Selective absence of cone outer segment β3-transducin immunoreactivity in hereditary cone degeneration (cd)

Kathryn E. Gropp, A. Szél, Jun C. Huang, Gregory M. Acland, Debora B. Farber, Gustavo D. Aguirre

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

We have used immunocytochemistry and in situ hybridization to examine the expression of photoreceptor specific genes in retinas of normal dogs and those affected with hereditary cone degeneration (cd), a rare autosomal recessive disorder that selectively affects cones. In the cd retina, cone disease begins early in life; cones are lost by extrusion of the nucleus into the inner segment, and later, by displacement of the nucleus, surrounded by a thin rim of cytoplasm, into the interphotoreceptor space. Two micrometer sections from the superior and inferior retinal meridians, extending from the optic disk to the ora serrata, were used for in situ hybridization with a bovine rod opsin and human red/green cone opsin cRNA probes, or were reacted with antibodies directed against photoreceptor-specific proteins and visualized with appropriate biotinylated antibodies. Antibodies against the following proteins were used: α- and β3-transducins, phosducin, α/β- and γ-phosphodiesterases, COS-1, and OS-2, opsin, S-antigen and IRBP. Immunoreactivity or hybridization labeling was evaluated in unstained sections; cone pathology was judged in adjacent Toluidine Blue-stained sections. With these methods it was possible to evaluate immunoreactivity or hybridization labeling and cone pathology at the single cell level. Both middle-(COS1) and short-(OS-2) wavelength-sensitive cones were present in controls and cd affected retinae at 2.2 months, and distinct transcripts of the red/green cone pigment gene were identified in the majority of cones in both normal and affected retinas at this age. However, β3-transducin immunoreactivity was completely absent from cd-affected cone outer segments. Both cone types were present but in reduced numbers in older animals (11.5 and 17 months), and no reactivity to β3-transducin was noted. No differences were found with the other antibodies used. The specific absence of β3-transducin immunoreactivity from the cone outer segments suggests a potential involvement of the β3-transducin gene or gene product in the disease process.

Original languageEnglish
Pages (from-to)285-296
Number of pages12
JournalExperimental Eye Research
Volume63
Issue number3
DOIs
Publication statusPublished - Sep 1996

Fingerprint

Transducin
Retina
Antibodies
Genes
In Situ Hybridization
Rod Opsins
Cone Opsins
Pathology
Retinal Cone Photoreceptor Cells
Opsins
Tolonium Chloride
Meridians
Complementary RNA
Optic Disk
Phosphoric Diester Hydrolases
Cytoplasm
Proteins
Immunohistochemistry
Dogs
Antigens

Keywords

  • β-transducin
  • animal model
  • cone degeneration
  • dog
  • hereditary disease
  • immunocytochemistry
  • in situ hybridization
  • photoreceptor
  • retina
  • rod monochromatism

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

Cite this

Selective absence of cone outer segment β3-transducin immunoreactivity in hereditary cone degeneration (cd). / Gropp, Kathryn E.; Szél, A.; Huang, Jun C.; Acland, Gregory M.; Farber, Debora B.; Aguirre, Gustavo D.

In: Experimental Eye Research, Vol. 63, No. 3, 09.1996, p. 285-296.

Research output: Contribution to journalArticle

Gropp, Kathryn E. ; Szél, A. ; Huang, Jun C. ; Acland, Gregory M. ; Farber, Debora B. ; Aguirre, Gustavo D. / Selective absence of cone outer segment β3-transducin immunoreactivity in hereditary cone degeneration (cd). In: Experimental Eye Research. 1996 ; Vol. 63, No. 3. pp. 285-296.
@article{326160b25a964399ac255a7f48b498d5,
title = "Selective absence of cone outer segment β3-transducin immunoreactivity in hereditary cone degeneration (cd)",
abstract = "We have used immunocytochemistry and in situ hybridization to examine the expression of photoreceptor specific genes in retinas of normal dogs and those affected with hereditary cone degeneration (cd), a rare autosomal recessive disorder that selectively affects cones. In the cd retina, cone disease begins early in life; cones are lost by extrusion of the nucleus into the inner segment, and later, by displacement of the nucleus, surrounded by a thin rim of cytoplasm, into the interphotoreceptor space. Two micrometer sections from the superior and inferior retinal meridians, extending from the optic disk to the ora serrata, were used for in situ hybridization with a bovine rod opsin and human red/green cone opsin cRNA probes, or were reacted with antibodies directed against photoreceptor-specific proteins and visualized with appropriate biotinylated antibodies. Antibodies against the following proteins were used: α- and β3-transducins, phosducin, α/β- and γ-phosphodiesterases, COS-1, and OS-2, opsin, S-antigen and IRBP. Immunoreactivity or hybridization labeling was evaluated in unstained sections; cone pathology was judged in adjacent Toluidine Blue-stained sections. With these methods it was possible to evaluate immunoreactivity or hybridization labeling and cone pathology at the single cell level. Both middle-(COS1) and short-(OS-2) wavelength-sensitive cones were present in controls and cd affected retinae at 2.2 months, and distinct transcripts of the red/green cone pigment gene were identified in the majority of cones in both normal and affected retinas at this age. However, β3-transducin immunoreactivity was completely absent from cd-affected cone outer segments. Both cone types were present but in reduced numbers in older animals (11.5 and 17 months), and no reactivity to β3-transducin was noted. No differences were found with the other antibodies used. The specific absence of β3-transducin immunoreactivity from the cone outer segments suggests a potential involvement of the β3-transducin gene or gene product in the disease process.",
keywords = "β-transducin, animal model, cone degeneration, dog, hereditary disease, immunocytochemistry, in situ hybridization, photoreceptor, retina, rod monochromatism",
author = "Gropp, {Kathryn E.} and A. Sz{\'e}l and Huang, {Jun C.} and Acland, {Gregory M.} and Farber, {Debora B.} and Aguirre, {Gustavo D.}",
year = "1996",
month = "9",
doi = "10.1006/exer.1996.0117",
language = "English",
volume = "63",
pages = "285--296",
journal = "Experimental Eye Research",
issn = "0014-4835",
publisher = "Academic Press Inc.",
number = "3",

}

TY - JOUR

T1 - Selective absence of cone outer segment β3-transducin immunoreactivity in hereditary cone degeneration (cd)

AU - Gropp, Kathryn E.

AU - Szél, A.

AU - Huang, Jun C.

AU - Acland, Gregory M.

AU - Farber, Debora B.

AU - Aguirre, Gustavo D.

PY - 1996/9

Y1 - 1996/9

N2 - We have used immunocytochemistry and in situ hybridization to examine the expression of photoreceptor specific genes in retinas of normal dogs and those affected with hereditary cone degeneration (cd), a rare autosomal recessive disorder that selectively affects cones. In the cd retina, cone disease begins early in life; cones are lost by extrusion of the nucleus into the inner segment, and later, by displacement of the nucleus, surrounded by a thin rim of cytoplasm, into the interphotoreceptor space. Two micrometer sections from the superior and inferior retinal meridians, extending from the optic disk to the ora serrata, were used for in situ hybridization with a bovine rod opsin and human red/green cone opsin cRNA probes, or were reacted with antibodies directed against photoreceptor-specific proteins and visualized with appropriate biotinylated antibodies. Antibodies against the following proteins were used: α- and β3-transducins, phosducin, α/β- and γ-phosphodiesterases, COS-1, and OS-2, opsin, S-antigen and IRBP. Immunoreactivity or hybridization labeling was evaluated in unstained sections; cone pathology was judged in adjacent Toluidine Blue-stained sections. With these methods it was possible to evaluate immunoreactivity or hybridization labeling and cone pathology at the single cell level. Both middle-(COS1) and short-(OS-2) wavelength-sensitive cones were present in controls and cd affected retinae at 2.2 months, and distinct transcripts of the red/green cone pigment gene were identified in the majority of cones in both normal and affected retinas at this age. However, β3-transducin immunoreactivity was completely absent from cd-affected cone outer segments. Both cone types were present but in reduced numbers in older animals (11.5 and 17 months), and no reactivity to β3-transducin was noted. No differences were found with the other antibodies used. The specific absence of β3-transducin immunoreactivity from the cone outer segments suggests a potential involvement of the β3-transducin gene or gene product in the disease process.

AB - We have used immunocytochemistry and in situ hybridization to examine the expression of photoreceptor specific genes in retinas of normal dogs and those affected with hereditary cone degeneration (cd), a rare autosomal recessive disorder that selectively affects cones. In the cd retina, cone disease begins early in life; cones are lost by extrusion of the nucleus into the inner segment, and later, by displacement of the nucleus, surrounded by a thin rim of cytoplasm, into the interphotoreceptor space. Two micrometer sections from the superior and inferior retinal meridians, extending from the optic disk to the ora serrata, were used for in situ hybridization with a bovine rod opsin and human red/green cone opsin cRNA probes, or were reacted with antibodies directed against photoreceptor-specific proteins and visualized with appropriate biotinylated antibodies. Antibodies against the following proteins were used: α- and β3-transducins, phosducin, α/β- and γ-phosphodiesterases, COS-1, and OS-2, opsin, S-antigen and IRBP. Immunoreactivity or hybridization labeling was evaluated in unstained sections; cone pathology was judged in adjacent Toluidine Blue-stained sections. With these methods it was possible to evaluate immunoreactivity or hybridization labeling and cone pathology at the single cell level. Both middle-(COS1) and short-(OS-2) wavelength-sensitive cones were present in controls and cd affected retinae at 2.2 months, and distinct transcripts of the red/green cone pigment gene were identified in the majority of cones in both normal and affected retinas at this age. However, β3-transducin immunoreactivity was completely absent from cd-affected cone outer segments. Both cone types were present but in reduced numbers in older animals (11.5 and 17 months), and no reactivity to β3-transducin was noted. No differences were found with the other antibodies used. The specific absence of β3-transducin immunoreactivity from the cone outer segments suggests a potential involvement of the β3-transducin gene or gene product in the disease process.

KW - β-transducin

KW - animal model

KW - cone degeneration

KW - dog

KW - hereditary disease

KW - immunocytochemistry

KW - in situ hybridization

KW - photoreceptor

KW - retina

KW - rod monochromatism

UR - http://www.scopus.com/inward/record.url?scp=0030249072&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0030249072&partnerID=8YFLogxK

U2 - 10.1006/exer.1996.0117

DO - 10.1006/exer.1996.0117

M3 - Article

VL - 63

SP - 285

EP - 296

JO - Experimental Eye Research

JF - Experimental Eye Research

SN - 0014-4835

IS - 3

ER -