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Thermostabilization, Expression, Purification, and Crystallization of the Human Serotonin Transporter Bound to S-citalopram
Published on: November 27, 2016
Serotonin transporter and receptor expression in osteocytic MLO-Y4 cells
M Bliziotes1, A Eshleman, B Burt-Pichat
1Portland VA Medical Center, Portland, OR 97239, USA. bliziote@ohsu.edu
Bone
|August 4, 2006
Summary
Bone cells, including osteocytes, utilize serotonin (5-HT) for communication. This neurotransmitter influences bone metabolism through functional receptors and transporters, impacting bone health.
Area of Science:
- Bone Biology
- Neuroendocrinology
- Cell Signaling
Background:
- Neurotransmitter regulation of bone metabolism is an emerging research area.
- Osteoblastic cells possess a functional serotonin (5-HT) signaling system.
- Previous work indicated osteoblasts respond to and regulate 5-HT uptake.
Purpose of the Study:
- To investigate the expression and function of the serotonin system in osteocytes.
- To determine if osteocytes synthesize, respond to, and transport serotonin.
- To explore the role of serotonin in bone cell communication.
Main Methods:
- Real-time RT-PCR and immunoblot analysis for receptor and transporter expression.
- Immunohistochemistry to localize proteins in bone cells.
- Radioligand binding assays to characterize transporter affinity.
- Serotonin uptake assays to quantify transport kinetics.
- Measurement of prostaglandin E2 (PGE2) release in response to 5-HT.
Main Results:
- Osteocytic cell line MLO-Y4 expresses serotonin transporter (5-HTT) and 5-HT1A/5-HT2A receptors.
- These proteins are also found in osteoblasts and osteocytes in rat tibia.
- MLO-Y4 cells exhibit high-affinity 5-HTT binding and serotonin uptake.
- Imipramine and fluoxetine potently inhibited 5-HTT binding and uptake.
- Serotonin stimulated PGE2 release from MLO-Y4 cells.
- Tryptophan hydroxylase, key for serotonin synthesis, is expressed in osteocytes and osteoblasts.
Conclusions:
- Osteocytes, like osteoblasts, express functional serotonin receptors and transporters.
- Osteocytes are capable of synthesizing serotonin.
- The serotonin system in bone cells plays a role in regulating bone metabolism.
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