Localization of kinesin superfamily proteins to the connecting cilium of fish photoreceptors

1996 ◽  
Vol 109 (4) ◽  
pp. 889-897 ◽  
Author(s):  
P.L. Beech ◽  
K. Pagh-Roehl ◽  
Y. Noda ◽  
N. Hirokawa ◽  
B. Burnside ◽  
...  
Keyword(s):  
Heavy Chain ◽  
Basal Body ◽  
Outer Segment ◽  
Rod Photoreceptors ◽  
Outer Segments ◽  
Immunological Tests ◽  
Peptide Antibody ◽  
N Terminus ◽  
Kinesin Superfamily ◽  
Rod Cell

Kinesin superfamily proteins (KIFs) are probable motors in vesicular and non-vesicular transport along microtubular tracks. Since a variety of KIFs have been recently identified in the motile flagella of Chlamydomonas, we sought to ascertain whether KIFs are also associated with the connecting cilia of vertebrate rod photoreceptors. As the only structural link between the rod inner segment and the photosensitive rod outer segment, the connecting cilium is thought to be the channel through which all material passes into and out of the outer segment from the rod cell body. We have performed immunological tests on isolated sunfish rod inner-outer segments (RIS-ROS) using two antibodies that recognize the conserved motor domain of numerous KIFs (anti-LAGSE, a peptide antibody, and anti-Klp1 head, generated against the N terminus of Chlamydomonas Klp1) as well as an antibody specific to a neuronal KIF, KIF3A. On immunoblots of RIS-ROS, LAGSE antibody detected a prominent band at approximately 117 kDa, which is likely to be kinesin heavy chain, and Klp1 head antibody detected a single band at approximately 170 kDa; KIF3A antibody detected a polypeptide at approximately 85 kDa which co-migrated with mammalian KIF3A and displayed ATP-dependent release from rod cytoskeletons. Immunofluorescence localizations with anti-LAGSE and anti-Klp1 head antibodies detected epitopes in the axoneme and ellipsoid, and immunoelectron microscopy with the LAGSE antibody showed that the connecting cilium region was particularly antigenic. Immunofluorescence with anti-KIF3A showed prominent labelling of the connecting cilium and the area surrounding its basal body; the outer segment axoneme and parts of the inner segment coincident with microtubules were also labelled. We propose that these putative kinesin superfamily proteins may be involved in the translocation of material between the rod inner and outer segments.

Movement of retinal along cone and rod photoreceptors

1994 ◽  
Vol 11 (2) ◽  
pp. 389-399 ◽  
Author(s):  
Jing Jin ◽  
Gregor J. Jones ◽  
M. Carter Cornwall
Keyword(s):  
Cell Body ◽  
Dark Adaptation ◽  
Visual Pigment ◽  
Outer Segment ◽  
Light Adaptation ◽  
Rod Photoreceptors ◽  
Rod Outer Segment ◽  
Rods And Cones ◽  
Outer Segments ◽  
Rod Cell

AbstractSingle isolated photoreceptors can be taken through a visual cycle of light adaptation by bleaching visual pigment, followed by dark adaptation when supplied with 11–cis retinal. Light adaptation after bleaching is manifested by faster response kinetics and a permanent reduction in sensitivity to light flashes, presumed to be due to the presence of bleached visual pigment. The recovery of flash sensitivity during dark adaptation is assumed to be due to regeneration of visual pigment to pre-bleach levels. In previous work, the outer segments of bleached, light-adapted cells were exposed to 11–cis retinal. In the present work, the cell bodies of bleached photoreceptors were exposed. We report a marked difference between rods and cones. Bleached cones recover sensitivity when their cell bodies are exposed to 11–cis retinal. Bleached rods do not. These results imply that retinal can move freely along the cone photoreceptor, but retinal either is not taken up by the rod cell body or retinal cannot move from the rod cell body to the rod outer segment. The free transfer of retinal along cone but not along rod photoreceptors could explain why, during dark adaptation in the retina, cones have access to a store of 11–cis retinal which is not available to rods. Additional experiments investigated the movement of retinal along bleached rod outer segments. The results indicate that retinal can move along the rod outer segment, but that this movement is slow, occurring at about the same rate as the regeneration of visual pigment.

scholarly journals MORPHOGENESIS OF THE RETINAL RODS

1956 ◽  
Vol 2 (4) ◽  
pp. 209-218 ◽  
Author(s):  
E. De Robertis
Keyword(s):  
Basal Body ◽  
Outer Segment ◽  
Surface Membrane ◽  
Apical Region ◽  
Outer Segments ◽  
Retinal Rods ◽  
Third Stage ◽  
Basal Portion ◽  
Rod Cell

The morphogenesis of the retinal rods has been studied with the electron microscope in white mice from birth up to the 16th day of age. Observations have been mainly concentrated on specimens of the 8th and 12th days and on the differentiation of the inner and outer segments of the retinal rods. In the morphogenesis of the outer segment three main stages have been considered. The first stage consists in the development of a primitive cilium projecting from a bulge of protoplasm which constitutes the primordium of the inner segment. A basal body, nine pairs of peripheral filaments, a surface membrane, and a matrix filled with a fine vesicular material have been recognized as components of the primitive cilium. The vesicles are called "morphogenetic material" because it is believed that they represent the macromolecular primordium of the rod sacs of the future outer segment. The second stage corresponds to the great enlargment of the apical region of the primitive cilium due to the rapid building up of the lamellar material of the rod sacs. The primitive rod sacs appear to be connected with the ciliary filaments. The basal portion of the primitive cilium remains undifferentiated and constitutes the connecting cilium of the adult rod (1). The third stage consists in the remodelling and reorientation of the rod sacs into their permanent transverse disposition. This process starts in the middle portion of the outer segments and proceeds towards both extremities which can be considered as zones of growth of the outer segment. The inner segment is at the beginning a bulge of protoplasm containing unoriented mitochondria, a basal body, a small Golgi zone, and numerous dense particles. Then this region becomes elongated and the different components assume the stratified disposition found in the adult (1). The demonstration that the entire outer segment of the rod cell is the result of the differentiation of a primitive cilium is discussed in view of the conflicting interpretations found in the literature. The possible macromolecular mechanisms that may be involved in the submicroscopic morphogenesis of the rod sacs are discussed and the possible role of the morphogenetic material is considered. The results described in this paper confirm and extend the interpretation of the submicroscopic morphology of the adult rod cell as presented in a previous paper (1).

The ageing photoreceptor

2007 ◽  
Vol 24 (2) ◽  
pp. 151-155 ◽  
Author(s):  
ALEXANDER CUNEA ◽  
GLEN JEFFERY
Keyword(s):  
Aging Process ◽  
Outer Segment ◽  
Retinal Disease ◽  
Segment Length ◽  
Rod Photoreceptor ◽  
Retinal Neurons ◽  
Rod Photoreceptors ◽  
Outer Segments ◽  
Age Related ◽  
Similar Density

With age many retinal neurons are lost. In humans the rod photoreceptor population in the perimacular region is subject to approximately 30% loss over life. Those that remain have been reported to suffer from extensive convolutions and localized swellings of their outer segments abnormally increasing their disc content and outer segment length. Here we examine quantitatively age-related changes in rat rod photoreceptors. The rat retina is ∼97% rod dominated. Here, aged rods showed significant reductions in outer segment length. The discs in their outer segments had a similar density, irrespective of whether they were young or old, however, in aged animals a higher proportion were misregistered. Surprisingly, in all of the tissue examined, we found no evidence for any convolution of outer segments or localized swelling as reported in humans, rather all remained straight. There are methodological differences between the research reported here and that undertaken on human retinae. There are also major differences in overall retinal architecture between humans and rodents that could contribute to differences in the aging process of individual cells. If it is the case that individual photoreceptors age differently in rodents compared to humans, it may pose significant problems for the use of this animal model in studies of ageing and age related outer retinal disease.

Na+-Ca2+ exchange in bovine rod outer segments requires and transports K+

1989 ◽  
Vol 257 (1) ◽  
pp. C153-C157 ◽  
Author(s):  
P. P. Schnetkamp ◽  
D. K. Basu ◽  
R. T. Szerencsei
Keyword(s):  
Outward Current ◽  
Rod Outer Segments ◽  
Rod Photoreceptors ◽  
K Channels ◽  
Outer Segments ◽  
Proton Current ◽  
Carbonyl Cyanide ◽  
First Time ◽  
K Release ◽  
Rod Cell

Intact outer segments isolated from bovine retinas (bovine ROS) display a high activity of Na+-Ca2+ exchange, and Na+-Ca2+ exchange appears to be the only functional ion transporter present. Here we demonstrate for the first time that Na+-Ca2+ exchange requires and transports K+ from the following observations. 1) Na+-Ca2+ exchange in bovine ROS required the simultaneous presence of K+ and Ca2+ on one side of the membrane and the presence of Na+ on the other side. 2) Na+-stimulated Ca2+ release from bovine ROS was accompanied by an equally large release of K+. We used the electrogenic protonophore carbonyl cyanide p-trifluoromethoxyphenylhydrazone (FCCP) as an added electrical shunt; in the intact rod cell, electrogenic Na+-Ca2+ exchange is shunted by K+ channels present in the rod inner segment. In the presence of FCCP, an inward Na+-Ca2+ exchange current was accompanied by an outward current of protons with a stoichiometry of 1 H+/Ca2+; in the absence of FCCP, no Na+-induced proton current was observed. Addition of FCCP did not uncouple Na+-induced K+ release from Na+-induced Ca2+ release. We conclude that Na+-Ca2+ exchange in bovine rod photoreceptors operates at an electrogenic stoichiometry of 4 Na+:(1 Ca2+ + 1 K+). In isolated ROS and in the absence of an external electrical shunt, Na+-Ca2+ exchange operated at an electroneutral stoichiometry of 3 Na+:(1 Ca2+ + 1 K+).

The photoreceptors and pigment epithelium of the larval Xenopus retina: morphogenesis and outer segment renewal

1978 ◽  
Vol 201 (1143) ◽  
pp. 149-167 ◽  
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Basal Body ◽  
Outer Segment ◽  
Pigment Epithelium ◽  
Ultrastructural Observation ◽  
Rod Outer Segments ◽  
Photoreceptor Outer Segments ◽  
Outer Segments ◽  
Constant Rate

Light microscopic autoradiography and electron microscopy were used to examine outer segment renewal and the development of photoreceptors and pigment epithelium in the larval Xenopus retina. Following the injection of [ 3 H]-leucine at stages 37/38–40 (when outer segments first develop) or 53–54 (when rod outer segments (r. o. s.) attain adult length), a band of label accumulated at the base of r. o. s. and was displaced sclerally with time, whereas label was diffusely distributed in cone outer segments (c. o. s.). By taking into account the change in shape of r. o. s. from conical to cylindrical around stage 46, and calculating outer segment growth (determined from the rate of band displacement) as volume of material added with time, we found a constant rate of membrane addition (1.59 μm/day) from the time of initial outer segment formation. The changes observed in r. o. s. length therefore indicate variations in the rate of disk shedding and phagocytosis, which is minimal before stage 46 and rises to 1.19 μm/day after stages 53–54. Ultrastructural observation showed that although all photoreceptor outer segments form by the repeated evagination of the plasma membrane of the connecting cilium, r. o. s. and c. o. s. are distinguishable by differences in membrane appearance even before they develop divergent membrane topologies. Fibrous granules near the basal body of young receptors may be precursors to the elongating ciliary microtubules. Clusters of cisternae observed near the ciliary base in photoreceptor inner segments may represent a stage in the transport of newly-synthesized opsin to the outer segment base.

Freeze-Fracture Replication of Frozen Outer Segments of Rat Retinal Rods

1969 ◽  
Vol 27 ◽  
pp. 392-393
Author(s):  
Thomas S. Leeson ◽  
C. Roland Leeson
Keyword(s):  
Electron Microscopy ◽  
Cross Section ◽  
Outer Segment ◽  
Freeze Fracture ◽  
X Ray Diffraction ◽  
X Ray ◽  
Outer Segments ◽  
Retinal Rods ◽  
Temperature Electron ◽  
Freeze Etching

Numerous previous studies of outer segments of retinal receptors have demonstrated a complex internal structure of a series of transversely orientated membranous lamellae, discs, or saccules. In cones, these lamellae probably are invaginations of the covering plasma membrane. In rods, however, they appear to be isolated and separate discs although some authors report interconnections and some continuities with the surface near the base of the outer segment, i.e. toward the inner segment. In some species, variations have been reported, such as longitudinally orientated lamellae and lamellar whorls. In cross section, the discs or saccules show one or more incisures. The saccules probably contain photolabile pigment, with resulting potentials after dipole formation during bleaching of pigment. Continuity between the lamina of rod saccules and extracellular space may be necessary for the detection of dipoles, although such continuity usually is not found by electron microscopy. Particles on the membranes have been found by low angle X-ray diffraction, by low temperature electron microscopy and by freeze-etching techniques.

scholarly journals TULP1 and TUB Are Required for Specific Localization of PRCD to Photoreceptor Outer Segments

2020 ◽  
Vol 21 (22) ◽  
pp. 8677
Author(s):  
Lital Remez ◽  
Ben Cohen ◽  
Mariela J. Nevet ◽  
Leah Rizel ◽  
Tamar Ben-Yosef
Keyword(s):  
Protein Interactions ◽  
Mammalian Cells ◽  
Outer Segment ◽  
Protein Protein Interactions ◽  
Photoreceptor Outer Segment ◽  
Photoreceptor Outer Segments ◽  
Outer Segments ◽  
Yeast Two Hybrid System ◽  
Specific Localization ◽  
Recruitment System

Photoreceptor disc component (PRCD) is a small protein which is exclusively localized to photoreceptor outer segments, and is involved in the formation of photoreceptor outer segment discs. Mutations in PRCD are associated with retinal degeneration in humans, mice, and dogs. The purpose of this work was to identify PRCD-binding proteins in the retina. PRCD protein-protein interactions were identified when implementing the Ras recruitment system (RRS), a cytoplasmic-based yeast two-hybrid system, on a bovine retina cDNA library. An interaction between PRCD and tubby-like protein 1 (TULP1) was identified. Co-immunoprecipitation in transfected mammalian cells confirmed that PRCD interacts with TULP1, as well as with its homolog, TUB. These interactions were mediated by TULP1 and TUB highly conserved C-terminal tubby domain. PRCD localization was altered in the retinas of TULP1- and TUB-deficient mice. These results show that TULP1 and TUB, which are involved in the vesicular trafficking of several photoreceptor proteins from the inner segment to the outer segment, are also required for PRCD exclusive localization to photoreceptor outer segment discs.

The 1D4 Antibody Labels Outer Segments of Long Double Cone But Not Rod Photoreceptors in Zebrafish

2012 ◽  
Vol 53 (8) ◽  
pp. 4943 ◽  
Author(s):  
Jun Yin ◽  
Jan Brocher ◽  
Bastian Linder ◽  
Anja Hirmer ◽  
Husvinee Sundaramurthi ◽  
...  
Keyword(s):  
Double Cone ◽  
Rod Photoreceptors ◽  
Outer Segments

scholarly journals Rhodopsin Gene Expression Determines Rod Outer Segment Size and Rod Cell Resistance to a Dominant-Negative Neurodegeneration Mutant

PLoS ONE ◽  
2012 ◽  
Vol 7 (11) ◽  
pp. e49889 ◽  
Author(s):  
Brandee A. Price ◽  
Ivette M. Sandoval ◽  
Fung Chan ◽  
Ralph Nichols ◽  
Ramon Roman-Sanchez ◽  
...  
Keyword(s):  
Gene Expression ◽  
Outer Segment ◽  
Dominant Negative ◽  
Cell Resistance ◽  
Rhodopsin Gene ◽  
Rod Outer Segment ◽  
Segment Size ◽  
Rod Cell

OTOİMMÜN ROMATOİD HASTALIKLARDA OTOANTİKORLAR

2021 ◽  
Vol 8 (17) ◽  
pp. 135-140
Author(s):  
Nuray GÜREL-POLAT
Keyword(s):  
Western Blot ◽  
Clinical Medicine ◽  
Standard Test ◽  
Immunological Tests ◽  
Peptide Antibody ◽  
Golden Standard ◽  
Organ Specific ◽  
Definition Of ◽  
Citrullinated Peptide ◽  
Rheumatoid Diseases

The diagnosis of autoimmune rheumatoid diseases can be made by combining parameters consisting of clinical, histopathological, laboratory and immunological tests. Among these parameters, autoantibodies are of great benefit in differential diagnosis, especially for patients with unclear clinical data. Immunofluorescence, ELISA, immunodiffusion, immunoprecipitation and Western blot can be used to identify autoantibodies. While autoantibodies like antinukleer antibody, double stranded deoksiribonukleic asid and anti neutrophils antibody are detected by immunofluorescence as a golden standard test, Anti-cyclic Citrullinated Peptide Antibody (CCP), antiphospholipid antibodies (anti-cardiolipid) are evaluated by ELISA and ENA group by immunoblot or western blot. Today the number of autoantibodies that can be detected is over 100. The indication of some of these autoantibodies are not known even today. The definition of antinuclear antibody group autoantibodies plays a crucial role for the diagnosis and treatment of systemic and organ-specific illnesses. With these methods, practical, fast and trustworthy results in clinical medicine and clinical immunology can be obtained.

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