Single cell RNAseq of murine tissues after, sham operation, myocaradial infarction (MI) or MI and sleep fragmentation (SF)

It remains unclear how sleep influences inflammatory pathways after myocardial infarction (SF). In this study, we relied on established murine models and assessed how sleep fragmentation (SF) alters transcriptional programing in the blood, heart, and brain after MI. Overall design: Female C57BL/6 mice 10-12 weeks of age were subjected to sham operation or permanent ligation of the left anterior descending coronary artery (MI). Half the mice subjected to MI were exposed to SF daily during the light period beginning immediately after surgical recovery (~1 hour after MI). The other half of the MI mice and all the sham mice were left undisturbed and allowed to sleep habitually. There were five mice in each of the three experimental groups: sham, MI, MI+SF. Three days after sham or MI operation, all mice were perfused with cold PBS. The hearts were collected from the MI and MI+SF mice; brains were collected from the sham and MI mice; blood was collected from mice in all experimental groups. From the hearts, the infarcted left ventricle area from MI mice or the corresponding uninfarcted tissue from sham mice was isolated, enzymatically digested, and subjected to MACS bead sorting to enrich for CD45+ leukocytes. Brains were enzymatically digested, subjected to a percoll gradient, and CD45+ leukocytes were also enriched for using MACS bead sorting. For the blood samples, whole blood was subjected red blood cell lysis, and the remaining leukocytes were collected. All single cell suspensions from the same tissues and experimental groups were pooled, washed, and subjected to scRNAseq.