Regulatory Peptides, 45 (1993) 31-35 31
© 1993 Elsevier Science Publishers B.V. All rights reserved 0167-0115/93/$06.00
REGPEP 01349
Transactivation of the rat oxytocin and vasopressin promoters by
nuclear hormone receptors
J. Peter H. Burbach, Roger A.H. Adan, Joke J. Cox and Sofia Lopes da Silva
Rudolf Magnus Institute, Department of Pharmacology, Utrecht University, Utrecht(The Netherlands)
Key words: Steroid hormone; Thyroid hormone; Retinoic acid; Estrogen; Gene regulation; Hormone
response element
Introduction
The regulation of the oxytocin (OT) and vaso-
pressin (VP) genes in the magnocellular neurons of
the supraoptic (SON) and paraventricular nuclei
(PVN) displays remarkable parallels. For instance,
the levels of OT mRNA and VP mRNA increase
similarly during postnatal development, hyperosmo-
lality, pregnancy and lactation [ 1]. It is believed that
the similarity in responsiveness resides in either com-
mon regulatory elements in the genes or in common
neural or hormonal inputs of the OT and VP neurons
[2]. Although there is no significant overall homol-
ogy between the 5'-flanking regions of the OT and
VP genes, both genes contain a number of TGACC
motifs [3]. Such sequences are often part of response
elements of nuclear hormone receptors. Steroid hor-
mones have been implicated in the regulation of OT
and VP systems [4]. We here summarize the results
of experiments aimed to determine and compare the
responsiveness of OT and VP genes to members of
the steroid/thyroid hormone receptor superfamily.
Correspondence to: J.P.H. Burbach, Rudolf Magnus Institute, De-
partment of Pharmacology, Utrecht University, Vondellaan 6,
3521 GD Utrecht, The Netherlands.
Transactivation
Since cultured cells with abundant expression of
the OT gene or VP gene are not available experi-
ments were performed in heterologous expression
systems, i.e., tumor cell lines without endogenous OT
or VP gene expression that were transfected with OT
or VP promoter-reporter gene constructs. Two types
of cell lines were used: cell lines with endogenous
hormone receptors, e.g., MCF-7 breast tumor cells
containing the estrogen receptor (ER) and thyroid
hormone receptors (TR) and P 19 embryocarcinoma
(EC) cells containing retinoic acid receptors (RAR)
and retinoid X receptors (RXR), or cell lines co-
transfected with plasmids expressing nuclear hor-
mone receptors.
The activity of the rat OT promoter (nucleotides
-361 to + 16) was stimulated by the ligand-activated
ER, TR~ and RAR~ and/~ [5-7]. The human OT
promoter was similarly responsive to these hormone
receptors [6-9]. In P19 EC cells the response to
estrogens was by far the strongest (approx. 200-300
fold) and to thyroid hormone the weakest (approx.
10-fold) when using the -361 tot + 16 region of the
rat OT gene coupled to luciferase (pROLUC). The
stimulation was about 10-fold when the endogenous