Identification of novel molecular components of the photoreceptor connecting cilium by immunoscreens

Autoren

Angelika Schmitt
Uwe Wolfrum

DOI

10.1006/exer.2001.1086

ISSN

0014-4835

Ausgabe der Veröffentlichung

6

Zeitschrift

Experimental Eye Research

Sprache

en

Paginierung

837 - 849

Datum der Veröffentlichung

2001

Status

Published

Herausgeber

Elsevier BV

Herausgeber URL

http://dx.doi.org/10.1006/exer.2001.1086

Datum der Datenerfassung

2021

Titel

Identification of Novel Molecular Components of the Photoreceptor Connecting Cilium by Immunoscreens

Ausgabe der Zeitschrift

73

Data source: Crossref

Abstract

The connecting cilium of photoreceptor cells is the only intracellular link between the morphologically, functionally and biochemically different compartments of the inner and outer segments. The non-motile modified cilium plays an important role in the organization and the function of photoreceptor cells, namely in delivery and turnover of enzymes and substrates of the visual transduction cascade, and the photosensitive membranes of the outer segment. The protein components of the cilium participate in the intracellular transport through the cilium, in the outer segment disk morphogenesis and in the maintenance of discrete membrane domains. In order to identify yet unknown cytoskeletal components of the connecting cilium, a combined biochemical and molecular biological strategy was applied. For this purpose, antibodies were raised against proteins of photoreceptor cell axonemes. Using this AX-4-antiserum, a rat retina cDNA expression library was immunoscreened and clones encoding partial sequences of (i) already known photoreceptor specific proteins; (ii) ubiquitously expressed proteins; (iii) clones with homologies to retinal ESTs; and (iv) clones coding for cytoskeletal proteins were isolated. Further analysis revealed that these candidate clones have homologies to Drosophila flightless I, mouse APC-binding protein EB2, human microtubule associated protein 4 (MAP4), human centrin 3, human cytoplasmic dynein intermediate chain 2C, and human dynamitin.The immunoscreening approach used here was successfully applied to isolate genes encoding yet unknown cytoskeletal proteins of photoreceptor cell axonemes. The obtained information will provide further insight into the role of the connecting cilium in photoreceptor cell function.

Addresses

Institute of Zoology, Johannes Gutenberg-University of Mainz, Mainz, Germany.

Autoren

A Schmitt
U Wolfrum

DOI

10.1006/exer.2001.1086

eISSN

1096-0007

Externe Identifier

PubMed Identifier: 11846514

Open access

false

ISSN

0014-4835

Ausgabe der Veröffentlichung

6

Zeitschrift

Experimental eye research

Schlüsselwörter

Cilia
Cytoskeleton
Animals
Mice, Inbred C57BL
Mice
Rats
Rats, Sprague-Dawley
Xenopus Proteins
Calcium-Binding Proteins
Adenomatous Polyposis Coli Protein
Microtubule-Associated Proteins
Drosophila Proteins
Cloning, Molecular
Gene Library
Photoreceptor Cells, Vertebrate
Dynactin Complex

Sprache

eng

Medium

Print

Paginierung

837 - 849

Datum der Veröffentlichung

2001

Status

Published

Datum der Datenerfassung

2002

Titel

Identification of novel molecular components of the photoreceptor connecting cilium by immunoscreens.

Sub types

Research Support, Non-U.S. Gov't
Journal Article

Ausgabe der Zeitschrift

73

Data source: Europe PubMed Central

Abstract

The connecting cilium of photoreceptor cells is the only intracellular link between the morphologically, functionally and biochemically different compartments of the inner and outer segments. The non-motile modified cilium plays an important role in the organization and the function of photoreceptor cells, namely in delivery and turnover of enzymes and substrates of the visual transduction cascade, and the photosensitive membranes of the outer segment. The protein components of the cilium participate in the intracellular transport through the cilium, in the outer segment disk morphogenesis and in the maintenance of discrete membrane domains. In order to identify yet unknown cytoskeletal components of the connecting cilium, a combined biochemical and molecular biological strategy was applied. For this purpose, antibodies were raised against proteins of photoreceptor cell axonemes. Using this AX-4-antiserum, a rat retina cDNA expression library was immunoscreened and clones encoding partial sequences of (i) already known photoreceptor specific proteins; (ii) ubiquitously expressed proteins; (iii) clones with homologies to retinal ESTs; and (iv) clones coding for cytoskeletal proteins were isolated. Further analysis revealed that these candidate clones have homologies to Drosophila flightless I, mouse APC-binding protein EB2, human microtubule associated protein 4 (MAP4), human centrin 3, human cytoplasmic dynein intermediate chain 2C, and human dynamitin.The immunoscreening approach used here was successfully applied to isolate genes encoding yet unknown cytoskeletal proteins of photoreceptor cell axonemes. The obtained information will provide further insight into the role of the connecting cilium in photoreceptor cell function.

Autoren

A Schmitt
U Wolfrum

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/11846514

DOI

10.1006/exer.2001.1086

ISSN

0014-4835

Ausgabe der Veröffentlichung

6

Zeitschrift

Exp Eye Res

Schlüsselwörter

Adenomatous Polyposis Coli Protein
Animals
Calcium-Binding Proteins
Cilia
Cloning, Molecular
Cytoskeleton
Drosophila Proteins
Dynactin Complex
Gene Library
Mice
Mice, Inbred C57BL
Microtubule-Associated Proteins
Photoreceptor Cells, Vertebrate
Rats
Rats, Sprague-Dawley
Xenopus Proteins

Sprache

eng

Country

England

Paginierung

837 - 849

PII

S0014-4835(01)91086-3

Datum der Veröffentlichung

2001

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2002

Titel

Identification of novel molecular components of the photoreceptor connecting cilium by immunoscreens.

Sub types

Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

73

Data source: PubMed

Identification of novel molecular components of the photoreceptor connecting cilium by immunoscreens

Tools