This review comprehensively explores the complex interplay between extracellular vesicles (ECVs)/exosomes and circadian rhythms, with a focus on the role of this interaction in hepatocellular carcinoma (HCC). Exosomes are nanovesicles derived from cells that facilitate intercellular communication by transporting bioactive molecules such as proteins, lipids, and RNA/DNA species. ECVs are implicated in a range of diseases, where they play crucial roles in signaling between cells and their surrounding environment. In the setting of cancer, ECVs are known to influence cancer initiation and progression. The scope of this review extends to all cancer types, synthesizing existing knowledge on the various roles of ECVs. A unique aspect of this review is the emphasis on the circadian-controlled release and composition of exosomes, highlighting their potential as biomarkers for early cancer detection and monitoring metastasis. We also discuss how circadian rhythms affect multiple cancer-related pathways, proposing that disruptions in the circadian clock can alter tumor development and treatment response. Additionally, this review delves into the influence of circadian clock components on ECV biogenesis and their impact on reshaping the tumor microenvironment, a key component driving HCC progression. Finally, we address the potential clinical applications of ECVs, particularly their use as diagnostic tools and drug delivery vehicles, while considering the challenges associated with clinical implementation.

The ability to manipulate neurons that express corticotropin‐releasing factor (CRF) or its receptors has led to many new discoveries about how CRF signaling modulates stress‐related behaviors in mice. However, similar advances in rats have been limited by the lack of reporter lines targeting CRF receptors, restricting access to behavioral paradigms that are more suitable for, or specific to, rats. Here, we provide an extensive neuroanatomical characterization of a recently generated CRFR1‐Cre rat. We show that Cre and tdTomato are expressed in a pattern that closely aligns with previously reported CRFR1 expression patterns, as determined by in situ hybridization in rats and by genetic labeling in CRFR1 transgenic mice. We detail expression patterns at multiple rostrocaudal levels to validate the use of this transgenic rat in multiple neural circuits that signal via CRF activation of CRFR1. Furthermore, we demonstrate that expression of Cre recombinase, combined with viral vector delivery, can be used to trace CRFR1‐expressing neurons and their axonal terminals in target regions. This study is intended to serve as a resource for examining the expression pattern of CRFR1 and as a validation of the CRFR1‐Cre rat for reliable use with Cre‐dependent viral approaches to manipulate and/or record the activity of CRFR1‐expressing neurons in distinct brain regions. We expect that the availability of the CRFR1‐Cre rat, along with an increasing number of transgenic rat lines, will further expand our understanding of how CRF transmission mediates stress responses by leveraging the behavioral strengths of the rat as a model system.

Background: Cannabis is widely used during pregnancy, increasing with cannabis legalization, and has been associated with altered neurodevelopment, yet the precise molecular mechanisms remain poorly understood. Aims: To assess whether utilizing fetal central nervous system-derived extracellular vesicles (fCNSEVs) isolated from maternal blood reflects cannabis-associated changes in the fetal brain. Methods: In a matched case-control study, maternal plasma (to isolate fCNSEVs) and paired fetal cortical tissue (9–18 weeks’ gestation) from pregnancies exposed or unexposed to cannabis were collected. We quantified protein (CB1R, D2R) and miRNA expression (using µ-paraflo ^TM microarray). Selected miRNAs were validated by qPCR. Bayesian Generalized Linear Models were used to assess exposure-group by sex effects. Results: Maternal cannabis use was associated with altered CB1R and D2R levels in the brain with opposite directional changes in fCNSEVs. 21 miRNAs were differentially expressed: 14 downregulated and 6 upregulated, following a sex-dependent pattern (males>>females), while miR-216a-5p follows the opposite pattern (females>>males). We confirmed concordant miRNA changes in fetal brain and fCNSEVs. Many of the identified molecules are associated with pathways linked to adverse neurodevelopmental outcomes. Conclusions: Our data suggest that fCNSEV-based assays may provide the ability to monitor fetal brain effects of maternal cannabis in ongoing pregnancies in clinical cohorts. A critical next step is to determine if alterations in fCNSEV protein and miRNA markers predict changes in fetal brain connectivity and neurodevelopment measures of executive function.

Animal welfare has evolved during the past decades to improve not only the quality of life of laboratory animals but also the quality and reproducibility of scientific investigations. Bibliometric analysis has become an important tool to complete the current knowledge with academic databases. Our objective was to investigate whether scientific research on cannibalism/infanticide is connected with maternal aggression towards the offspring in laboratory animals. To carry out our research, we performed a specific search for published articles on each concept. Results were analyzed in the opensource environment RStudio with the package Bibliometrix. We obtained 228 and 134 articles for the first search (cannibalism/infanticide) and the second search (maternal aggression towards the pups) respectively. We observed that the interest in infanticide cannibalism started in the 1950s, while researchers started showing interest in maternal aggression towards the pups 30 years later. Our analyses indicated that maternal aggression had better citations in scientific literature. In addition, although our results showed some common features (e.g., oxytocin or medial preoptic area in the brain), we observed a gap between cannibalism/infanticide and maternal aggression towards the pups with only 18 published articles in common for both the searches. Therefore, we recommend researchers to combine both concepts in further investigations in the context of cannibalism for better dissemination and higher impact in laboratory animals’ welfare research. Highlights - Cannibalism/Infanticide and maternal aggression have been investigated separately. - Maternal aggression has a higher impact on scientific literature. - Combining both topics may increase cannibalism/infanticide impact.

Alcohol relapse is associated with corticotropin-releasing factor (CRF) signaling and altered reward pathway function, though the precise mechanisms remain unclear. Here, using both mice and rats, we investigated how CRF modulates cholinergic interneurons (CINs) in the dorsal striatum, a region critical in mediating cognitive flexibility and action selection. Using monosynaptic and retrograde circuit tracing, we identified direct inputs from CRF-expressing (CRF ⁺ ) neurons in the central amygdala (CeA) and bed nucleus of the stria terminalis (BNST) to dorsal striatal CINs. We showed that CINs express CRF receptor 1 (CRFR1) and established their functional connectivity with CeA/BNST CRF ⁺ projections. Functional recordings revealed that CRF enhanced CIN excitability and promoted acetylcholine release in the dorsal striatum. However, acute alcohol exposure and withdrawal attenuated the excitatory effect of CRF on CIN firing, suggesting a mechanism by which alcohol disrupts CRF-dependent neuromodulation. These findings reveal a previously unrecognized CRF-CIN pathway linking the extended amygdala to the dorsal striatum and provide new insight into how CRF and alcohol interact to impair striatal function. This work highlights CRF signaling as a potential target for understanding stress-induced changes to the reward pathway.