title: Human postmortem lacrimal and submandibular glands stored in RNAlaterare suitable for molecular, biochemical, and cell biological studies.
abstract: Gene expression and protein analysis studies require high-quality human tissue which is a challenge and difficult to obtain through live human biopsies. Human postmortem lacrimal gland (LG) and submandibular gland (SMG) tissues have the potential to provide an invaluable source for studying the mechanisms involved in LG and SMG dysfunction. Therefore, we aimed to test the suitability of post-mortem LG and SMG for molecular, biochemical, and cell biological studies.
  
  
  
  LG and SMG tissue from healthy donors was collected and immediately placed in RNAlatersolution and then shipped overnight at 4 °C. After receipt, each gland was divided into three pieces for RNA, protein, and histological analysis, respectively. Total RNA isolated from each LG and SMG was analyzed for RNA integrity using an Agilent Bioanalyzer and reverse transcription-PCR (RT-PCR). For histology, tissues were embedded in paraffin and stained with hematoxylin and eosin. For protein analysis, lysates were prepared and processed for sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting.
  
  
  
  When the LG and SMG samples were preserved in RNAlater, the RNA integrity number (RIN) values from the LG and SMG were >7.0 from all three donors, while the RNAs from tissue not preserved in RNAlaterwere of poorer quality. The gene and/or protein expression of E-cadherin, aquaporin 5, alpha-smooth muscle actin (α-SMA), β-actin, and GAPDH was preserved in all samples. In addition, histological analyses showed normal tubuloacinar structures of all glands with serous and mucous producing acini within lobules interspersed with adipose fat.
  
  
  
  In this study, we determined that RNA, protein, and histological sections obtained from postmortem human LG and SMG tissue preserved in RNAlaterwere of high quality. This would provide a viable source of human LG and SMG tissue suitable for studies of diseases that affect these glands, such as Sjögren's syndrome.
authors: Department of Comprehensive Care, Tufts University School of Dental Medicine, Boston, MA.
authors: Dillon
authors: D
authors: Hawley
authors: Department of Comprehensive Care, Tufts University School of Dental Medicine, Boston, MA.
authors: Hema
authors: H
authors: Aluri
authors: Department of Comprehensive Care, Tufts University School of Dental Medicine, Boston, MA.
authors: Helene
authors: H
authors: Armaos
authors: Department of Comprehensive Care, Tufts University School of Dental Medicine, Boston, MA.
authors: Gina
authors: G
authors: Kim
authors: Department of Comprehensive Care, Tufts University School of Dental Medicine, Boston, MA.
authors: Claire
authors: C
authors: Kublin
authors: Department of Comprehensive Care, Tufts University School of Dental Medicine, Boston, MA; Department of Ophthalmology, Tufts University School of Medicine, Boston, MA.
authors: Driss
authors: D
authors: Zoukhri
mesh_terms: D000199:Actins / Q000378:metabolism; D000369:Aged, 80 and over; D015703:Antigens, CD; D051402:Aquaporin 5 / Q000378:metabolism; D001344:Autopsy; D015153:Blotting, Western; D015820:Cadherins / Q000378:metabolism; D015925:Cryopreservation; D004591:Electrophoresis, Polyacrylamide Gel; D005136:Eye Proteins / Q000302:isolation & purification* / Q000378:metabolism; D005260:Female; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / Q000378:metabolism; D006801:Humans; D007765:Lacrimal Apparatus / Q000737:chemistry* / Q000378:metabolism; D008875:Middle Aged; D009926:Organ Preservation; D019188:Organ Preservation Solutions; D012313:RNA / Q000302:isolation & purification* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D013363:Submandibular Gland / Q000737:chemistry* / Q000378:metabolism; D014019:Tissue Donors
chemical_list: C541116:ACTA2 protein, human; C496036:AQP5 protein, human; D000199:Actins; D015703:Antigens, CD; D051402:Aquaporin 5; C513384:CDH1 protein, human; D015820:Cadherins; D005136:Eye Proteins; D019188:Organ Preservation Solutions; D012313:RNA; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+)