Co-expression of tyrosine hydroxylase and GTP cyclohydrolase I in arginine vasopressin-synthesizing neurons of the human supraoptic nucleus demonstrated by laser microdissection and real-time PCR

Dimitra P Kontostavlaki, Maria T Panayotacopoulou, Jacqueline A Sluijs, Unga A Unmehopa, Inge Huitinga, Elly M Hol, Dick F Swaab

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Tyrosine hydroxylase (TH), the first and limiting enzyme for catecholamine synthesis, has been identified immunohistochemically (IHC) in human neurosecretory neurons where it is found to colocalize with vasopressin (AVP) or oxytocin. TH expression shows striking interindividual variability and appears to depend on neuronal activation. Since GTP cyclohydrolase I (GCHI), the first enzyme for tetrahydrobiopterin synthesis, the essential cofactor of TH, and aromatic L-amino acid decarboxylase (AADC) have so far not been detected in neurosecretory neurons, the functional role of TH in catecholamine synthesis is still questionable. Our purpose was to investigate in postmortem hypothalamus whether GCHI and AADC mRNAs are co-expressed with TH in human AVP-synthesizing neurons. Total RNA was extracted from laser microdissected TH-IHC-identified neurons as well as from dissected parts of the dorsolateral supraoptic nucleus (dl-SON) of 12 control subjects, i.e. without known neurological, psychiatric or endocrinological illness. GCHI, AADC and TH mRNA expression was determined by real-time PCR. Our results showed that GCHI mRNA is co-expressed with TH in almost all cases that had a considerable number of TH-immunoreactive (TH-IR) neurosecretory neurons. A positive correlation was found between TH-immunohistochemical intensity and the presence of GCHI mRNA. AADC mRNA expression was detected only in microdissected areas of dl-SON in 2 cases that showed an increased number of TH-IR neurons. The co-expression of GCHI with TH indicates that TH is indeed active in human neurosecretory neurons. The apparent limited expression of AADC indicates that dopamine might be produced in human neurosecretory neurons under activation of the hypothalamoneurohypophyseal system, although the possibility that L-dopa is the final product cannot be excluded.

Original languageEnglish
Pages (from-to)386-95
Number of pages10
JournalNeuroendocrinology
Volume84
Issue number6
DOIs
Publication statusPublished - 2006

Keywords

  • Aged
  • Aged, 80 and over
  • Arginine Vasopressin
  • Aromatic-L-Amino-Acid Decarboxylases
  • Female
  • GTP Cyclohydrolase
  • Gene Expression Regulation
  • Humans
  • Male
  • Microdissection
  • Middle Aged
  • RNA, Messenger
  • Reverse Transcriptase Polymerase Chain Reaction
  • Supraoptic Nucleus
  • Tyrosine 3-Monooxygenase
  • Journal Article
  • Research Support, Non-U.S. Gov't

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