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Immortalization of neuroendocrine pinealocytes from transgenic mice by targeted tumorigenesis using the tryptophan
1Laboratory of Molecular Neurobiology, Cornell University Medical College, W.M. Burke Medical Research Institute, White Plains, NY 10605, USA.
Abstract:
Tryptophan hydroxylase (TPH) is the first enzyme in both serotonin and melatonin biosynthesis in neuroendocrine cells of the pineal gland. The lack of immortalized neuroendocrine pineal cell lines has been a major obstacle to the study of the tissue-specific and circadian regulation of TPH gene expression in the pineal gland. Previously, we demonstrated that a 6.1 kb 5' upstream region of the mouse TPH gene directs the restricted expression of a lacZ reporter gene to the pineal gland and the raphe nuclei of transgenic mice. Therefore, to develop TPH-expressing pineal cell lines we first established transgenic mice carrying a construct consisting of 6.1 kb of 5' flanking region fused to the SV40 T-antigen. These animals developed highly invasive pineal tumors and died at 12-15 weeks of age. The pineal tumors obtained from the transgenic mice were utilized to establish the immortalized pinealocyte-derived cell lines. These cells express two marker enzymes, TPH and serotonin N-acetyltransferase (NAT). In pineal gland TPH and NAT expressions have been known to be regulated during circadian cycle. The two established cell lines therefore promise to be a valuable in vitro model system for the study of the rhythmic nature of the pineal function at molecular level in mammal.
Insights
Researchers developed immortalized pineal cell lines from tumors in transgenic mice. These cell lines express tryptophan hydroxylase (TPH) and serotonin N-acetyltransferase (NAT), offering a new model for studying pineal gland function.
Area of Science:
- Neuroscience
- Molecular Biology
- Genetics
Background:
- Tryptophan hydroxylase (TPH) is crucial for serotonin and melatonin synthesis in pineal gland neuroendocrine cells.
- Lack of immortalized pineal cell lines hinders research into TPH gene regulation and circadian rhythms.
- Previous studies identified a 6.1 kb 5' upstream region of the mouse TPH gene for restricted expression.
Purpose of the Study:
- To develop immortalized neuroendocrine pineal cell lines for studying TPH gene regulation.
- To establish an in vitro model for investigating circadian regulation of pineal function.
- To overcome limitations in studying the mammalian pineal gland's molecular mechanisms.
Main Methods:
- Generated transgenic mice with a construct linking the TPH 5' flanking region to SV40 T-antigen.
- Established immortalized pineal cell lines from tumors developed in these transgenic mice.
- Confirmed expression of TPH and serotonin N-acetyltransferase (NAT) in the derived cell lines.
Main Results:
- Transgenic mice developed invasive pineal tumors, enabling cell line establishment.
- The derived cell lines express key pineal enzymes TPH and NAT.
- These cell lines provide a viable in vitro system for studying pineal gland molecular regulation.
Conclusions:
- Successfully established immortalized pinealocyte-derived cell lines expressing TPH and NAT.
- These cell lines serve as a valuable model for in vitro research on pineal gland circadian rhythms.
- Facilitates future studies on the molecular mechanisms of pineal function in mammals.
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