Protocol for altering connexin hemichannel function in primary chicken lens fiber cells using high-titer retroviral RCAS(A) infection

Yu Du, Francisca M. Acosta, Jianping Zhang, Yuxin Tong, Yumeng Quan, Sumin Gu, Jean X. Jiang

Research output: Contribution to journalArticlepeer-review

Abstract

Connexins (Cxs) play a crucial role in maintaining lens transparency. Here, we present a protocol for altering Cx hemichannel (HC) function in primary chicken lens fiber cells using high-titer retroviral replication competent avian sarcoma-leukosis virus long terminal repeat with splice acceptor (A) infection. We describe steps for incubating eggs, isolating lenses, culturing cells, preparing reagents, and infecting cells. We then detail cell treatment and detection of apoptosis and death. This protocol can assess protein kinase A, HC activity, and increased glutathione transport for protecting lens fiber cells against oxidative stress. For complete details on the use and execution of this protocol, please refer to Liu et al.,1 Riquelme et al.,2 Shi et al.,3 Jiang,4 and Rath et al.5

Original languageEnglish (US)
Article number102564
JournalSTAR Protocols
Volume4
Issue number4
DOIs
StatePublished - Dec 15 2023

Keywords

  • Cell Biology
  • Cell-Based Assays

ASJC Scopus subject areas

  • General Neuroscience
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology

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