Biology subjects

Radwanska, K.

Publications and source records attributed to Radwanska, K..

10 recordsLinked to original sources

bioRxiv2026

Estrogen receptor alpha regulates alcohol craving in females: convergent evidence from mouse models and human genetics

Alcohol craving and consumption fluctuate across the reproductive cycle in females with alcohol use disorder (AUD), suggesting that estrogen signaling contributes to disease vulnerability. Here, we investigated the role of estrogen receptor alpha (ER; Esr1; ESR1) in alcohol seeking using complementary mouse and human approaches, as this receptor was previously identified as a risk factor for AUD. Female mice were characterized in an IntelliCage-based multidimensional AUD paradigm that stratifies individuals into AUD-prone and AUD-resistant phenotypes. Transcriptomic profiling of the amygdala revealed that Esr1 is a top transcription factor for differentially expressed genes in mice drinking alcohol, and the estrogen signaling pathway was deregulated specifically in AUD-prone mice. Although alcohol exposure did not alter overall Esr1/ER mRNA or protein abundance, both transcript and protein levels positively correlated with cue-induced alcohol seeking, indicating that inter-individual variation in ER signaling predicts relapse-like behavior. Causal manipulations confirmed a functional role of ER. Local knockdown of Esr1 in the basolateral amygdala reduced excitatory synaptic transmission, attenuated alcohol motivation, cue-induced seeking, and relapse drinking, and impaired cue-associated memory recall without affecting anxiety-like behavior. Similarly, ovariectomy decreased amygdala ER expression, altered synaptic protein markers, and reduced alcohol-seeking behaviors, supporting regulation by endogenous ovarian hormones. Extending these findings to humans, ESR1 gene polymorphisms (rs6902771, rs11155819 and rs6557171) were associated with the probability of alcohol binge drinking and alcohol consumption days as well as craving and loss of control in real world in a longitudinal clinical cohort, while ESR1 mRNA blood levels were increased in women with AUD diagnosis. Together, these convergent molecular, circuit, behavioral, and genetic data identify ER signaling in the amygdala as an important modulator of alcohol-seeking behavior induced by alcohol cue and relapse vulnerability, highlighting estrogen pathways as potential therapeutic targets and markers for AUD.

neuroscience↗

bioRxiv2026

Spatial learning-specific remodeling of the hippocampal palmitoylome

Protein S-palmitoylation is a reversible lipid modification that regulates protein trafficking, membrane association, and synaptic signaling, yet its role in learning-induced neuronal plasticity remains incompletely understood. Here, we investigated how spatial learning remodels the hippocampal palmitoylome in rats trained in the Morris water maze using short-term (STT; one session, 15 trials; probe at 1 h) or long-term (LTT; four sessions over four days; probe at 24 h) paradigms. Palmitoylated proteins were profiled by acyl-biotin exchange coupled with tandem mass tag labeling and LC-MS/MS. We identified 5,260 proteins, including 763 palmitoylated species. Spatial learning induced extensive remodeling of protein S-palmitoylation, with markedly greater changes after STT than LTT. Using yoked controls, we distinguished palmitoylation changes associated with learning the hidden platform location from those induced by general behavioral experience. Comparison of trained and yoked animals identified 186 and 62 differentially palmitoylated proteins after STT and LTT, respectively, whereas yoked animals also exhibited extensive changes relative to naive controls, demonstrating that behavioral experience alone substantially reshapes the hippocampal palmitoylome. Functional enrichment revealed that STT preferentially engaged pathways related to synaptic transmission, cytoskeletal remodeling, GTPase signaling, and cellular metabolism, whereas LTT was associated with protein translation and synaptic organization. Site-specific analysis identified numerous previously unreported palmitoylation sites. Hierarchical analysis further identified diacylglycerol lipase- (DAGLA) as the only protein whose palmitoylation consistently reflected both general Morris water maze experience and learning the hidden platform location across both paradigms. Together, these findings establish S-palmitoylation as a dynamic regulator of experience-dependent hippocampal plasticity.

neuroscience↗

bioRxiv2024

Terminal nucleotidyltransferase Tent2 microRNA tailing regulates excitatory/inhibitory balance in the hippocampus

One of the post-transcriptional mechanisms regulating the stability of RNA molecules involves the addition of non-templated nucleotides to their 3 ends, a process known as RNA tailing. To systematically investigate the physiological consequences of terminal nucleotidyltransferase TENT2 absence on RNA 3 end modifications in the mouse hippocampus we developed a new Tent2 knockout mouse. Electrophysiological measurements revealed increased excitability in Tent2 KO hippocampal neurons, and behavioral analyses showed decreased anxiety and improved fear extinction in these mice. At the molecular level, we observed a significant contribution of TENT2 to the monoadenylation of various classes of miRNAs, but found no effect of the enzymes loss on the total poly(A) tail length of mRNAs, as measured by Direct Nanopore RNA sequencing. Alterations in monoadenylation of a large population of microRNAs affected the overall mRNA abundance, particularly transcripts related synaptic transmission, which were downregulated in the hippocampus of Tent2 knockout mice. These changes explain the observed behavioral and electrophysiological alterations. Our data thus establish a link between TENT2-dependent microRNA tailing and the balance of inhibitory and excitatory neurotransmission.

neuroscience↗

bioRxiv2024

Projections from thalamic nucleus reuniens to medial septum enable extinction of remote fear memory

Aversive experiences lead to the formation of long-lasting memories. Despite the need to better understand how enduring fear memories can be attenuated, the underlying brain circuits remain largely unknown. In this study, employing a combination of genetic manipulations, neuronal circuit mapping, and chemogenetics in mice, we identify a new projection from the thalamic nucleus reuniens (RE) to the medial septum (MS), and show that this circuit is involved in the extinction of remote (30-day old), but not recent (1-day old), fear memories. These findings provide the first functional description of the RE[->]MS circuit and highlight the significance of the thalamo-septal regions in memory organization as a function of memory age, a phenomenon known as systems consolidation.

neuroscience↗

bioRxiv2024

Projections from thalamic nucleus reuniens to hippocampal CA1 area enable context value updating

The ability to extinguish contextual fear in a changing environment is crucial for animal survival. Recent data support the role of the thalamic nucleus reuniens (RE) and its projections to the dorsal hippocampal CA1 area (RE[->]dCA1) in this process. However, it remains poorly understood how RE impacts dCA1 neurons during contextual fear extinction (CFE). Here, we reveal that the RE[->]dCA1 pathway contributes to extinction of contextual fear by affecting CFE-induced molecular remodeling of excitatory synapses. Anatomical tracing and chemogenetic manipulation in mice demonstrate that RE neurons form synapses and regulate synaptic transmission in the stratum oriens (SO) and lacunosum-moleculare (SLM) of the dCA1 area, but not in the stratum radiatum (SR). We also observe CFE-specific structural changes of excitatory synapses and expression of the synaptic scaffold protein, PSD-95, in both strata innervated by RE, but not in SR. Interestingly, only the changes in SLM are specific for the dendrites innervated by RE. To further support the role of the RE[->]dCA1 projection in CFE, we demonstrate that brief chemogenetic inhibition of the RE[->]dCA1 pathway during a CFE session persistently impairs the formation of CFE memory and CFE-induced changes of PSD-95 levels in SLM. Thus, our data indicate that RE participates in CFE by regulating CFE-induced molecular remodeling of dCA1 synapses.

animal behavior and cognition↗

bioRxiv2023

Identification of neuronal ensembles involved in remote fear memory extinction impairments

Remote memory consolidation, extinction, and its impairments have been of interest to researchers for years, especially due to its clinical relevance in patients with emotional disorders, like PTSD (Post Traumatic Stress Disorder), anxiety, and phobias. Its neuronal substrates are key elements to understand the persistent nature of remote memory and open a new perspective to novel therapeutic approaches for human fear-related disorders. While the majority of reports investigate the mechanisms and ensembles of recent fear memory extinction (hours to days following conditioning), only a few refer to the remote time point (i.e. weeks after conditioning), usually describing successful memory extinction. The neuronal correlates of impaired remote fear memory extinction were yet beyond the scope. Here we present selective impairment of contextual remote fear memory extinction in alpha calcium/calmodulin-dependent protein kinase II (CaMKII) autophosphorylation-limited mice (T286A+/-). To map brain regions involved in this phenomenon, we applied screening of c-Fos expression, a neuroplasticity marker, across 23 brain areas following contextual fear conditioning and extinction of recent (1-day old) and remote (30-days old) fear memory in WT and T286A+/- mice. Following impaired remote fear memory extinction in T286A+/- mice, we found upregulated c-Fos expression in the entorhinal cortex (ENT), nucleus reuniens (RE), centromedial (CM), mediodorsal (MD), anterodorsal (AD) thalamic nuclei, and medial septum (MS), compared to WT animals performing normal remote fear memory extinction. Thus our data suggest that CaMKII-autophosphorylation-dependent c-Fos expression in these areas controls distant contextual fear extinction and may shed light on these brain regions as potential targets for therapeutic strategies against emotional disorders such as PTSD.

animal behavior and cognition↗

bioRxiv2023

Phosphorylation of PSD-95 at Serine 73 in dCA1 is required for extinction of contextual fear

The updating of contextual memories is essential for survival in a changing environment. Accumulating data indicate that the dorsal CA1 area (dCA1) contributes to this process. However, the cellular and molecular mechanisms of contextual fear memory updating remain poorly understood. Postsynaptic density protein 95 (PSD-95) regulates the structure and function of glutamatergic synapses. Here, using dCA1-targeted genetic manipulations in vivo, combined with ex vivo 3D electron microscopy and electrophysiology, we identify a novel, synaptic mechanism that is induced during attenuation of contextual fear memories and involves phosphorylation of PSD-95 at Serine 73 in dCA1. Our data provide the proof that PSD-95-dependent synaptic plasticity in dCA1 is required for updating of contextual fear memory.

neuroscience↗

bioRxiv2022

The molecular fingerprint of stress resilience

Stress resilience is the ability of neuronal networks to maintain their function despite the stress exposure. Using a mouse model we here investigate stress resilience phenomenon. To assess the resilient and anhedonic behavioral phenotypes developed after the induction of chronic unpredictable stress, we quantitatively characterized the structural and functional plasticity of excitatory synapses in the hippocampus using a combination of proteomic, electrophysiological, and imaging methods. Our results indicate that stress resilience is an active and multifactorial process manifested by structural, functional, and molecular changes in synapses. We reveal that chronic stress influences palmitoylation of synaptic proteins, whose profiles differ between resilient and anhedonic animals. The changes in palmitoylation are predominantly related with the glutamate receptor signaling thus affects synaptic transmission and associated structures of dendritic spines. We show that stress resilience is associated with structural compensatory plasticity of the postsynaptic parts of synapses in CA1 subfield of the hippocampus. One Sentence SummaryCompensatory remodeling of dendritic spines at the structural and molecular levels underlies stress resilience.

neuroscience↗

bioRxiv2020

PSD-95 in dorsal CA1 contributes to the persistence of fear memory

The ability to extinguish fearful memories is essential for survival. Accumulating data indicate that the dorsal CA1 area (dCA1) contributes to this process. However, the cellular and molecular basis of fear memory extinction remains poorly understood. Postsynaptic density protein 95 (PSD-95) regulates the structure and function of glutamatergic synapses. Here, using dCA1-targeted genetic and chemogenetic manipulations in vivo combined with PSD-95 immunostaining and 3D electron microscopy ex vivo, we demonstrate that phosphorylation of PSD-95 at serine 73 PSD-95(S73) is necessary for contextual fear extinction-induced expression of PSD-95 and synaptic plasticity. Moreover, PSD-95(S73) phosphorylation is not necessary for fear memory formation and recall but is required for extinction of contextual fear. Overall, our data shows how PSD-95-dependent synaptic plasticity in the hippocampus contributes to the persistence of fear memories.

neuroscience↗

bioRxiv2019

Contribution of PSD-95 protein to reward location memory

The molecular mechanisms involved in formation of memory are still poorly understood. We focus here on the function of post-synaptic density protein 95 (PSD-95) and its phosphorylation by CaMKII in spontaneous learning about reward location in female mice. We show that formation of reward location memory leads to downregulation of PSD-95 protein in dendritic spines of the stratum radiatum, area CA1, and selective shrinkage of dendritic spines that contain PSD-95. ShRNA-driven, long-term downregulation of PSD-95 in the area CA1 decreases precision of memory. Autophosphorylation deficient CaMKII mutant mice (CaMKII:T286A) need more time than wild-type animals to learn the location of reward. The same impairment is observed after CA1-targeted overexpression of CaMKII phosphorylation-deficient form of PSD-95 (PSD-95:S73A). In contrast to young adult mice, in aged animals reward location learning affects only spines that lack PSD-95. The frequency and size of the spines without PSD-95 are increased, while shRNA targeted to PSD-95 affects neither speed of learning nor precision of memory indicating alternative mechanisms to support successful memory formation in old mice. Altogether, our data suggest that dynamic regulation of PSD-95 expression is a mechanism that accelerates learning and improves precision of reward location memory in young mice. The function of PSD-95 in memory processes changes in aged animals.

neuroscience↗