May 2006
Volume 47, Issue 13
ARVO Annual Meeting Abstract  |   May 2006
Comparison of Functional Properties of Single M– and S–Cones from Wt And Gt–/– Mice
Author Affiliations & Notes
  • S.S. Nikonov
    Ophthalmology, University of Pennsylvania, Philadelphia, PA
  • J. Lem
    Ophthalmology & Cardiology, Tufts–New England Med Ctr, Boston, MA
  • E.N. Pugh, Jr.
    Ophthalmology, University of Pennsylvania, Philadelphia, PA
  • Footnotes
    Commercial Relationships  S.S. Nikonov, None; J. Lem, None; E.N. Pugh, None.
  • Footnotes
    Support  NIH Grant EY02660, RPB Foundation
Investigative Ophthalmology & Visual Science May 2006, Vol.47, 4313. doi:
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      S.S. Nikonov, J. Lem, E.N. Pugh, Jr.; Comparison of Functional Properties of Single M– and S–Cones from Wt And Gt–/– Mice . Invest. Ophthalmol. Vis. Sci. 2006;47(13):4313.

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      © ARVO (1962-2015); The Authors (2016-present)

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Purpose: : To compare functional properties of single S(UV)– and M– cones in WT and Gαt–/–mice.

Methods: : Cones of WT and Gαt–/– (transducin alpha knockout) mice were studied with suction pipette recordings. Cells were maintained at 35–37 C and stimulated with steps and flashes of light of varied intensity and wavelength. Rod activity was suppressed with 501 nm light.

Results: : Most mouse cones co–express S– and M–opsins, with M–opsin expression following a dorsal–ventral gradient. Responses of cones from the most ventral region (n=8, WT) were S–dominant, from the dorsal region M–dominant (n=5, WT; n=4, Gαt–/–). WT M–dominant cones (n=8) were from 4 to 9 times more sensitive to 501 nm than to 361 nm light, indicating low levels (if any) of S–opsin co–expression. The kinetics of S– and M–opsin driven responses of individual cones were nearly identical for all cones. WT M–dominant cones appear to be slightly faster and less sensitive than S–dominant cones, with response time to peak (tpeak), dominant recovery time constant (τ D), and half–saturating flash intensity (Q1/2) as follows: tpeak = 63 ± 6 ms, τ D = 69 ± 22 ms, Q1/2=7,900 ± 1,700 photons µm–2 (M–dominant); tpeak = 73 ± 5 ms, τ D = 73 ± 10 ms, Q1/2=5,400 ± 1,500 photons µm–2 (S–dominant). The responses of M– (n=6) and S–dominant (n=8) WT cones to steps of light at lower light levels followed a hyperbolic response vs. intensity relation with I1/2 ∼ 100,000 – 200,000 photons s–1 µm–2. However, increasing the background intensity up to 3,500,000 photons s–1 µm–2 did not cause complete suppression of the circulating current in 5 out of 6 M–dominant cones. Calculations suggest this escape from saturation is caused by reduction in light capture due to bleaching of the M–pigment. Even after bleaching up to 90% of the M–pigment, in absence of light the amplitude of the circulating current remained essentially unchanged from its pre–bleach level. S– and M–cones of Gαt–/– mice were slower and more sensitive than WT cones, with tpeak and τ D ∼ 100 ms, Q1/2 ∼ 4,000 photons µm–2 and I1/2 ∼ 30,000 photons s–1 µm–2.

Conclusions: : Pigment bleaching appears to be one of the mechanisms allowing cones to escape saturation by a background light. Very large amounts of bleached pigment per se produce little (if any) suppression of the circulating current of M–dominant mouse cones. The difference in kinetics and sensitivity between WT and Gαt–/– cones may be explained by adaptation to rod suppressing background; however, the difference in step sensitivity of WT and Gαt–/– cones suggests that functional rods may be required for fully normal cone light adaptation behavior.

Keywords: photoreceptors • electrophysiology: non-clinical • opsins 

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