Expression of amyloidogenic ser84 variant of human transthyretinc1tr in transgenic mice: Immunoelectron microscopic, immunohistochemical, and reverse transcription-polymerase chain reaction(RT-PCR) studies

M. T. Hull, R. P. Waits, T. Uemichi, S. R. Zeldenrust, Loren Field, M. P. Gofaeen, M. D. Season

Research output: Contribution to journalArticle

Abstract

Transgenic mice expressing amyloidogenic human Ser84 variant of TTR were produced by pronuclear injection of Ser84 construct and oviduct transfer. RTPCR demonstrated TTR mRNA in liver, pancreas, stomach, and choroid plexus. ImmuDohistochemicaJ studies showed localization of TTR in hepatocytes in the liver, islet cells in the pancreas, isolated neuroendocrine cells in the stomach, and epithelial cells in the choroid plexus. Within the islets, there was considerable cell to cell variation in reaction intensity. Inununoreactivity was uniformly intense in gastric neuroendocrine cells and moderate in choroid plexus cells. Because islet cells showed such cell to cell variability in TTR reactivity, immunoelectron microscopy was carried out on pancreas which was fixed in 4 % paraformaldehyde and . 1 % glutaraldehyde and men embedded in LR White resin. Reaction intensity was strong in Alpha cells, moderate in Beta cells, and weak io Delta cells. In each cell the TTR was locallized predominantly to the cytoplasm immediately adjacent to the neurosecretory granules. Acinar and ductal cells were negative. Although amyloid has yet to be identified in tissues, mice over 12 months of age have not been examined. This transgenic model should be useful in the study of Ser84 TTR expression, and it may prove useful in the study of Ser84 TTR related amyloidosis.

Original languageEnglish
JournalFASEB Journal
Volume10
Issue number3
StatePublished - 1996

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Polymerase chain reaction
Transcription
Liver
Transgenic Mice
Reverse Transcription
reverse transcriptase polymerase chain reaction
genetically modified organisms
Polymerase Chain Reaction
mice
Glutaral
Amyloid
Choroid Plexus
Microscopic examination
cells
Tissue
choroid plexus
Pancreas
Neuroendocrine Cells
Stomach
Messenger RNA

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Cell Biology

Cite this

Expression of amyloidogenic ser84 variant of human transthyretinc1tr in transgenic mice : Immunoelectron microscopic, immunohistochemical, and reverse transcription-polymerase chain reaction(RT-PCR) studies. / Hull, M. T.; Waits, R. P.; Uemichi, T.; Zeldenrust, S. R.; Field, Loren; Gofaeen, M. P.; Season, M. D.

In: FASEB Journal, Vol. 10, No. 3, 1996.

Research output: Contribution to journalArticle

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abstract = "Transgenic mice expressing amyloidogenic human Ser84 variant of TTR were produced by pronuclear injection of Ser84 construct and oviduct transfer. RTPCR demonstrated TTR mRNA in liver, pancreas, stomach, and choroid plexus. ImmuDohistochemicaJ studies showed localization of TTR in hepatocytes in the liver, islet cells in the pancreas, isolated neuroendocrine cells in the stomach, and epithelial cells in the choroid plexus. Within the islets, there was considerable cell to cell variation in reaction intensity. Inununoreactivity was uniformly intense in gastric neuroendocrine cells and moderate in choroid plexus cells. Because islet cells showed such cell to cell variability in TTR reactivity, immunoelectron microscopy was carried out on pancreas which was fixed in 4 {\%} paraformaldehyde and . 1 {\%} glutaraldehyde and men embedded in LR White resin. Reaction intensity was strong in Alpha cells, moderate in Beta cells, and weak io Delta cells. In each cell the TTR was locallized predominantly to the cytoplasm immediately adjacent to the neurosecretory granules. Acinar and ductal cells were negative. Although amyloid has yet to be identified in tissues, mice over 12 months of age have not been examined. This transgenic model should be useful in the study of Ser84 TTR expression, and it may prove useful in the study of Ser84 TTR related amyloidosis.",
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AU - Hull, M. T.

AU - Waits, R. P.

AU - Uemichi, T.

AU - Zeldenrust, S. R.

AU - Field, Loren

AU - Gofaeen, M. P.

AU - Season, M. D.

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