title: X-gal staining of canine skin tissues: A technique with multiple possible applications.
abstract: BACKGROUND
  Estimation of β-galactosidase (βgal) activity in human cells and tissues indicate its possible use as a marker of senescence.
  
  
  OBJECTIVE
  This study was done to detect senescence-associated βgal (SA-βgal) activity in canine skin tissue by using its substrate 5-bromo-4-chloro-3-indolyl β-D-galactosidase (X-gal).
  
  
  METHODS
  Skin samples were collected through rapid necropsy process. The X-gal staining was done by altering different factors of the staining procedure like pH of the reagents and incubation time. Further, effect of tissue preservation procedure was also evaluated.
  
  
  RESULTS
  Typical X-gal staining was detected in old dogs' skin samples and it was detectable both at pH 6 and pH 7.3. The cells present in the inner lining of the hair follicles and sebaceous glands are the major cells that have high SA-βgal activity. The X-gal staining intensity was more prominent in tissues preserved in liquid nitrogen at -196°C than in -80°C freezer. Prolonged incubation period increased the intensity of staining.
  
  
  CONCLUSIONS
  This study indicates possibility of X-gal staining in canine tissues and opens an avenue for further in-depth studies that might be useful for different research and clinical studies like determination of dog's approximate age.
authors.affiliation: Department of Veterinary Pathology, Orissa University of Agriculture and Technology, Bhubaneswar, India.
authors.forename: Soumyaranjan
authors.initials: S
authors.lastname: Pati
authors.affiliation: Institute of Life Sciences, Department of Translational Research and Technology Development, Nalco Square, Bhubaneswar, Orissa, India.
authors.forename: Sumeet
authors.initials: S
authors.lastname: Jain
authors.affiliation: Department of Veterinary Pathology, Orissa University of Agriculture and Technology, Bhubaneswar, India.
authors.forename: Monalisa
authors.initials: M
authors.lastname: Behera
authors.affiliation: Department of Veterinary Pathology, Orissa University of Agriculture and Technology, Bhubaneswar, India.
authors.forename: Aditya Prasad
authors.initials: AP
authors.lastname: Acharya
authors.affiliation: Department of Veterinary Pathology, Orissa University of Agriculture and Technology, Bhubaneswar, India.
authors.forename: Susen K
authors.initials: SK
authors.lastname: Panda
authors.affiliation: Institute of Life Sciences, Department of Translational Research and Technology Development, Nalco Square, Bhubaneswar, Orissa, India.
authors.forename: Shantibhusan
authors.initials: S
authors.lastname: Senapati
keywords: X-gal; canine; senescence; skin; β-galactosidase
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