Sexual maturation and fertility were assessed in fourteen cotton-top tamarin. (Saguinus oedipus) females under various social conditions. Six tamarin females.
BIOLOGY
OF
The
REPRODUCTION
37,
Endocrinology
618-627
(1987)
of Puberty
(Saguinus
Tamarins
TONI
and
Reproductive
oedipus)
E. ZIEGLER,2’3’4
under Varying
ANNE and
SAVAGE,3’4
CHARLES
in Female
Social
GUENTHER
Cotton-Top
Conditions’ SCHEFFLER,4
T. SNOWDON3’4
Department Wisconsin
Functioning
of Psychology3
Regional University
and
Primate Research of Wisconsin
Madison,
Wisconsin
Center4
53706
ABSTRACT Sexual maturation under various social while mone of
living (LH) the
with their families. and estrone-conjugates
six
females
ovulatory
LH
nancy.
Two
family there ception
were
peak of
six
were assessed in fourteen tamarin females (20-28
Hormonal (E1 C).
removed
occurred
the
Six
from
as early
females
profiles A rapid
their
were
and
days
housed
in
elevated LH and male. These findings
cycling. Sexual mo of age when
after
paired
with
pairing
in total
of
age
with
maturation, mean LH an
E1 C levels, but cyclicity indicate that removal
or puberty, occurs and E1 C levels began
unrelated
male,
elevated
findings indicate that a suppression reproductive suppression does not environment
and
activity.
An
development
isolation
stimulation
acceleration and
by of
paired
with
puberty
an
unrelated occurred
social
environment
of
was not female
of LH
occurs process male when
and
unrelated
male.
in conception
for
30
following
E1 C may
days
tamarin
were
a female
In
one
and their
horfour
female,
an
full-term
removal
pregfrom
the
female
primates
USPHS
10
and
11
induce
mo
of age.
families, from
normal
from
her
with an ovarian 15 and 17 paired at 9 Our
but that the family
reproductive family
grants
MH
35,215
Snowdon and RR 00,167 to the Wisconsin Regional Primate Center. 2Reprinr requests: Dr. Toni E. Ziegler, Dept. Psychology, W. Johnson St., University of Wisconsin, Madison, WI 53706.
1970; Dunbar, females may
early
in
to
behavioral
and
1980). Subordinate have reduced fertility endocrine
the
group
consists
of a stable
a breeding female, an unrelated monogamous pair bond may sets of offspring, and possibly
this the has
to CT. Research
female family.
1202
618
females Epple
(Dawson, and
Low-
be manifested in lower incidences sustained pregnancy, conception,
of ovulation and normal Callitrichid species (marmosets
social
or in
changes.
ovarian and
even
In
1987. 5, 1987. by
to removed
ered fertility may of infant survival,
1984; Evans and Hodges, 1984), and Katz, 1984; French et a!., 1984; talapoins (Bowman et a!., 1978; Keverne et al., 1984; Abbott et a!., (Drickamer, 1974), and baboons
supported
by
paired initiate
families between is removed and
living with their Both removal
was
(Rowe!!, low-ranking
can
1978;
were
be seen
was not
a male.
and April 10, February
the female alone does
necessary
tamarin
or
Abbott, (Epple 1984), 1979; macaques
until family
tamarins maturation.
1981; tamarins Tardif, Keverne, 1986),
studies
an
resulted
in cotton-top of sexual
response
‘These
(Saguinus oedipus) females a suppression of fertility
and
observed from the
have a pronounced effect on their fertility. Lowered fertility in subordinates has been reported in several primate species, including marmosets (Abbott et al.,
Accepted Received
tamarin showed
in female tamarins living with their to increase. However, when a female
levels
of fertility affect the
ofa
INTRODUCTION
The
age)
demonstrated low, acyclic levels of urinary luteinizing onset of ovarian and pituitary cyclicity occurred when
families
as eight
cotton-top mo of
and prior to pairing. Gradual increases in hormone concentrations occurred during isolation; however, was no ovarian cyclicity until each female was paired with an unrelated male. In all six females, conoccurred before or as a result of the third ovulatory cycle. Partial isolation of a 36-mo-old female
resulted unrelated
mo
and fertility conditions.
Katz,
1977; 1984;
nucleus
cyclicity. tamarins), comprising
male with which a be formed, successive other unrelated male Neyman,
1977;
Izawa,
Terborgh,
1984).
Within
social system, all other females are subordinate breeding female. In captivity, this subordination been associated with suppression of fertility offspring who In the common
remain caged with marmoset (Callithr’ix
to
their natal jacchus),
in
PUBERTY
Evans
and
in
Hodges
urinary
offspring but
(1984)
a
rapid
females
reported
pregnanediol-glu remained were
of
paired
no
family
with
males.
back
resulted
tamarins
cyclicity
under
have
(Saguinusfuscicollis)
in urinary
their families 1984). Also, been reported
estrogens
until 25 cotton-top to have
when
who
paired age
with (15
and
an
mo)
families assessed
were unrelated
than
(21 by
peaks
with
families
at an
remained
in Tardif’s of plasma
indicative
and
When
removed from the family, common marmosets have ovulated by 13 mo of age (Abbott and Hearn, 1978), and saddle-back tamarins have conceived as early as seven (Epple of
mo of age when paired with an older adult male and Katz, 1980). Whether this is an acceleration
puberty
suppression the
a release
of
luteal
phase
of
their
effect tion.
the
family-induced
have the
tamarin, and therefore, urinary indicative of ovulation (Ziegler progesterone and pregnanediol
been
cycle
families, social
1)
and
to
3)
determine
and
environment
to
has
MATERIALS
the
LH
2) to removed
determine
on
in
urinary
what
sexual
matura-
AND METHODS
subjects,
14 cotton-top
tamarin
and third generation were raised in large,
a breeding except Ann, in
male who
and was
months
of
as follows: 2.3 mH),
captive-born. stable families
female with hand-reared.
females
families, upon pairing their social conditions.
females,
with The
All of the containing
their Table
studied
were
offspring, 1 lists the
while
in
their
an unrelated male, housing conditions
and were
large family cages (1.8 mW X 3.0 mL X male-female pair cages (0.85 mW X 1.5 mL
2.3 mH), and isolation cages (0.64 mWX 0.64mL 2.44 mH). All cages contained an elaborate system of branches, ropes, and a nest box. (For further X X
details
of
caging,
et al.,
1985).
husbandry,
and
diet,
see
Snowdon
fertility
shown in
estrone
and
monitoring
to family induced fertility suppression, fertility when female offspring are
Sampling
is unclear. estrogen peaks
Urinary
in
or
by
cotton-
ages
of
age
their
study was progesterone
of ovulation.
various
response assess from
and
Female
at
levels
second females
earlier
with
studied
environments
LH
to further our its relationship
in Callitrichids.
were
social
The
family
was undertaken suppression and
Subjects
of fertility of young found that
the
matured
who
mo). Maturity increased levels
progesterone
from
male
daughters
has
several
619
maturation
tamarins
(Epple and Katz, (S. oedipus) have levels of urinary
suppression or puberty, (1984)
removed
sexual
top
exhibited
remain
estrogens when they remain with their natal for up to 40 mo of age (French et a!., 1984).
daughters
to
the
Saddle-
not
they
mo of age tamarins low, acyclic
It is unclear what effect the has on the sexual maturation, daughter Callitrichids. Tardif
female
(1984)
conditions.
TAMARINS
of age,
50% of marmoset over 12 mo of age family. However, no
these
COTTON-TOP
The present study knowledge of fertility
when
Abbott
shown that in approximately families, one adult female offspring will ovulate while living with the pregnancies
to 25 mo
occurred
IN
pattern
while
up
cyclicity
FERTILITY
cyclic
curonide
in the
onset
AND
the
Early
to occur first
cotton-top
estrogen peaks are not et al., 1987). To date, have been undetectable
void
female
Techniques morning of the without
Details of the been described
urine
(EMU)
morning
and
restraint, collection (Ziegler
was was
obtained
isolation and
eta!.,
collected
as the from
or
each
handling.
storage technique have 1987) with the following
in cotton-top tamarins’ urine (Brand, 1981; Ziegler, unpublished data). Measuring urinary luteinizing hormone (LH) activity, therefore, is important to the
modifications. Urine was collected in polypropylene containers (rodent cages, American Scientific Products, McGraw Park, IL) to prevent steroids from
assessment of ovulatory activity in cotton-top tamarms, especially during the suppression of estrogen cyclicity that occurs when female offspring remain in their natal family. Since Tardif (1984) and French et
adhering
a!. (1984) progesterone female important
have and
offspring to
found urinary that
determine
measurable estrogens,
remain the
in the
levels of respectively, natal
gonadotropic
serum
family, excretion
indicative of ovulation as well. LH is also an important indicator of sexual maturation of the hypothalamus and pituitary (Terasawa et a!., 1983).
it is
the
container.
Prior
to freezing,
were centrifuged (1000 X g for 5 mm) and into two aliquots for steroid and protein mination. To the aliquots for gonadotropin mination,
in
to
0.52
mol/l
glycerol
St. Louis, MO) was added to molecules while frozen (Livesey was collected daily or a minimum week for all social conditions. attempted increased
daily for scent marking
newly often
(Sigma
Chemical
samples divided deterdeterCo.,
stabilize the peptide et a!., 1983). Urine of three times per Urine collection was paired subjects, but prevented collection.
ZIEGLER
620 TABLE
1. Age
in mo
of female
offspring
at each Social
social
ET
AL.
condition.
condition
Subject
Family
Isolation
Paired
Sibling
#in
Lew Kla Pip Hal Yas Zie Hdy Ann Lau Kim Ren Cu Ant Sea
20-27 20-27 23-26 20-26 25-28 25-28
-
-
27 27 26 27 29 29
twin(Kla) twin(Lew) twin(male) twin(male) twin(Zie) twin(Yas)
8 8 6 8 8 8
-
36-39
-
-
39 10
-
-
12-17 13-16 11-19 13-19 15-19 11-19
-
-
-
-
-
-
-
-
-
-
-
-
aD
Indicates
death
-
28-29 28-29
handreared twin(male) twin(male) twin(male) twin(male) trip(male,Da) twin(male)
Family
Condition
7 4
1b 1 1 1 i&iIc I&II II 1J1d III III III Ill III
5
III
-
6
5 5
of infant.
bCOndition
I females
were
CConditiOn
II females
were
dCOndition
Ill
females
housed
with
isolated
were
their
and
families
housed
prepubertal
at the
and
then
in individual beginning
paired cages
with
before
of sampling
and
Assays
an unrelated pairing lived
male.
with with
To
an unrelated their
families
male. or with
an unrelated
determine
ovulation are not luteinizing
male.
and
pregnancy
(since
A method for measuring estrone-conjugates by radioimmunoassay (RIA) directly in the developed and validated. Estrone is known
(E1 C) urine was to be the
estrogen peaks immunoreactive
predominate (Hodges and
tamarins the endo-
of Toivola et a!. (1978) as a mixed heterologous RIA. This procedure correlated closely with bioactive measurement of LH/CG (r> 0.80 in over 300 samples)
urinary Eastman,
estrogen 1984)
in cotton-top and reflects
crine events of parturition, estrogen cyclicity, pregnancy (Ziegler et al., 1987). Estrone-3-glucuronide (E1 G) rabbit antiserum (developed and donated Dr.
J. K. Hodges)
form 105% 1.08%
measured
by
in its conjugated
and showed the following cross-reactivities: estrone, 49.6% estrone-3-sulfate, 3.05% estriol, estradiol3-glucuronide, 0.92% estradiol1 7j3,
0.7 3% estriol-3-glucuronide, than 0.005% with all obtained from Sigma standards, (Arlington dilutions placement
estrone
and
and
3H-E1
0.68% estradiol, and less other steroids tested. E1 G, Chemical Co., was used for G,
Heights, IL), of cotton-top curve parallel
obtained
from
nadotropin
(LH/CG)
indicative of ovulation), hormone/chorionic go-
was
measured
using
the
method
and has been described elsewhere (Ziegler et a!., 1987). The intrassay coefficients of variation were 11.87% (n=28) and 17.6% (n=4) and the interassay coefficient of variation was 13.5 3% (n=6). hormone
Urinary
for variable concentration
urine
hormone expressed
concentrations
output (Tietz,
were
corrected
by measuring creatinine 1976) and dividing Cr
concentration. per mg of Cr.
Hormone
(Cr) into
values
were
Amersham
was used as tracer. Serial tamarin urine gave a disto that obtained with E1 G
Determination
of
Hormonal
Cycles
profiles
and
are
Pregnancies
shown
in
Figure
1
for
standards. The sensitivity of the assay was 4 pg/tube and the intraand interassay coefficients of variation were 6.25% and 4.49% (n=34) respectively. This method of measuring E1 correlated well with our previous method (Ziegler et al., 1987) of enzyme
urinary pattern estrogens Hormonal
LH and E1 C in a cycling female. The typical seen in cotton-top tamarins of elevated occurring after the LH peak is indicated. profiles during ovarian cycles and preg-
nancies
have
hydrolyses, extraction, column individual RIA of E1 during the three female cotton-top tamarins
Ziegler
et
r=0.96, n19; one tamarin
r=0.88, (r=0.91,
n=27) n=88).
and
separation, ovulatory (r=0.98, during
and cycle of n=29;
pregnancy
in
females we found
been al.
(1987).
for urinary prominent
described
by
By estrogen LH/CG
increases in E1 and E2 peak. Urinary estrogens
the
method
monitoring
adult
found
in
breeding
and LH/CG excretion, peaks with corresponding
on the continued
day
of to
the LH/CG increase and
PUBERTY
AND
IN COTTON-TOP
FERTILITY
1500
fertility. One urinary 50
1250
I’ I
II
were
30
then
I
C
W
10
250
15
25 Days
FIG. 1. Urinary concentrations estrone-conjugate (E,C) in a yr of age, during ovarian cycling.
at high decreased
conception occurred
35 of
45
sampling
of
luteinizing cotton-top
single
Cr
55
=
hormone tamarin
(LH) female,
and
5
for
cyclicity.
and
colony
room.
then 20
LH/CG
peak.
increase again shortly high for approximately reached their maximum sharply
183
15 days, occurred. If
increase days after
The
in
estrogens
after this. 80 days, levels at
at parturition.
is approximately Experimental
an
contact
LH/CG the pen-
began
I. Six family
length
days.
first oldest
female
sampling
birth
to
twins
during
the
period. Two sets of twin females and two females with twin males were studied. All six were older than this
removed
is the
from
age at which
their
colony to maximize not obtained on natal family past
natal
tamarins
family
reproduction. female offspring 28 mo of age.
and
are normally paired
in our
Control data were living with their However, family-
induced fertility suppression was not age-dependent; our colony has had four females who had failed to show signs of ovarian activity prior to pairing with a Three family
female up to
with
offspring had remained with their 36 mo of age with no overt signs of
each
was
other
for two Condition
families. pregnant for one
gave
contact then
of the subjects were studied cage. All subjects were the
an unrelated were
other
studied
or any under
male
monitored
male
for mo III.
two
and
mo
other
from During
her family but the study, this with other but no
cages visual,
tamarins.
Urine
female
during
tamarin.
partial isolation.
housed alone in a room auditory and olfactory,
LH/CG remained and the estrogens midpregnancy and
female offspring and were housed in large The adult breeding female of each family was throughout the entire sampling period, except who
female was that allowed collected
Conditions
Condition in their natal
with
Hormones
This female had been removed remained unpaired for 18 mo.
was
gestation
with
female
to
The
paired
mo after removal from the family, but prior to pairing with an unrelated male. These two subjects were in total isolation, with no auditory, olfactory or visual
creatinine.
levels for approximately if conception had not
occurred, approximately
ovulatory
male. natal
monitored
estrogen
four to five months following pairing. Condition II. Females were isolated from conspecifics to determine if reproductive cycling could be initiated by removal from the natal family, or if
Another
mo;
was no
the stimulus of an unrelated male was needed. Two of the above-mentioned females were isolated for one
0
24
females
showed
for
20
500 -J
dropped
and
removed
in another
E
remained and then
these
estrogens
I’
o
750
of
621
Another female remained housed with her female twin until she was 42 mo of age and showed no cyc!icity in urinary estrogens (French et a!., 1984). Four of the six subjects studied in their natal family
40
9
1000
c5
TAMARINS
partial
isolation
and
after pairing with an unrelated male. Since the first six females had low,
acyclic, but measurable, levels of hormones living with their families (age 20-28 mo),
while six ad-
ditional females (see Table 1) were monitored at an earlier age while still housed in their families to determine when hormonal levels began to increase. These females were monitored from 11 to 19 mo of age. A mean monthly concentration of hormone was determined for each female, and the total mean ± SE was determined for all six females for each month. An additional female was paired with an unrelated male at 9 mo of age and studied for two mo to determine if early pairing accelerated sexual maturation,
in contrast
to young
their families. The mean month was determined.
females ±
who
remained
SE hormone
level
with for
each
RESULTS Condition I. All six females showed essentially the same pattern of low, acyclic concentrations of E1 C and LH while living with their families (see Figures 2 and 3). For the six subjects, the mean level was 1.22 tg/mgCr ± 0.36 SE for E1 C and 258.81 ng/mgCr ± 61.84
SE
for
LH.
There
was
no
ovarian
cycling
of
622
ZIEGLER
ET
AL.
3000
2500
2000
1500
1000
500
0 Dl
E
0
0)
3000
() w 2500
2000
1500
1000
500
20
21
22
23
24
25
26
Age FIG. 2. Hormonal profiles of urinary luteinizing and paired phases. Concentration of hormones was due to illness and she was remated 12 days later.)
E1 C.
All
female
offspring
living
in
as shown
in Figure
cycling female, and they occurred indicate ovulation. In female Lew, were elevated during the 26th
28
29
30
families
had
1 in a normally
too frequently to basal levels of LH month, but E1 C
31
32
33
in months
hormone (LH) and estrone-conjugate (E,C) plotted against age in months. Ov = ovulatory
fluctuating levels of LH, with peaks occurring at ovulatory levels of cycling females. None of the LH peaks were associated with corresponding increases in E1 C concentration,
27
remained mo of declined. Upon
for female twins, KIa LH peak, Cr = creatinine.
low. This same age
for
pairing
increased
50
an unrelated
dramatically
Lew, (KIa’s
during the family mate was removed
profile was seen in Hal at 20
approximately with
and
for
days male,
until
levels
E1 C and
all females.
The
LH
mean
concentration of E1 C after pairing was 11.62 jig/mgCr ± 2.02 SE and for LH was 1171.60 ng/mgCr ± 496.64 SE
for
the
six
subjects.
Hormonal
concentrations
PUBERTY
AND
FERTILITY
IN
COTTON-TOP
Isolated
Family
TAMARINS
623
Paired
Pregnancy
4I
C.) 40
E
35
0
w .3Q 25
20
15
10
25
26
28
27
29
Age FIG. 3. Hormonal isolated, and paired during paired phase.
TABLE
profiles phases.
2. Reproductive
of urinary luteinizing Infants = birth of twins
data
Days first Subject
LH
Lew KIa Pip Hal Yas Zie
8 37 45 33 18 70 aConception bAbortion sDay
is defined occurred
1 is defined
in days
six cotton-top
Days first peak
to peak
by the when
as the
from
number
female day
the
was females
hormone to the
of days transferred were
(LII) breeding
tamarin
30
in
from LH to
following
second
Days from second LH peak to third
-
-
26 32 21
-
-
-
24
-
pairing
fertile
to another paired
with
LH
peak.
colony. a male.
LH
=
33
34
35
luteinizing
(E, C) for female LH peaks, Cr
with
an unrelated
=
twins, Yas creatinine.
and Zie, during the Note larger hormone
family, scale
male.
Gestation length & # of infants
27 25
to the
32
months
and estrone-conjugate female, ov = ovulatory
females
31
hormone.
Conceptiona
born
8 64 105 80 18 94
186, 187, 72, 182, 184, 184,
single single abortionb single twins twins
ZIEGLER
624 were
statistically
greater
after
the
females
were
than before (one-tailed, t=4.95, p
