Establishment of an appropriate animal model for lacritin studies: Cloning and characterization of lacritin in monkey eyes

T. Nakajima, R. D. Walkup, A. Tochigi, Thomas (Tom) Shearer, M. Azuma

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Lacritin is a mitogen of human salivary gland cells as well as a stimulator of human corneal epithelial cells. It is expected to be an important factor in maintaining the surrounding ocular surface. The monkey would be a relevant animal model in which to study the role of lacritin in ophthalmic physiology and pathology. However, to our knowledge, no cDNA cloning or functional analysis of monkey lacritin has been performed. Thus, the purposes of this study were: (1) to clone the monkey ortholog of lacritin; (2) to characterize lacritin in tears from several species; and (3) to determine the tissues where lacritin is produced and secreted. cDNA for lacritin from rhesus macaque contained 547 bp, with 411 bp in an open reading frame (ORF) encoding a protein of 137 amino acids. Monkey lacritin showed 89% amino acid homology with human lacritin; one amino acid was deleted in all three monkey strains. The predicted MW of mature lacritin was 12.2 kDa, and the isoelectric point was 4.99. Lacritin showed anomalous migration at approximately 21.0 kDa on SDS-PAGE, as confirmed by immunoblotting and amino acid sequencing. Similar to native lacritin in monkey tears, a 21 kDa band was also detected in human tears. In contrast, no lacritin was observed at a similar position on SDS-PAGE in rat, rabbit and dog tears. In the monkey, lacritin mRNA was expressed highly in the lacrimal gland, moderately in the conjunctiva and the meibomian gland, and weakly in corneal epithelium. In primates, lacritin was produced in the lacrimal gland and secreted into tear fluid. These results suggest that lacritin might be important for the maintenance of the ocular surface in higher animals, such as monkeys and humans.

Original languageEnglish (US)
Pages (from-to)651-658
Number of pages8
JournalExperimental Eye Research
Volume85
Issue number5
DOIs
StatePublished - Nov 2007

Fingerprint

Haplorhini
Organism Cloning
Animal Models
Tears
Lacrimal Apparatus
Amino Acids
Polyacrylamide Gel Electrophoresis
Ocular Physiological Phenomena
Complementary DNA
Meibomian Glands
Corneal Epithelium
Conjunctiva
Protein Sequence Analysis
Isoelectric Point
Salivary Glands
Macaca mulatta
Mitogens
Immunoblotting
Primates
Open Reading Frames

Keywords

  • lacritin
  • monkey
  • tear film

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems

Cite this

Establishment of an appropriate animal model for lacritin studies : Cloning and characterization of lacritin in monkey eyes. / Nakajima, T.; Walkup, R. D.; Tochigi, A.; Shearer, Thomas (Tom); Azuma, M.

In: Experimental Eye Research, Vol. 85, No. 5, 11.2007, p. 651-658.

Research output: Contribution to journalArticle

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