cGMP accumulation causes photoreceptor degeneration in CNG channel deficiency.

1. As you may know, Cyclic-nucleotide gated channels are a family of channels which bind to and
are activated by cyclic nucleotides, and manipulate ion concentrations within the cell. The very
first cyclic-nucleotide-gated ion channels to be discovered were in rod photoreceptors within the
retina. It is known that these play a fundamental role in phototransduction, the process which
enables light energy to produce a change in the electrical potential across the cell membrane,
enabling a signal to be produced and sent to the brain.

2. When your eyes are closed, or when it’s very dark, this channel is actively bound to cGMP, and
thus remains open, with constant influx of sodium and calcium ions. When your eyes are open
and able to perceive colour, cGMP is released and there is no movement of ions through the
channel.

3. Specifically, upon photon absorption, the rhodopsin receptor becomes active, which activates a
G-protein-coupled receptor, which in turn activates photoreceptor phosphodiesterase PDE6.
PDE6 hydrolyses diffusible cGMP resulting in free cytoplasmic cGMP decline. This causes
surface membrane cyclic-nucleotide gated ion channels to close, effectively hyperpolarizing the
cell. This leads to eventual termination of glutamate release at the synaptic terminal by further
processes. The lack of glutamate then acts as a signal which is further processed by other neurons
in the retina before being transmitted to higher centres in the brain.

4. The described CNG is a tetramer, consisting of 3 CNGA3 subunits and 1 CNGB3 gene.

5. Mutations in either gene causes achromatopsia, or total colorblindness due to loss of function,
causing degeneration and deficiency of these channels.

5. In rod and cone cells, cGMP synthesis is conducted by an enzyme called retinal guanylyl cyclase,
or retGC, whereas cGMP is degraded by the PDE6 enzyme as mentioned previously. The retGC
enzyme is first activated by regulating proteins called calcium binding guanylyl cyclase-activating
proteins, or GCAPs, which respond to low concentrations of intracellular calcium. From the last
slide, we know that a reduction in calcium can be a direct result of light-activated PDE6
hydrolysing cGMP, causing subsequent closure of the cGMP channels.

However, high rates of cGMP synthesis or low rates of hydrolysis have been found to cause cGMP
accumulation, which leads to photoreceptor death and thus degeneration of CNG channels.
Accumulation of cGMP is primarily due to mutations affecting PDE6 or retGC activity.
Mutations in PDE6 subunits may render it unable to hydrolyse cGMP, causing overaccumulation.
Mutations in GCAPs (or the proteins which activate retGC) causing the enzyme to activate
without light or an active retGC, can also incur cGMP accumulation, leading to cyclic-nucleotide
gated channel degeneration and thus totalcolorblindness.

6. Essentially what the current experiment aimed to explore was the mechanism of photoreceptor
degeneration in mice with either of two channel deficiency mutations; a mutation in the CNGA3
subunit of the channel itself, or a knockout line with no rod expression and a cone-dominant
background. This was done by maintaining mice in low light conditions, extracting retinal cGMP,
and making comparisons in cGMP levels.