Three-dimensional architecture of murine rod outer segments determined by cryoelectron tomography

Nickell, Stephan; Park, Paul Shin-Hyun; Baumeister, Wolfgang; Palczewski, Krzysztof

doi:10.1083/jcb.200612010

Cited by 197 publications

(254 citation statements)

References 82 publications

Supporting

Mentioning

226

Contrasting

Order By: Relevance

“…An interesting candidate is the cGMP phosphodiesterase, which may span the interdiscal space (Fig. 4 C and D) and is present at 1:310 relative to rhodopsin (52), so that it has a surface density on pairs of discs of 250 μm −2 , intriguingly close to that of the as-yet unidentified "spacers" (49).…”

Section: Estimating the Spring Constant Of An Elementary Rod Os Elasticmentioning

confidence: 99%

“…Freeze-fracture and cryoelectron tomographic investigations have revealed macromolecular structures that appear to link adjacent disc membranes (48,49). Because most proteins can act as elastic elements (50), it is reasonable to hypothesize that such disc-to-disc "linkers" serve as springs that resist swelling, affording some protection of the rod OS from stimulus-induced osmotic overload.…”

Section: Estimating the Spring Constant Of An Elementary Rod Os Elasticmentioning

confidence: 99%

See 1 more Smart Citation

In vivo optophysiology reveals that G-protein activation triggers osmotic swelling and increased light scattering of rod photoreceptors

Zhang

Zawadzki

Goswami

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

111

164

View full text Add to dashboard Cite

The light responses of rod and cone photoreceptors have been studied electrophysiologically for decades, largely with ex vivo approaches that disrupt the photoreceptors' subretinal microenvironment. Here we report the use of optical coherence tomography (OCT) to measure light-driven signals of rod photoreceptors in vivo. Visible light stimulation over a 200-fold intensity range caused correlated rod outer segment (OS) elongation and increased light scattering in wildtype mice, but not in mice lacking the rod G-protein alpha subunit, transducin (Gα t ), revealing these responses to be triggered by phototransduction. For stimuli that photoactivated one rhodopsin per Gα t the rod OS swelling response reached a saturated elongation of 10.0 ± 2.1%, at a maximum rate of 0.11% s −1. Analyzing swelling as osmotically driven water influx, we find the H 2 O membrane permeability of the rod OS to be (2.6 ± 0.4) × 10 −5 cm·s, comparable to that of other cells lacking aquaporin expression. Application of Van't Hoff's law reveals that complete activation of phototransduction generates a potentially harmful 20% increase in OS osmotic pressure. The increased backscattering from the base of the OS is explained by a model combining cytoplasmic swelling, translocation of dissociated G-protein subunits from the disc membranes into the cytoplasm, and a relatively higher H 2 O permeability of nascent discs in the basal rod OS. Translocation of phototransduction components out of the OS may protect rods from osmotic stress, which could be especially harmful in disease conditions that affect rod OS structural integrity.osmotic stress | phototransduction | optical coherence tomography | intrinsic optical signals | photoreceptor waveguiding

show abstract

Section: Estimating the Spring Constant Of An Elementary Rod Os Elasticmentioning

confidence: 99%

Section: Estimating the Spring Constant Of An Elementary Rod Os Elasticmentioning

confidence: 99%

In vivo optophysiology reveals that G-protein activation triggers osmotic swelling and increased light scattering of rod photoreceptors

Zhang

Zawadzki

Goswami

et al. 2017

Proc. Natl. Acad. Sci. U.S.A.

111

164

View full text Add to dashboard Cite

show abstract

“…The rod outer segment ͑rod OS͒ is packed with hundreds of membrane sacs, called disks, on the surface of which the initial biochemical reactions of visual transduction take place. 2,3 The disks, with dimensions of about 1 m diam, 4,5 are regularly stacked on top of each other with a repeat distance of 300 Å. 4 The visual pigment rhodopsin ͑Rh͒, an integral membrane protein, accounts for about 90% 6 of total disk protein content.…”

mentioning

confidence: 99%

“…2,3 The disks, with dimensions of about 1 m diam, 4,5 are regularly stacked on top of each other with a repeat distance of 300 Å. 4 The visual pigment rhodopsin ͑Rh͒, an integral membrane protein, accounts for about 90% 6 of total disk protein content. Photoreceptor disks are continuously renewed through formation of new disks at the base of the outer segment, displaced distally along the length of the outer segment, and eventual detachment and phagocytosis occurs by adjacent pigment epithelial cells.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Confocal laser scanning microscopy of retinal rod outer segment intact disks: new labeling technique

et al. 2007

View full text Add to dashboard Cite

Abstract. Vertebrate retinal rod outer segment disks house the proteins involved in the phototransduction cascade that converts light into neuronal signal. We develop a technique for the immunofluorescent labeling of osmotically intact isolated rod outer segment disks for confocal laser scanning microscopy imaging. Osmotically intact Ficoll-flotation isolated bovine disks are directly labeled with antibodies in solution. For the first time, osmotically intact single disks can be visualized. Thus, imaging of purified disks, based on advanced optical techniques, may serve as a powerful complement to other methods in studies on phototransduction. In fact, even though much is known about the rod outer segment photoresponse, some unanswered questions remain, particularly about ATP supply, light adaptation, and morphogenesis. Oct. 3, 2007. Confocal laser scanning microscopy ͑CLSM͒ is a valuable tool for obtaining high-resolution images and 3-D reconstructions. 1 Vertebrate rod photoreceptors, the neurons responsible for visual phototransduction, mediate vision at low-light intensities using a modified cilium ͑called the outer segment͒ for the detection of light. The rod outer segment ͑rod OS͒ is packed with hundreds of membrane sacs, called disks, on the surface of which the initial biochemical reactions of visual transduction take place. 2,3 The disks, with dimensions of about 1 m diam, 4,5 are regularly stacked on top of each other with a repeat distance of 300 Å. 4 The visual pigment rhodopsin ͑Rh͒, an integral membrane protein, accounts for about 90% 6 of total disk protein content. Photoreceptor disks are continuously renewed through formation of new disks at the base of the outer segment, displaced distally along the length of the outer segment, and eventual detachment and phagocytosis occurs by adjacent pigment epithelial cells. 7 Several microscopy techniques have been used to analyze the morphological organization of rods and to identify the membrane proteins in rod OS. For example, measurements of the ellipsoid mitochondria density, sizes, and shapes of inner segments were conducted by electron microscopy and Nomads differential interference contrast ͑NDIC͒ imaging. 8 Studies on the stability of Rh in native membranes were done by single-molecule force spectroscopy. 9 Moreover, a recent progress in modeling the native conformation of Rh was based on topography of the disk membranes recorded by an atomic force microscope ͑AFM͒. 9 CLSM was only used on whole retina or rod OS to observe the protein distribution in the disk incisures. 10 Therefore, even though rod imaging has been achieved in many ways, to the best of our knowledge, until now no study has been reported regarding the application of immunofluorescence for confocal optical imaging of isolated intact disks. In fact, even though fluorescent optical imaging is among the most widely used approach for studying cells in vivo, it is not easily applicable for subcellular fractions. Apart from the classic inclusion methods, it is difficult to isolate an...

show abstract

Serial Block Face‐Scanning Electron Microscopy: A Method to Study Retinal Degenerative Phenotypes

2014

Self Cite

View full text Add to dashboard Cite

Retinal degenerative conditions can vary in their clinical presentation and often present with subtle phenotypic features before the onset of clinically overt disease. To capture these isolated events that precipitate disease, large representative areas of the retina must be imaged at high resolution. Compared to light microscopic methods, traditional electron microscopy can provide images at sufficient resolution to detect subtle pathologic changes in the retina, but are limited to the area being surveyed. The advent of serial block face-scanning electron microscopy (SBFSEM) provides the resolution needed with the unprecedented advantage of imaging large volumes of retinal tissue. Furthermore, automation of SBF-SEM bypasses errors from manual sectioning and can produce reliable serial sections as thin as 25 nanometers. Moreover, the three-dimensional structures generated can highlight cellular connectivity and interactions in the retina and reveal pathological changes. Using SBF-SEM, we have identified subtle phenotypic features in mouse models of various human retinal dystrophies. This method will allow researchers to identify and monitor the time course of these pathologies. Here, we summarize the SBF-SEM methodology and its application to mouse models of retinal degeneration.

show abstract

Three-dimensional architecture of murine rod outer segments determined by cryoelectron tomography

Cited by 197 publications

References 82 publications

In vivo optophysiology reveals that G-protein activation triggers osmotic swelling and increased light scattering of rod photoreceptors

In vivo optophysiology reveals that G-protein activation triggers osmotic swelling and increased light scattering of rod photoreceptors

Confocal laser scanning microscopy of retinal rod outer segment intact disks: new labeling technique

Serial Block Face‐Scanning Electron Microscopy: A Method to Study Retinal Degenerative Phenotypes

Contact Info

Product

Resources

About