October 1997
Volume 38, Issue 11
Free
Articles  |   October 1997
Immunochemical localization of the Batten disease (CLN3) protein in retina.
Author Affiliations
  • M L Katz
    Mason Eye Institute, University of Missouri School of Medicine, Columbia 65212, USA.
  • C L Gao
    Mason Eye Institute, University of Missouri School of Medicine, Columbia 65212, USA.
  • M Prabhakaram
    Mason Eye Institute, University of Missouri School of Medicine, Columbia 65212, USA.
  • H Shibuya
    Mason Eye Institute, University of Missouri School of Medicine, Columbia 65212, USA.
  • P C Liu
    Mason Eye Institute, University of Missouri School of Medicine, Columbia 65212, USA.
  • G S Johnson
    Mason Eye Institute, University of Missouri School of Medicine, Columbia 65212, USA.
Investigative Ophthalmology & Visual Science October 1997, Vol.38, 2375-2386. doi:
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      M L Katz, C L Gao, M Prabhakaram, H Shibuya, P C Liu, G S Johnson; Immunochemical localization of the Batten disease (CLN3) protein in retina.. Invest. Ophthalmol. Vis. Sci. 1997;38(11):2375-2386.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

PURPOSE: Batten disease, also known as juvenile ceroid-lipofuscinosis and CLN3, is an autosomal recessively inherited disorder that results in blindness due to retinal degeneration. The CLN3 gene has been identified, but the function of the protein that this gene encodes is unknown. Experiments were conducted to determine where the CLN3 protein is localized in the mouse retina. Localization should provide a clue in evaluating potential functions of this protein. METHODS: Using oligonucleotide primers based on the reported human CLN3 cDNA sequence, the mouse cDNA nucleotide sequence was determined from products of the reverse transcriptase-polymerase chain reaction and 3' rapid amplification of cDNA ends. A synthetic 20-amino-acid peptide corresponding to an internal hydrophilic region of the predicted amino acid sequence of the mouse CLN3 protein was used to immunize rabbits. The resulting antiserum was used in immunoblot analysis of mouse retina homogenates and in electron microscopic immunocytochemical labeling of mouse retina sections. RESULTS: The peptide antibody labeled a single protein band of approximately 50 kDa on immunoblots of mouse retina homogenates. No labeling was detected with homogenates from human retinas. The antibody specifically labeled mitochondria of Müller cells and inner retinal neurons. Little labeling was observed in mitochondria of the photoreceptor cells. Mitochondria of other cell types, including the retinal pigment epithelium and choroidal cells, were not labeled. CONCLUSIONS: The retinal CLN3 protein appears to be localized almost exclusively in the mitochondria, but was detected only in certain cell types. Batten disease is characterized by massive lysosomal accumulations of a small inner mitochondrial membrane protein (subunit c of ATP synthase). The mitochondrial localization of the CLN3 protein suggests that it may play a role in the normal processing of subunit c.

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