The long-term objectives of this application are to elucidate the mechanisms that control light-dependent ion channels in photoreceptors, and the modulation of the photoresponse by ambient illumination. The proposed experiments take advantage of the unique double retina of organisms in which both evolutionary lines of visual receptors are represented: (1) rhabdomeric, which depolarize to light and utilize the phosphoinositide cascade for transduction, and (ii) ciliary, which hyperpolarize to light and, like vertebrate rods, rely on a cGMP-based cascade. Dissociated cells of both classes are exceptionally suitable for recording light-dependent whole-cell and single-channel currents, intracellular dialysis, and fluorescence measurements of cytosolic calcium. In rhabdomeric photoreceptors a light-regulated PLC is critically involved in excitation, but the IP3/Ca branch of the cascade has proved insufficient to account for phototransduction, and the importance of other elements of the PLC pathway has been gaining recognition. One component that was recently identified as a key signaling molecule in other systems is PIP2 itself. Proposed studies will examine its role in visual transduction. The pivotal role of Ca in light adaptation of rhabdomeric photoreceptors is undisputed, but its mode of action remains poorly understood. Although Ca-regulated molecules such as PKC and calmodulin have been implicated, direct physiological evidence is lacking. We will further our studies linking PKC activation to desensitization, and systematically test the involvement of CaM in light adaptation, especially the acceleration of the response. It is also becoming clear that some additional modulatory pathways operate independently of Ca. Ciliary photoreceptors allow these processes to be studied in isolation, because neither influx nor internal mobilization of Ca are coupled to photoexcitation. We have uncovered two novel Ca-independent regulatory schemes: (i) cGMP, in addition to controlling light-sensitive channels, is also implicated in negative feedback regulation of the cascade. (ii) Photoresponse kinetics during light adaptation is modulated by photopigment state (which, owing to its bi-stability, can be readily manipulated); we will probe the underlying mechanisms and their possible dependency on cAMP. Finally, we will further investigate the gating of the light-dependent conductance of ciliary photoreceptors, testing a recent conjecture that links it to the inactivation process of voltage-dependent K channels, to which cyclic nucleotide gated channels are related.

Agency
National Institute of Health (NIH)
Institute
National Eye Institute (NEI)
Type
Research Project (R01)
Project #
5R01EY007559-16
Application #
6706973
Study Section
Visual Sciences C Study Section (VISC)
Program Officer
Mariani, Andrew P
Project Start
1988-03-01
Project End
2007-02-28
Budget Start
2004-03-01
Budget End
2007-02-28
Support Year
16
Fiscal Year
2004
Total Cost
$354,547
Indirect Cost
Name
Boston University
Department
Physiology
Type
Schools of Medicine
DUNS #
604483045
City
Boston
State
MA
Country
United States
Zip Code
02118
Gomez, Maria del Pilar; Nasi, Enrico (2005) On the gating mechanisms of the light-dependent conductance in Pecten hyperpolarizing photoreceptors: does light remove inactivation in voltage-dependent K channels? J Gen Physiol 125:455-64
del Pilar Gomez, Maria; Nasi, Enrico (2005) A direct signaling role for phosphatidylinositol 4,5-bisphosphate (PIP2) in the visual excitation process of microvillar receptors. J Biol Chem 280:16784-9
del Pilar Gomez, Maria; Nasi, Enrico (2005) Calcium-independent, cGMP-mediated light adaptation in invertebrate ciliary photoreceptors. J Neurosci 25:2042-9
Piccoli, Giuseppe; Del Pilar Gomez, Maria; Nasi, Enrico (2002) Role of protein kinase C in light adaptation of molluscan microvillar photoreceptors. J Physiol 543:481-94
Gomez, M P; Nasi, E (2000) Light transduction in invertebrate hyperpolarizing photoreceptors: possible involvement of a Go-regulated guanylate cyclase. J Neurosci 20:5254-63
Nasi, E; del Pilar Gomez, M (1999) Divalent cation interactions with light-dependent K channels. Kinetics of voltage-dependent block and requirement for an open pore. J Gen Physiol 114:653-72
Gomez, M P; Nasi, E (1997) Light adaptation in Pecten hyperpolarizing photoreceptors. Insensitivity to calcium manipulations. J Gen Physiol 109:371-84
Gomez, M P; Nasi, E (1997) Antagonists of the cGMP-gated conductance of vertebrate rods block the photocurrent in scallop ciliary photoreceptors. J Physiol 500 ( Pt 2):367-78
Gomez, M D; Nasi, E (1996) Ion permeation through light-activated channels in rhabdomeric photoreceptors. Role of divalent cations. J Gen Physiol 107:715-30
del Pilar Gomez, M; Nasi, E (1995) Activation of light-dependent K+ channels in ciliary invertebrate photoreceptors involves cGMP but not the IP3/Ca2+ cascade. Neuron 15:607-18

Showing the most recent 10 out of 18 publications