Dibyendu Chakraborty1, Shannon M Conley1, Marci L DeRamus2, Steven J Pittler2, Muna I Naash3. 1. Department of Cell Biology University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, United States. 2. Department of Vision Sciences, University of Alabama at Birmingham, Birmingham, Alabama, United States. 3. Department of Biomedical Engineering, University of Houston, Houston, Texas, United States.
Abstract
PURPOSE: The beta subunit of the rod cyclic nucleotide gated channel B1 (CNGB1) contains a proline/glutamic acid-rich N-terminal domain (GARP), which is also present in rods as a non-membrane-bound protein (GARP1/2). GARP2 and CNGB1 bind to retinal degeneration slow (RDS), which is present in the rims of rod and cone outer segment (OS) layers. Here we focus on the importance of RDS/GARP complexes in OS morphogenesis and stability. METHODS: Retinal structure, function, and biochemistry were assessed in GARP2-Tg transgenic mice crossed onto rds+/+, rds+/-, and rds-/- genetic backgrounds. RESULTS: GARP2 expression decreased in animals with reduced RDS levels. Overexpression of GARP2 led to abnormalities in disc stacking in GARP2-Tg/rds+/+ and the accumulation of abnormal vesicular structures in GARP2-Tg/rds+/- OS, as well as alterations in RDS-ROM-1 complex formation. These abnormalities were associated with diminished scotopic a- and b-wave amplitudes in GARP2-Tg mice on both the rds+/+ and rds+/- backgrounds. In addition, severe defects in cone function were observed in GARP2-Tg mice on all RDS backgrounds. CONCLUSIONS: Our results indicate that overexpression of GARP2 significantly exacerbates the defects in rod function associated with RDS haploinsufficiency and leads to further abnormalities in OS ultrastructure. These data also suggest that GARP2 expression in cones can be detrimental to cones. RDS/GARP interactions remain under investigation but are critical for both OS structure and function.
PURPOSE: The beta subunit of the rod cyclic nucleotide gated channel B1 (CNGB1) contains a proline/glutamic acid-rich N-terminal domain (GARP), which is also present in rods as a non-membrane-bound protein (GARP1/2). GARP2 and CNGB1 bind to retinal degeneration slow (RDS), which is present in the rims of rod and cone outer segment (OS) layers. Here we focus on the importance of RDS/GARP complexes in OS morphogenesis and stability. METHODS: Retinal structure, function, and biochemistry were assessed in GARP2-Tg transgenic mice crossed onto rds+/+, rds+/-, and rds-/- genetic backgrounds. RESULTS: GARP2 expression decreased in animals with reduced RDS levels. Overexpression of GARP2 led to abnormalities in disc stacking in GARP2-Tg/rds+/+ and the accumulation of abnormal vesicular structures in GARP2-Tg/rds+/- OS, as well as alterations in RDS-ROM-1 complex formation. These abnormalities were associated with diminished scotopic a- and b-wave amplitudes in GARP2-Tg mice on both the rds+/+ and rds+/- backgrounds. In addition, severe defects in cone function were observed in GARP2-Tg mice on all RDS backgrounds. CONCLUSIONS: Our results indicate that overexpression of GARP2 significantly exacerbates the defects in rod function associated with RDShaploinsufficiency and leads to further abnormalities in OS ultrastructure. These data also suggest that GARP2 expression in cones can be detrimental to cones. RDS/GARP interactions remain under investigation but are critical for both OS structure and function.
Authors: Simon M Petersen-Jones; Nathaniel Pasmanter; Laurence M Occelli; Janice R Querubin; Paige A Winkler Journal: Doc Ophthalmol Date: 2022-09-15 Impact factor: 1.854