Development of factors to convert frequency to rate for β-cell replication and apoptosis quantified by time-lapse video microscopy and immunohistochemistry

Yoshifumi Saisho, Erica Manesso, Tatyana Gurlo, Chang Jiang Huang, Gianna M. Toffolo, Claudio Cobelli, Peter C. Butler

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

obstacle to development of methods to quantify (β-cell turnover from pancreas tissue is the lack of conversion factors for the frequency of (β-cell replication or apoptosis detected by immunohistochemistry to rates of replication or apoptosis. We addressed this obstacle in islets from 1-mo-old rats by quantifying the relationship between the rate of (β-cell replication observed directly by time-lapse video microscopy (TLVM) and the frequency of (β-cell replication in the same islets detected by immunohistochemistry using antibodies against Ki67 and insulin in the same islets fixed immediately after TLVM. Similarly, we quantified the rate of (β-cell apoptosis by TLVM and then the frequency of apoptosis in the same islets using TdT-mediated dUTP nick-end labeling and insulin. Conversion factors were developed by regression analysis. The conversion factor from Ki67 labeling frequency (%) to actual replication rate (%events/h) is 0.025 ± 0.003 h-1. The conversion factor from TdT-mediated dUTP nick-end labeling frequency (%) to actual apoptosis rate (%events/h) is 0.41 ± 0.05 h -1. These conversion factors will permit development of models to evaluate (β-cell turnover in fixed pancreas tissue.

Original languageEnglish
Pages (from-to)E89-E96
JournalAmerican Journal of Physiology - Endocrinology and Metabolism
Volume296
Issue number1
DOIs
Publication statusPublished - 2009 Jan 1

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Keywords

  • Conversion factor
  • Insulin
  • Ki67
  • TdT-mediated dUTP nick-end labeling

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Physiology
  • Physiology (medical)

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