The general goal of this proposal is elucidation of the mechanism by which light generates the receptor potential in photoreceptors. This process can be divided into early stages involving the light- sensitive molecule rhodopsin, intermediate stages involving chemical amplification, and the final stage in which the chemical message affects ion channels, thereby generating the receptor potential. An aspect of the early steps, the average number of G- proteins activated by a single isomerized rhodopsin, will be investigated in experiments involving agents which block or stimulate G-protein. In addition the possibility of using peptides to inhibit specific reactions involving rhodopsin will be explored by dialyzing such peptides into tiger salamander rods or Limulus photoreceptors. The main questions involving intermediate events have to do with the new hypothesis that a rise in cGMP mediates the excitation of invertebrate photoreceptors. This will be studied in biochemical assays of the effect of light on guanylate cyclase activity, phosphodiesterase activity, and the concentration of cGMP in the living squid eye. Physiological methods will be used to study the effect of putative activators and inhibitors of enzymes involved in cyclic nucleotide metabolism. The general goal is to determine which enzyme is responsible for light-dependent changes in cGMP and how this enzyme might be controlled. Finally, anatomical methods will be used to identify the structural basis for the very inhomogeneous sensitivity to injected cGMP. Ongoing work on the light- activated channels in Limulus using the patch clam method will be continued. Preliminary results indicate that these channel events have two sizes. We will attempt to determine if these are due to two different states of the same channel or to two different channel types. These channels will also be studied in excised patches where it is possible to test whether substances of interest can have direct effects on the channel.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Method to Extend Research in Time (MERIT) Award (R37)
Project #
5R37EY001496-15
Application #
3483899
Study Section
Visual Sciences B Study Section (VISB)
Project Start
1977-06-01
Project End
1992-05-31
Budget Start
1988-06-01
Budget End
1989-05-31
Support Year
15
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Brandeis University
Department
Type
Schools of Arts and Sciences
DUNS #
616845814
City
Waltham
State
MA
Country
United States
Zip Code
02454
Segev, Ronen; Schneidman, Elad; Goodhouse, Joe et al. (2007) Role of eye movements in the retinal code for a size discrimination task. J Neurophysiol 98:1380-91
Garger, Alexander V; Richard, Edwin A; Lisman, John E (2004) The excitation cascade of Limulus ventral photoreceptors: guanylate cyclase as the link between InsP3-mediated Ca2+ release and the opening of cGMP-gated channels. BMC Neurosci 5:7
Chen, F H; Baumann, A; Payne, R et al. (2001) A cGMP-gated channel subunit in Limulus photoreceptors. Vis Neurosci 18:517-26
Garger, A; Richard, E A; Lisman, J E (2001) Inhibitors of guanylate cyclase inhibit phototransduction in limulus ventral photoreceptors. Vis Neurosci 18:625-32
Fain, G L; Lisman, J E (1999) Light, Ca2+, and photoreceptor death: new evidence for the equivalent-light hypothesis from arrestin knockout mice. Invest Ophthalmol Vis Sci 40:2770-2
Richard, E A; Ghosh, S; Lowenstein, J M et al. (1997) Ca2+/calmodulin-binding peptides block phototransduction in Limulus ventral photoreceptors: evidence for direct inhibition of phospholipase C. Proc Natl Acad Sci U S A 94:14095-9
Goldring, M A; Lisman, J E (1994) Multi-step rhodopsin inactivation schemes can account for the size variability of single photon responses in Limulus ventral photoreceptors. J Gen Physiol 103:691-726
Shin, J; Richard, E A; Lisman, J E (1993) Ca2+ is an obligatory intermediate in the excitation cascade of limulus photoreceptors. Neuron 11:845-55
Kahana, A; Robinson, P R; Lewis, L J et al. (1992) ATP-independent deactivation of squid rhodopsin. Vis Neurosci 9:595-602
Erickson, M A; Robinson, P; Lisman, J (1992) Deactivation of visual transduction without guanosine triphosphate hydrolysis by G protein. Science 257:1255-8

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