Background Regulation of transcript structure generates transcript isoform and splicing diversity and plays an important role in health and disease. Differences in isoform expression account for substantial effect sizes and genetic enrichments in neuropsychiatric disorders such as ASD and schizophrenia; however, a systematic characterization of isoform expression in the human brain is lacking as most isoforms are yet to be directly profiled at the tissue or cellular level. Methods Here, we performed deep single-molecule real-time (SMRT) isoform long-read sequencing of prefrontal cortex (PFC), cerebellum (CBL), hippocampus (HIP) and dorsal anterior cingulate cortex (dACC) from >5 0 individuals and 7 pools, totaling > 60 libraries with > 100 SMRT cells and > 250 M long-reads. Results Our map of > 350k isoforms greatly expands the transcriptional landscape of brain-expressed genes with > 150k novel unannotated complete isoforms (54%), extends known coding and noncoding gene annotations, and uncovers hundreds of novel or reconstructed gene loci that can show cell-type specific expression. Orthogonal data, including, ribosome sequencing and mass-spectrometry data from > 280 prefrontal cortical samples, 5’-end TSS profiling, and nanopore long-read sequencing, were used to further characterize the structure of novel isoforms and open reading frames. We further leveraged PsychENCODE single-nuclei transcriptomic and chromatin data to annotate cell-type-specific regulatory interactions for novel isoforms, reconstructed loci, and novel genes. Multi-platform analysis of single-nuclei full-length transcripts across PFC, HIP and striatum further revealed that isoform diversity seen in brain tissues is often due to underlying cell-type-specific patterns. Finally, we leveraged our reference map of full-length isoforms to characterize medically relevant genes, assess the functional consequence of rare genetic variants from a compendium of disease studies, and to improve quantitative trait analyses and refine the prioritization of candidate risk genes. Discussion Taken together, we provide a comprehensive resource to characterize dysregulation of isoform expression in neuropsychiatric disorders. Disclosure Nothing to disclose.

Recent research has shown expression of clock genes in peripheral tissue explants, targeting multiple pathways leading to the entrainment of circadian rhythms. Temporal variations are not solely regulated by photoperiod, but factors such as maternal feed availability can entrain fetal circadian clock. Currently, a paucity of information exists for clock gene expression and short-term temporal transcript abundance in the bovine placenta, which is essential for proper offspring development. Therefore, the objective of this study was to determine the effect of early to mid-gestational nutrient restriction on clock genes, angiogenic factors, and nutrient sensing genes mRNA transcript abundance in placental explants during a 24 h period. Placentomes from adequately fed and nutrient restricted heifers were collected via Cesarean section at day 180 of gestation; separated into caruncular and cotyledonary tissue and placed in culture media for a 24 h period. The mRNA transcript abundance of clock genes (ARNTL, CRY1, and PER2), angiogenic factors (HIF1A and VEGFA), and nutrient sensing genes (NAMPT and NR3C1) was determined every 4 h. Clock genes were expressed in caruncular and cotyledonary explant tissue. The caruncular explant transcript abundance of the clock genes was not influenced by time (P > 0.05); while ARNTL abundance decreased over time in the cotyledon explant (P < 0.05). A main effect of time was observed for HIF1A, VEGFA, and NR3C1 in the caruncular tissue (P < 0.05). Although, angiogenic factors and nutrient sensing genes in cotyledonary tissue displayed evident temporal variation in transcript abundance (P < 0.05). Nutrient restriction did not alter (P > 0.15) mRNA transcript abundance of clock genes, angiogenic factors, or nutrient sensing genes in either caruncular or cotyledonary tissue. Interestingly, these data may indicate limited transmission and synchronization of maternal and fetal temporal variations in transcript abundance. These findings demonstrate that multiple timepoint collections are needed in future studies due to the innate existence of temporal oscillations observed in the bovine placenta.

Background and Aims: Intraplaque hemorrhage (IPH) is an important feature of plaque vulnerability. Patients with a history of transient ischemic attack or stroke are treated with platelet aggregation inhibitors to prevent secondary events, but it also has adverse effects. This study investigates the effect of antiplatelet agents on the formation of IPH.