GABAergic regulation of Locus coeruleus activity in necdin-deficient mice, an animal model of Prader-Willi syndrome

TL;DR

This study explores how a genetic deficiency in necdin affects brain activity in mice linked to Prader-Willi syndrome, focusing on GABA signaling in the locus coeruleus.

Contribution

The study identifies GABAB receptor dysfunction and astrocytic dysregulation as novel mechanisms underlying LC hypoactivity in a PWS mouse model.

Findings

Necdin-deficient mice show impaired GABAB receptor-mediated tonic inhibition in locus coeruleus neurons.

Astrocytes from necdin-deficient mice exhibit increased proliferation and higher GABA secretion.

GABAA receptor function remains intact despite LC hypoactivity in the PWS model.

Abstract

Prader-Willi syndrome (PWS) is a neurodevelopmental disorder caused by loss of paternally expressed genes on chromosome 15q11–13, including NDN, which encodes necdin. Necdin deficiency has been linked to impaired visuospatial memory, social recognition, and stress regulation—features also seen in PWS. Previous work showed that necdin-deficient (Ndn + m/−p) mice exhibit reduced activity of noradrenergic neurons in the locus coeruleus (LC), a nucleus essential for arousal and stress responses. However, the mechanisms underlying LC hypoactivity remain unclear. Because GABAergic signaling is critical for LC excitability, this study examined the role of GABAA ​and GABAB ​receptor-mediated inhibition in Ndn + m/−p mice.

Electrophysiological recordings from brainstem slices of wild-type (WT) and Ndn + m/−p mice were used to measure spontaneous firing rates (SFRs) of LC noradrenergic neurons. The effects of bicuculline (GABAA​ antagonist) and CGP54626 (GABAB​ antagonist) were tested. Whole-cell patch-clamp recordings assessed receptor-mediated currents. Western blotting quantified receptor subunit expression in peri-LC tissue. Immunocytochemistry and ELISA examined GABAB​ receptor expression and GABA release in cultured astrocytes.

LC-NE neurons in Ndn + m/−p mice exhibited significantly reduced baseline SFR compared with WT. Bicuculline did not alter firing in either genotype, whereas CGP54626 significantly increased SFR in WT but not in Ndn + m/−p neurons, indicating impaired GABAB receptor-mediated tonic inhibition. Whole-cell patch-clamp experiments confirmed intact GABAA receptor-mediated inward currents in both genotypes, while no GABAB receptor-mediated phasic currents were detected. Western blot analysis revealed comparable expression of GABAA receptor α2 subunit and GABABR1 in peri-LC tissue between WT and Ndn + m/−p mice, suggesting functional rather than expression-level deficits. GFAP-positive cell density in the LC region was unchanged in vivo; however, in astrocyte cultures, Ndn + m/−p astrocytes exhibited greater proliferation by DIV 19 and consistently secreted higher levels of GABA, with significant elevations at later culture stages.

Necdin deficiency selectively disrupts GABAB receptor-mediated tonic inhibition of LC-NE neurons while preserving GABAA receptor function. Elevated astrocytic proliferation and GABA release may further enhance ambient inhibition, contributing to LC hypoactivity and the neurobehavioral phenotypes of PWS. These findings identify GABAB receptor dysfunction and astrocytic dysregulation as potential mechanistic targets for therapeutic intervention in PWS.

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