Language：English   Publisher：Circulation Research  

BACKGROUND: The internal milieu of the body is controlled by a system of interoceptors coupled to motor outflows that drive compensatory adaptive responses. These include the arterial chemoreceptors, best known for sensing arterial oxygen. In cardiometabolic diseases, such as essential hypertension, the carotid bodies (CB) exhibit heightened reflex sensitivity and tonic activity without an apparent stimulus. The mechanisms behind CB sensitization in these conditions are not well understood. METHODS: Guided by functional genomics, a range of functional assays is used to interrogate downstream intracellular and interorgan signaling pathways involved in arterial chemosensory function. RESULTS: Here, we report the presence of the MC4R (melanocortin 4 receptor) in the mammalian CB and show its elevated expression in experimental hypertension. We demonstrate that melanocortin agonists activate arterial chemosensory cells, modulating CB chemosensory afferent drive to influence chemoreflex-evoked sympathetic and ventilatory activity. Transcriptional analysis of hypertensive CB implicates the activation of the Mash1 (mammalian achaete-scute homolog 1; Ascl1) regulatory network in driving elevated Mc4r expression. CONCLUSIONS: Collectively, our data indicate a primarily pathophysiological role of melanocortin signaling in arterial chemosensation, contributing to excess sympathetic activity in cardiometabolic disease.

DOI： 10.1161/CIRCRESAHA.125.326394

Web of Science

Scopus

PubMed

Language：English  

DOI： 10.1016/j.cmet.2024.02.010

PubMed

Language：Japanese   Publisher：Human Genetics  

CRISPR-Cas9 are widely used for gene targeting in mice and rats. The non-homologous end-joining (NHEJ) repair pathway, which is dominant in zygotes, efficiently induces insertion or deletion (indel) mutations as gene knockouts at targeted sites, whereas gene knock-ins (KIs) via homology-directed repair (HDR) are difficult to generate. In this study, we used a double-stranded DNA (dsDNA) donor template with Cas9 and two single guide RNAs, one designed to cut the targeted genome sequences and the other to cut both the flanked genomic region and one homology arm of the dsDNA plasmid, which resulted in 20–33% KI efficiency among G0 pups. G0 KI mice carried NHEJ-dependent indel mutations at one targeting site that was designed at the intron region, and HDR-dependent precise KIs of the various donor cassettes spanning from 1 to 5 kbp, such as EGFP, mCherry, Cre, and genes of interest, at the other exon site. These findings indicate that this combinatorial method of NHEJ and HDR mediated by the CRISPR-Cas9 system facilitates the efficient and precise KIs of plasmid DNA cassettes in mice and rats.

DOI： 10.1007/s00439-020-02198-4

Open Access

Scopus

PubMed

Event date： 2024.3

Language：English   Presentation type：Symposium, workshop panel (public)  

Event date： 2023.12

Language：Japanese   Presentation type：Symposium, workshop panel (public)  

Venue：東京都健康長寿医療センター研究所 会議室  

Event date： 2023.3

Language：English   Presentation type：Poster presentation  

Event date： 2022.9

Language：English   Presentation type：Oral presentation (invited, special)  

Country：Australia  

Audience： General

Type：Seminar, workshop