 This study examined the role of alternative isoforms of a gene in the cochlea, providing new insights into how they affect biological functions. The researchers used multiple methods including single cell INasec, RTBCR, Tanger sequencing, and mass spectrometry to identify and verify unannotated transcripts. They then focused on one gene, Otoferlin, which is important for synaptic transmission in inner hair cells. By deleting the canonical isoform, they discovered a shorter version of the gene that can support normal hearing thresholds but with reduced sustained exocytosis of inner hair cells. Further analysis showed that this shorter isoform is also associated with auditory phenotypes and IHC functions. This study provides valuable insight into the roles of alternative isoforms in the cochlea and their potential impact on auditory function. This article was authored by Huahui Liu, Hongchao Liu, Leng Hao Wang, and others.