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Distinguishing L from M photopigment coding sequences by hybridization to novel locked nucleic acid (LNA) oligonucleotide probes

Published online by Cambridge University Press:  03 July 2008

CHRISTINA PETTAN-BREWER ,
LI FU  and
SAMIR S. DEEB
CHRISTINA PETTAN-BREWER*
Affiliation:
Division of Medical Genetics, Department of Medicine, University of Washington, Seattle, Washington
LI FU
Affiliation:
Division of Medical Genetics, Department of Medicine, University of Washington, Seattle, Washington
SAMIR S. DEEB
Affiliation:
Division of Medical Genetics, Department of Medicine, University of Washington, Seattle, Washington Department of Genome Sciences, University of Washington, Seattle, Washington
*
Address correspondence and reprint requests to: Christina Pettan-Brewer, Division of Medical Genetics, Box 357720, University of Washington, Seattle, WA 98195. E-mail: kcpb@u.washington.edu
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Abstract

Many attempts have been made over the years to distinguish human and primate L (long-wavelength sensitive) from M (middle-wavelength sensitive) cone photoreceptors using either immunohistochemistry or in situ hybridization. These attempts have been unsuccessful due to the very high degree of identity between the sequences of the L and M proteins and encoding mRNAs. The recent development of chemically modified oligonucleotide probes, referred to as locked nucleic acid (LNA) probes, has shown that they hybridize with much greater affinity and specificity to the target nucleic acid. This has greatly increased the potential for differentiating L from M cones by in situ hybridization. We have designed LNA oligonucleotide probes that are complementary to either the L or M coding sequences located in exon 5 of the Macaca nemestrina L and M pigment genes. We have shown that the LNA-M and LNA-L probes hybridize specifically to their respective target nucleic acid sequences in vitro. This result strongly suggests that these probes would be instrumental in rapidly distinguishing L from M cone in the entire retina, and in defining the cone mosaic during development and in adults.

Keywords

Locked nucleic acid (LNA)Cone pigmentsPrimate
Type
Research Article
Information
Visual Neuroscience , Volume 25 , Issue 3 , May 2008 , pp. 283 - 287
Copyright
Copyright © Cambridge University Press 2008

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