Divergent Population-Specific Effects of Chronic Ethanol Exposure on Excitability and Inhibitory Transmission in Male and Female Rat Central Amygdala.

The central nucleus of the amygdala (CeA) is implicated in alcohol use disorder (AUD) and AUD-associated plasticity. The CeA is a primarily GABAergic nucleus that is subdivided into lateral and medial compartments with genetically diverse subpopulations. GABAA receptors are heteromeric pentamers with subunits conferring distinct physiological characteristics. GABAA receptor signaling in the CeA has been implicated in ethanol-associated plasticity; however, population-specific changes in inhibitory signaling and subunit expression remain unclear. Here, we combined electrophysiology with single-cell gene expression analysis of population markers and GABAA receptor subunits to examine population-specific changes in inhibitory control in male and female rats following chronic ethanol exposure. We found that chronic ethanol exposure and withdrawal produced global changes in GABAA receptor subunit expression at the transcript and protein levels, increased excitability in female CeA neurons, and increased inhibitory synaptic transmission in male CeA neurons. When we examined CeA neurons at the single-cell level we found heterogenous populations, as previously reported. We observed ethanol-induced increases in excitability only in somatostatin neurons in the CeA of females, decreases in excitability only in the protein kinase C delta (PKCd) population in males, and ethanol-induced increases in inhibitory transmission in male PKCd and calbindin 2-expressing CeA neurons. There were no population-specific differences in GABAA receptor (Gabr) subunits in males but reduced GabrA5 expression in female somatostatin neurons. Collectively, these findings suggest that defined CeA populations display differential ethanol sensitivity in males and females, which may play a role in sex differences in vulnerability to AUD or expression of AUD pathology.SIGNIFICANCE STATEMENT The CeA is involved in the effects of ethanol in the brain; however, the population-specific changes in CeA activity remain unclear. We used recordings of CeA neuronal activity and single-cell gene expression to examine population-specific changes in inhibitory control in male and female rats following chronic ethanol exposure and found sex- and population-specific effects of chronic ethanol exposure and withdrawal. Specifically, female CeA neurons displayed increased excitability in the somatostatin CeA population, whereas male CeA neurons displayed increased inhibitory control in both PKCd and calbindin populations and decreased excitability in the PKCd population. These findings identify CeA populations that display differential sensitivity to ethanol exposure, which may contribute to sex differences in vulnerability to alcohol use disorder.

GDNF-transfected macrophages produce potent neuroprotective effects in Parkinson's disease mouse model.

The pathobiology of Parkinson's disease (PD) is associated with the loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc) projecting to the striatum. Currently, there are no treatments that can halt or reverse the course of PD; only palliative therapies, such as replacement strategies for missing neurotransmitters, exist. Thus, the successful brain delivery of neurotrophic factors that promote neuronal survival and reverse the disease progression is crucial. We demonstrated earlier systemically administered autologous macrophages can deliver nanoformulated antioxidant, catalase, to the SNpc providing potent anti-inflammatory effects in PD mouse models. Here we evaluated genetically-modified macrophages for active targeted brain delivery of glial cell-line derived neurotropic factor (GDNF). To capitalize on the beneficial properties afforded by alternatively activated macrophages, transfected with GDNF-encoded pDNA cells were further differentiated toward regenerative M2 phenotype. A systemic administration of GDNF-expressing macrophages significantly ameliorated neurodegeneration and neuroinflammation in PD mice. Behavioral studies confirmed neuroprotective effects of the macrophage-based drug delivery system. One of the suggested mechanisms of therapeutic effects is the release of exosomes containing the expressed neurotropic factor followed by the efficient GDNF transfer to target neurons. Such formulations can serve as a new technology based on cell-mediated active delivery of therapeutic proteins that attenuate and reverse progression of PD, and ultimately provide hope for those patients who are already significantly disabled by the disease.

The primacy of physicochemical characterization of nanomaterials for reliable toxicity assessment: a review of the zebrafish nanotoxicology model.

Engineered nanomaterials (ENMs) have become increasingly prevalent in the past two decades in academic, medical, commercial, and industrial settings. The unique properties imbued with nanoparticles, as the physiochemical properties change from the bulk material to the surface atoms, present unique and often challenging characteristics that larger macromolecules do not possess. While nanoparticle characteristics are indeed exciting for unique chemistries, surface properties, and diverse applications, reports of toxicity and environmental impacts have tempered this enthusiasm and given cause for an exponential increase for concomitant nanotoxicology assessment. Currently, nanotoxicology is a steadily growing with new literature and studies being published more frequently than ever before; however, the literature reveals clear, inconsistent trends in nanotoxicological assessment. At the heart of this issue are several key problems including the lack of validated testing protocols and models, further compounded by inadequate physicochemical characterization of the nanomaterials in question and the seminal feedback loop of chemistry to biology back to chemistry. Zebrafish (Danio rerio) are emerging as a strong nanotoxicity model of choice for ease of use, optical transparency, cost, and high degree of genomic homology to humans. This review attempts to amass all contemporary nanotoxicology studies done with the zebrafish and present as much relevant information on physicochemical characteristics as possible. While this report is primarily a physicochemical summary of nanotoxicity studies, we wish to strongly emphasize that for the proper evolution of nanotoxicology, there must be a strong marriage between the physical and biological sciences. More often than not, nanotoxicology studies are reported by groups dominated by one discipline or the other. Regardless of the starting point, nanotoxicology must be seen as an iterative process between chemistry and biology. It is our sincere hope that the future will introduce a paradigm shift in the approach to nanotoxicology with multidisciplinary groups for data analysis to produce predictive and correlative models for the end goal of rapid preclinical development of new therapeutics into the clinic or insertion into environmental protection.