279

Relationships between prolactin, LH and broody behaviour

in bantam hens
P. J. Sharp, M. C. Macnamee, R. J. Sterling, R. W. Lea and H. C. Pedersen*
AFRC Institute of Animal Physiologyand GeneticsResearch, Edinburgh ResearchStation, Roslin,
Midlothian EH25 9PS
*Department of Zoology, Universityof Trondheim, N-7055 Dragvoll, Norway
(M. C. Macnameeis now at EndocrinologyDepartment, Bourn Hall Clinic, Bourn Hall, Bourn,
Cambridge CB3 7TR)
(R. W. Lea is now at School of Applied Biology,LancashirePolytechnic,Preston PRI 2TQ)
RECEIVED 21 December 1987

ABSTRACT
The interactions between broody behaviour and
changesin concentrations of plasma prolactin and LH
were investigatedin bantam hens. Adoption of newly
hatched chicks caused incubating hens to leave their
nests and prevented plasma prolactin decreasing as
rapidly as in hens deprivedof their nests and not given
chicks. Further, the hens allowed to rear chickscame
back into lay later (P < 0-001) than the hens not
allowed chicks. Plasma prolactin decreased and
plasma LH increased in hens deprived of their nests:
these changes were reversedwhen the hens re-nested.
The changes in plasma LH and prolactin in nest-
deprived and re-nesting birds were not always syn-
chronous; this was particularly clear immediately
after nest deprivation when the increasein plasma LH
preceded the decreasein the plasma prolactin. Readi-
ness to incubate disappeared between 48 and 72h
after nest deprivation and corresponded with the
time when plasma prolactin decreased to baseline
values. Administration of ovine prolactin depressed
(P <0.01) the initial increasein plasma LH after nest
deprivation, but repeated administration of prolactin
for up to 72h failed to suppress plasma LH to the
values found in incubating hens. Repeated adminis-
tration of ovineprolactin at 5-to 8-hintervalsfor 72h
maintained readiness -to incubate in nest-deprived
hens.
It is concluded that the secretion of prolactin in
broody hens is facilitated by the presence of chicks
and that increasedconcentrationsof plasma prolactin
maintain incubation behaviour. In incubating hens
the secretionof LH and prolactin may be partly regu-
lated independently. In addition, LH secretion may
also be inhibited by increasedplasma prolactin.
J. Endocr. (1988)118, 279-286

INTRODUCTION 1984) but not in precocial (Goldsmith & Williams,

The maintenance of high concentrations of plasma
prolactin in birds incubating eggs appears to be a
direct result of incubation behaviour. Removal of the
nest results in a rapid decreasein plasma prolactin (EI
Halawani, Burke & Dennison, 1980; Lea & Sharp,
1982; Goldsmith, Burke & Prosser, 1984; Ramsey,
Goldsmith & Silver, 1985;Lea, Vowles& Dick, 1986)
which is reversed if the nest is returned and the bird
resumes incubation in the turkey (E1Halawani et aL
1980)and canary (Goldsmith et al. 1984)but not in
the ring dove (R. W. Lea, unpublished observation).
The presence of chicks in altricial (Buntin, 1979;
Dawson & Goldsmith, 1982; Silverin & Goldsmith,
J. Endocr. (1988) 118, 279-286 © 1988 Journal of Endocrinology Ltd
0022-0795/88/0118-0279 $02.00/0
1980;Wentworth, Proudman, Opelet al. 1983;Hector
& Goldsmith, 1985)speciesmaintains high levels of
plasma prolactin. It is not known in precocial species
whether plasma prolactin is as depressed after the
hatching of young as after the removalof the nest and
eggs.
The increasedconcentration of plasma prolactin in
birds incubating eggs appears to maintain readiness
to incubate and to depress plasma gonadotrophins.
Thus, injections of prolactin maintain incubation in
ring doves (Lehrman & Brody, 1964;Janik & Buntin,
1985; Lea et al. 1986) and stimulate incubation in
turkeys(ElHalawani, Silsby,Behnke& Fehrer, 1986).
However, the role of prolar ':n in the maintenance of
Printed in Great Britain
280 P. J. SHARP and others • Hormones and broody behaviour
incubation has been questioned by the observation
that the initiation of re-nesting in nest-deprived
turkeys and canariesdoesnot appear to depend on the
maintenance of plasma prolactin levelsabove baseline
(EI Halawani et al. 1980;Goldsmith et al. 1984).
A depressiveeffectof prolactin on gonadotrophin
secretionis suggestedby the association betweenhigh
concentrations of plasma prolactin and low concen-
trations of plasma luteinizing hormone (LH) in
broody birds (e.g.Sharp, Scanes,Williamset al. 1979;
Burke & Dennison, 1980; Goldsmith & Williams,
1980;Goldsmith, Edwards, Koprucu & Silver, 1981;
Lea, Dods, Sharp & Chadwick, 1981; Dawson &
Goldsmith, 1982;Goldsmith et al. 1984).The finding
in bantam hens that plasma prolactin increasesbefore
plasma LH falls at the onset of incubation, and that
treatment of broody bantam hens with anti-prolactin
serum increasesplasma LH, suggeststhat the inverse
relationship betweenplasma prolactin and LH may be
causal (Lea et al. 1981).This view is consistent with
the ability of exogenousprolactin to induce ovarian
regression in poultry (e.g. Opel & Proudman, 1980)
and ring doves (Buntin & Tesch, 1985; Janik &
Buntin, 1985).However, there is evidencein broody
mallards (Goldsmith & Williams, 1980),turkeys (E1
Halawani, Silsby, Fehrer & Behnke, 1983),canaries
(Goldsmith et al. 1984)and doves(R. W. Lea, unpub-
lished observation) that changes in plasma LH con-
centrations are not always associated with inverse
changesin plasma prolactin.
This series of experiments in bantam hens was
designed to increase our understanding of the inter-
actions betweenbroody behaviour, prolactin and LH.
The possibility that the presence of young chicks
might influenceplasma prolactin after hatching was
investigated, as was the putative role of prolactin in
the maintenance of incubation behaviour. Finally, an
attempt was made to determine whether injections of
prolactin suppressthe concentration of plasma LH.
MATERIALS AND METHODS
Laying bantam hens, 18- to 30 months old, from the
-
laboratory breedingflock, were kept in groups of two
to six in floor pens (4 x 1m) with nest boxes or indi-
vidually in wire battery cages. The lighting schedule
was 14h light:10h darkness (14L:10D;lights on from
07.00-21.00h) and the birds had free access to food
and water.
Experiment 1
This experiment used broody hens to determine the
effectof the presenceof chicks on the concentrations
of plasma prolactin and LH. These hormones were
measured in 11 bantam hens throughout the period
.1".Endocr. (1988) 118, 279-286
they were brooding chicks and compared with pro-
lactin and LH concentrations when the birds were
laying or incubating. The rate of decrease in plasma
prolactin after hatch was compared with that in the
incubating hens deprived of their nestsin experiments
2 and 3. The onset of broodiness was synchronizedas
closelyas possibleby transferring 20 layingbirds with
previous experienceof broodiness from a stimulatory
(14L:10D) to a non-stimulatory (8L:16D) daylength
for 6 weeks to put them out of lay. They were trans-
ferredback to 14L:10Dto stimulatethe resumptionof
lay and kept in floor pens with nest boxes. Eggswere
removedfor 5-6 weeksafter comingback into lay, but
subsequently the birds were allowed to accumulate
clutchesof eggsto encourage broodiness.The 11hens
selectedfor the study became broody between 12and
16days after being allowed to accumulateclutchesof
eggs. After the birds had been incubating for 18-21
days, all the eggswere replaced with day-old bantam
chicks hatched from eggs which had been incubated
artificially.The normal length of incubation in ban-
tam hens is 21 days. Each hen was given four chicks.
This was done to make it easier to take blood samples
at precise intervals after the birds began to brood
chicks.Blood sampleswere taken directlyfrom a wing
veinat 3-to 4-dayintervalsstarting after the birds had
resumed laying and ending after they had incubated
eggs,reared chicksand started layingagain.
Experiment2
This experiment was carried out to determine the
temporal relationships between changes in plasma
prolactin and LH in incubating birds deprivedof their
nest for 24 or 48 h. A second purpose was to establish
whether there are differencesin plasma prolactin con-
centrations between birds which re-nest after nest
deprivation and those that do not. The hens had been
incubatingfor between8 and 14 days.The experimen-
tal birds were transferred at 08.00h from their nests
and placed one bird to a cage, in wire battery cages,
for 24h (sevenhens) or 48 h (ten hens) and were then
returned to their nests. Serial blood samples were
taken directly from a wing vein. Birds deprived of
their nests for 24 or 48 h were sampled at 0, 3, 6, 12,
18,24, 27, 30, 36and 48 h or 0, 6, 12,24, 36,42,48, 51,
54,60 and 72h respectively,after being transferred to
cages.Groups of fivecontrol birds incubatingeggsin
their nest boxeswere sampled at the same time.
Experiment3
This experimentwas carried out to determinewhether
the administration of ovine prolactin would maintain
readinessto incubate in incubating bntams deprived
of their nests for 72h. A 72-h nest Aeprivationperiod

was chosen becausethe result from experiment2 indi-
cated that 50% of the hensdid not re-nestafter 48h of
nest deprivation. It was anticipated that a further 24-h
nest deprivation would result in the disappearance of
readinessto incubate in the majority of the hens.
The hens had been incubating eggsfor 24+ 3 (s.D.)
days. They were transferred to battery cages, and
eight experimentalhens were injectedi.p. at 5- to 8-h
intervals with 25 IU ovine prolactin (Lot 123F-0320;
Sigma Chemical Company, Poole, Dorset, U.K.) in
1ml of an alkaline saline solution (0.15mol NaC1/1
made alkaline with 0.01mol Na0H/; pH 8.5) begin-
ning 3 h after transfer to cages. Eight control hens
were injected at the same time with alkaline saline
vehicle. In total, each bi-rd received 12 injections
spread over 72h. Blood samples were taken at 12-h
intervals to demonstrate that the treatment had
resulted in the expected decrease in plasma con-
centrations of chicken prolactin. The effect of i.p.
injections of ovine prolactin on plasma levelsof this
hormone was investigatedin experiment4. After the
last injection,at 08.00h, the birds werereturned to the
floor pens containing their nest boxes. The birds were
then observed continuously for 1h, and spot checks
were made 12, 54, 78 and 102h later to observe
nestingbehaviour.
Experiment4
This experimentwas carried out to determinewhether
injectionsof ovineprolactin depressthe increasein the
concentration of plasma LH seen after nest depri-
vation. Groups of eight experimentaland eight con-
trol hens, which had been incubatingeggsfor between
8 and 13 days, were transferred to battery cages for
24h. The experimental birds were injected i.p. with
25IU ovine prolactin (Lot 123F-0320;Sigma)at the
time of transfer and at 6, 12, 18 and 22h thereafter.
Control birds were injectedat the same timeswith the
alkaline saline vehicle. Blood samples were taken
from a wingvein immediatelybefore each injectionof
ovine prolactin for measurement of ovine prolactin
and chicken LH.
Additional information on the effectof ovine pro-
lactin on plasma LH was obtained by measuring
plasma LH and chicken prolactin levels in samples
taken in experiment3.
Hormone assays
Plasma concentrations of chicken prolactin and
LH were measured using the homologous assays
described by Lea, Sharp & Chadwick (1982) and
Sharp, Dunn & Talbot (1987) respectively. Ovine
prolactin was measured as described by McNeilly&
Andrews (1974) in a single assay. The intra-assay
coefficientof variation was 4.2%. Prolactin and LH
Hormones and broody behaviour • P. J. SHARP and others 281
concentrations for each experimentwere measured in
single assays to eliminate interassay variation. The
inter- and intra-assay coefficientsof variation for the
chicken prolactin assay were 4.5 and 3%, and for the
LH assay 6 and 4% respectively.
Statistics
Friedman's analysis of variance and Wilcoxon's tests
were used for serial samples unless otherwise stated
(Conover, 190). Comparisons between groups were
made using the Mann—WhitneyU test.
RESULTS
Effect of the presenceof chicks on concentrationsof
plasma prolactinandLH
At the onset of incubation the concentration of
plasma LH decreasedwhile that of plasma prolactin
increased (Fig. 1). After the eggs were replaced
by chicks (designated day 0 from hatch) the con-
centration of plasma prolactin began to decrease
(P < 0.05)within 3 days (Fig. 1).However, it was still
50% above baselinebetween6 and 9 days after hatch
and did not fall to baselinevaluesuntil 10days before
the resumption of egg production. At this time the
plasma concentration of LH began to increase
(P < 0-05).The hensbeganto lay 30+ 2 days after they
had been givennewlyhatched chicks.
The hens took a closeinterest in the newlyhatched
chicks, brooding them, encouragingthem to feed and
drink, and defending them against intruders. As the
chicks grew, this intensematernal response gradually
declined, so that 2-3 weeks after hatching brooding
was infrequent and the chicks were spending more
time away from their mothers.
In experiments in which incubating hens were
deprived of their nests the plasma concentration of
prolactin fell more rapidly than in incubating hens
given day-old chicks (Figs 2b, 3a and 4b). In hens
deprived of their nests, plasma prolactin returned to
baseline values after 36-48 h and had decreased to
50% above baseline after about 18-24h (Figs 2, 3
and 4). Control hens deprived of their nests in the
experiment shown in Fig. 4 came back into lay after
12+ 3 days. This was significantlyearlier (P < 0-001;
Student's t-test) than in the hens allowed to rear
chicks.
Relationshipsbetweenplasma prolactinand LH in
nest-deprivedandre-nestinghens
Plasma concentrations of prolactin decreased signifi-
cantly (P <0.05) after 12h of nest deprivation in all
hens deprived of their nests for 24h (Fig. 2b) and in
J. Endocr. (1988) 118, 279-286
282 P. J. SHARP and others • Hormones and broody behaviour
90
J_
60
1 (p.g/1)
, Plasma
30
-40 -20 0 +20
Time since hatch (days)
-20 6 ' +20
Time since onset of
lay (days)
FIGURE 1. Changesin the concentrationsof plasmaprolactin
(0) and LH (•) (means + s.E.M.) in 11bantam hensduring
a broody cycle.Layinghenswereallowedto accumulate
clutchesof eggs,incubatethem, rear chicksand return to
lay. After the henshad beenincubatingfor 18-21days,the
eggswerereplacedwithday-oldbantam chicks(day0 from
hatch). The henswerelayinguntil about 25daysbefore
hatch: the onsetof incubationbehaviouriscorrelatedwith
the steepincreasein plasmaprolactin and the decreasein
plasma LH at 18-21daysbeforehatch. The hensresumed
laying30+ 2 daysafter beingprovidedwithday-oldchicks.
six out of ten hens deprived of their nests for 48 h (Fig.
3a). In the latter, plasma prolactin had re'turned to
baseline values 48 h after nest deprivation (Fig. 3b).
This observation was confinned in a subsequent
experiment in which nest-deprived hens were injected
with ovine prolactin or saline vehicle (Fig 4b).
Plasma concentrations of LH increased significantly
(P <0-05) within 3 h of nest deprivation and reached
maximum values after 6 h (Fig. 2b). Plasma LH was
also found to be significantly increased (P < 0-001) 6 h
after nest deprivation in hens deprived of their nests
for 48 h (Fig. 3b) or deprived of their nests and
injected with saline vehicle (Fig. 4a).
All the hens deprived of their nests for 24 h resumed
nesting immediately after being allowed access to their
nests. Re-nesting was associated with an immediate
decrease in plasma LH, which was significant
(P < 0-01) after 3 h, and an increase in plasma pro-
lactin which was significant (P < 0.05) after 6 h (Fig.
2b). Concentrations of plasma LH and prolactin
returned to values characteristic of incubating hens
12 h after re-nesting (Figs 2b, 3a and b).
J. Endocr, (1988) 118, 279-286
105 1.5
3
90 1.0
75 0•5
(lig/1)
LH 3
LH
2
Plasma
30 1
t nest removed t nest returned
0
0 24 48
Time (h)
FIGURE 2. Changesin the concentrationsof plasmaprolactin
(0) and LH (•) (means + s.E.M.) in incubatingbantam
hens (a) not deprived(n=5) or (b)deprivedof their nests
(n= 7)for 24h.
Of the ten hens deprived of their nests for 48 h, six
resumed nesting when given the opportunity. Changes
in plasma concentrations of LH and prolactin were
plotted separately for the birds that re-nested and
those which did not re-nest (Fig. 3). Plasma concen-
trations of prolactin fell significantly (P < 0-05) during
the 48-h period of nest deprivation in both groups.
However, in birds that re-nested plasma prolactin was
higher than in those which did not re-nest, at the start
of the experiment, when the nests were removed
(P < 0.05) and again when the nests were returned 48 h
later (P < 0.001) (Fig. 3a). Concentrations of plasma
LH and changes in plasma LH during the 48-h period
of nest deprivation were similar in hens which
re-nested and in those which did not (Fig. 3b).
Maintenanceof readinessto incubatein nest-deprived
hens treated with ovine prolactin
Five out of eight hens deprived of their nests for 72 h
and injected with ovine prolactin returned to their
nests within 10 min of being allowed to do so. One of
the eight control hens injected with saline vehicle also
re-nested within 10 min. After 1 h of being allowed to
re-nest, seven of the eight nest-deprived hens injected
with prolactin had re-nested. The hen which had been
injected with prolactin and did not re-nest after 1 h

120
J.
90
I 11
60
c7. 1..
.....................
30
4 (b)
3 Previous studies (for review see Goldsmith, 1983)
...... ),„ ...............
LH 2 ' maintained after hatching in species with altricial
- —6—
Plasma
t nest removed nest returned
0
0 24 48
Time (h)
FIGURE 3. Changesin the concentrationof (a) plasmapro-
lactin and (b) plasma LH in incubatingbantam hensnot
deprived(11;n=5) or deprivedof their nestsfor 48 h (open
symbols).Hens deprivedof theirnestsfor 48h weresubdi-
videdinto those whichre-nested(0; n= 6) and thosewhich
did not re-nest(A; n=4). Valuesrepresentmeans + S.E.M.
raised her feathers and emittedcallscharacteristicof a
broody hen when approached. Twelve hours after
being allowed to re-nest this hen had returned to its
nest but in contrast with the sevenother hens injected
with prolactin, left her nest when an observer placed
his hand under her. Sevenof the eight hens injected
with prolactin and one of the eight control hens
injectedwith salinevehiclewereincubatingeggs54,78
and 102h after being allowedto re-nest.
Effect of injectionof ovineprolactinon concentrations
of plasma LH in hens deprivedof their nests
Plasma concentrations of LH increased significantly
(P <0-01) in hens deprived of their nests irrespective
of whether they were injectedwith ovine prolactin or
alkaline saline vehicle(Fig. 4). A peak in the concen-
tration of LH was observedbetween 6 and 12h after
nest removal in the control hens but not in hens
injectedwith prolactin. A similar peak in plasma LH
was also observed in the 48-h (Fig. 3b) but not in the
24-h (Fig. 2b) nest deprivation experiments. The
absence of a peak concentration of plasma LH for
12h after nest deprivation in hens injectedwith ovine
prolactin resed in plasma concentrations of LH
being significantlylowerthan in control hens (Fig. 4).
Hormones and broodv behaviour P. J. SHARP and others 283
Repeated injectionsof ovine prolactin for more than
12h after nest deprivation failed to depress the con-
centration of plasma LH below that in the control
hens (Fig. 4).
The repeated injections of ovine prolactin resulted
in circulating concentrations of more than about
300gg/1(Fig. 4a). The concentration of chicken pro-
lactin decreased progressivelyto the baseline value
over 36h, irrespective of whether the hens were
injectedwith ovine prolactin or alkaline salinevehicle
(Fig. 4b).
DISCUSSION
suggest that concentrations of plasma prolactin are
young but not in those with precocial young. The
implication that precocial young do not affect pro-
lactin secretion in their mothers is not supported by
72 the present study. The fall in plasma prolactin
observedafter incubatingbantam hens wereprovided
with day-old chicks was not as rapid as in bantam
hens deprived of their nests and not provided with
chicks.This associationbetweenpartially maintained
concentrations of plasma prolactin and broody
behaviour could be causal sinceinjection of prolactin
inducesmaternalbehaviourin domesticfowl(Burrows
& Byerly, 1938; Nalbandov, 1945; Nalbandov &
Card, 1945;Saeki&Tanabe, 1955;Opel& Proudman,
1980).Further, in willow ptarmigan, administration
of prolactin enhances the intensity of parental care
(Pedersen, 1988). However, it is likely that the
presence of chicks does not stimulate prolactin
secretion but tends to maintain it. This can be
deduced from the observation that plasma prolactin
does not increase in laying or non-laying hens in
which parental behaviour was induced by forced
adoption of newly hatched chicks (Richard-Yris,
Leboucher,Chadwick& Garnier, 1987).
The relative importance of visual, auditory and
tactile stimuli provided by the chicks in maintaining
brooding behaviour is unknown. Bantam hens will
'incubate' sittingon an empty nest if they are provided
with a clutch of eggswhich they can seebut not touch
(Steen & Parker, 1981). This observation demon-
strates the importanceof visualstimuliin maintaining
incubation behaviour, and it is possible that newly
hatched chicks also provide a visual stimulus which
maintains parental behaviour. On the other hand
tactile contact between hen and chicks has been
shown to be necessaryin the domestic hen to main-
tain full maternal brooding behaviour (Maier, 1963;
Richard-Yris & Leboucher, 1986). Since the fall in
plasma prolactin during the 2-3 weeksafter hatch was
J. Endocr. (1988) 118, 279-286
284 P. J. SHARP and others • Hormones and broody behaviour

(ps/1)
2
LI-I

?-
Plasma

0 H HHHH
300

72. 200 1;
)

•- (1) 0
cp>100

0
0 12 24 0 12 24 36 48 60 72
Time (h)
FIGURE 4. Effect of repeated i.p. injections of 25 IU ovine prolactin (0) or saline
vehicle (•) in groups of eight incubating bantam hens for (a) 24 h or (b) 72 h after
nest deprivation on concentrations of plasma LH and ovine or chicken (ch)
prolactin. The arrows indicate the times of injection of ovine prolactin or vehicle.
The values represent means + S.E.M. *P< 0.05, **P< 0.001 compared with
saline-injected control birds (Student's t-test).

associated with a progressive reduction in brooding
behaviour, it is possible that prolactin secretion is
maintained in hens with chicks by tactile stimuli.
The rapid fall in plasma prolactin after nest depri-
vation and the increase after renesting confirms the
finding in other species that prolactin secretion is
maintained by incubation behaviour (see Introduc-
tion). The 48- to 72-hperiod required after nest depri-
vation for the bantam hens to lose their readiness to
incubate is similar to that required by nest-deprived
turkey hens (ElHalawani et al. 1980)and corresponds
with the time required for plasma prolactin to return
to baselinevalues.It thus appears that in thesespecies
readinessto incubate cannot be maintained after con-
centrations of plasma prolactin have returned to these
values.
The temporal relationship betweenplasma LH and
prolactin after removal and return of nests suggests
that changes in plasma LH are not always inversely
related to concentrations of plasma prolactin. Most
striking was the steep increase in plasma LH which
J. Endocr. (1988)118, 279-286
preceded the fall in plasma prolactin after nest depri-
vation (Figs 2b and 3). A similar relationship was
seen in incubating turkeys treated with the anti-
serotoninergic drug, p-chlorophenlyalanine, to dis-
rupt broodiness (EI Halawani et al. 1980). These
observations,togetherwith the asynchronouschanges
in plasma LH or prolactin seen at the onset of
incubation or during re-nesting in other species
(Goldsmith & Williams,1980;Goldsmith et al. 1984),
suggests that the secretion of these hormones is not
entirely interdependent.
The observation that the administration of pro-
lactin attenuated the intitial increase in plasma LH
after nest deprivation is consistent with an inhibitory
action of prolactin on LH release. A similar conclu-
sion was drawn from the finding that the adminis-
tration of anti-prolactin serum to incubating bantam
hens results in an increase in plasma LH (Lea et al.
1981).An inhibitory action of increased plasma pro-
lactin on gonadotrophin secretionis also suggestedby
the fact that hens allowed to rear chicks returned to

lay significantlylater (30+ 2 days versus 12+ 3 days;
P <0.001) than hens deprived of their nests and not
given chicks. It seems that the maintenance of
increased concentrations of plasma prolactin in hens
brooding chicks may have delayed the resumption of
ovarian folliculogenesis.
The failure of repeated administration of ovine
prolactin for 72h to depress plasma LH to the values
found in incubating hens argues against high concen-
trations of plasma prolactin being the only factor
responsible for inhibiting LH release in these birds.
It is possible that the failure of ovine prolactin to
suppressconcentrations of plasma LH to those found
in incubating hens is due to differences in species
specificity between chicken and ovine prolactin.
Alternatively, the blood—brainbarrier might have
restricted the access of the administered ovine pro-
lactin to target sites in the brain (Buntin & Ruzychi,
1987).It is probable that prolactin enters the brain via
the choroid plexusas has beendemonstrated in the rat
by Walsh, Slaby & Posner (1987).
The maintenance of readiness to incubate in nest-
deprived bantam hens after repeated injections of
prolactin confirms similar observations in the ring
dove (Lehrman & Brody, 1964;Janik & Buntin, 1985;
Lea et al. 1986).Similarly,El Halawani et al. (1986)
showed that nesting behaviour induced in ovari-
ectomized turkeys by treatment with oestrogen and
progesterone is advanced to incubation behaviour
after the administration of prolactin.
The contrast between the marked effects of ovine
prolactin on the maintenanceof incubation behaviour
and its modestly depressive effect on the concen-
tration of plasma LH suggestsdifferent mechanisms
of action. Studies on the ring dove show that intra-
cerebroventricular injections of ovine prolactin
depress testicular weight (Buntin & Tesch, 1985)and
plasma LH (Buntin, Lea & Figge, 1988)but do not
maintain readiness to incubate (Buntin & Tesch,
1985).It seemsthat prolactin acts, at least in part, on
peripheral tissues to maintain incubation behaviour.
A similar differencein the sites of action of prolactin
in the bantam hen may explain the contrast between
its marked effecton incubation behaviour but modest
effecton LH secretion.
Previous attempts to show that prolactin plays a
role in incubation behaviour in the domestic fowl
have produced conflicting results. Early studies
showed that injections of mammalian prolactin
induces incubation behaviour in laying hens (Riddle,
Bates & Lahr, 1935;Saeki & Tanabe, 1955),but this
was not confirmed in more recent work using better
purifed preparations of ovine prolactin (Opel &
Proudman, 1980;R. W. Lea and P. J. Sharp, unpub-
lished obserw tions). Nesting behaviour normally
occurs in laying hens around the time of oviposition
Hormones and broody behaviour • P. J. SHARP and dthers 285
and is initiatedby progesteroneactingon a neural sub-
strate previouslyprimed with oestrogen (Wood-Gush
& Gilbert, 1973, 1975).It seems that at the onset of
incubation an increasein plasma prolactin facilitates
this steroid-dependent behaviour. The initial depen-
dencyof incubation behaviour on an ovarian steroid-
primed neural substrate seems to disappear after the
behaviour becomes fully established. Thus, well-
established incubation behaviour is not disrupted
after ovariectomy(Sharp, Sterling& Pedersen, 1986).
It seemsthat the successor failure of earlier attempts
to demonstrate a role for prolactin in incubation
behaviour in domesticfowlmay reflectappropriate or
inappropriate balances between circulating ovarian
steroids and the dose of prolactin administered. In
addition, it is also likelythat the onset of incubation is
influencedby poorly defined non-hormonal environ-
mental factors suchas the availabilityof an acceptable
nest site. It is not surprising, therefore, that a role for
prolactin in incubation behaviour can be more readily
demonstrated in nest-deprived hens in which the
behaviour is not directly dependent on ovarian
steroidsor undefinedenvironmentalstimuli.
ACKNOWLEDGEMENTS
We thank Mr R. T. Talbot and Mr S. Anderson for
help with the immunoassays and Drs A. Chadwick
and A. S. McNeillyfor the supply of reagents for the
prolactin radioimmunoassay.
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