title: Activation and quenching of the phototransduction cascade in retinal cones as inferred from electrophysiology and mathematical modeling.
abstract: OBJECTIVE
  To experimentally identify and quantify factors responsible for the lower sensitivity of retinal cones compared to rods.
  
  
  METHODS
  Electrical responses of frog rods and fish (Carassius) cones to short flashes of light were recorded using the suction pipette technique. A fast solution changer was used to apply a solution that fixed intracellular Ca2+ concentration at the prestimulus level, thereby disabling Ca2+ feedback, to the outer segment (OS). The results were analyzed with a specially designed mathematical model of phototransduction. The model included all basic processes of activation and quenching of the phototransduction cascade but omitted unnecessary mechanistic details of each step.
  
  
  RESULTS
  Judging from the response versus intensity curves, Carassius cones were two to three orders of magnitude less sensitive than frog rods. There was a large scatter in sensitivity among individual cones, with red-sensitive cones being on average approximately two times less sensitive than green-sensitive ones. The scatter was mostly due to different signal amplification, since the kinetic parameters of the responses among cones were far less variable than sensitivity. We argue that the generally accepted definition of the biochemical amplification in phototransduction cannot be used for comparing amplification in rods and cones, since it depends on an irrelevant factor, that is, the cell's volume. We also show that the routinely used simplified parabolic curve fitting to an initial phase of the response leads to a few-fold underestimate of the amplification. We suggest a new definition of the amplification that only includes molecular parameters of the cascade activation, and show how it can be derived from experimental data. We found that the mathematical model with unrestrained parameters can yield an excellent fit to experimental responses. However, the fits with wildly different sets of parameters can be virtually indistinguishable, and therefore cannot provide meaningful data on underlying mechanisms. Based on results of Ca2+-clamp experiments, we developed an approach to strongly constrain the values of many key parameters that set the time course and sensitivity of the photoresponse (such as the dark turnover rate of cGMP, rates of turnoffs of the photoactivated visual pigment and phosphodiesterase, and kinetics of Ca2+ feedback). We show that applying these constraints to our mathematical model enables accurate determination of the biochemical amplification in phototransduction. It appeared that, contrary to many suggestions, maximum biochemical amplification derived for "best" Carassius cones was as high as in frog rods. On the other hand, all turnoff and recovery reactions in cones proceeded approximately 10 times faster than in rods.
  
  
  CONCLUSIONS
  The main cause of the differing sensitivity of rods and cones is cones' ability to terminate their photoresponse faster.
authors: Institute for Evolutionary Physiology and Biochemistry, Russian Academy of Sciences.
authors: Luba
authors: L
authors: Astakhova
authors: Institute for Evolutionary Physiology and Biochemistry, Russian Academy of Sciences.
authors: Michael
authors: M
authors: Firsov
authors: Institute for Evolutionary Physiology and Biochemistry, Russian Academy of Sciences.
authors: Victor
authors: V
authors: Govardovskii
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002347:Carps; D006152:Cyclic GMP / Q000378:metabolism; D007700:Kinetics; D008027:Light; D055537:Light Signal Transduction / Q000502:physiology*; D008564:Membrane Potentials / Q000502:physiology*; D008954:Models, Biological; D009928:Organ Specificity; D018408:Patch-Clamp Techniques; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D055668:Photochemical Processes; D011895:Rana ridibunda; D017949:Retinal Cone Photoreceptor Cells / Q000166:cytology / Q000502:physiology*; D017948:Retinal Rod Photoreceptor Cells / Q000166:cytology / Q000502:physiology*; D013997:Time Factors
chemical_list: D010727:Phosphoric Diester Hydrolases; D006152:Cyclic GMP; D002118:Calcium