Comp. Biochem. Physiol. Vol. 116B
, No. 2, pp. 269–277, 1997
ISSN 0305-0491/ 97/$17.00
Copyright 1997 Elsevier Science Inc.
Seasonal Levels of Reproductive Hormones and Their
Relationship to the Antler Cycle of Male and Female
Reindeer (Rangifer tarandus
George A. Bubenik,a Dieter Schams,b
Robert J. White,c Janice Rowell,c John Blake,d and Ludek Bartosd
Department of Zoology, University of Guelph, Guelph, Ontario, Canada, N1G 2W1; Institut fuer
Physiologie, Technische Universitaet Munchen, D-85350 Freising-Weihenstephan, Germany; Institute
of Arctic Biology, University of Alaska, Fairbanks, AK 99775, U.S.A.; Research Institute of Animal
Production, Department of Biology, Ethology Group, CZ-104 00 Praha 10-Uhrineves, Czech Republic
Seasonal levels of LH, FSH, testosterone (T), estradiol, progesterone (P), and prolactin (PRL)
were determined in the plasma of five adult bulls, and five barren and four pregnant cows of Alaskan reindeer
), which were sampled every 3 weeks for 54 weeks. The male reproductive axis was sequentially
activated; LH peaked in May–June (2 ng/ml), FSH in June (51 ng/ml), and T in September (11.8 ng/ml). LH
levels in females reached a maximum in both groups at the end of August (the beginning of the rut). Seasonal
variation in FSH was minimal in pregnant cows, but exhibited one elevation (41 ng/ml) in barren ones in
November. T levels in cows remained at barely detectable levels. The decrease of T values observed in both
groups in December and March was not significant. PRL peaked in May in cows (135 ng/ml pregnant, 140 ng/
ml non-pregnant) and in June in bulls (92 ng/ml). Estradiol was highest in bulls in the rut (August), in non-
pregnant cows in January and in pregnant cows in April, shortly before parturition. P levels in the pregnant
cows rose from September and peaked (9 ng/ml) shortly before parturition in April. In the non-pregnant females
P values increased and decreased several times before peaking (5 ng/ml) in March. In the males, the variation
of T and estradiol levels correlated relatively well with the antler cycle but in the females the variation of neither
estradiol, progesterone nor T appeared to be related to mineralization or casting of antlers. Copyright
Elsevier Science Inc.
comp biochem physiol 116B;2:269–277, 1997.
LH, FSH, testosterone, prolactin, estradiol, progesterone, reindeer, pregnancy, seasonality, antler
dominant factor in determination of individual phases ofthe antler cycle, such as the growth period and the time of
Seasonal variation of reproductive hormones [LH, FSH, tes-
polishing of the velvet and casting of antlers (9,19,24,42).
tosterone (T), and prolactin (PRL)] has been investigated
In the male reindeer this correlation between T levels and
frequently in various cervids of temperate regions [white-
the antler cycle is less pronounced. Although the growth
tailed deer (9,10,28); roe deer (42); red deer (2,18); Pere Da-
of male reindeer antlers occurs also during the period of low
vid's deer (25); fallow deer (3)]. Conversely, studies of the
concentrations of plasma T (58), out-of-phase polishing of
reproductive hormones in the boreal cervids, e.g., moose,
antlers can be induced by administration of large doses of
reindeer, and caribou are relatively rare (4,23,26,40,57).
exogenous T (39) and antler casting will follow shortly after
Reindeer/caribou (Rangifer tarandus
) is the most northern
castration (50), the timing of the individual phases of the
cervid and the only one in which the females carry antlers.
antler cycle does not entirely relate to the variation of T
Whereas the regulation of the antler cycle in male deer of
in plasma (57). In addition, unlike in most other cervids,
temperate zones has been studied extensively (7,21,49), and
antlers are produced in both sexes, the pedicles and antlers
is well understood, this is not the case for male and female
developed well before puberty and gonadectomy does not
prevent the growth and subsequent replacement of antlers
In most cervids, the seasonal variation of T is the pre-
(23,50,59). In addition, long-term administration of an an-drogen receptor blocker cyproterone acetate which causes
Address reprint requests to
: Dr. G. A. Bubenik, Dept. Zoology, University
quick casting of antlers in other cervids (7,19), was not ef-
of Guelph, 50 Stone Rd.-E., Guelph, Ontario, Canada, N1G 2W1. Tel.
fective in the reindeer male or female calves or yearlings
(519) 824-4120; Fax (519) 767-1656; E-mail: email@example.com.
Received 6 February 1996; accepted 14 June 1996.
(G. Bubenik et al.
, unpublished observation). Therefore,
G. A. Bubenik et al
based on the above data, Lincoln and Tyler (23) concluded
individual animal temperament. Rutting bulls are exqui-
that in males ‘‘the secretion of T by testes, although not
sitely sensitive to xylazine and seem to be unable to properly
essential for the growth and seasonal replacement of antlers,
clear ketamine (J. Blake, unpublished data), therefore they
has an overriding influence of the timing of the antler
were immobilized using only 0.1–0.2 mg/kg of xylazine hy-
drochloride. Drugs were administered via hand injection,
In the female reindeer the regulation of the antler cycle
pole syringe, or blow pipe. The method of administration
has been investigated only rarely (22,23). The results of
was determined by animal temperament and human safety
these studies indicate that in intact females ovarian steroids,
consideration with the primary intent to minimize excite-
possibly estradiol, participate in antler mineralization. In
ment. After blood collection and antler measurement the
ovariectomized animals the hardening of antlers may have
reindeer received an intravenous injection of yohimbine
been induced by androstenedione, which is most likely of
(0.125 mg/kg) to reverse the xylazine.
adrenal origin (23).
Once immobilized, three consecutive samples of blood
In view of the many gaps which exist in the elucidation
(35 ml each) were drawn from the jugular vein into heparin-
of the endocrine regulation of the antler cycle in the male
ized tubes at 10 min intervals. The blood was centrifuged
and female reindeer we decided to perform a detailed inves-
within 4 hr of collection and plasma was separated and fro-
tigation of the seasonal changes of reproductive hormones
zen at 220°C. Life history events were recorded for each
in relationship to antler cycles of both sexes. As the timing
animal throughout the study period. The antlers were usu-
of casting of antlers differs in pregnant (PG) and non-preg-
ally polished within a two-day period and mostly both at
nant (NP) cows (26), our present study aimed to investigate
the same time. The casting was also mostly synchronous; in
each group separately.
cases of a desychronous casting an average date of bothdrops was recorded. As the date of initiation of antler re-growth we have chosen the period when a complete re-epi-
MATERIAL AND METHODS
thelization of the pedicle wound occurred. This was fol-
Animals and Sampling Procedure
lowed almost immediately by a vigorous antler growth. The
Experimental protocols for this study were approved by the
data of parturition were recorded daily for the PG group.
University of Alaska Fairbanks campus-wide animal welfarecommittee (IACUC #92-016).
Fourteen adult reindeer (Rangifer tarandus
) were housed
outside at the Large Animal Research Station and at the
The concentrations of LH, FSH, testosterone, prolactin, es-
Biological Reserve, Institute of Arctic Biology, Fairbanks,
tradiol, and progesterone were determined in plasma in du-
AK (latitude 66°N). The animals were divided into 3
plicate by radioimmunoassays, according the techniques de-
groups: 2- to 6-year-old bulls (n 5 5), 2- to 5-year-old non-
scribed in previous papers. For each hormone all samples
pregnant cows (n 5 4), and 2- to 5-year-old pregnant cows
were assayed at the same time.
(n 5 5). The females that were selected to become pregnant
LH was determined by a homologous bovine assay with
were kept with bulls during the entire rutting season and
no crossreactivity to other pituitary hormones (43,44). The
some time thereafter. The females selected to be non-preg-
reference preparation was a bovine pituitary LH-DSA. The
nant were kept apart from bulls in a separate enclosure ap-
sensitivity was 0.05 ng/tube; intra-assay coefficient of varia-
proximately 300 m away. All animals were kept in large
tion (CV) averaged 8.6%, and interassay variation was
paddocks with constant access to pasture grasses. They also
11.5–14%. The reference preparation was a pure bovine pi-
received a diet composed of 85% textured pelleted reindeer
tuitary extract prepared in our laboratory (LH-DSA with a
feed (dry matter values ranged from 15–15.6% digestible
biological activity of 1.0 times NIH-LH-S1). The assay was
protein, 9.6–10.8% acid detergent fiber, 77.5–78.5%
validated for reindeer plasma by recovery studies of 4 differ-
TDN), and 15% chopped grass hay (dry matter values
ent concentrations added to plasma and was on average 97.5
ranges from 5–9% digestible protein, 38.6–41% acid deter-
6 6.2%. Dilution curves of reindeer plasma with a high LH
gent fiber, 56.8–57.9% TDN). Consumption in reindeer
content run parallel to bovine standard.
varies seasonally (56) and ranged from approximately 1–4
FSH was determined by a system where pure ovine FSH
kg/animal/day. Consumption of grass pasture was not mea-
was used in labeling, and the antisera against ovine FSH
sured. All animals were provided with ad libitum
was produced in guinea pigs (43,46). As a reference prepara-
the summer and snow in the winter.
tion we used USDA-bFSH-B1. The sensitivity was 5 ng/
Every 3 weeks, from June 5, 1992 to June 18, 1993, each
tube and intra-assay CV was 7.5%, an interassay CV was
animal was immobilized using 1–2 mg/kg xylazine hydro-
12–18%. Prolactin was determined by a homologous bovine
chloride (Anased, Lloyd Labs. Shenandoah, IA, U.S.A.)
assay. The antisera produced in rabbits exhibited no cross-
and 1.5 mg/kg of ketamine hydrochloride (Ketaset, Aveco
reactivity to other pituitary hormones. The reference prepa-
Co., Fort Dodge, IA, U.S.A.). Doses varied depending on
ration used was USDA-bPRL-B1 (biological activity 0.49 3
Reproductive Hormones in Reindeer
1). The sensitivity was 0.05 ng/tube, intra-assay
Statistical values for analyzed hormones
CV was 8.4%, and the interassay CV was 14%. Mean recov-
ery of added FSH to reindeer plasma (4 different concentra-
tions) was 95.6 6 4.2% and the dilution curve of the rein-
deer plasma run parallel to the standard.
PRL was determined by a homologous bovine assay (45).
The antisera produced in rabbits exhibited no crossreactiv-ity to other pituitary hormones. The reference preparation
used was USDA-bPRL-B1 (biological activity 24.1 i.u./mg).
Validation experiment showed a mean recovery of added
bovine prolactin to reindeer plasma of 98.1 6 4.1% andthe dilution curves of reindeer plasma run parallel to the
Testosterone was determined by a highly sensitive en-
zyme immunoassay (EIA) after extraction with tertiarybutylmethylether/petrolether 30/70 (V/V) (16). The sensi-
tivity was 50 pg/ml. The interassay CV was 7.5–12% and
the interassay CV was 3.5–5.9% respectively.
Estradiol-17 beta was determined after some modification
of the original method (55) by EIA, 2 ml of plasma was
extracted with 5 ml of tertiar-butyl-methylether, evaporated
and redissolved in 0.5 ml of assay buffer and evaluated in
50 ul. The sensitivity was 0.1 pg/tube. The interassay CV
was in range between 7.9–13.5% and the intra-assay CV
was 4.1–7.2%, respectively.
Progesterone was evaluated directly in duplicate using 20
ul samples of blood plasma by an EIA in microtitre plates
based on the double antibody technique (35). The sensitiv-ity was 200 pg/ml. The interassay CV was in the range of7.2–13.1% and the intra-assay CV was between 3.1–6.2%,respectively.
There were overall differences for all hormones (P
The determination of hormone concentrations was per-
0.001). All hormones differed by Category (P
, 0.001) and
formed individually in all three samples and the statistical
, 0.001). Age differences were found in some cases
calculations were then performed on all data.
but trends were unclear and inconsistent (Table 1).
Residual correlation indicated significant relationship be-
tween gonadotropins and sexual hormones mostly in males.
Fewer correlations were found for females (Table 2).
The data on seasonal changes in plasma concentrations ofhormones (except those of progesterone) were subjected
Antler Cycles, Rut, and Parturition
to the General Linear Models Procedure for UnbalancedANOVA (SAS). Classes were ‘‘Order of sampling'' (one to
The range of dates of antler polishing, the beginning of ant-
three), ‘‘Category'' (males, NP and PG cows), ‘‘Date'' (19
ler growth, the casting, and the parturition are presented in
dates of sampling), and ‘‘Age'' (Males 2, 3, or 5 and older,
Females 2, 4 or 5 and older). Progesterone was analyzed forfemales only. The classes Date and Age were applied in the
LH, T, and FSH in Bulls
model as Nested effects within the class Category. Least-square means (LSMEANS) were computed for each class
In bulls, LH reached peak levels (1.98 ng/ml) in May (late
and differences between classes were tested by t
March vs April, P
, 0.001, April vs May, P
, 0.001) andthe minimum (,0.1 ng/ml) was observed in January (lateJune vs July, P
, 0.001, July vs early August P
Mean T levels began to increase at the time when LH levels
were already declining (Fig. 1). A steep increase of T fol-
No differences (P
. 0.05) were observed based upon Order
lowed in August, achieving peak levels (11.8–11.5 ng/ml)
of sampling. Table 1 shows results of the GLM analysis.
from the end of August until mid-September (late August
G. A. Bubenik et al
TABLE 2. Residual correlation coefficients between selected hormones by category
centrations of LH, FSH, and T were observed in the twomost aggressive bulls.
LH, FSH, and T in Cows
In pregnant cows peak LH concentrations (0.84 ng/ml)were detected at the end of August, the beginning of therut, and then declined to levels below 0.1 ng/ml. In the NPcows LH reached also a peak in August but, due to a large(pre-ovulatory) peak (7.3 ng/ml) detected in one female,the LSMEANS reached 2.3 ng/ml. A second LH peak ob-served in that group in November matched a similar peakof FSH (both LH peaks being significantly higher than anyother value P
, 0.001) (Fig. 1). The pregnant group exhib-ited a FSH peak (25 ng/ml) in mid-November (Fig. 1). Sim-ilarly, FSH concentrations in the NP group were almostidentical to FSH values in the PG group except for a sharppeak (40 ng/ml) detected in November (this FSH peak wassignificantly higher than any other value P
, 0.001) (Fig.
1). Variations of T levels in females (,0.05–0.3 ng/ml) wasnot related to the antler cycle either in the PG or in theNP females. Interestingly, in both female groups T levels ex-hibited a very similar seasonal pattern with an unexplainedsharp decrease in December and May (Fig. 1, insert).
Peak estradiol (E2) levels in bulls (3.5 pg/ml) were reachedduring the rut in mid-August (P
, 0.001) (Fig. 2).
In NP cows E2 reached a maximum (2 pg/ml) in January
FIG. 1. Least-square means (
6S.E.) of seasonal levels of LH,
and in the PG females the peak concentration (3 pg/ml)
FSH and testosterone in the adult reindeer males and the
non-pregnant and pregnant females. Testosterone insert:
was detected in April, shortly before the parturition (P
values in females.
0.001). Unlike in males, E2 levels in females varied indepen-dently from the levels of testosterone (Table 2).
The elevation and decline of E2 in bulls correlated with
vs early August (P
, 0.001), the peak period of the rut. The
the mineralization and casting period of antlers, but a simi-
FSH levels began to rise significantly in April and achieved
lar relationship was not seen in the females.
maximum (51 ng/ml) in late May (April vs early May, P
, 0.001, early May vs late May, P
, 0.001) and June (Fig.
1). In males, T levels correlated relatively well with the
development of antlers. The polishing occurred at the timeof rising T levels and the antlers were cast at the time of
In the PG females, progesterone (P) concentrations reached
minimal T concentrations. Interestingly, the highest con-
peak levels (8.8 ng/ml) at the end of March, about one
Reproductive Hormones in Reindeer
TABLE 3. The range of dates of antler polishing, antler casting, new antler growth, and parturition
New antler growth
Aug. 8–Sep. 1 (Aug. 19)
Dec. 7–Jan. 17 (Dec. 27)
Feb. 5–Mar. 20 (Mar. 3)
Aug. 30–Sep. 15 (Sep. 9)
Mar. 26–Apr. 9 (Apr. 2)
Apr. 22–Apr. 28 (Apr. 25)
Apr. 4–May 1 (Apr. 24)
Apr. 25–May 4 (Apr. 29)
Apr. 4–May 9 (Apr. 23)
NP 5 non-pregnant; PG 5 pregnant.
and bulls in May and June (Fig. 2). Both female groupsreached almost identical peak concentrations (135 NP vs140 ng/ml PG) as compared to the significantly lower peakof bulls (92 ng/ml). One female that gave birth 3 weeksbefore the rest of the group exhibited an early elevationof prolactin. However, during the late lactation period theconcentrations in both female groups were very similar.
In previous studies, the seasonal levels of reproductive hor-mones investigated in reindeer of both sexes followed a pat-tern established earlier in other cervids. However, there wasoften little information available about the relationship be-tween the peripheral levels of sexual steroids and the indi-vidual phases of the reindeer antler cycle.
LH, FSH, T.
In bulls, the sequential activation of the re-
productive system starts with an almost simultaneous eleva-tion of LH and FSH in April. LH achieved peak levels inMay, about one month before FSH (Fig. 1). Similar to val-ues reported by Stokkan and co-workers (51), LH valuesdecline from May on and were already below 1 ng/ml dur-ing the rut. As in other cervids (9,12,49,53), T exhibitedpeak levels in August and September (i.e., the rutting sea-son), more than 3 months after the LH maximum was de-tected. LSMEANS peak levels of T in our study (11.8 ng/ml) were in the range similar to 10 ng/ml observed by Stok-
FIG. 2. Least-square means (
6S.E.) of seasonal levels of es-
kan and co-workers (51). Highest T levels (60 ng/ml) re-
tradiol, progesterone and prolactin in the adult reindeer
ported by Whitehead and McEwan (57) were also compara-
males and the non-pregnant and pregnant females.
ble with the highest concentration observed in one of ourbulls (41 ng/ml).
Generally, the highest individual concentrations of LH,
month before parturition. After parturition P
FSH, and, subsequently, T were detected in the most aggres-
to non-detectable levels (,0.2 ng/ml) (P
sive bulls, exhibiting also the largest antlers. These observa-
In NP females, P concentrations increased and decreased
tions are in accordance with our study in white-tailed deer,
several times before reaching peak levels (4.8 ng/ml) in
where highest T and LH levels were detected in the most
March (Fig. 2). However, statistically the early March peak
aggressive bucks of prime breeding age (10). Similar correla-
did not differ from the February values.
tion between T concentrations and the fighting rank wasreported in reindeer by Stokkan and co-workers (51).
The increase of T levels observed during the rapidly de-
Prolactin concentration exhibited a typical seasonal varia-
clining concentrations of LH is mostly due to an altered
tion with significant peak levels detected in all cows in May
sensitivity of the reproductive system to the stimulation by
G. A. Bubenik et al
GnRH. In the early stages of the reproductive activation a
Tyler (23) concluded that E2 is the main regulator of the
small dose of GnRH induces a rapid elevation of LH but a
antler cycle in the female reindeer and adrenal androgen
small increase of T; in the later stages, the same dose GnRH
androstenedione may be the secondary steroid involved in
will cause a massive increase of T but a small one of LH
antlerogenesis. As the relationship between the testoster-
one and antler cycle in the male reindeer is less pronounced
Unlike in other cervids, some males did not finish the
than in other cervids, it could be speculated that in addition
polishing of antlers before T levels reached a seasonal maxi-
to T, estradiol could play a role of a secondary steroid in-
mum of 11.8 ng/ml (see Fig. 1). Conversely, the casting of
volved in the male reindeer antlerogenesis.
antlers occurred in some bulls more than 6 weeks after adecline of T levels to a baseline (,0.5 ng/ml). Similar delay
in antler casting after a decline of T to basal levels (lastingfor 2–3 months) was observed also in caribou by Whitehead
LH, FSH, T.
In females, the evaluation of hormonal data
and McEwan (57). This differs from the observations in
in relationship to their reproductive activity is difficult, as
white-tailed deer, where T levels detected during the time
the 3 week period between the samples is too long to detect
of antler cleaning (around 1.5 ng/ml) are similar to levels
rapid changes in the fluctuation of reproductive hormones.
detected during the time of antler casting (7). In addition,
In all cows, peak levels of LH were detected during the rut
antler casting was not achieved with an intramuscular ad-
(28 August); however, the presence of 2 peaks of LH in the
ministration of 100 or 200 mg/animal/week of androgen re-
NP cows may indicate that subsequent estrouses occurred
ceptor blocker cyproterone acetate given to yearling male
in that group. This assumption is further supported by the
and female reindeer (G. Bubenik et al.
, unpublished obser-
data on FSH where in the NP cows several oscillations were
vation). These data indicate that androgen receptors in
detected before peak levels were observed on November 6
reindeer antlers are relatively slow to respond to rapid
changes of circulating androgen levels.
The much higher elevation of average LH levels observed
The onset of new antler growth varied considerably
in the non-pregnant group was largely due to a massive in-
within our bull group (February 5–March 20) but it usually
crease of LH (7.2 ng/ml) observed in one cow. Such a rapid
started about 8 weeks after casting (December 7–January
(preovulatory) increase of LH which lasts only several hours
20). As expected, the two more aggressive bulls cast the
(3,33) is difficult to detect and was probably missed in most
antlers earlier than the three tamer ones. Our casting dates
of our other animals.
were in variance with the data of Lincoln and Tyler (23)
Seasonal variation of testosterone levels in both groups
who reported the casting of antlers in their bulls already in
of females was remarkably similar, with both groups exhib-
November–December but the new antler growth began
iting a non-significant trend with a decrease in December
only 3–4 months later, in April. This difference could be
and May. The time course of T was not in any case related
either due to the more northern latitude of norvegian rein-
to the milestones of antlerogenesis, such as the polishing or
deer (69.2°N), different nutritional conditions or due to the
casting of antlers (Fig. 1, insert).
genetic differences between groups. Persisting individualdifferences in the casting dates (up to 2 months apart) have
Seasonal levels of estradiol stayed below 1
been reported in white-tailed deer (6).
pg/ml for most of the year. Only in the pregnant cows,LSMEANS of E2 rose to 3 pg/ml at the beginning of parturi-
Maximum male estradiol levels (3.5 ng/ml),
tion, three weeks after peak concentration of progesterone
reached during the peak of the rut, were surprisingly higher
were detected (Fig. 2). This slow elevation of E2 was in
than peak levels observed in pregnant females (3.0 ng/ml)
agreement with Baksi and Newbrey (4) who reported that
detected shortly before parturition in April (Fig. 2). Simi-
E2 concentrations in pregnant reindeer cows in January
larly, concentrations of estradiol in the feces of caribou bulls
were only 50% higher than values detected in June. Similar
were reported to be comparable to levels of non-pregnant
slow rise in E2 levels in Rangifers
was also reported by Mess-
cows (27). A significant correlation found between T and
ier and co-workers (27) who noted that until day 150 of
E2 may indicate that, similar to stallions (36), reindeer tes-
pregnancy no difference was detected between E2 of preg-
tes are aromatizing considerable amount of T into estradiol.
nant and non-pregnant cows. Conversely, the rapid eleva-
Whether estrogens are involved in the mineralization of
tion of E2 shortly before parturition is in agreement with
reindeer antlers has not yet been investigated, however ex-
data reported by Blom and co-workers (5) in reindeer cows,
perimental evidence indicates that E2 is 10–503 more po-
Plotka and co-workers (32) in white-tailed does, and by
tent in the mineralization of antlers than T (14,30). In addi-
Kelly and co-workers (18) in red deer hinds. The variation
tion, growing antlers exhibit estradiol receptors (20) and an
in LSMEANS of E2 concentration in our study (1–3 pg/
experimental blockade of E2 receptors interrupts the miner-
ml) was very similar to data of Lincoln and Tyler (23) who
alization process in the compact antler bone mantle, re-
reported average E2 values varying seasonally between 2–
sulting in severe reduction of its thickness (8). Lincoln and
6 pg/ml. However, probably because of species differences,
Reproductive Hormones in Reindeer
variations in laboratory techniques and because of an infre-
PG and NP caribou during the first 50 days of pregnancy.
quent sampling intervals, our peak values of E2 (3 pg/ml)
Finally, P levels were also found elevated in NP cows in the
were much lower than the ones reported by Plotka and co-
study of McEwen and Whitehead (26).
workers (31) in the white-tailed deer (180 pg/ml) or by
It could be argued that a sharp drop in progesterone levels
Kelly and co-workers (18) in the red deer (35 pg/ml).
at the end of pregnancy may be related to antler casting.
The rapid decline of E2 levels, which correlated with the
However, whereas there is plenty of evidence about the role
time of parturition was usually followed by the casting of
of estradiol in the antler cycle (as mentioned above), there
antlers (Fig. 2). Similar to reports given by McEwen and
is little evidence that progesterone levels can decisively in-
Whitehead (26) and Lincoln and Tyler (23), the NP cows
fluence antler mineralization and retention. Whereas Jac-
cast antlers earlier than the pregnant ones (Table 3). Based
zewski (15) reported partial or complete velvet shedding
on the results in ovariectomized cows, Lincoln and Tyler
and retention of antlers in some castrated red deer stags
(23) concluded that ovarian E2 is the major steroid regulat-
treated with large doses of P, neither Waldo and Wislocki
ing the antler cycle in the female reindeer. This appears
(54) in white-tailed deer nor Goss (13) in sika deer were
to be true in some females, which cast their antler after
able to induce velvet shedding in castrated bucks treated
parturition. However, in our non-pregnant females no de-
with P. Furthermore, McEwen and Whitehead (26) ob-
cline of E2 was observed before their casting (Fig. 1) and in
served antler casting in captive pregnant reindeer from Jan-
the pregnant group two cows cast their antlers several days
uary until March (the period of high P) whereas in the wild
before parturition. As on few occasions, we have observed
females the casting occurred mostly after calving in June or
antler casting in the pregnant cows more than 2 weeks be-
July (the period of very low P). Therefore, it appears that P
fore parturition and McEwan and Whitehead (26) reported
does not play a crucial role in the antler cycle of the female
a casting period 2 months before the calving, it was con-
cluded that a sudden drop in E2 levels is probably not theonly factor initiating the casting of antlers in the female
PROLACTIN IN MALES AND FEMALES.
Prolactin levels ex-
hibited a typical seasonal curve with peak levels observedin May and June and trough concentrations detected from
The seasonal variations of progesterone
September until March (12). In both female groups, maxi-
concentrations detected in our pregnant cows (Fig. 2) con-
mum concentration exceeded that of the bulls (Fig. 2). This
firmed the expected time course with highest levels (9 ng/
is in agreement with PRL data on white-tailed deer and
ml), observed shortly before parturition (Table 3) [for re-
red deer, where peak female levels were higher than ones
view, see (38)]. Similarly to some bovids, ovaries are the
observed in males (18,47). Time courses of PRL levels in
essential source of P in the pregnant cervid female (34).
the individual cows indicate that the period of parturition/
Peak P levels (9 ng/ml) detected in pregnant reindeer, were
lactation coincides with the increase of PRL levels, however
higher than 5 ng/ml reported in pregnant red deer (2), 4.6
a continuation of lactation in ruminants does not appar-
ng/ml found in white-tailed deer (31) or 3.2 ng/ml observed
ently require peak PRL concentrations (17). This was con-
in roe deer by Schams et al.
(43). However, they were very
firmed in our study, as the sharp decline in PRL levels was
similar to 10.9 pg/ml reported in reindeer by Blom and co-
observed in both the lactating as well as the non-lactating
workers (5) or 8.4 ng/ml observed in roe deer by Sempere
cows. However, this finding was in contrast to data of Adam
(48). Whereas in red deer P concentrations were main-
and co-workers (1) who, later in the lactation period, re-
tained high for most of the second and third trimester and
ported higher PRL concentrations in the plasma of lactating
calving occurred only after P dropped precipitously (2,18),
hinds, as compared to the non-lactating ones.
in our reindeer P levels rose steadily until peak levels in
Because in some cervids new antler growth coincides
March, some 3–4 weeks before parturition and then de-
with PRL increase in the spring, Lincoln and Tyler (23)
clined sharply. Similar steady rise in P values during the
suggested that PRL may be involved in the reinitiation of
pregnancy, followed by parturition and a subsequent rapid
antler growth. This hypothesis is not supported by our data
decline, was also reported by Blom and co-workers (5).
as a new antler growth was observed in some reindeer bulls
In contrast, the peak levels in the NP cows were not only
already in February, the time of minimal levels of PRL de-
smaller (6.0 ng/ml) but also exhibited an irregular variation
tected in our study (see Fig. 2). In addition, new antler
(Fig. 2). The time course of P levels in our NP females may
growth in tropical rusa deer is initiated during the decline
be the result of seasonal polyestrous as demonstrated by
of PRL levels (53) and in the subtropical Eld's deer, PRL
Plotka and co-workers (33) in white-tailed deer and re-
levels rise when the antlers are hard (29).
cently by Ropstadt and co-workers (37) in reindeer. Simi-
In conclusion, our studies indicate that in reindeer, the
larly, during the first trimester, peak P levels in NP white-
individual milestones of their antler cycle are less related
tailed does were not different from concentrations recorded
to the seasonal variation of circulating levels of sexual hor-
in the pregnant ones (31). Similar data were reported by
mones than in most other cervids. In that case, either other
Messier and co-workers (27) who investigated P in feces of
hormones than those investigated in this study are involved
G. A. Bubenik et al
of progesterone on the antler cycle in red deer (Cervus ela-
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berger, E.; Schams, D. Testosterone, luteinizing hormone(LH) and follicle stimulating hormone (FSH) in peripheral
We would like to thank Don Hartbauer and the staff at the Animal
plasma of bulls: levels from birth through puberty and short-
Quarters and Bill Hauer and the staff at the Large Animal Research
term variations. Zentbl. Vet. Med. A. 23:793–803;1976.
Station, University of Alaska, Fairbanks for their assistance in mainte-
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nance and sampling of the reindeer, Peter Bubenik and Yoko Imai for
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Animal Hormone Program, Beltsville, MD, U.S.A.) for the generous
Plasma concentrations of LH, prolactin, oestradiol and pro-
gift of pituitary hormones. The project was supported by NSERC Can-
gesterone in female red deer (Cervus elaphus
) during preg-
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say for bovine follicle-stimulating hormone and its application
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Advanced Software Protection Now Diego Bendersky1,2, Ariel Futoransky1, Luciano Notarfrancesco1, Carlos Sarraute1,3, and Ariel Waissbein1,3 1 Corelabs, Core Security Technologies; 2 Departamento de Computaci´on, FCEyN, Universidad de Buenos Aires (UBA), 3 Departamento de Matem´atica, FCEyN, UBA Argentina. Abstract. We introduce a novel software-protection method, which canbe fully implemented with today's technologies, that provides traitortracing and license-enforcement functionalities, and requires no addi-tional hardware nor inter-connectivity other than those needed to ex-ecute the respective protected software.In  authors introduce the secure triggers and show that it is secure,modulo the existence of an ind-cpa secure block cipher. In this work,we present a framework for license enforcement and fingerprinting ofcomputer programs in which these capabilities are coupled with securetriggers to produce a secure software protection system.