The Insight of Chronobiology with an Emphasis on the Nightly Surge of Serotonin Output in the Rat Pineal Gland and

the Role of TPH1

Asamanja Chattoraj, PhD
Scientist-D; Animal Bioresources Programme
INSTITUTE OF BIORESOURCES & SUSTAINABLE DEVELOPMENT (IBSD) DEPARTMENT OF BIOTECHNOLOGY, GOVERNMENT OF INDIA Takyelpat, Imphal 795001, Manipur (India) Email: achattoraj.ibsd@nic.in asamanja.chattoraj@gmail.com
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What is chronobiology?
Biological rhythm persisting as a fundamental property of biologic entities under various conditions of presumed constancy in environmental factors. The formal study of biological temporal rhythms such as daily, tidal, weekly, seasonal, and annual rhythms is called chronobiology. A circadian rhythm is an endogenously driven roughly 24-hour cycle in biochemical, physiological, or behavioural processes. Circadian rhythms have been widely observed, in plants, animals, fungi and cyanobacteria. The term "circadian" comes from the Latin circa, meaning "around", and diem or dies, meaning "day". Although circadian rhythms are endogenous ("built-in", self-sustained), they are adjusted (entrained) to the environment by external cues called zeitgebers, the primary one of which is daylight.
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Pineal Gland and its Indoleamines

The pineal gland is a small organ, shaped like a pine cone (hence its name). It is located on the midline, attached to the posterior end of the roof f the third ventricle in the brain. It is an endocrine gland and the important secretory products are melatonin (N-acetyl-5methoxytryptamine) and serotonin (5-hydroxytryptamine). Synthesis of melatonin is rhythmic, regardless of whether the animal is diurnal or nocturnal. Serotonin is the precursor of melatonin, it is constitutively high during the daytime, increases at early night and finally decreases precipitously during the later half of night when melatonin levels are high.
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Travelling of Light into the Mammalian Pineal Gland

Pathway of Indoleamines Biosynthesis in The Pineal Gland

Importance of Melatonin and Serotonin

Serotonin: Neurotransmitter Control of appetite Sleep Memory and learning Temperature regulation Mood Behavior (including sexual and hallucinogenic behavior) Cardiovascular function Muscle contraction Endocrine regulation Depression Melatonin:

Sleep-Wakefulness Cycle Reproduction Hibernation Hypertension Cell Proliferation Seasonal Affective Disorders Immune-Response Jet-lag Scavenger of Free Radicals Aging

Existing Information
The pineal gland of all vertebrates produces 5-HT.
The synthesis of 5-HT from tryptophan requires two enzymes: a) Tryptophan Hydroxylase 1(TPH1) which limits the rate of 5-HT production and

b) Aromatic Amino Acid Decarboxylase, constitutively expressed at high levels.

TPH1 enzyme activity displays a two fold increase in the night pineal gland of rats. A number of studies points to an essential role of -adrenargic signalling in the regulation of TPH1 activity. In vitro studies also pointed out the role of cAMP and PKA in the activity of TPH1. PKA phosphorylates TPH1 in vitro at serine 58 residue.

It remains unknown , whether PKA elevates TPH1 activity by increasing its protein stability and whether TPH1 is phosphorylated in vivo at the S58 site.

Aims of the Present Study

The regulation of nocturnal increase of 5-HT total output in the night pineal gland.
The role of cAMP signaling in 5-HT synthesis in vivo. The role of TPH1 phosphorylation in the regulation of TPH1 protein content.

TPH1 mRNA Expression is NOT Under Circadian Control In Rat Pineal

DARK

LIGHT

Night-time TPH1 mRNA Expression is INSENSIBLE to Light Exposure

1: No Light 2: 10 mints Light 3: 24 hr Light

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Measurement of Pineal Indoleamines by Microdialysis

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5-HT Total Output and TPH1 Protein Levels Display Dramatic Circadian Rhythms

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Cyclic AMP Signaling in the Pineal Gland Stimulates the Total Output of 5-HT in vivo

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TPH1 is Phosphorylated at the S58 in the Night Pineal Gland

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TPH1-S58 Phosphorylation by PKA Increases TPH1 Protein Content

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Light Pulses at Night Suppress the Nocturnal Increase of both 5HT Total Output and TPH1 Protein Levels in the Living Pineal Gland

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Conclusion and Summary

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Current Research
Melatonin as Bioindicator for Oocyte maturation in carp. Fish Gut Microflora and Melatonin: A Chronobiological Study on Indian Major Carp

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The Group

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Acknowledgement
I am grateful to my mentor Prof. S.K. Maitra, Department of Zoology, Visva-Bharati, West-Bengal, INDIA, for everything I have learned and still learning from him. I am thankful to Dr Jimo Borjigin, MIP, University of Michigan, USA For helping me in my post-doctoral studies. Financial Support from United States Air force and NIH, USA, are thankfully acknowledged.

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