Expression of the vasopressin and oxytocin genes in rats occurs in mutually exclusive sets of hypothalamic neurons

Mohr, Evita; Bahnsen, Ulrich; Kiessling, Christiane; Richter, Dietmar

doi:10.1016/0014-5793(88)81003-2

Cited by 109 publications

(50 citation statements)

References 22 publications

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“…The common consensus sequence (CYTTGACC) exhibits the conserved 5 bp motif TGACC, which is known to constitute one half of the estrogen-as well as the thyroid hormone-responsive element (1 9). The human and the rat oxytocin gene promoter contain a sequence with a nearly perfect match to the estrogen-responsive element (15,17). This sequence has been shown to be involved in estrogen-dependent --174, -174 to -167, -159 to -152, -108 to -101, -88 to -81, T A H L~ I .…”

Section: S H Ementioning

confidence: 99%

Mapping of the Bovine Oxytocin Gene Control Region: Identification of Binding Sites for Luteal Nuclear Proteins in the 5′ Non‐Coding Region of the Gene

Walther¹,

Wehrenberg²,

Brackmann³

et al. 1991

J Neuroendocrinology

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In view of the small number of hormone-producing cells, the factors regulating oxytocin gene expression in the classic site of synthesis, in the magnocellular neurons of the hypothalamus, have not yet been characterized. In the early bovine corpus luteurn there is a tissue-specific oxytocin expression involving many more cells. This tissue therefore was chosen as a experimental system to identify deoxyribonucleic acid elements and nuclear proteins involved in the regulation of oxytocin gene expression. 3.2 kb from the 5' non-coding region of the bovine oxytocin gene have been sequenced and subcloned fragments used as probes for gel retardation and footprinting experiments. Binding sites for luteal as well as more ubiquitous proteins were detected in the oxytocin promoter region and in an artiodactyl-specific dispersed repeated deoxyribonucleic acid element. A binding site in the promoter region with a superficial similarity to an estrogen-responsive element (-159 to -152) was shown not to bind this steroid hormone receptor but to bind two nuclear proteins alternatively. One is a luteal protein, the other a more general transcription factor belonging to the steroid hormone receptor superfamily and similar, if not identical to the COUP protein. This alternative binding of a tissue-and phase-specifically expressed protein or an ubiquitous factor to the same site in the oxytocin promoter suggests a role for these two proteins in the transient up-regulation and subsequent down-regulation of the oxytocin gene during the differentiation of the bovine corpus luteum.

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Section: S H Ementioning

confidence: 99%

Mapping of the Bovine Oxytocin Gene Control Region: Identification of Binding Sites for Luteal Nuclear Proteins in the 5′ Non‐Coding Region of the Gene

Walther¹,

Wehrenberg²,

Brackmann³

et al. 1991

J Neuroendocrinology

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“…There are a number of variations on this method, but most procedures employ radiolabelled complementary DNA or R N A probes which can only be visualized by autoradiography. Autoradiographic procedures used on their own are not well suited t o studies of the possible co-existence or co-expression of two mRNAs in one cell and in order to d o this either a combination of radiolabelled and non-radiolabelled complementary probes (2), o r a combination of two non-radiolabelled probes which can be detected by different coloured reporter molecules has to be used (3). In an initial study using two non-radioactive in situ probes ( 3 ) we found that the majority (> 95%) of magnocellular neurosecretory cells in osmotically-stressed rats contained only one transcript (either oxytocin or vasopressin).…”

mentioning

confidence: 99%

Evidence for the Co‐Expression of Oxytocin and Vasopressin Messenger Ribonucleic Acids in Magnocellular Neurosecretory Cells: Simultaneous Demonstration of Two Neurohypophysin Messenger Ribonucleic Acids by Hybridization Histochemistry

Kiyama¹,

Emson²

1990

J Neuroendocrinology

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A novel enzyme-labelled oligonucleotide probe specific for oxytocin messenger ribonucleic acid (mRNA) and a radiolabelled oligonucleotide probe specific for vasopressin mRNA were used together to visualize both oxytocin and vasopressin mRNA's in hypothalamus sections from salt loaded (2% saline) rats. The results demonstrate that the majority of magnocellular neurons contain only oxytocin or vasopressin mRNA, however a small number of neurons, 1% to 2%, contained both oxytocin and vasopressin transcripts.In situ hybridization histochemistry is a method which can be used to demonstrate the cellular localization of an mRNA signal (1). There are a number of variations on this method, but most procedures employ radiolabelled complementary DNA or R N A probes which can only be visualized by autoradiography. Autoradiographic procedures used on their own are not well suited t o studies of the possible co-existence or co-expression of two mRNAs in one cell and in order to d o this either a combination of radiolabelled and non-radiolabelled complementary probes (2), o r a combination of two non-radiolabelled probes which can be detected by different coloured reporter molecules has to be used (3). In an initial study using two non-radioactive in situ probes ( 3 ) we found that the majority (> 95%) of magnocellular neurosecretory cells in osmotically-stressed rats contained only one transcript (either oxytocin or vasopressin). However, there were a small number of cells where we could not be certain using our two colour detection system, whether the cells might possibly contain both transcripts.In order to clarify this important point we have now used a combination of a [35S]radiolabelled antisense vasopressin probe specific for the glycopeptide sequence of vasopressin neurophysin not present in the oxytocin mRNA and an alkaline phosphataselabelled antisense oligonucleotide probe specific for a 5' sequence of oxytocin which did not detect vasopressin mRNA (for details see legends to Figs. 1 and 2). In double-labelling experiments the two specific probes were applied simultaneously to hypothalamic sections from control and osmotically-stressed male rats (2% saline in the drinking water for 2 weeks). After an appropriate stringent washing of the sections, the sections were first incubated in the alkaline phosphatase substrate to visualize the sites of oxytocin mRNA expression. Once the colour development was The strategy for demonstrating two mRNAs in a single section. The sequences of the probes were selected to avoid possible cross-reaction as the two genes (vasopressin, AVP or oxytocin, OT) have a considerable degree of sequence similarity especially in the regions coding for the hormone and neurophysins. The OT probe sequence was selected from the 5'-part of the signal peptide, and the AVP sequence was derived from the sequence coding for the first 12 amino-acids of the glycopeptide (not present in the OT transcript). In order to demonstrate the two mRNAs, two different labelling methods were used. Here we employed a nov...

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“…Based on biochemical, morphological and physiological criteria, the HNS neuronal population has been historically divided into two distinct phenotypes, the OT and VP neurons [53, 54]. The two neuronal phenotypes in rats have also been discriminated by their distinct electrical properties [44, 45, 46, 47].…”

Section: Ot and Vp Gene Expression In The Hypothalamusmentioning

confidence: 99%

Transgenesis and the Study of Expression, Cellular Targeting and Function of Oxytocin, Vasopressin and Their Receptors

Young

Gainer²

2003

Neuroendocrinology

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The neuropeptides oxytocin and vasopressin and the neurons in the hypothalamus that synthesize them have been a rich source for the exploration and understanding of both the brain and the endocrine system. Because of their large size and compact nuclear organization the magnocellular neurons of the hypothalamoneurohypophysial system have traditionally attracted scientists using state-of-the-art techniques, including the subject of this review, transgenesis. We discuss the role of transgenics in deciphering gene elements necessary for the appropriate expression of oxytocin and vasopressin and to deliver exogenous genes, such as green fluorescent protein, selectively to secretory granules in the neurons in the hypothalamoneurohypophysial system. Finally, we review the studies of mice whose genes for oxytocin and, most recently, for the oxytocin and vasopressin receptors have been knocked out through homologous recombination.

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Expression of the vasopressin and oxytocin genes in rats occurs in mutually exclusive sets of hypothalamic neurons

Cited by 109 publications

References 22 publications

Mapping of the Bovine Oxytocin Gene Control Region: Identification of Binding Sites for Luteal Nuclear Proteins in the 5′ Non‐Coding Region of the Gene

Mapping of the Bovine Oxytocin Gene Control Region: Identification of Binding Sites for Luteal Nuclear Proteins in the 5′ Non‐Coding Region of the Gene

Evidence for the Co‐Expression of Oxytocin and Vasopressin Messenger Ribonucleic Acids in Magnocellular Neurosecretory Cells: Simultaneous Demonstration of Two Neurohypophysin Messenger Ribonucleic Acids by Hybridization Histochemistry

Transgenesis and the Study of Expression, Cellular Targeting and Function of Oxytocin, Vasopressin and Their Receptors

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