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Biology subjects

Savic, N.

Publications and source records attributed to Savic, N..

4 recordsLinked to original sources

The nucleus accumbens shell regulates hedonic feeding via a rostral hotspot

The medial nucleus accumbens shell (medNAcSh) is increasingly recognized as a key node in the regulation of hedonic feeding, exerting strong control over reward consumption through its projections to downstream structures. Recent studies showed that one of the primary cellular mediators of these effects are dopamine 1 receptor positive striatal projection neurons (D1-SPNs). Specifically, population D1-SPN activity gets inhibited during reward consumption and such inhibition is necessary and sufficient to authorize reward consumption, independent of metabolic need. Anatomically, the medNAcSh spans a 1 to 1.5 mm distance along the rostro-caudal axis in mice and previous studies have reported functional gradients along this axis in behavioral control. For instance, classical pharmacological studies have suggested that rostral rather than caudal medNAcSh regulates appetitive behavior. However, the cellular, circuit, and molecular mechanisms underlying this topographical gradient remain unknown. Here we hypothesized that D1-SPNs contribute to this gradient by regulating hedonic feeding via a specific hotspot in the rostral medNAcSh. Using population level calcium monitoring with fiber photometry in mice, we show that rostral medNAcSh D1-SPNs demonstrate inhibitory responses during reward consumption, while caudal D1-SPNs do not. Consistently, optogenetic stimulation of rostral medNAcSh D1-SPNs inhibits consumption, while stimulation of caudal D1-SPNs had minimal effects, confirming the existence of a functional rostro-caudal gradient. Importantly, we observed no differences between rostral and caudal D1-SPNs in their behavioral or activity responses to aversive stimuli, suggesting that the D1-SPN gradient is specific to appetitive contexts. To investigate potential molecular correlates of this functional gradient, we leveraged open-source anatomy datasets and performed fluorescent in situ hybridization, identifying Stard5 and Peg10 as markers enriched in the rostral and caudal medNAcSh, respectively. Finally, we developed a novel Stard5-Flp driver mouse line, which allowed us to target the rostral hotspot selectively. We demonstrated that Stard5+ neurons recapitulate activity patterns of rostral medNAcSh D1-SPNs. Together, these findings establish a spatially confined rostral subregion of the medNAcSh as a critical regulator of reward consumption and introduce Stard5 as a molecular tool for its manipulation - offering new opportunities for intervention in dysregulated eating beyond classical homeostatic circuits.

neuroscience↗

Intrinsic disorder specifies discreet functions of nucleoplasmin-like histone chaperones

Nucleoplasmin (NPM) histone chaperones regulate distinct processes in the nucleus and nucleolus. While intrinsically disordered regions (IDRs) are hallmarks of NPMs, it is not clear if all NPM functions require these unstructured features. We assessed the importance of IDRs in yeast NPM-like proteins and found that regulation of rDNA copy number, and genetic interactions with the nucleolar RNA surveillance machinery require the highly conserved FKBP prolyl isomerase domain, but not the NPM domain or IDRs. By contrast, transcriptional silencing in the nucleus requires IDRs. We demonstrate that multiple lysines in poly-acidic serine/lysine motifs of IDRs are required for both lysine polyphosphorylation and NPM-mediated transcriptional silencing. These results demonstrate that NPM-like proteins do not rely on IDRs for all chromatin-related functions.

molecular biology↗

Genetic dissection of the redundant and divergent functions of histone chaperone paralogs in yeast

Gene duplications increase organismal robustness by providing freedom for gene divergence or by increasing gene dosage. The yeast histone chaperones Fpr3 and Fpr4 are paralogs that can assemble nucleosomes in vitro, however the genomic locations they target and their functional relationship is poorly understood. We refined the yeast synthetic genetic array (SGA) approach to enable the functional dissection of gene paralogs. Applying this method to Fpr3 and Fpr4 uncovered their redundant and divergent functions: while Fpr3 is uniquely involved in chromosome segregation, Fpr3 and Fpr4 co-operate on some genes and are redundant on others where they impact gene expression and transcriptional processivity. We find that the TRAMP5 RNA exosome is essential in{Delta} fpr3{Delta}fpr4 yeast and leverage this information to identify Fpr3/4 target loci. Amongst these are the non-transcribed spacers of ribosomal DNA where either paralog is sufficient to establish chromatin that is both transcriptionally silent and refractory to recombination. These data provide evidence that Fpr3 and Fpr4 have shared chromatin-centric functions, especially at nucleolar rDNA. However, their distinct genetic interaction profiles show they also have evolved separate functions outside of the nucleolus.

genetics↗

Covalent linkage of the DNA repair template to the CRISPR/Cas9 complex enhances homology-directed repair

The CRISPR/Cas9 targeted nuclease technology allows the insertion of genetic modifications with single base-pair precision. The preference of mammalian cells to repair Cas9-induced DNA double-strand breaks via non-homologous end joining (NHEJ) rather than via homology-directed repair (HDR) however leads to relatively low rates of correctly edited loci. Here we demonstrate that covalently linking the DNA repair template to Cas9 increases the ratio of HDR over NHEJ up to 23-fold, and therefore provides advantages for clinical applications where high-fidelity repair is needed.

molecular biology↗