PMID- 37738121
OWN - NLM
STAT- MEDLINE
DCOM- 20240311
LR  - 20240311
IS  - 2666-1667 (Electronic)
IS  - 2666-1667 (Linking)
VI  - 4
IP  - 4
DP  - 2023 Dec 15
TI  - Protocol for altering connexin hemichannel function in primary chicken lens fiber 
      cells using high-titer retroviral RCAS(A) infection.
PG  - 102564
LID - S2666-1667(23)00531-2 [pii]
LID - 10.1016/j.xpro.2023.102564 [doi]
LID - 102564
AB  - 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).
CI  - Copyright (c) 2023 The Authors. Published by Elsevier Inc. All rights reserved.
FAU - Du, Yu
AU  - Du Y
AD  - Department of Biochemistry and Structural Biology, University of Texas Health 
      Science Center, San Antonio, TX 78229-3900, USA; Department of Ophthalmology, 
      Lanzhou University Second Hospital; Second Clinical School, Lanzhou University, 
      Lanzhou, Gansu 730000, China. Electronic address: duyuyanke@163.com.
FAU - Acosta, Francisca M
AU  - Acosta FM
AD  - Department of Biochemistry and Structural Biology, University of Texas Health 
      Science Center, San Antonio, TX 78229-3900, USA.
FAU - Zhang, Jianping
AU  - Zhang J
AD  - Department of Biochemistry and Structural Biology, University of Texas Health 
      Science Center, San Antonio, TX 78229-3900, USA.
FAU - Tong, Yuxin
AU  - Tong Y
AD  - Department of Biochemistry and Structural Biology, University of Texas Health 
      Science Center, San Antonio, TX 78229-3900, USA.
FAU - Quan, Yumeng
AU  - Quan Y
AD  - Department of Biochemistry and Structural Biology, University of Texas Health 
      Science Center, San Antonio, TX 78229-3900, USA.
FAU - Gu, Sumin
AU  - Gu S
AD  - Department of Biochemistry and Structural Biology, University of Texas Health 
      Science Center, San Antonio, TX 78229-3900, USA.
FAU - Jiang, Jean X
AU  - Jiang JX
AD  - Department of Biochemistry and Structural Biology, University of Texas Health 
      Science Center, San Antonio, TX 78229-3900, USA. Electronic address: 
      jiangj@uthscsa.edu.
LA  - eng
GR  - F32 DK134051/DK/NIDDK NIH HHS/United States
GR  - R01 EY012085/EY/NEI NIH HHS/United States
PT  - Journal Article
DEP - 20230921
PL  - United States
TA  - STAR Protoc
JT  - STAR protocols
JID - 101769501
RN  - 0 (Connexins)
SB  - IM
MH  - Animals
MH  - *Connexins/genetics/metabolism
MH  - Chickens
MH  - Retroviridae/genetics/metabolism
MH  - *Lens, Crystalline/metabolism
MH  - Epithelium/metabolism
PMC - PMC10519848
OTO - NOTNLM
OT  - Cell Biology
OT  - Cell-Based Assays
COIS- Declaration of interests The authors declare no competing interests.
EDAT- 2023/09/22 18:42
MHDA- 2024/03/11 06:43
PMCR- 2023/09/21
CRDT- 2023/09/22 12:35
PHST- 2023/04/29 00:00 [received]
PHST- 2023/07/05 00:00 [revised]
PHST- 2023/08/18 00:00 [accepted]
PHST- 2024/03/11 06:43 [medline]
PHST- 2023/09/22 18:42 [pubmed]
PHST- 2023/09/22 12:35 [entrez]
PHST- 2023/09/21 00:00 [pmc-release]
AID - S2666-1667(23)00531-2 [pii]
AID - 102564 [pii]
AID - 10.1016/j.xpro.2023.102564 [doi]
PST - ppublish
SO  - STAR Protoc. 2023 Dec 15;4(4):102564. doi: 10.1016/j.xpro.2023.102564. Epub 2023 
      Sep 21.