Histopathologic and immunocytochemical analysis of the retina and ocular tissues in Batten disease

Ophthalmology. 2000 Sep;107(9):1746-53. doi: 10.1016/s0161-6420(00)00264-5.

Abstract

Purpose: To describe the pathophysiologic features of retinal degeneration in Batten disease (juvenile neuronal ceroid lipofuscinosis [JNCL]) caused by mutations in the CLN3 gene.

Study design: Comparative human tissue study.

Materials: The retina and other ocular tissues of a 22-year-old man with JNCL were compared with the same tissues of a healthy 30-year-old man. DNA from whole blood and RNA from retina were used for genotype analysis.

Methods: The retinas, corneas, conjunctiva, and ciliary body were processed for histopathologic and immunofluorescence analysis. Genomic DNA was subjected to polymerase chain reaction (PCR) and nucleotide sequence analyses. Reverse transcriptase/PCR and sequence analysis were performed on retinal RNA.

Results: The JNCL donor was heterozygous for a approximately 1 kb deletion in CLN3, as found in most JNCL patients. The other allele had a single base pair deletion in exon 6 that resulted in a frame shift. Gross pathology of the JNCL retina resembled that in retinitis pigmentosa, including deposits of bone spicule pigment. Histopathologic studies revealed loss of neurons from all retinal layers. Immunofluorescence labeling with antibodies to rhodopsin, recoverin, and cone opsin demonstrated degenerate rods and cones with short outer segments in the far periphery. Autofluorescent lipopigment granules were prominent in ganglion cells and some cells of the inner nuclear layer, but not in the photoreceptors. The retinal pigment epithelium (RPE) had fewer lipofuscin granules than the control specimen. Increased numbers of lipofuscin granules were found in the epithelia of the ciliary body and conjunctiva, but not in the cornea of the JNCL eye.

Conclusions: Immunofluorescence studies revealed degenerate rods and cones in the far periphery. Lipofuscin granules were decreased in the RPE, consistent with loss of photoreceptor outer segments. The novel finding that degenerate photoreceptors did not contain autofluorescent inclusions suggests that granule accumulation may not precede photoreceptor degeneration in JNCL. The presence of normal photoreceptor proteins in the degenerate rods and cones suggests that these cells may be capable of functional regeneration if a therapy for Batten disease is developed.

Publication types

  • Case Reports
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Cell Cycle
  • Ciliary Body / metabolism
  • Ciliary Body / pathology*
  • Conjunctiva / metabolism
  • Conjunctiva / pathology*
  • Cornea / metabolism
  • Cornea / pathology*
  • Cyclins*
  • Eye Proteins / metabolism
  • Female
  • Fluorescent Antibody Technique, Indirect
  • Genotype
  • Humans
  • Lipofuscin / metabolism
  • Male
  • Membrane Glycoproteins / genetics
  • Microscopy, Fluorescence
  • Molecular Chaperones / genetics
  • Neuronal Ceroid-Lipofuscinoses / genetics
  • Neuronal Ceroid-Lipofuscinoses / metabolism
  • Neuronal Ceroid-Lipofuscinoses / pathology*
  • Retina / metabolism
  • Retina / pathology*
  • Retinal Degeneration / genetics
  • Retinal Degeneration / metabolism
  • Retinal Degeneration / pathology*
  • Saccharomyces cerevisiae Proteins*

Substances

  • CLN3 protein, S cerevisiae
  • CLN3 protein, human
  • Cyclins
  • Eye Proteins
  • Lipofuscin
  • Membrane Glycoproteins
  • Molecular Chaperones
  • Saccharomyces cerevisiae Proteins