scholarly journals KINETICS OF ROD OUTER SEGMENT RENEWAL IN THE DEVELOPING MOUSE RETINA

1973 ◽  
Vol 58 (3) ◽  
pp. 650-661 ◽  
Author(s):  
Matthew M. LaVail
Keyword(s):  
Outer Segment ◽  
Photoreceptor Cells ◽  
Mouse Retina ◽  
Rod Photoreceptor ◽  
Adult Level ◽  
Rod Outer Segment ◽  
Equilibrium Length ◽  
Kinetics Of ◽  
Late Stages ◽  
Adult Rate

The kinetics of rod outer segment renewal in the developing retina have been investigated in C57BL/6J mice. Litters of mice were injected with [3H]amino acids at various ages and killed at progressively later time intervals. Plastic 1.5 µm sections of retina were studied by light microscope autoradiography. The rate of outer segment disk synthesis, as judged by labeled disk displacement away from the site of synthesis, is slightly greater than the adult level at 11–13 days of age; it rises to more than 1.6 times the adult rate between days 13 and 17, after which it falls to the adult level at 21–25 days. The rate of disk disposal, as measured by labeled disk movement toward the site of disposal, is less than 15% of the adult level at 11–13 days of age; it rises sharply to almost 70% of the adult level by days 13–15 and then more gradually approaches the adult rate. The net difference in rates of synthesis and disposal accounts for the rapid elongation of rod outer segments in the mouse between days 11 and 17 and the subsequent, more gradual elongation to the adult equilibrium length reached between days 19 and 25. The changing rate of outer segment disk synthesis characterizes the late stages of cytodifferentiation of the rod photoreceptor cells.

scholarly journals Raman Spectroscopic Imaging of Cholesterol and Docosahexaenoic Acid Distribution in the Retinal Rod Outer Segment

2011 ◽  
Vol 64 (5) ◽  
pp. 611 ◽  
Author(s):  
Zachary D. Schultz
Keyword(s):  
Docosahexaenoic Acid ◽  
Outer Segment ◽  
Rana Catesbeiana ◽  
Spectroscopic Imaging ◽  
Photoreceptor Cells ◽  
Integral Membrane Protein ◽  
Rod Photoreceptor ◽  
Rod Outer Segment ◽  
Cellular Processes ◽  
Retinal Rod

Raman vibrational spectroscopic imaging was performed on retinal rod cells isolated from bullfrogs (Rana catesbeiana). The Raman spectra enable determination of the lipid and protein rich rod outer segment (ROS) from the nucleus and inner segment of the cell. Peak fitting analysis of spectra obtained from individual rod photoreceptor cells show characteristic vibrational modes that can be associated with cholesterol and docosahexaenoic acid-containing lipids. These results provide direct observations of biomolecular gradients in the rod photoreceptor cells, which, thus far, have been based on indirect detergent extracts and histochemical analysis with indicators such as filipin. The detected biomolecules are associated with regulation of the integral membrane protein rhodopsin, and methods capable of direct observation of these biomolecules offer new routes to exploring their role in the regulation of cellular processes.

scholarly journals Radioimmunocytochemical localization of retinal S-antigen with monoclonal antibodies.

1984 ◽  
Vol 32 (8) ◽  
pp. 834-838 ◽  
Author(s):  
N D Das ◽  
R J Ulshafer ◽  
Z S Zam ◽  
V R Leverenz ◽  
H Shichi
Keyword(s):  
Monoclonal Antibodies ◽  
Outer Segment ◽  
Antigenic Site ◽  
Photoreceptor Cells ◽  
Apical Region ◽  
Rod Photoreceptor ◽  
Inner Limiting Membrane ◽  
Silver Grains ◽  
Homogeneous Preparation ◽  
Labeling Pattern

Two monoclonal antibodies (RSA1/83 and RSA2/83) were developed against a homogeneous preparation of bovine retinal S-antigen. The two hybridomas produced by mouse X mouse hybrid myeloma cells secrete immunoglobulin G. Indirect autoradiography on glutaraldehyde-fixed preparations of bovine explants was used to locate the antigenic site. Antibody RSA1/83 recognizes the antigen primarily in the apical region of the rod outer segment, while antibody RSA2/83 located the antigen both in the outer and inner segments of the rod photoreceptor cells. A distinct band of silver grains also appeared along the inner limiting membrane with both antibodies. Control explants showed no specific labeling pattern over the various retinal compartments.

scholarly journals Immunocytochemical localization of opsin in the cell membrane of developing rat retinal photoreceptors.

1984 ◽  
Vol 98 (5) ◽  
pp. 1788-1795 ◽  
Author(s):  
I Nir ◽  
D Cohen ◽  
D S Papermaster
Keyword(s):  
Plasma Membrane ◽  
Outer Segment ◽  
Plasma Membranes ◽  
Photoreceptor Cells ◽  
Rod Photoreceptor ◽  
Rod Photoreceptors ◽  
Retinal Photoreceptors ◽  
Retinal Rod ◽  
Ros Formation ◽  
Developing Rat

Mature retinal rod photoreceptors sequester opsin in the disk and plasma membranes of the rod outer segment (ROS). Opsin is synthesized in the inner segment and is transferred to the outer segment along the connecting cilium that joins the two compartments. We have investigated early stages of retinal development during which the polarized distribution of opsin is established in the rod photoreceptor cell. Retinas were isolated from newborn rats, 3-21 d old, and incubated with affinity purified biotinyl-sheep anti-bovine opsin followed by avidin-ferritin. At early postnatal ages prior to the development of the ROS, opsin is labeled by antiopsin on the inner segment plasma membrane. At the fifth postnatal day, as ROS formation begins opsin was detected on the connecting cilium plasma membrane. However, the labeling density of the ciliary plasma membrane was not uniform: the proximal cilium was relatively unlabeled in comparison with the distal cilium and the ROS plasma membrane. In nearly mature rat retinas, opsin was no longer detected on the inner segment plasma membrane. A similar polarized distribution of opsin was also observed in adult human rod photoreceptor cells labeled with the same antibodies. These results suggest that some component(s) of the connecting cilium and its plasma membrane may participate in establishing and maintaining the polarized distribution of opsin.

scholarly journals Dysfunction of Heterotrimeric Kinesin-2 in Rod Photoreceptor Cells and the Role of Opsin Mislocalization in Rapid Cell Death

2010 ◽  
Vol 21 (23) ◽  
pp. 4076-4088 ◽  
Author(s):  
Vanda S. Lopes ◽  
David Jimeno ◽  
Kornnika Khanobdee ◽  
Xiaodan Song ◽  
Bryan Chen ◽  
...  
Keyword(s):  
Cell Death ◽  
Outer Segment ◽  
Photoreceptor Cell ◽  
Photoreceptor Cells ◽  
Cell Loss ◽  
Rod Photoreceptor ◽  
Retinal Degenerations ◽  
Cell Counts ◽  
And Function

Due to extensive elaboration of the photoreceptor cilium to form the outer segment, axonemal transport (IFT) in photoreceptors is extraordinarily busy, and retinal degeneration is a component of many ciliopathies. Functional loss of heterotrimeric kinesin-2, a major anterograde IFT motor, causes mislocalized opsin, followed by rapid cell death. Here, we have analyzed the nature of protein mislocalization and the requirements for the death of kinesin-2-mutant rod photoreceptors. Quantitative immuno EM showed that opsin accumulates initially within the inner segment, and then in the plasma membrane. The light-activated movement of arrestin to the outer segment is also impaired, but this defect likely results secondarily from binding to mislocalized opsin. Unlike some other retinal degenerations, neither opsin–arrestin complexes nor photoactivation were necessary for cell loss. In contrast, reduced rod opsin expression provided enhanced rod and cone photoreceptor survival and function, as measured by photoreceptor cell counts, apoptosis assays, and ERG analysis. The cell death incurred by loss of kinesin-2 function was almost completely negated by Rho−/−. Our results indicate that mislocalization of opsin is a major cause of photoreceptor cell death from kinesin-2 dysfunction and demonstrate the importance of accumulating mislocalized protein per se, rather than specific signaling properties of opsin, stemming from photoactivation or arrestin binding.

scholarly journals A Computational Pipeline for Functional Gene Discovery

2021 ◽  
Author(s):  
Aolani Colon ◽  
Rishabh Hirday ◽  
Ami Patel ◽  
Amrita Poddar ◽  
Emma Tuberty-Vaughan ◽  
...  
Keyword(s):  
Candidate Genes ◽  
Outer Segment ◽  
Gene Discovery ◽  
Immunohistochemical Analysis ◽  
Transcriptome Profiling ◽  
Photoreceptor Cells ◽  
Functional Gene ◽  
Mouse Retina ◽  
Computational Pipeline ◽  
Photoreceptor Outer Segment

Abstract Many computational pipelines exist for the detection of differentially expressed genes. However, computational pipelines for functional gene detection are rarely exist. We developed a new computational pipeline for functional gene identification from transcriptome profiling data. Key features of the pipeline include clustering optimization by gap statistics, gene ontology analysis for each cluster, and literature analysis for functional gene discovery. By leveraging this pipeline on RNA-seq datasets of mouse retinal development studies, we identified 14 candidate genes involved in the formation of the photoreceptor outer segment. The expression of top three candidate genes (Pde8b, Laptm4b, and Nr1h4) in the outer segment of the developing mouse retina were experimentally validated by immunohistochemical analysis. This computational pipeline can accurately predict novel functional gene for a specific biological process, e.g., the outer segment development of the photoreceptor cells in the mouse retina. This pipeline is also applicable to functional gene discovery for any other biological processes and in any other organs and tissues.

Tubulin in bovine retinal rod outer segments

1992 ◽  
Vol 103 (1) ◽  
pp. 157-166
Author(s):  
D.F. Matesic ◽  
N.J. Philp ◽  
J.M. Murray ◽  
P.A. Liebman
Keyword(s):  
Plasma Membrane ◽  
Outer Segment ◽  
Photoreceptor Cells ◽  
High Salt ◽  
Immunofluorescent Staining ◽  
Rod Outer Segments ◽  
Rod Outer Segment ◽  
Retinal Rod ◽  
Additional Role ◽  
Retinal Rod Outer Segments

Bovine rod outer segment (ROS) preparations contain a major 58 kDa protein doublet that was identified by immunoblot as tubulin. Quantification by gel densitometry showed that the total amount of tubulin was 5- to 10-fold higher than that attributable to the rod axoneme, suggesting additional role(s) for tubulin in photoreceptor cells. Approximately 20% of this nonaxonemal tubulin (15% of total tubulin) is tightly associated with outer segment membranes. This fraction remains membrane-associated after extensive low- or high-salt washing, requiring detergents or protein denaturants for release from ROS membranes. Unlike ROS soluble tubulin it associates tightly with liposomes upon detergent solubilization and reconstitution. The ROS membrane-associated tubulin is highly enriched in isolated ROS plasma membrane fractions compared to the total outer segment membrane pool and can be localized to the plasma membrane but not to disks by immunofluorescent staining, suggesting a possible role in the structure or electrophysiology of the rod outer segment plasma membrane.

scholarly journals Localization of peripherin/rds in the disk membranes of cone and rod photoreceptors: relationship to disk membrane morphogenesis and retinal degeneration.

1992 ◽  
Vol 116 (3) ◽  
pp. 659-667 ◽  
Author(s):  
K Arikawa ◽  
L L Molday ◽  
R S Molday ◽  
D S Williams
Keyword(s):  
Plasma Membrane ◽  
Retinal Degeneration ◽  
Outer Segment ◽  
Photoreceptor Cells ◽  
Growth Stages ◽  
Rod Outer Segments ◽  
Rod Photoreceptor ◽  
Outer Segments ◽  
Disk Membrane ◽  
Membrane Morphogenesis

The outer segments of vertebrate rod photoreceptor cells consist of an ordered stack of membrane disks, which, except for a few nascent disks at the base of the outer segment, is surrounded by a separate plasma membrane. Previous studies indicate that the protein, peripherin or peripherin/rds, is localized along the rim of mature disks of rod outer segments. A mutation in the gene for this protein has been reported to be responsible for retinal degeneration in the rds mouse. In the present study, we have shown by immunogold labeling of rat and ground squirrel retinas that peripherin/rds is present in the disk rims of cone outer segments as well as rod outer segments. Additionally, in the basal regions of rod and cone outer segments, where disk morphogenesis occurs, we have found that the distribution of peripherin/rds is restricted to a region that is adjacent to the cilium. Extension of its distribution from the cilium coincides with the formation of the disk rim. These results support the model of disk membrane morphogenesis that predicts rim formation to be a second stage of growth, after the first stage in which the ciliary plasma membrane evaginates to form open nascent disks. The results also indicate how the proteins of the outer segment plasma membrane and the disk membranes are sorted into their separate domains: different sets of proteins may be incorporated into membrane outgrowths during different growth stages of disk morphogenesis. Finally, the presence of peripherin/rds protein in both cone and rod outer segment disks, together with the phenotype of the rds mouse, which is characterized by the failure of both rod and cone outer segment formation, suggest that the same rds gene is expressed in both types of photoreceptor cells.

scholarly journals Diurnal, localized exposure of phosphatidylserine by rod outer segment tips in wild-type but not Itgb5-/- or Mfge8-/- mouse retina

2012 ◽  
Vol 109 (21) ◽  
pp. 8145-8148 ◽  
Author(s):  
L. Ruggiero ◽  
M. P. Connor ◽  
J. Chen ◽  
R. Langen ◽  
S. C. Finnemann
Keyword(s):  
Outer Segment ◽  
Mouse Retina ◽  
Wild Type ◽  
Rod Outer Segment

scholarly journals Determinants shaping the nanoscale architecture of the mouse rod outer segment

2021 ◽  
Author(s):  
Matthias Pöge ◽  
Julia Mahamid ◽  
Sanae S Imanishi ◽  
Jürgen M Plitzko ◽  
Krzysztof Palczewski ◽  
...  
Keyword(s):  
Outer Segment ◽  
Molecular Mechanisms ◽  
Structural Information ◽  
Structural Basis ◽  
Radius Of Curvature ◽  
Rod Photoreceptor ◽  
Molecular Architecture ◽  
Structural Determinants ◽  
Rod Outer Segment ◽  
Wide Range

The unique membrane organization of the rod outer segment (ROS), the specialized sensory cilium of rod photoreceptor cells, provides the foundation for phototransduction, the initial step in vision. ROS architecture is characterized by a stack of identically shaped and tightly packed membrane disks loaded with the visual receptor rhodopsin. A wide range of genetic aberrations compromise ROS ultrastructure, impairing photoreceptor viability and function. Yet, the structural basis giving rise to the remarkable long range order of ROS membrane stacks and the molecular mechanisms underlying genetically inherited diseases remain elusive. Here, cryo-electron tomography (cryo-ET) performed on native ROS at molecular resolution provides insights into key structural determinants of ROS membrane architecture. Our data reveal the existence of two molecular connectors/spacers which likely contribute to the nanometer scale precise stacking of the ROS disks. We further show that the extreme radius of curvature at the disk rims is enforced by a continuous supramolecular assembly composed of peripherin-2 (PRPH2) and rod outer segment membrane protein 1 (ROM1) tetramers. We suggest that, together these molecular assemblies constitute the structural basis of the highly specialized ROS functional architecture. Cryo-ET therefore provides novel quantitative and structural information on the molecular architecture in ROS and insights into possible mechanisms underlying pathologies of certain PRPH2 mutations leading to blindness.

scholarly journals A computational pipeline for functional gene discovery

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Aolani Colon ◽  
Rishabh Hirday ◽  
Ami Patel ◽  
Amrita Poddar ◽  
Emma Tuberty-Vaughan ◽  
...  
Keyword(s):  
Candidate Genes ◽  
Outer Segment ◽  
Gene Discovery ◽  
Immunohistochemical Analysis ◽  
Transcriptome Profiling ◽  
Photoreceptor Cells ◽  
Functional Gene ◽  
Mouse Retina ◽  
Computational Pipeline ◽  
Photoreceptor Outer Segment

AbstractMany computational pipelines exist for the detection of differentially expressed genes. However, computational pipelines for functional gene detection rarely exist. We developed a new computational pipeline for functional gene identification from transcriptome profiling data. Key features of the pipeline include batch effect correction, clustering optimization by gap statistics, gene ontology analysis of clustered genes, and literature analysis for functional gene discovery. By leveraging this pipeline on RNA-seq datasets from two mouse retinal development studies, we identified 7 candidate genes involved in the formation of the photoreceptor outer segment. The expression of top three candidate genes (Pde8b, Laptm4b, and Nr1h4) in the outer segment of the developing mouse retina were experimentally validated by immunohistochemical analysis. This computational pipeline can accurately predict novel functional gene for a specific biological process, e.g., development of the outer segment and synapses of the photoreceptor cells in the mouse retina. This pipeline can also be useful to discover functional genes for other biological processes and in other organs and tissues.

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