Antisense oligonucleotide experiments elucidate the essential role of titin in sarcomerogenesis in adult rat cardiomyocytes in long-term culture

• DOI: 10.1242/jcs.113.21.3851
• Publication Date: 2021-04-25T18:34:29Z
• Authors: Veronika Person; Sawa Kostin; Keisuke Suzuki; Siegfried Labeit; Jutta Schaper
• Keywords: Sarcomeres; 0303 health sciences; Base Sequence; Myocardium; Muscle Proteins; Heart; Oligonucleotides, Antisense; Rats; 03 medical and health sciences; Animals; Connectin; RNA, Messenger; Rats, Wistar; Protein Kinases; Cells, Cultured; DNA Primers

An essential role of titin as a molecular ruler in sarcomerogenesis has been frequently discussed. In this study, we tested the hypothesis that expression is prerequisite for thick filament incorporation into sarcomeres by using an antisense oligonucleotide approach to interfere with translation de-/redifferentiation model adult rat cardiomyocytes (ARC) long-term culture. As first step, growth pattern ranging from rod shape round and later spreading cells cell surface area ARC were quantitatively evaluated standardized. This represents basis experiments interfering sarcomere formation three different phosphorothioate oligonucleotides (S-ODN) at dosage 10 microM specific mRNA. Presence fluorescein labeled S-ODN indicated cellular uptake both, random S-ODN, induced significant increase size compared control untreated ARC. At days 12 16 culture, treatment resulted reduced disturbance myosin sarcomeres, evident diffuse labeling significantly decreased regular cross-striation (control 75%, day 20%, 14%) shown laser scanning confocal microscopy. Cellular integrity presence alpha-actinin was not disturbed. These findings provide evidence template therefore sarcomerogenesis.