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Oxytocin transgenic mice

D Murphy1, M Y Ho

  • 1Neuropeptide Laboratory, Institute of Molecular and Cell Biology, Singapore, Republic of Singapore.

Advances in Experimental Medicine and Biology
|January 1, 1995
PubMed
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Bovine oxytocin constructs were tested in transgenic mice. A specific construct (bOT6.5) proved toxic, while another (bOT3.5) unexpectedly expressed oxytocin in the hypothalamus, suggesting downstream sequences repress hypothalamic expression.

Area of Science:

  • Neuroendocrinology
  • Molecular Biology
  • Genetics

Background:

  • Oxytocin (OT) plays crucial roles in social behavior, reproduction, and stress response.
  • Understanding the regulation of OT gene expression is vital for deciphering its physiological functions.
  • Transgenic mouse models are powerful tools for studying gene regulation and function.

Purpose of the Study:

  • To investigate the regulatory elements controlling bovine oxytocin gene expression in transgenic mice.
  • To identify sequences responsible for tissue-specific expression, particularly in the hypothalamus.
  • To assess the impact of different construct lengths on transgene expression and viability.

Main Methods:

  • Construction and microinjection of various bovine oxytocin gene constructs into mouse embryos.

Related Experiment Videos

  • Generation of transgenic mice and analysis of transgene integration and expression.
  • Quantitative analysis of transgene RNA levels using in situ hybridization.
  • Assessment of transgene response to physiological stimuli like salt-loading, pregnancy, and lactation.

    • Main Results:

    • The bOT6.5 construct (3 kbp upstream, 2.6 kbp downstream) was toxic to mouse embryos, preventing founder identification.
    • The bOT construct (0.6 kbp upstream, 2.6 kbp downstream) showed expression in lung and testicular Sertoli cells, but not the hypothalamus.
    • The bOT3.5 construct (0.6 kbp upstream, 1.9 kbp downstream) exhibited expression in lung, testis, and surprisingly, the hypothalamus (SON and PVN).
    • Salt-loading increased bOT3.5 transgene RNA in the SON, mirroring endogenous OT response.
    • Transgene RNA levels in the hypothalamus increased during late pregnancy and lactation.

    Conclusions:

    • Downstream sequences in the bovine oxytocin gene (0.7 kbp present in bOT, absent in bOT3.5) appear to mediate hypothalamic repression.
    • The bOT3.5 construct, lacking this repressor element, allows for hypothalamic expression.
    • Hypothalamic expression of bOT3.5 in SON and PVN is regulated by physiological states such as salt balance and reproductive status.
    • These findings provide insights into the complex regulation of oxytocin gene expression.