main PointsO_LINicotine regulates astrocytes in vivo: acute exposure boosts complexity; chronic exposure diminishes it. C_LIO_LIAKT2, expressed in astroglia, modulates morphological changes in response to nicotine and nicotine-dependent conditioned place preference. C_LI

neuroscience↗