Cholangiocyte apoptosis is an early event during induced metamorphosis in the sea lamprey, Petromyzon marinus L

Laura A. Boomer, Seth Bellister, Linda L. Stephenson, Stanley D. Hillyard, Joseph D. Khoury, John H. Youson, John R. Gosche

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Background: Research in biliary atresia has been hindered by lack of a suitable animal model. Lampreys are primitive vertebrates with distinct larval and adult life cycle stages. During metamorphosis the biliary system of the larval lamprey disappears. Lamprey metamorphosis has been proposed as a model for biliary atresia. We have begun to explore cellular events during lamprey metamorphosis by assessing for cholangiocyte apoptosis. Materials and Methods: Sea lamprey larvae were housed under controlled environmental conditions. Premetamorphic larvae were induced to undergo metamorphosis by exposure to 0.01% KClO4. Animals were photographed weekly, and the stage of metamorphosis was assigned based upon external features. Livers were harvested and processed for routine histology and immunohistochemistry. DNA fragmentation was detected using deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assays and cholangiocytes were identified with antibodies to cytokeratin-19. Percent TUNEL+ cholangiocytes at different stages of metamorphosis was determined. Results: The percentage of TUNEL+ cholangiocytes was 10% in premetamorphic (stage 0) lamprey (n = 6), 51% at stage 1 (n = 6), 40% at stage 2 (n = 5), 18% at stage 3 (n = 5), and 9% stage 4 (n = 4). Routine hemotoxylin and eosin stained paraffin-embedded tissue sections revealed frequent apoptotic bodies at stages 3 and 4 of metamorphosis without histologic evidence of necrosis. Conclusions: DNA fragmentation is identified at the earliest stages of metamorphosis during induced metamorphosis in lampreys. Additional studies are necessary to validate this potentially valuable animal model.

Original languageEnglish (US)
Pages (from-to)114-120
Number of pages7
JournalJournal of Pediatric Surgery
Volume45
Issue number1
DOIs
StatePublished - Jan 1 2010
Externally publishedYes

Fingerprint

Petromyzon
Lampreys
Apoptosis
Biotin
Biliary Atresia
DNA Fragmentation
Transferases
Larva
Animal Models
Keratin-19
DNA Nucleotidylexotransferase
Biliary Tract
Hematoxylin
Eosine Yellowish-(YS)
Life Cycle Stages
Paraffin
Vertebrates
Histology
Necrosis
Immunohistochemistry

Keywords

  • Apoptosis
  • Biliary atresia
  • Cholangiocyte
  • Lamprey
  • Metamorphosis

ASJC Scopus subject areas

  • Surgery
  • Pediatrics, Perinatology, and Child Health

Cite this

Cholangiocyte apoptosis is an early event during induced metamorphosis in the sea lamprey, Petromyzon marinus L. / Boomer, Laura A.; Bellister, Seth; Stephenson, Linda L.; Hillyard, Stanley D.; Khoury, Joseph D.; Youson, John H.; Gosche, John R.

In: Journal of Pediatric Surgery, Vol. 45, No. 1, 01.01.2010, p. 114-120.

Research output: Contribution to journalArticle

Boomer, Laura A. ; Bellister, Seth ; Stephenson, Linda L. ; Hillyard, Stanley D. ; Khoury, Joseph D. ; Youson, John H. ; Gosche, John R. / Cholangiocyte apoptosis is an early event during induced metamorphosis in the sea lamprey, Petromyzon marinus L. In: Journal of Pediatric Surgery. 2010 ; Vol. 45, No. 1. pp. 114-120.
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title = "Cholangiocyte apoptosis is an early event during induced metamorphosis in the sea lamprey, Petromyzon marinus L",
abstract = "Background: Research in biliary atresia has been hindered by lack of a suitable animal model. Lampreys are primitive vertebrates with distinct larval and adult life cycle stages. During metamorphosis the biliary system of the larval lamprey disappears. Lamprey metamorphosis has been proposed as a model for biliary atresia. We have begun to explore cellular events during lamprey metamorphosis by assessing for cholangiocyte apoptosis. Materials and Methods: Sea lamprey larvae were housed under controlled environmental conditions. Premetamorphic larvae were induced to undergo metamorphosis by exposure to 0.01{\%} KClO4. Animals were photographed weekly, and the stage of metamorphosis was assigned based upon external features. Livers were harvested and processed for routine histology and immunohistochemistry. DNA fragmentation was detected using deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assays and cholangiocytes were identified with antibodies to cytokeratin-19. Percent TUNEL+ cholangiocytes at different stages of metamorphosis was determined. Results: The percentage of TUNEL+ cholangiocytes was 10{\%} in premetamorphic (stage 0) lamprey (n = 6), 51{\%} at stage 1 (n = 6), 40{\%} at stage 2 (n = 5), 18{\%} at stage 3 (n = 5), and 9{\%} stage 4 (n = 4). Routine hemotoxylin and eosin stained paraffin-embedded tissue sections revealed frequent apoptotic bodies at stages 3 and 4 of metamorphosis without histologic evidence of necrosis. Conclusions: DNA fragmentation is identified at the earliest stages of metamorphosis during induced metamorphosis in lampreys. Additional studies are necessary to validate this potentially valuable animal model.",
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T1 - Cholangiocyte apoptosis is an early event during induced metamorphosis in the sea lamprey, Petromyzon marinus L

AU - Boomer, Laura A.

AU - Bellister, Seth

AU - Stephenson, Linda L.

AU - Hillyard, Stanley D.

AU - Khoury, Joseph D.

AU - Youson, John H.

AU - Gosche, John R.

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AB - Background: Research in biliary atresia has been hindered by lack of a suitable animal model. Lampreys are primitive vertebrates with distinct larval and adult life cycle stages. During metamorphosis the biliary system of the larval lamprey disappears. Lamprey metamorphosis has been proposed as a model for biliary atresia. We have begun to explore cellular events during lamprey metamorphosis by assessing for cholangiocyte apoptosis. Materials and Methods: Sea lamprey larvae were housed under controlled environmental conditions. Premetamorphic larvae were induced to undergo metamorphosis by exposure to 0.01% KClO4. Animals were photographed weekly, and the stage of metamorphosis was assigned based upon external features. Livers were harvested and processed for routine histology and immunohistochemistry. DNA fragmentation was detected using deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) assays and cholangiocytes were identified with antibodies to cytokeratin-19. Percent TUNEL+ cholangiocytes at different stages of metamorphosis was determined. Results: The percentage of TUNEL+ cholangiocytes was 10% in premetamorphic (stage 0) lamprey (n = 6), 51% at stage 1 (n = 6), 40% at stage 2 (n = 5), 18% at stage 3 (n = 5), and 9% stage 4 (n = 4). Routine hemotoxylin and eosin stained paraffin-embedded tissue sections revealed frequent apoptotic bodies at stages 3 and 4 of metamorphosis without histologic evidence of necrosis. Conclusions: DNA fragmentation is identified at the earliest stages of metamorphosis during induced metamorphosis in lampreys. Additional studies are necessary to validate this potentially valuable animal model.

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