L. Martini (Eds.

)
Progress in Brain Research, Vol. 181
ISSN: 0079-6123
Copyright Ó 2010 Elsevier B.V. All rights reserved.

CHAPTER 7

Physiological significance of the rhythmic secretion

of hypothalamic and pituitary hormones

Earn-Hui Gan and Richard Quinton

Endocrine Research Group, Institute of Human Genetics, University of Newcastle-on-Tyne, Newcastle-upon-Tyne,

United Kingdom

Abstract: The various hypothalamic–pituitary–end-organ/gland axes are central to the regulation of

mammalian homeostasis. These have a core role in integrating the response of both endocrine and
nervous systems to external and internal stimuli, by means of multi-level signalling through negative
and positive feedback loops. The content of these hormonal signals is overwhelmingly conveyed in a
rhythmic secretory pattern (frequency modulation of signal) that is energetically more efficient in
transmitting neuroendocrine signals than the alternatives (modulation of signal by amplitude or by total
area-under-curve). These rhythmic neuroendocrine secretions are individually distinct but the majority
display a common feature of low-level basal secretion with superimposed pulsatile rhythms.
The underlying mechanisms contributing to this unique rhythmic secretion are complicated and
incompletely understood, but are beginning to be better defined as a result of several elegant studies
performed in recent years. In some cases, signal transduction in the target tissue is critically dependent
upon a pulsatile input, but in others the observed pulsatility is a downstream echo of obligate pulsatility
exhibited by a higher-level control hormone. Thus, the gonads are presented with a pulsatile
gonadotrophin signal, not because this is essential to gonadotrophin action (the same level of
stimulation can be elicited by a continuous input), but as a downstream consequence of pulsatile
GnRH-mediated stimulation of pituitary gonadotrophs. By contrast, rhythmicity of signal is embedded
at all levels of the hypothalamo–pituitary–adrenal axis.
Hypothalamic–pituitary rhythmic secretions are influenced by various internal and external inputs such
as age, gender, sleep and wakefulness, food intake, light (photoperiod) or exposure to stress.
Understanding the physiological significance of the rhythmic secretion of hypothalamic and pituitary
hormones has the potential to provide insights into disease mechanisms, to validate diagnostic tests and,
ultimately, to help develop novel therapeutic interventions. This chapter will overview the physiological
basis of rhythmic secretion of hypothalamic and pituitary hormones, principally in humans and, by
reference to specific examples, describe the various feedback loops and internal and external stimuli
that precisely determine these neuroendocrine secretory patterns.

Keywords: rhythmic secretory patterns GH TSH GnRH photoperiod

Corresponding author.
Tel.: þ44-191-2824635; Fax: þ44-191-2820129
E-mail: richard.quinton@ncl.ac.uk

DOI: 10.1016/S0079-6123(08)81007-2 111

112
Introduction
Oscillating and pulsatile signals in neuroendo-
crine axes have been demonstrated in virtually
all mammalian species studied, from rodents to
human (Gudmundsson and Carnes, 1997). In
mammals, this observable rhythm is generated
by an endogenous circadian clock located within
the suprachiasmatic nucleus (SCN) of the
hypothalamus (Cermakian and Boivin, 2009;
Klein et al., 1991; Moore and Silver, 1998). This
time-controlling mechanism is shown to be an
intrinsic property at the cellular level, with circa-
dian rhythmicity being constitutively exhibited in
vitro by SCN neurons (Cermakian and Boivin,
2003, 2009; Ko and Takahashi, 2006). This prop-
erty results from the differential expression of
various clock genes (e.g. Per1-3, Cry1-2) that
have been isolated based on sequence homology
with known clock genes in Drosophila melanoga-
ster. Their associated protein products, including
neuropeptides, transcription factors and meta-
bolic enzymes, contribute to both negative and
positive feedback loops in neuroendocrine axes
(Cermakian and Boivin, 2009).
In this chapter, we will focus on patterns
and control mechanisms of rhythmic hormone
secretion, particularly in humans, with specific
reference to growth hormone (GH), adrenocorti-
cotropic hormone (ACTH), thyroid-stimulating
hormone (TSH), prolactin (PRL) and oxytocin.
The gonadotropic axis and its hypothalamic reg-
ulator are discussed elsewhere in this volume.
However, before beginning our description, it is
necessary to define a number of key terms and
definitions. The ‘diurnal’ (or perhaps more accu-
rately, ‘ultradian’) rhythm is a 24-hour periodicity
that reflects the superimposition of external and
internal stimuli on the endogenous (circadian)
signal. Endogenous neurosecretory rhythms are
defined as ‘circadian’, because they typically
cycle over a period of around (but never exactly)
24 hours. The two principal environmental cues
that act to resynchronise the endogenous circa-
dian rhythms are the light–dark and rest–activity
cycles. A rhythmic signal can encode information
in three principal ways: through the pulse
frequency (frequency modulation – as in the
‘FM radio’ signal), the integrated pulse amplitude
(amplitude modulation – as in the ‘AM radio’
signal) or through the total area-under-curve. Of
these, the former is typically most energetically
efficient (Goldbeter et al., 2000).
Neurohumoral regulation of growth hormone
secretion
GH is essential for promoting somatic growth
from the end of the second year of life until pub-
erty (in man), by regulating protein synthesis,
lipolysis and skeletal growth. It also has a homeo-
static role in adult life (Davidson, 1987; Furuhata
et al., 2002). GH secretion is predominantly pul-
satile in healthy young adults and carries signifi-
cant physiological signals to target tissues by
determining the GH-receptor turnover, mode of
second-messenger signalling, cell-specific gene
expression and distinct metabolic responses
(Achermann et al., 1999; Giustina and Veldhuis,
1998; Veldhuis and Bowers, 2009; Veldhuis et al.,
2009a).
Relationship between pulsatile GH secretion and
IGF-1 level
In humans, 85% of GH secretion is pulsatile,
with up to 10 pulses of GH secretion in a 24-
hour period (Ionescu and Frohman, 2006; Stolar
and Baumann, 1986). Many of the physiological
effects of GH are mediated indirectly through
stimulation of insulin-like growth factor 1 (IGF-1)
secretion by the liver and other target tissues.
The circulating IGF-1 level strongly correlates
with the pattern of rhythmic GH secretion
(Veldhuis and Bowers, 2009). The attenuation
of GH pulse-dependent STAT5b (signal transdu-
cer and activator of transcription 5B) signalling
is shown to associate with the diminution of
IGF-1 concentration, which in turn leads to
growth failure (Davey et al., 1999; Veldhuis
and Bowers, 2009). Most studies have demon-
strated that mean GH concentration, as reflected
by pulse amplitude, best correlates with
serum IGF-1 concentration (i.e. amplitude
 113

25

20

15
GH(mU/L)

10

0
14:00

21:20
12:00

14:40

16:00
16:40

20:40

22:40

00:40

04:40

06:00
12:40
13:20

18:00
18:40

03:20

05:20

06:40

08:00
15:20

17:20

20:00

23:20

01:20
02:00

04:00

07:20

08:40
19:20

00:00
22:00

02:40
Time

Fig. 1. Twenty-hour GH profile in a normal 18-year-old girl.

modulation of signal), though a raised basal GH amplifying signals, GHRH (growth-hormone-releas-

secretion (reflecting ‘area-under-curve’), as ing hormone) and GHS (growth hormone secretago-
observed for instance in patients with acromegaly, gue or ghrelin), the inhibitory factor somatostatin
clearly also has a role (Ionescu and Frohman, (SRIF), and feedback-inhibition from circulating
2006; Veldhuis et al., 1995). In man, pulsatile GH IGF-1 (Bluet-Pajot et al., 1998; Frohman, 1996; Fur-
secretion peaks in the evening and displays a dis- uhata et al., 2002; Shuto et al., 2002; Veldhuis and
tinctive composite of low-frequency volleys and Bowers, 2009). The interplay between episodic facil-
high-frequency secretory bursts (Fig. 1) (Farhy itative drive by GHRH, intermittent suppression by
and Veldhuis, 2003). Multi-burst volleys recur central neural SRIF and reversible negative feed-
every 1.5–2.5 hours in humans, with rapid discrete back by both systemic and local central nervous sys-
GH pulses arising every 30–60 minutes within an tem GH secretion and peripherally derived IGF-1
individual volley (Farhy and Veldhuis, 2003; result in discrete GH pulses (Farhy and Veldhuis,
Gevers et al., 1998; Giustina and Veldhuis, 1998; 2004; Giustina and Veldhuis, 1998; Mueller et al.,
Hartman et al., 1991, 1992). 1999). Several other potent neuromodulators prob-
ably also influence the GH axis, including
endogenous opioids, neuropeptide Y, galanin, choli-
Neuropeptide interactions in the generation nergic and adrenergic neurotransmitters, gamma-
of GH pulses aminobutyric acid and sex steroids (Veldhuis, 1998).
Ghrelin, the endogenous ligand of the GHS receptor,
GH pulses are instituted by a multi-signal interaction is secreted dominantly by the gastrointestinal tract
between various neuropeptides, including the and is the only known circulating orexigen.
114
Although not essential for GH secretion, it does
seem to amplify GHRH-mediated GH pulses
(Farhy and Veldhuis, 2005). Differential timing
of the release of these neuropeptides, along with
superimposed variation reflecting age and sexual
dimorphism, contributes to the observed pulsati-
lity of GH secretion (Farhy et al., 2002; Haus,
2007).
Control of GH secretion: lessons from animal
studies
Peaks and troughs of GH secretion in rodents
have been shown to result from a complex inter-
active network. This comprises (1) time-delayed
negative feedback to stimulate SRIF secretion
from the periventricular nucleus (PEV) of the
hypothalamus into the hypophysial portal circula-
tion, which in turn antagonises GH release from
the anterior pituitary (Farhy and Veldhuis, 2004;
Robinson, 1991), and (2) a GHRH–SRIF oscilla-
tor in the arcuate nucleus (ARC) of the hypotha-
lamus (Fig. 2) (Farhy and Veldhuis, 2004; Giustina
and Veldhuis, 1998; Mueller et al., 1999). SRIF
receptor is expressed by both (ARC) GHRH-
positive neurons and (PEV and ARC) SRIF-
Arcuate Nucleus
SRIF
GHRH
Pituitary
Peripheral
GH
IGH-1
Stimulatory effects
Inhibitory effects
Fig. 2. Modulation of pulsatile GH secretion: schema of interaction between hypothalamus (arcuate and periventricular nuclei),
pituitary and peripheral tissues.
positive neurons (Bertherat et al., 1992; Farhy
and Veldhuis, 2004). There is presumed to be a
time-delayed, short-latency, reciprocal interaction
between the actions of GHRH and SRIF to create
a damped intra-arcuate oscillator (Farhy and
Veldhuis, 2004).
An unequal feedback latency exists between the
systemic GH input to (PEV) SRIF secretion (long
loop feedback) and (ARC) GHRH secretion to
drive SRIF-mediated inhibition of GH secretion
(short loop feedback), leading to prolonged inter-
volley duration and short intra-volley intervals
(Farhy and Veldhuis, 2004). It is postulated that
the systemic GH pulses stimulate (PEV) SRIF-
dependent inhibition of somatotrope GH release,
by reducing (1) GHRH secretion into portal blood
and (2) intra-hypothalamic GHRH feed-forward
on (ARC) SRIF secretion, leading to an inter-
volley interval of reduced GH secretion (Farhy
and Veldhuis, 2004).
Sexual dimorphism in the pattern of pulsatile GH
secretion
The reduction in circulating GH and hypothalamic
(PEV) SRIF concentration during an inter-pulse
Periventricular
Nucleus
SRIF

trough disinhibits the putative GHRH–SRIF oscil-
lator, thereby triggering the high-frequency,
amplitude-damped GHRH and GH secretory
bursts demonstrated in male rats. In females, a
decline in GH-induced (PEV) SRIF release will
attenuate (PEV) SRIF-enforced suppression and
disinhibition of coupled (ARC) GHRH–SRIF
interactions, leading to discrete, high-frequency
and low-amplitude GH pulses with only
occasional volley-like complexes (Clark et al.,
1986, 1987; Farhy and Veldhuis, 2004; Pincus
et al., 1996). This is in keeping with a study in
young human adults showing that, in the fasting
state, women secrete several-fold more GH in
bursts than men, and that the somatosergic
outflow in women is opposed by greater feed-
forward by both GHRH and GHS (Soares-
Welch et al., 2005).
Sexual dismorphism of the human GH
secretory pattern is also demonstrated during
sleep. In men, the highest pulse occurs after
sleep onset within the first phase of slow-wave
sleep and accounts for the highest secretory out-
put per 24 hours – even up to 70% (Haus, 2007;
Ho et al., 1987). By contrast, females exhibit a
wider distribution of GH pulses throughout the
day, with the sleep-onset-entrained GH surge
accounting for a smaller fraction of the total
24-hour secretion (Haus, 2007; Ho et al., 1987).
These observations help explain the alterations
in GH secretion observed with sleep disorders,
among shift workers or in trans-meridian travel-
lers (Cauter et al., 1998; Haus, 2007).
Effects of ageing on pulsatile GH secretion
GH secretion is reduced in older men and
women (Veldhuis, 2008). In comparison with
the volley-like clusters of prominent pulses
observed in pubertal children, the elderly dis-
play frequent, isolated and low-amplitude GH
bursts. This results in a reduced circadian mean
GH concentration by selectively reducing pulsa-
tile GH secretion (Farhy and Veldhuis, 2004;
Hartman et al., 1991; Martha et al., 1992;
Veldhuis et al., 2000). Young adults also exhibit
a notably higher GH secretory response to
115
lowering of IGF-1 levels, compared with elderly
subjects (Veldhuis, 2008; Veldhuis et al., 2006),
which implies an impaired hypothalamic–pitui-
tary drive to GH secretion with ageing, possibly
via diminished responsiveness to GHRH and
increased responsiveness to SRIF (Haus, 2007;
Martin et al., 1997).
Potential estrogen-independent factors
contributing to the depletion of GH in older
woman includes catecholamines, thyroxines,
cortisol, free fatty acids, inflammatory cytokines
and regulatory peptides (Veldhuis 2008;
Veldhuis et al., 2005). In men, testosterone
increases pulsatile GH secretion by augmenta-
tion of pulse amplitude (Gentili et al., 2002;
Iranmanesh et al., 1998; Veldhuis, 2008). How-
ever, GH pulse frequency, GH elimination
kinetic or hepatic action of GH to stimulate
IGF-1 production is not affected by age or tes-
tosterone (Gentili et al., 2002; Muniyappa et al.,
2007; Veldhuis et al., 2006). Hence, the full
extent of the interplay between testosterone
and age in affecting GH secretory burst size is
still not entirely clear.
Effects of developmental stage and pubertal status
on GH pulsatile secretion
Developmental stage and the physiological
changes in response to adiposity, aerobic capacity
and ageing have significant effects on GH secre-
tion. Smaller GH pulses, as quantified by less GH
secreted per burst (which diminishes the mean
GH concentration), are observed in normal mid-
childhood, in association with low aerobic capa-
city, in young adults with hypogonadism and in
obesity (Lang et al., 1987; Lieman et al., 2001;
Veldhuis et al., 2009; Weltman et al., 1994). The
first day of infant life and the period correspond-
ing to Tanner stages IV–V of puberty represent
periods of dominant amplitude modulation of sig-
nal, being associated with significant augmenta-
tion of the GH pulses with no change in GH pulse
frequency, GH half life or GH basal release
(Gentili et al., 2002; Shah et al., 1999; Veldhuis
et al., 2006).
116
Pulsatile GH secretion can be stimulated by
fasting or by GHRH/GHS or estradiol, but is
diminished by factors associated with food intake,
abdominal visceral fat and ageing (Lang et al.,
1987; Vahl et al., 1997; Veldhuis and Bowers,
2003; Veldhuis et al., 2009b). Veldhuis found a
strong negative impact of computed tomography-
estimated abdominal visceral fat on fasting and
GHRH/GHS-stimulated pulsatile GH secretion in
an estrogen-enriched milieu (Veldhuis et al.,
2009).
Unresolved or poorly understood issues
Recent human experiments have demonstrated
interesting and distinct findings in response to
different patterns of GH or GHRH infusion. For
instance, the observed pulsatility of GH secretion
in post-menopausal women is maintained under
continuous stimulation with GHRH and growth
hormone-releasing peptide 2 (GHRP-2) (Haus,
2007; Veldhuis et al., 2002, 2006). Likewise, con-
tinuous GHS infusion in adults for 1–30 days
resulted in augmented GH secretory burst activ-
ity, but with unchanged GH pulse frequency and
circadian rhythmicity (Haus, 2007; Shah et al.,
1999). Whereas bolus GH infusion enhanced lipo-
lysis to a greater extent than continuous delivery,
it was less effective at increasing the IGF-1 con-
centration (Haus, 2007; Jorgensen et al., 1991).
Little is known about the physiological signifi-
cance of tonic basal GH secretion, but an increase
is observed experimentally in mice with knockout
of the type 1 SRIF receptor and clinically in
humans with acromegaly (Hartman et al., 1994;
Veldhuis et al., 2009). It is impaired with advan-
cing age and by exogenous estrogen administra-
tion (Veldhuis et al., 2009).
Regulation of corticotrophic function and action
Activation of the hypothalamus–pituitary–adre-
nal (HPA) axis involves the pulsatile secretion of
corticotropin-releasing hormone (CRH) and of
arginine vasopressin (AVP) from the PEV. CRH
triggers the release of ACTH from pituitary
corticotrophs; ACTH in turn upregulates adre-
nal cortical enzymatic activity, principally con-
version of cholesterol to pregnenolone (the first
and rate-limiting step in the steroid biosynthetic
pathway), leading to increased cortisol secretion
(Crofford et al., 2004). Cortisol is a key mediator
of vascular integrity, immune response, energy
balance and metabolism (Baigent, 2001; Richard
et al., 2000). Diurnal variation in (PEV) CRH
and ACTH secretion is regulated by the endo-
genous circadian clock located in the SCN, but is
also modulated by direct inhibitory feedback
from circulating glucocorticoids, and by a few
inhibitory and excitatory pathways from the
hippocampus, amygdale, stria terminalis, brain
stem nucleus and other brain centres that
allow HPA activity to vary according to the
prevailing psychological, physical and immuno-
logical stressors (Lightman et al., 2002).
Patterns of pulsatile hormone release within the
hypothalamo–pituitary–adrenal axis
The HPA axis exhibits a mixture of basal and
pulsatile hormone release. The ultradian pulse
pattern (spikes within 24 hours) is superimposed
upon the underlying circadian rhythms and is
entrained to the light–dark and sleep–wake
cycles (Czeisler, 1995; Veldhuis et al., 1990,
2009). The corticotropic axis receives time infor-
mation via oscillator neurons in the SCN that
input to (ARC) CRH-ergic neurons, which then
release CRH into the portal system in a periodic
and pulsatile manner (Haus, 2007). ACTH is
then secreted in a pulsatile pattern superimposed
on a circadian variation in response to the pulsa-
tile CRH secretion, followed by corresponding
pulses of cortisol secretion into the adrenal
veins (Haus, 2007).
There is also some evidence for intrinsic
pulsatility of ACTH secretion by pituitary corti-
cotrophs, independent of hypothalamic inputs
(Gambacciani et al., 1987; Gudmundsson and
Carnes, 1997). In vitro studies suggest that CRH
may predominantly regulate sustained HPA axis
activation, whereas AVP is mainly responsible for
acute, short-term stimulation (Gudmundsson and

Carnes, 1997; Watanabe et al., 1989). HPA axis
activity varies according to stress and mealtimes,
with specific alterations in the timing, intensity
and duration of any stress stimulus resulting in
widely varying patterns of ACTH pulsatile activ-
ity across the day (Crofford et al., 2004; Dallman
et al., 1992). Studies also demonstrate acute inhi-
bition of ACTH-mediated cortisol secretion with
the onset of sleep (Haus, 2007; Weibel et al.,
1995).
Periodicity of ACTH secretion
About 12–18 pulses per 24-hour span of ACTH
have been documented and a diurnal variation
is seen, with trough ACTH secretion in the
evening and the peak just before awakening in
the early morning (Crofford et al., 2004; Haus,
2007). The corresponding cortisol peak in the
early morning (around 07.00–08.00 hour) is
followed by a gradual decline during the
daytime and a ‘quiet period’ of lower adrenal
responsiveness, with continuous, small-amplitude
pulses throughout the evening and early night
hours (Haus, 2007). The existence of this ‘quiet
period’ when the cortisol secretory response to
sustained ACTH pulses is relatively blunted indi-
cates, perhaps, an intrinsic circadian property-
variable ACTH responsiveness within the
adrenal cortex (Haus, 2007; Haus and Touitou,
1994). Overall, the percentage pulsatile ACTH
secretion predicts the mean daily cortisol
concentration, suggesting that pulsatility of
pituitary ACTH secretion is functionally impor-
tant to (adrenal) target tissue responsiveness,
rather than merely echoing pulsatility of
upstream hypothalamic hormones (Leng and
Brown, 1997).
Effects of ageing, gender and body adiposity on
ACTH secretion
A recent study on the impact of age, gender
and body mass upon ACTH dynamics showed
that basal, pulsatile and total 24-hour secretory
rates were higher in men than women (Veldhuis
117
et al., 2009). This is in keeping with previous
studies showing higher concentrations of ACTH
in men than in women, both before and after
psychological stress exposure, and consistent
with the postmortem observation that CRH-
immunoreactive neurons are more abundant in
older men than older women (Bao and Swaab,
2007; Traustadottir et al., 2003; Uhart et al.,
2006; Veldhuis et al., 2009). Cortisol–ACTH
feedback-synchrony deteriorates with age in
both men and women, and cortisol secretory
patterns are more irregular with advancing
age. Nevertheless, age does not seem to be
a significant influence on the basal, pulsatile
or total concentration of ACTH secretion
(Leng and Brown, 1997). The possible mechan-
isms of age-related decline in the inhibition of
the hypothalamo–corticotroph unit remain
speculative.
Increased body mass index (BMI) is also
shown to be associated with enhanced pulsatile,
basal and total 24-hour ACTH (Leng and Brown,
1997). The explanation for this positive relation-
ship between ACTH secretion and BMI is
unclear, but might be due to obesity-modulated
amplification of ACTH responsiveness (Katz
et al., 2000; Kopelman et al., 1988, Veldhuis
et al., 2009). The possible effects of hyperinsuline-
mia, increased adrenergic outflow and circulating
cytokines or adipokines require further study
(Leng and Brown, 1997; Solano et al., 2001; Vice-
nnati et al., 2006).
The glucocorticoid ultradian rhythm is
maintained across the blood–brain barrier
Using rapid-sampling in vivo microdialysis, Droste
et al. demonstrated that the basal ultradian
rhythm for corticosterone in male and female
rats (around 1.2 pulses/hour) is maintained across
the blood–brain barrier, allowing brain exposure
to glucocorticoids to modulate emotion, motiva-
tion and cognition. However, peak levels of corti-
costerone secreted in response to stress are
delayed by 20 minutes in the brain extracellular
fluid compared to plasma, even though
clearance occurs concurrently. This results in a
118
smaller exposure of the brain to stress-induced
corticosterone than would be predicted from
plasma hormone concentrations (Droste et al.,
2008, 2009).
Regulation of thyrotroph function
Thyroid hormone secretion is necessarily very
tightly controlled, being essential for energy
homeostasis and basal heat production (Morley,
1981). Thyrotrophin-releasing hormone (TRH)
stimulates the release of both TSH and PRL
from the anterior pituitary and demonstrates a
light-dark-dependent circadian rhythm (Covarru-
bias et al., 1988; Haus, 2007; Martino et al.,
1985). It also interacts with other circadian per-
iodic neurotransmitters such as 5-HT (5-hydro-
xytryptamine), dopamine and noradrenaline
(Bennett et al., 1989; Haus, 2007). This pulsatile
pattern of TRH secretion is biologically impor-
tant, because pituitary thyrotrophs are more
responsive to intermittent than continuous
TRH stimulation, in terms of TSH secretion
(Haus, 2007; Spencer et al., 1980). Pulsatility of
TSH secretion is not correlated with serum
thyroid hormone levels, implying that TSH
pulses are predominantly TRH driven (Brabant
et al., 1991).
Ultradian pattern and regulation of TSH
secretion
TSH secretion is characterised by a diurnal
variation with superimposed bursts (spiky secre-
tion) (Roelfsema et al., 2009b). It demonstrates a
series of discrete pulses with an average fre-
quency of 0.4 pulses/hour in normal men and
women (Brabant et al., 1990; Haus, 2007; Nicolau
and Haus, 1994). These pulses show an unequal
distribution, with clustering of pulses during the
evening and night hours and consequent fusion
of pulses resulting in a nocturnal rise in pulse
amplitude. Hence, peak TSH concentration is
between 2300 and 0400 hours, with subsequent
decline to a nadir between 1200 and 1400
(Brabant et al., 1990; Greenspan et al., 1986;
Haus, 2007; Mantzoros et al., 2001). TSH pulsa-
tile secretion is regulated through interplay
between an apparent hypothalamic oscillator
(modulated by inhibitory influences from dopa-
mine, SRIF and other inhibitory neurotransmit-
ters), direct stimulation from hypothalamic TRH
and the negative feedback effect of circulating
thyroid hormones (T3 and T4) (Mariotti, 2006;
Roelfsema et al., 2009b). Sleep deprivation aug-
ments the circadian variation in TSH secretion
attributed to changes in pulse amplitude, but not
pulse frequency. Similarly, resumption of sleep
(or fasting) suppresses TSH secretion by redu-
cing pulse amplitude (Brabant et al., 1990; Romi-
jin et al., 1990).
Effects of ageing and adiposity on ultradian TSH
secretion
Studies to date have shown no evidence for sexual
dimorphism of TSH concentration, TSH pulsatile
or basal secretion, TSH pattern regularity, or TSH
diurnal release. The relationship between diurnal
TSH secretion and body mass index has not been
studied extensively, but some studies have shown
increased TSH secretion in obese women com-
pared with normal-weight, age-matched controls.
This difference was reduced by weight reduction
or administration of bromocriptin (Kok et al.,
2005; Roelfsema et al., 2009a). The
adipocyte-derived hormone leptin demonstrates
a pattern of ultradian fluctuation that is synchro-
nous with TSH, and it may turn out to be an
important modulator of the thyrotroph axis via a
stimulating effect on TRH synthesis and release
(Lechan and Fekete, 2006; Roelfsema et al.,
2009b).
TSH secretion and the circadian rhythm remain
intact in the elderly but a blunted nocturnal peak
has been found. This is mainly due to a decrease in
TSH pulse amplitude, as higher daytime trough
levels result in higher mean concentration
(Barreca et al., 1985; Greenspan et al., 1991;
Haus, 2007; van Coevorden et al., 1989). In com-
parison with children or young adults, elderly sub-
jects showed a slight phase delay in the circadian
peak time of TSH and more marked phase

delay in T3, T4 and rT3, suggesting a delayed
response of the thyroid gland to TSH stimulation
in the elderly (Haus, 2007; Nicolau and Haus,
1994).
Regulation of lactotroph function
The regulation of PRL secretion is complex and
not fully understood. Release is regulated by var-
ious stimulatory and inhibitory secretagogues
involving the hypothalamus, peripheral peptide
modulators and sex hormones. Factors identified
include TRH, dopamine, oxytocin, vasoactive
intestinal peptides (VIPs), estrogen and growth
factor, such as EGF (epidermal growth factor)
and FGF-2 (fibroblast growth factor-2) (Haus,
2007; Iranmanesh et al., 1999; Tolis and Franks,
1979). In addition, external stimuli such as nipple
stimulation, emotional stress and physical exercise
are contributory factors in PRL secretion (Hauffa,
2001). PRL is primarily under inhibitory neural
control of the hypothalamus via tonic suppression
mediated by dopamine, which then exerts nega-
tive feedback on the ARC (Hauffa, 2001). PRL
concentration rises during pregnancy and postpar-
tum period (high estrogen states), but the 24-hour
secretory pattern is maintained (Tay et al., 1996).
Nursing reflexes dominate the pulsatile secretion
during postpartum, with PRL pulses follow suck-
ling episodes and usual nocturnal rise only appar-
ent after cessation of breast-feeding (Haus, 2007;
Tay et al., 1996).
Effect of light–dark and sleep–wake cycles on
prolactin secretion
PRL release exhibits basal, episodic (ultradian
and pulsatile) and circadian rhythm in human
and animal studies (Iranmanesh et al., 1999;
Veldhuis et al.,1990). The normal secretory pat-
tern of PRL comprises intermittent pulses,
occurring every two to three hours with vari-
able amplitude (Haus, 2007; Veldhuis et al.,
1992). The highest plasma PRL concentration
occurring late at night coincides with rapid eye
movement (REM) sleep in both men and non-
119
lactating, non-pregnant women (Haus, 2007;
Obal et al., 1994; Roky et al., 1995). This
night-time elevation results from an increase in
basal PRL secretion and pulse amplitude, but
not in pulse frequency (Veldhuis et al., 1990).
However, during nursing, nipple stimulation
becomes the main regulator of circulating
plasma PRL, with reflex elevation of both
PRL and oxytoxin levels in lactating women
(Haus, 2007; Leng and Brown, 1997).
PRL pulse amplitude is also enhanced during
sleep periods, irrespective of the time of the day,
with immediate offset of active PRL secretion
upon awakening (Haus, 2007; Reppert and Wea-
ver, 2001). On the other hand, there is modest
elevation in PRL concentration around the time
of the usual sleep onset, even when subjects
remain awake (Haus, 2007). These findings sug-
gest that PRL is mainly governed by a circadian
rhythm with superimposed pulsatile secretion
modulated by the sleep–wakefulness pattern;
peak PRL concentration occurs when sleep
onset coincides with circadian rhythmicity
(Haus, 2007; Spiegel et al., 1999; Waldstreicher
et al., 1996).
Effect of ageing on prolactin secretion
Ageing is associated with a significant decre-
ment in circulating PRL concentrations, with
reduced secretory burst mass, pulse amplitude
and decline in the maximal PRL secretory rate
achieved within each pulse, without alteration
of pulse duration (Iranmanesh et al., 1999).
This has been shown in studies involving older
men and estrogen-unreplaced post-menopausal
women (Iranmanesh et al., 1999; Maddox et al.,
1991). These phenomena could be explained by
the accumulation of PRL-derived amyloid in
the human ageing pituitary gland and reduction
in PRL-binding sites in the choroid plexus,
hippocampus and hypothalamus in elderly indi-
viduals, contributing to a decline in lactotroph
cell responsiveness to available stimulating hor-
mones and/or a reduction in lactotroph cell
mass (Di Carlo et al., 1992; Goya et al., 1991;
Iranmanesh et al., 1999; Westermark et al.,
120
1997). The bioactivity of circulating PRL may
also fall in elderly men and women (Iranma-
nesh et al., 1999; Maddox et al., 1991). In addi-
tion there was also greater ‘disorderliness’ with
higher approximate entropy of PRL release
seen in the older men suggesting lack of robust-
ness in the synchrony of the regulation of PRL
secretion (Iranmanesh et al., 1999).
Interaction between pulsatile secretion of
prolactin and LH
Many studies have showed synchrony between
spontaneous pulsatile luteinizing hormone (LH)
and PRL secretion, mediated through
gonadotropin-releasing hormone (GnRH), in
both normal or hypogonal women, with a strong
positive relationship in pulse frequency and
amplitude (Casper and Yen, 1981; Mais and
Yen, 1986; Masaoka et al., 1988). In menstruat-
ing women, the pulse frequency of both LH and
PRL is markedly lower and the pulse amplitude
notably higher during luteal phase, compared
with the follicular phase (Filicori et al., 1986;
Masaoka et al., 1988). PRL levels increase fol-
lowing infusion of GnRH or agonist, and inter-
mittent administration of GnRH has elicited
pulsatile release of PRL in the luteal phase (Cas-
per and Yen, 1981; Masaoka et al., 1988), con-
firming that, under some conditions, GnRH can
act as a PRL-releasing factor, either via direct
action on pituitary lactotrophs, or at a hypotha-
lamic level, whereby cyclic attenuation of dopa-
mine activity by episodic discharge of GnRH
results in pulsatile release of both PRL and LH
(Masaoka et al., 1988).
Physiological significance of pulsatile oxytocin
secretion
Along with AVP (which does not exhibit pulsatile
secretion), oxytoxin is synthesised in the magno-
cellular neuroendocrine cells of the supraoptic
nucleus (SON) and paraventricular nucleus
(PVN) of the hypothalamus and stored in secre-
tory granules in the axon terminals that constitute
the posterior pituitary (Rubin et al., 1978).
Oxytocin has an important role in lactation and
normal progression of labour in human (Leng and
Brown, 1997). Oxytocin neurons discharge action
potential bursts in response to the suckling of
hungry rat pups, at an interval of 5–10 minutes,
resulting in synchronised firing and, hence, pulsa-
tile oxytocin secretion (Wakerley and Ingram,
1993). The pulse of oxytocin then enters the
blood as a bolus in high concentration to induce
myoepithelial contraction and milk let-down
(Bicknell, 1988; Leng and Brown, 1997). Gradual
increases in oxytocin levels or continuous infusion
of oxytocin elicit progressive desensitisation of
the mammary receptors (Bicknell, 1988; Leng
and Brown, 1997). Hence, pulses of oxytocin are
optimally efficient in triggering lactation with
brief, synchronised and high-frequency discharge
from oxytocin cells (Leng and Brown, 1997). Oxy-
tocin also has a facilitatory role in parturition;
experimental inhibition of oxytocin action inter-
rupts parturition and full restoration action
requires restoration of normal oxytocin secretory
pattern (Leng and Brown, 1997). However, both
oxytocin knock-out mice and oxytocin-deficient
women can have normal labour.
Discussion
The phenomenon of oscillatory secretion is not
exclusive to neuroendocrine axes and is widely
found in various biological systems such as periph-
eral inter-cellular signalling (e.g. insulin secretion
from beta-islet cells) and intra-cellular messengers
[e.g. cyclic adenosine monophosphate (AMP)
secretion] (Li and Goldbetet, 1992). Pulsatility
appears to be a common and fundamental means
of biological communication and organisation that
is even present in a primitive organism such as
slime mould (Dictyostelium discoideum). Slime
mould is noted to respond to an external pulse of
cyclic AMP every five minutes, but not to a con-
tinuous stimulus or to a shorter pulse interval
(Martiel and Goldbeter, 1987).
Biological oscillations were once considered to
be the results of breakdown of effective self-reg-
ulation (Savageau, 1976), but subsequent work

has clearly evidenced their physiological advan-
tages. Biological oscillation potentially confer
five functional benefits in respect of (1) temporal
organisation, entrainment and synchronisation,
(2) spatial organisation, (3) prediction of repeti-
tive events, (4) energy efficiency of frequency
encoding of information and (5) precision of con-
trol (Rapp, 1987). Energy efficiency was first
demonstrated in oscillatory insulin-driven glycoly-
sis, with a reduction in 5–10% of free energy
requirements (Rehmus et al., 1981) and other stu-
dies have demonstrated a close relationship
between a specific pattern of periodic stimulation
and the optimal responsiveness of the target cells
in various cellular systems.
Periodic stimuli were found to be more efficient
than random or chaotic signals in a study based on
a model on receptor desensitisation, with cellular
responsiveness being more sensitive to variations
in the time interval between pulses than to varia-
tions in the duration of each pulse (Li and Gold-
betet, 1992). The key to maintaining high
responsiveness is to ensure adequate recovery
time, so not surprisingly, continuous stimulation
induces a decrease in the responsiveness of target
cells – due to receptor desensitisation – that in
turn protects target cells from the adverse conse-
quences of over-stimulation. Hence, pulsatile sig-
nals appears to be the optimal mode of
communication with downstream targets by
achieving a balance between two distinct
demands: (1) a reasonable ligand-free time to
elapse for the receptor to resensitise up to a suffi-
cient level and (2) time elapsed before the next
stimulus arrives should not be so long as to overly
constrain the number of significant responses that
the receptor can generate in a given period (Li
and Goldbetet, 1992). However, because of differ-
ences in receptor kinetics and on/off times, not all
receptor systems demonstrate desensitisation.
Biological oscillations enable the frequency
encoding of physiological information. A constant
hormone concentration in the fluid surrounding a
specialised secretory cell can result in sustained
periodic oscillation of that cell’s membrane poten-
tial. Moreover, the frequency of the oscillation can
be tuned according to the extracellular hormone
concentration, further enhancing the precision of
121
control (Berridge and Rapp, 1979). Hence, altera-
tions in hormone concentration arising from pul-
satility elicit corresponding change in membrane
oscillator frequency that tends to optimise the
physiological consequences (Rapp, 1987).
In view of various physiological advantages in
biological oscillations, it is not surprising to find
that ultradian and circadian rhythms, which have
been known about in hypothalamic–pituitary–
end-organ axes for decades, appear to be a com-
mon phenomenon in mammalian species. This
physiological rhythm of hormonal secretion is
demonstrated throughout human life span from
the newborn period, through childhood, puberty,
pregnancy and lactation, to ageing, and may
exhibit sexual dimorphism (Hauffa, 2001).
Pulsatility in the neuroendocrine axes involves
a dynamic interaction via non-linear, time-
delayed, dose-responsive feedback and feed-
forward activities (Veldhuis et al., 2001). Each
neuroendocrine axis exhibits unique time-depen-
dent secretory patterns to presumptively achieve
homeostasis. Hence, neuroendocrine disease
could be attributed to the disruption of multi-
level, inter-nodal communication or within a con-
trol locus itself (Pincus et al., 1999; Veldhuis et al.,
2001). The use of frequent hormone sampling
techniques in conjunction with sophisticated new
mathematical tools for the analysis of the beha-
vior of these integrated biological networks has
enabled us to gain some insight into the patho-
physiology of human disease associated with
altered pulsatility (Veldhuis et al., 2001). In addi-
tion, novel parameters describing the overall
orderliness of neurohormone secretion, such as
approximate entropy (ApEn) may facilitate the
identification of disease-specific alterations of
hormone pulsatile patterns, potentially leading
to targeted biopharmaceutical modification of
pulsatile hormonal secretion as a form of disease
therapy in the future.
Abbreviations
ACTH adrenocorticotropic hormone
AMP adenosine monophosphate
ARC arcuate nucleus
122

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