Journal of Animal Research
  • Year: 2021
  • Volume: 11
  • Issue: 2

Identification of Differentially Expressed Tear Proteins after Corneal Reconstruction with DPSIS Graft in Dogs Suffering from Keratoconjunctivitis Sicca

  • Author:
  • Sowbharenya Chelladurai1, Kiranjeet Singh1, Gopinathan Aswathy1,*, Ravi Kant Agrawal2, Ajay Kumar3, Pawan Kumar4, Monalisa Sahoo4
  • Total Page Count: 11
  • Published Online: Sep 21, 2022
  • Page Number: 273 to 283

1Division of Surgery, ICAR-IVRI, Izatnagar, Bareilly, Uttar Pradesh, India

2Division of Livestock Products technology, ICAR-IVRI, Izatnagar, Bareilly, Uttar Pradesh, India

3Division of Biochemistry, ICAR-IVRI, Izatnagar, Bareilly, Uttar Pradesh, India

4Division of Pathology, ICAR-IVRI, Izatnagar, Bareilly, Uttar Pradesh, India

*Corresponding author: A Gopinathan; E-mail: aswathykiran77@gmail.com

Online Published on 21 September, 2022.

Abstract

Differentially expressed tear proteins after corneal reconstruction with decellularized porcine small intestinal sub mucosa (DPSIS) in Keratoconjunctivitis sicca (KCS) affected dogs was explored through Matrix Assisted Laser Desorption/Ionization Mass spectrometry (MALDI-TOF/MS) followed by Peptide mass fingerprinting (PMF). Anterior lamellar keratoplasty followed by DPSIS graft was performed in seven dogs with corneal ulcer (group A) and in nine dogs with corneal ulcer as a complication of KCS (group B). Tear was also collected from 12 healthy dogs with no ocular pathology. PMF based protein identification was performed on tryptic peptides of tear film trapped in major bands of ID-SDS PAGE (in duplicate) using MALDI-TOF/ MS. PMF revealed 432 unique tear proteins from major gel bands of DPSIS treated dogs (group A and B) and normal tear from healthy dogs. Based on MOWSE score, relative abundance of proteins in both groups were compared with that in normal tear. Proteins USP37, KRT1, Major allergen Can F1, CCDC39, PRKCD, Serum albumin, TERT and CD163 were found to be significantly (p<0.001) down-regulated in group A and group B compared to normal tear from healthy dogs. Protein INVS was found significantly up regulated (p<0.001) in Group B post DPSIS graft compared to pre-operative tear. ZNF252 was found significantly up regulated (p<0.001) in Group A tear film post-operatively compared to pre-operative tear. Intrinsic cellular stress disrupting DNA replication and cell division, cellular senescence and apoptosis were found as key events underlying corneal pathology in KCS which needs consideration while attempting surgical reconstruction of cornea using DPSIS graft.

• Differentially expressed tear proteins post- DPSIS graft in KCS affected dogs were elucidated.

• Intrinsic cellular stress disrupting DNA replication and cell division was the key event underlying corneal pathology in KCS affected dogs.

Keywords

Tear Proteome, MALDI-TOF/MS, DPSIS graft, Keratoconjunctivitis sicca, Dogs