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Isolation, Fixation, and Immunofluorescence Imaging of Mouse Adrenal Glands
08:37

Isolation, Fixation, and Immunofluorescence Imaging of Mouse Adrenal Glands

Published on: October 2, 2018

Differential expression of somatostatin receptor subtype-related genes and proteins in non-functioning and

Hanna Pisarek1, Roman Krupiński, Robert Kubiak

  • 1Department of Neuroendocrinology, Medical University of Łódź, Łódź 91-425, Poland. hanna.pisarek@umed.lodz.pl

Molecular Medicine Reports
|July 2, 2011
PubMed

Insights

Investigating somatostatin receptor (SSTR) expression in adrenal adenomas using immunohistochemistry (IHC) and PCR revealed discrepancies between mRNA and protein levels. This highlights that mRNA detection alone cannot predict SSTR protein presence for therapy.

Area of Science:

  • Endocrinology
  • Oncology
  • Molecular Biology

Background:

  • Adrenocortical adenomas exhibit variable somatostatin receptor (SSTR) subtype expression, crucial for somatostatin (SST) analog therapy efficacy.
  • Immunohistochemistry (IHC) and polymerase chain reaction (PCR) are common methods to assess SSTR expression, but mRNA and protein levels may not always correlate.

Purpose of the Study:

  • To investigate SSTR expression in adrenal adenomas using IHC.
  • To compare IHC findings with real-time PCR data for SSTR expression.
  • To determine if functioning and non-functioning adenomas differ in SSTR expression patterns.

Main Methods:

  • Surgical removal of 15 adrenocortical adenomas (9 non-functioning, 6 functioning).
  • Semiquantitative scoring of IHC reaction intensity by two independent observers.
  • Real-time PCR with SYBR-Green fluorescence monitoring to assess SSTR mRNA expression.

Main Results:

  • In non-functioning adenomas, IHC and PCR compatibility for SSTR 1 and 2 was 62.5%, for SSTR 4 and 5 was 50%, and for SSTR 3 was 37.5%.
  • In functioning adenomas, 100% compatibility was observed for SSTR 2 and 66.6% for SSTR 5.
  • Significant variability in concordance between mRNA and protein expression was noted across different SSTR subtypes and adenoma function.

Conclusions:

  • Adrenocortical adenoma receptor gene and protein expression are not always synchronized.
  • Messenger RNA detection alone is insufficient to predict the presence of SSTR protein, a key target for SST analog therapy.

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