Role of guanylyl cyclase modulation in mouse cone phototransduction

A negative phototransduction feedback in rods and cones is critical for the timely termination of their light responses and for extending their function to a wide range of light intensities. The calcium feedback mechanisms that modulate phototransduction in rods have been studied extensively. However, the corresponding modulation mechanisms that enable cones to terminate rapidly their light responses and to adapt in bright light, properties critical for our daytime vision, are still not understood. In cones, calcium feedback to guanylyl cyclase is potentially a key step in phototransduction modulation. The guanylyl cyclase activity is modulated by the calcium-binding guanylyl cyclase activating proteins (GCAP1 and GCAP2). Here, we used single-cell and transretinal recordings from mouse to determine how GCAPs modulate dark-adapted responses as well as light adaptation in mammalian cones. Deletion of GCAPs increased threefold the amplitude and dramatically prolonged the light responses in dark-adapted mouse cones. It also reduced the operating range of mouse cones in background illumination and severely impaired their light adaptation. Thus, GCAPs exert powerful modulation on the mammalian cone phototransduction cascade and play an important role in setting the functional properties of cones in darkness and during light adaptation. Surprisingly, despite their better adaptation capacity and wider calcium dynamic range, mammalian cones were modulated by GCAPs to a lesser extent than mammalian rods. We conclude that a disparity in the strength of GCAP modulation cannot explain the differences in the dark-adapted properties or in the operating ranges of mammalian rods and cones

Substrate binding induces structural changes in cytochrome P450cam

X-ray structures of ferric cytochrome P450cam partially complexed with the substrate (+)-camphor to two different extents were determined at 1.30–1.35 Å resolution, revealing the protein structures of the substrate-free and substrate-bound forms

The Na+/Ca2+, K+ exchanger 2 modulates mammalian cone phototransduction

Calcium ions (Ca(2+)) modulate the phototransduction cascade of vertebrate cone photoreceptors to tune gain, inactivation, and light adaptation. In darkness, the continuous current entering the cone outer segment through cGMP-gated (CNG) channels is carried in part by Ca(2+), which is then extruded back to the extracellular space. The mechanism of Ca(2+) extrusion from mammalian cones is not understood. The dominant view has been that the cone-specific isoform of the Na(+)/Ca(2+), K(+) exchanger, NCKX2, is responsible for removing Ca(2+) from their outer segments. However, indirect evaluation of cone function in NCKX2-deficient (Nckx2(−/−)) mice by electroretinogram recordings revealed normal photopic b-wave responses. This unexpected result suggested that NCKX2 may not be involved in the Ca(2+) homeostasis of mammalian cones. To address this controversy, we examined the expression of NCKX2 in mouse cones and performed transretinal recordings from Nckx2(−/−) mice to determine the effect of NCKX2 deletion on cone function directly. We found that Nckx2(−/−) cones exhibit compromised phototransduction inactivation, slower response recovery and delayed background adaptation. We conclude that NCKX2 is required for the maintenance of efficient Ca(2+) extrusion from mouse cones. However, surprisingly, Nckx2(−/−) cones adapted normally in steady background light, indicating the existence of additional Ca(2+)-extruding mechanisms in mammalian cones

Scanning Tunneling Microscopy Measurements of the Local Work Function around Steps on the Au/Cu(111) Surface(STM-local states)

With a scanning tunneling microscope we have taken local work function images of the Au/Cu(111) surface. The local work function measured from large Au-covered terraces is in good agreement with the result obtained by non-local techniques, and the local work function around step edges is significantly lower than that on terraces. The experimental details of the local work function lowering around a step can be reproduced nicely with a simple simulation if a dipole moment row with a proper linear density is put at the center of the step and if the influence of the topographic change of the step due to formation of the dipole moment is taken into account. It is demonstrated that STM has the ability to reveal the details of how local work function varies around steps and other defects on metal surfaces

Ground-state phase diagram and magnetic properties of a tetramerized spin-1/2 J_1-J_2 model: BEC of bound magnons and absence of the transverse magnetization

We study the ground state and the magnetization process of a spin-1/2 $J_1$-$J_2$ model with a plaquette structure by using various methods. For small inter-plaquette interaction, this model is expected to have a spin-gap and we computed the first- and the second excitation energies. If the gap of the lowest excitation closes, the corresponding particle condenses to form magnetic orders. By analyzing the quintet gap and magnetic interactions among the quintet excitations, we find a spin-nematic phase around $J_1/J_2\sim -2$ due to the strong frustration and the quantum effect. When high magnetic moment is applied, not the spin-1 excitations but the spin-2 ones soften and dictate the magnetization process. We apply a mean-field approximation to the effective Hamiltonian to find three different types of phases (a conventional BEC phase, ``striped'' supersolid phases and a 1/2-plateau). Unlike the BEC in spin-dimer systems, this BEC phase is not accompanied by transverse magnetization. Possible connection to the recently discovered spin-gap compound (CuCl)LaNb2O7 is discussed.Comment: 18pages, 17figures; title changed, typos correcte

Physiological Properties of Rod Photoreceptor Cells in Green-sensitive Cone Pigment Knock-in Mice

Rod and cone photoreceptor cells that are responsible for scotopic and photopic vision, respectively, exhibit photoresponses different from each other and contain similar phototransduction proteins with distinctive molecular properties. To investigate the contribution of the different molecular properties of visual pigments to the responses of the photoreceptor cells, we have generated knock-in mice in which rod visual pigment (rhodopsin) was replaced with mouse green-sensitive cone visual pigment (mouse green). The mouse green was successfully transported to the rod outer segments, though the expression of mouse green in homozygous retina was ∼11% of rhodopsin in wild-type retina. Single-cell recordings of wild-type and homozygous rods suggested that the flash sensitivity and the single-photon responses from mouse green were three to fourfold lower than those from rhodopsin after correction for the differences in cell volume and levels of several signal transduction proteins. Subsequent measurements using heterozygous rods expressing both mouse green and rhodopsin E122Q mutant, where these pigments in the same rod cells can be selectively irradiated due to their distinctive absorption maxima, clearly showed that the photoresponse of mouse green was threefold lower than that of rhodopsin. Noise analysis indicated that the rate of thermal activations of mouse green was 1.7 × 10−7 s−1, about 860-fold higher than that of rhodopsin. The increase in thermal activation of mouse green relative to that of rhodopsin results in only 4% reduction of rod photosensitivity for bright lights, but would instead be expected to severely affect the visual threshold under dim-light conditions. Therefore, the abilities of rhodopsin to generate a large single photon response and to retain high thermal stability in darkness are factors that have been necessary for the evolution of scotopic vision

Spatial Variation of Local Work Function of the Au/Cu(111) and Pd/Cu(111) Systems(STM-local states)

Spatial variation of local work function has been investigated for the Au/Cu(111) and Pd/Cu(111) systems using scanning tunneling microscopy (STM). While the measured work function of the Au adlayer is in the range between the Au(111) and Cu(111)\u27s values consistent with the results obtained by others, the work function on the Pd adsorbate shows a marked difference. The work function of the first Pd layer exceeds that of the Pd(111) surface and it increases further on its second layer, suggesting an overshooting (possibly oscillatory) behavior, which has never been reported experimentally

Long-Term Outcome of Endoscopic Balloon Dilation in Obstructive Gastrointestinal Crohn's Disease: A Prospective Long-Term Study

Background The short- and long-term results of balloon dilation therapy in Crohn's patients with non-anastomotic obstructive gastrointestinal lesions are investigated