Seasonal changes in plasma leptin concentration related to antler cycle in Iberian red deer stags

The aim of this study is to describe the leptin cycle in male Iberian red deer (Cervus elaphus hispanicus) and relate it to antler and testosterone cycles. An additional aim is to assess the relationship between the plasma leptin concentration during antlers’ growth and their final size. Therefore, blood from 21 Iberian red deer males was sampled monthly to analyse leptin and testosterone. At the same time the deer were weighed and their body condition was assessed. The length of antlers was measured every 2 weeks and, after casting, their final length and perimeters were taken. Leptin showed a seasonal cycle, with a peak in June that decreased as testosterone increased. Low values were observed in autumn, winter and early spring. The relationship observed between leptin and body mass or body condition score was different in spring, when plasma testosterone concentration is low, than in autumn, when testosterone increases. Leptin peak amplitude was positively related to final antler size. In conclusion, the relationship between leptin and body mass and body condition score changes through the year, possibly due to the influence of androgens and photoperiod. There was a positive relationship between plasma concentration of leptin during antler growth and final antler length.     

Changes in food intake,body weight,gonads and plasma concentrations of thyroxine,luteinizing hormone and testosterone in captive male buntings exposed to natural daylengths at 29° N

We have investigated the seasonal changes in food intake, body weight, gonadal volume and plasma concentrations of thyroxine, luteinizing hormone and testosterone in male blackheaded bunting (Emberiza melanocephala) in captivity under natural daylengths at 29° N. The cycles in food intake, body weight and testis size in buntings appeared to be phase related. While the changes in body weight and testicular size were parallel to each other and correspond to the increasing daylengths of spring and early summer, cycle in food intake was almost antiphase to the cycles in body weight and testicular growth and development. Furthermore, buntings showed a distinct seasonal cycle in plasma concentrations of thyroxine, luteinizing hormone and testosterone. It is suggested that these seasonal cycles in buntings are endogenously programmed and their entrainment to the environmental photoperiod ensures the occurrence of different physiological functions at temporally fixed time of the year.     

Endocrine control of antler growth in red deer stags

Observations of body weight, testis size, antler status, plasma testosterone and prolactin were made on 12 red deer stags during their first 2 years of life. Six of the stags were fed to appetite throughout the study (Group A) and 6 were fed a 70% restricted diet during each winter (Group B). In addition 6 of the stags , 3 from each group, were studied in more detail; LH and testosterone were measured either after a single injection of LH-RH or in samples taken at frequent intervals over a period of 8 or 24 h. During the study the stags became sexually mature, developed first their pedicles and then antlers and showed at least one complete cycle of casting and regrowth of the antlers . The stags in Group A developed their testes and pedicles about 2 months earlier than did those in Group B. Pedicle initiation was associated with increasing plasma testosterone levels in response to changes in LH secretion, and antler development occurred when testosterone levels were low or decreasing. Cleaning of the velvet was associated with high levels of plasma testosterone. Antler casting occurred when plasma testosterone concentrations were low or undetectable and prolactin levels were high or increasing. The relationship between LH and testosterone varied during the study; in spring when the testes and antlers were growing, relatively high levels of LH were associated with only small peaks of testosterone, yet in summer, when antler growth was complete and the antlers were clean of velvet, low LH concentrations were associated with large peaks of testosterone.     

Abnormal testes in reindeer, Rangifer tarandus.

In a sample of reindeer from South Georgia, 4 males were abnormal in that 1 had bilateral cryptorchid testis, 2 each had 1 cryptorchid testis and 1 had a vestigial testis. The antlers of the cryptorchid males were small, but the antler cycle itself was hardly affected.     

Evidence that the seasonal antler cycle of adult Reeves' muntjac (Muntiacus reevesi) is not associated with reproductive quiescence

Data were obtained from post-mortem investigations of 190 culled and road-killed muntjac bucks between 1967 and 1989. Although adult bucks (i.e. those having undergone at least one antler cycle) have a synchronous annual antler cycle, unlike temperate-zone cervids there was little seasonal variation in testis size or activity, or in the size and activity of the epididymidis or accessory reproductive glands. Spermatogenesis was not abated when the antlers were in velvet and year-round fertility was achieved without additional sperm storage. There was little seasonal change in plasma testosterone concentrations in samples obtained from captive and free-living bucks although castration caused antler casting and prevented mineralization. Hence the data are equivocal as to the role of steroids in driving the antler cycle; experimental work on this species would be valuable in examining the mechanisms which regulate the antler cycle.     

Annual changes in testicular development and plasma sex steroids in the captive male dojo loach <Emphasis Type="Italic">Misgurnus anguillicaudatus</Emphasis>

Testicular development in the captive male dojo loach Misgurnus anguillicaudatus was examined monthly in relation to the levels of plasma sex steroids [testosterone (T), 11-ketotestostrone (11-KT), and 17,20β-dihydroxy-4-pregnen-3-one (DHP)]. On the basis of testicular histology, the annual gonadal cycle was found to be divisible into 3 periods: the recovery and proliferation period, which mainly consists of early spermatogenic testis from August to November (reproductive phase I); the preparation period for the next spawning period, which mainly consists of late spermatogenic testis from December to April (reproductive phase II); and the mature period, characterized by a high proportion of mature testis from May to July (reproductive phase III). Individual variability in testicular development was high, and continuous spermatogenesis was observed throughout the year. High levels of plasma T, 11-KT, and DHP were observed during reproductive phase III. 11-KT began to increase in February, while T was present at low levels in reproductive phase II. These results suggest that the physiologically active season of testis development for breeding in the dojo loach is from May to July, although spermatogenesis occurs throughout the year.     

Basic properties and annual changes of follicle-stimulating hormone receptors in the testis of horseshoe bats, Rhinolophus ferrumequinum

The unique reproductive patterns, delayed fertilization in females, and asynchrony between spermatogenesis and mating behavior in males are well documented in bats living in temperate latitudes. The present study was undertaken to examine follicle-stimulating hormone (FSH) receptors in the testis of bats, Rhinolophus ferrumequinum, during the annual reproductive cycle. Male bats were captured at natural roosting sites and testicular preparations were subjected to a radioligand binding assay for FSH receptors. The weight of paired testes increased considerably in the spermatogenic period and decreased from the mating to hibernation periods. Meiotic division in the testis was observed in the spermatogenic period but not the mating period. Serum testosterone concentrations increased in the spermatogenic period and rapidly decreased in the mating period. The binding of FSH was specific for mammalian FSHs and detected primarily in the testis. Scatchard plot analyses of the binding of FSH to bat testicular preparations showed straight lines, suggesting the presence of a single class of binding sites. The affinities (equilibrium association constant) of FSH receptors were consistent throughout the annual reproductive cycle. The specific binding per unit weight of testis and total binding in the paired testes were highest in the mating period and in the spermatogenic period, respectively, among reproductive periods. The accumulation of cyclic adenosine 3', 5'-monophosphate to FSH stimulation was higher in the spermatogenic period than in the hibernation period. These findings suggest that testicular function of bats is associated with seasonal changes in the number of binding sites, while the number per target cell and the activation of adenylate cyclase led by FSH-receptor complex considerably decreases in the hibernation period.     

Seasonal changes in fecal testosterone concentrations and their relationship to the reproductive behavior, antler cycle and grouping patterns in free-ranging male Pampas deer (Ozotoceros bezoarticus bezoarticus)

The purpose of this study was to validate noninvasive endocrine monitoring techniques for Pampas deer and to evaluate seasonal changes in testicular steroidogenic activity and their correlation to reproductive behavior, antler cycle and group size. Thus, fecal samples, behavioral data and observations of antler status were collected at monthly intervals during 1 year from free-ranging Pampas deer stags (three radio-collared individuals and 15 random individuals) living in Emas National Park, Brazil (18 degrees S latitude). Fecal steroids were extracted using 80% methanol and steroid concentrations were quantified by a commercial enzyme immunoassay (EIA). Fecal testosterone concentrations peaked in December-January (summer), March (early autumn) and in August-September (winter-spring), with minimal values from April-July. Reproductive behavior had two peaks, the first in December-January, characterized by predominately anogenital sniffing, flehmen, urine sniffing, chasing and mounting behavior, and the second peak in July-September (behavior primarily related to gland marking). There were significant correlations between fecal testosterone and reproductive behavior (r=0.490), and between fecal testosterone and antler phases (r=0.239). Antler casting and regrowth occurred under low testosterone concentrations, whereas velvet shedding was associated with high concentrations of testosterone. We inferred that Pampas deer stags exhibited a seasonal cycle that modulated sexual behavior and the antler cycle, and we concluded that fecal steroid analysis was a practical and reliable non-invasive method for the evaluation of the endocrine status of free-ranging Pampas deer.     

Effect of antiandrogen cyproterone acetate on the development of the antler cycle in Southern pudu (Pudu puda)

The antler cycle of pudu is similar to other cervids, but unlike most boreal deer, male Southern pudu (Pudu puda) exhibits two seasonal peaks of LH and testosterone. In that respect, pudu is similar to roe deer. Whereas the antler cycle in some deer species, such as roe deer or white-tailed deer, is very sensitive to variation of testosterone, in other cervids, such as fallow deer or reindeer, a blockade of androgens with cyproterone acetate (CA) has little or no effect on the timing of the antler casting. In order to test the sensitivity of pudu antlers to variations of androgens, CA (administered 2x weekly at 50 mg/buck) was injected intramuscularly for 3 weeks in 5 adult male pudu, starting February 19 (late summer). Four other males of similar age served as controls. The experiment was performed at the University of Concepcion, Chile, latitude 36.6 degreeS. Blood samples were taken once a week between January 19 and April 3. In CA-treated bucks, the antlers were cast approximately 3 weeks after the initiation of CA treatment and a new antler growth began almost immediately. The antlers reached about 5 cm in length, before ceasing to grow at the end of April, when they became mineralized and were subsequently polished. CA had no effect on the already declining levels of LH. Plasma levels of testosterone in controls increased from February 15, whereas in CA-treated bucks remained depresses until March 21. It is concluded that similarly to white-tailed deer, the antler cycle of Southern pudu is very sensitive to manipulation of androgen levels.     

Effect of advancing births on testosterone until 2.5 years of age and puberty in Iberian red deer (Cervus elaphus hispanicus)

This study aimed to determine the age of attainment of puberty, as well as the pattern of reproductive seasonality in males of Iberian red deer and the relationship of these factors with antler development. A second aim was to examine the effect of the advance of births on the variables mentioned. Fourteen males of Iberian deer were used, five of them born 2.5 months before the others (early group). Male deer were blood sampled from birth until 2.5 years of age to assess the time course of plasma testosterone. The study also included measurements of testicular and neck circumference. As in most cervids, concentration of plasma testosterone was associated to the phases of antler development. Early development affected testosterone titres, since the most precocious males in antler growth also showed higher levels of testosterone during the first and second year of age. In the same way, results suggested that the advance of calving may result in puberty attainment in the first autumn of its life. However, this would require an advance of calving greater than the one we induced since puberty in the Iberian deer is usually attained during second autumn of life at 15-16 months of age. Finally, calving advance also affected antler development. Thus, two of the five males whose birth was advanced, showed two antler cycles in their first year of life.     

Testosterone and FSH have independent,synergistic and stage-dependent effects upon spermatogenesis in the rat testis

Summary Adult rats were hypophysectomized and treated with ethane dimethanesulphonate (EDS) selectively to eliminate the Leydig cells in the testis. By removing the source of endogenous gonadotrophins and androgens, the subsequent effects on the seminiferous epithelium were studied after 20 days of treatment with vehicle, or FSH (2x50 g/day) or a low dose of testosterone (0.6 mg testosterone esters every 3rd day) alone or in combination. Compared to vehicle-treated hypophysectomized rats with Leydig cells, testis weight in saline-treated hypophysectomized rats treated with EDS declined by 50%, spermatogenesis was disrupted severely and only 18% of the tubules contained spermatids, these being confined to stages I–VI of the spermatogenic cycle. Treatment with either FSH or testosterone esters alone significantly (P<0.01) increased testis weight compared to vehicle-treated hypophysectomized rats treated with EDS and 40% of tubules contained spermatids either at stages I–VI after FSH, or at all stages I–XIV after testosterone treatment. Treatment with FSH and testosterone esters together maintained testis weights approximately 20% above vehicle-treated hypophysectomized controls; over 70% of the seminiferous tubules contained spermatids and there was a marked stimulation of spermatogenesis at all stages of the spermatogenic cycle. The results suggest, that in the absence of the pituitary gland and the Leydig cells, FSH alone partially supports spermatogenesis up to the development of round spermatids whereas testosterone is capable of maintaining spermatid development at all 14 stages of the cycle. When FSH and testosterone were administered in combination, the effects upon spermatogenesis were far greater than the response expected if their individual effects were simply additive. It is therefore concluded that FSH may play a role in normal spermatogenesis and that this role is essentially that of augmenting the response of the testis to testosterone. The biochemical mechanisms via which this might occur are discussed and hypophysectomized rats treated with EDS used in the present studies should provide a useful approach for their identification.     

Abolition of seasonal testis changes in the Ile-de-France ram by short light cycles: relationship to luteinizing hormone and testosterone release

The purpose of this study was to examine the relationships between testis weight and the luteinizing hormone (LH) and testosterone releases in rams subjected to 14 successive 2-mo artificial light cycles in which daylength increased from 8 to 16 h in one month and decreased from 16 to 8 h the following month. Testis weights were measured fortnightly. Serial bleedings were performed during 6 to 8 h the last three 2-mo light cycles, when daylengths were 8, 12 (increasing), 16 and 12 h (decreasing) and plasma LH and testosterone were measured by radioimmunoassay. The overall mean testis weight, continuously high and equal to 314 g per testis, was not correlated with daylength. Mean LH levels and LH pulse height varied significantly according to daylength (P < 0.05 and P < 0.01, respectively) and were maximal on short days (8 h). Both mean and maximal testosterone levels presented variations according to daylength (P = 0.05). LH and testosterone pulsatility were quite high (equivalent to 7 to 10.3 pulses/day); however, mean plasma testosterone levels remained low. It is proposed that in rams under 2-mo light cycles, frequent but short stimulation of LH release by decreasing daylength phases allows maximal testis weight, while the interruption of LH stimulation by increasing daylengths prevents overstimulation of testosterone that can inhibit the LH release by negative feedback. Rams a under 2-mo light regimen presented a persistent high testis weight, indicating that seasonality had been broken down in the Ile-de-France male which is normally a marked seasonal breeder.     

Seasonal changes in the reproductive organs and plasma and pituitary hormone content of the male Bennett's wallaby (Macropus rufogriseus vufogriseus)

In a number of species of seasonally breeding marsupial, the male is fertile throughout the year but there is a marked seasonal change in weight of the accessory sexual glands. In this study, body weight, prostate, epididymis and testis weights and plasma concentrations of testosterone, LH and prolactin and pituitary content of LH and prolactin were determined in male Bennett's wallabies shot at 1–2 month intervals over a period of 17 months. There was a highly significant increase in prostate weight which was coincident with the breeding season for this species. A small but significant increase in testis weight was also observed but epididymis weight remained unchanged. Plasma testosterone concentrations were significantly increased at a time coincident with the increase in prostate weight. Plasma prolactin and LH concentrations were low in most animals and remained unchanged during the study. In contrast, pituitary prolactin and LH contents showed highly significant changes, with increasing and peak hormone content preceding maximum prostate weight and plasma testosterone concentrations by several months. While these latter results suggest a role for prolactin and LH in the seasonal control of the reproductive organs in the male wallaby, a more intensive study of the pattern of secretion of these hormones and possibly more sensitive hormone assays are required to understand their relative roles in regulating the annual cycle of prostate growth.     

Seasonal variations of LH, prolactin, androstenedione, testosterone and testicular FSH binding in the male blue fox (Alopex lagopus)

The seasonal changes in testicular weight in the blue fox were associated with considerable variations in plasma concentrations of LH, prolactin, androstenedione and testosterone and in FSH-binding capacity of the testis. An increase in LH secretion and a 5-fold increase in FSH-binding capacity were observed during December and January, as testis weight increased rapidly. LH levels fell during March when testicular weight was maximal. Plasma androgen concentrations reached their peak values in the second half of March (androstenedione: 0.9 +/- 0.1 ng/ml: testosterone: 3.6 +/- 0.6 ng/ml). A small temporary increase in LH was seen in May and June after the breeding season as testicular weight declined rapidly before levels returned to the basal state (0.5-7 ng/ml) that lasted until December. There were clear seasonal variations in the androgenic response of the testis to LH challenge. Plasma prolactin concentrations (2-3 ng/ml) were basal from August until the end of March when levels rose steadily to reach peak values (up to 13 ng/ml) in May and June just before maximum daylength and temperature. The circannual variations in plasma prolactin after castration were indistinguishable from those in intact animals, but LH concentrations were higher than normal for at least 1 year after castration.     

Effects of short photoperiod on ATPase activities in the testis of the immature Siberian hamster.

The effects of changes in photoperiod length upon body weight; spleen, thymus, and testis weights; testis protein content; testis cation pump enzyme activities; and plasma testosterone were studied in the developing Siberian hamster, Phodopus sungorus. Male hamsters were exposed to a cycle of 16L:8D (long-day), until Day 18 when half were switched to a 10L:14D (short-day) cycle, until killed 0, 2, 4, 7, 10, 12, or 15 days later. Body weight and relative testis weight (expressed as percentage of body weight) increased steadily during the first week of exposure. After 10 days, the long-day hamsters consistently weighed more (p less than 0.05). Relative testis weights in the short-day group began to decrease (p less than 0.005) within 10 days and continued to decline. Testis homogenate K(+)-pNPPase- (as a measure of Na+,K(+)-ATPase) and Mg(2+)-pNPPase-specific activities closely paralleled testis weight, with the short-day animals (p less than 0.05) differing after 10 days. Plasma testosterone levels remained below adult levels through exposure Day 15, but were relatively lower (p less than 0.05) in the short-day group after 10 days. Spleen weights were similar for the long- and short-day groups. The short-day group had larger thymus weights after 12 days (p less than 0.05), but thymus enzyme activities did not differ between the two groups. We conclude that cation pump activities in the Siberian hamster testis are significantly affected by changes in photoperiod length.     

LH and testosterone responses to GnRH in red deer (Cervus elaphus) stags kept in a manipulated photoperiod

Six red deer stags from age 4 months were kept in a light-proof room under an artificial photoperiod consisting of 5.5 cycles of alternate 2-month periods of 16 h light and 8 h dark (16L:8D) and 8L:16D. At 2 or 3 weekly intervals from 10 months of age through 4 cycles, the stags were anaesthetized with xylazine and challenged i.v. with 10 micrograms GnRH. Blood samples were withdrawn immediately before and 10 and 60 min after injection. LH and testosterone concentrations were measured in all samples by RIA. Antler status was recorded daily. Peak LH values on each sampling day occurred in the sample taken 10 min after GnRH stimulation while peak testosterone occurred in the sample taken at 60 min. There were 4 cycles of LH and testosterone secretion accompanied by 4 antler cycles in the stags. The highest LH responses were detected during short days (8L:16D), and the highest testosterone responses were detected around the time of the change from short to long days. The responses of both hormones were lowest at the end of periods of long days or the beginning of short days. The increased pituitary LH response to GnRH was evident 4 weeks after the change to short days which are stimulatory for gonadal development. Antler casting occurred at the end of long days and cleaning at the end of short days. It is considered that antler cycles were due to the ability of the stags to vary release of LH and testosterone in response to changes in the artificial photoperiod.     

Compensatory testicular hypertrophy in the lizard Anolis carolinensis

In reproductively responsive, male Anolis carolinensis undergoing artificially induced testicular recrudescence, unilateral orchidectomy of the left testis produced compensatory hypertrophy of the remaining testis. Testosterone inhibited this compensatory testicular hypertrophy on a weight basis, but did not reduce the rate of spermatogenic development. These results suggest that there is a mechanism of testosterone feedback in Anolis carolinensis that controls gonadotropin secretion during the recrudescent phase. In reproductively thermorefractory lizards, unilateral orchidectomy had no effect on the remaining testis. Administration of exogenous follicle-stimulating hormone to refractory animals increased testicular weight and stage of spermatogenic development. Sensitivity to gonadotropin, as well as failure of unilateral orchidectomy to produce compensatory hypertrophy in refractory male anoles, suggests that the control of the refractory period in A. carolinensis results from physiological mechanisms in the pituitary gland or brain rather than in the testis.     

Secondary sex characteristics and related physiological values in male fallow deer (Dama dama L.) and their relationships to changes in the annual cycle of daylengths: frequency alterations to 4- and 3-month photoperiodic cycles, and subsequent re-synchronisation under natural conditions.

Five adult male fallow deer were maintained in a barn with artificial light control. In a previous experiment, three 6-month photoperiodic cycles entrained morphogenetic and associated physiological values that revealed typical relationships to the antler cycle. Presented here, the light cycle was accelerated to three 4- and one 3-month photoperiods in the same group. Each artificial photoperiod generally resulted not only in an almost complete antler cycle but also in an entire cycle of seasonal fluctuations in neck girth. Increases in plasma levels of alkaline phosphatase (AP), total-, LDL- and HDL-cholesterol, were generally entrained and the maxima revealed positive correlations with antler formation, but the relationships slightly diverged. In neck girth and creatinine, positive correlations to the hard antler period as well as to each other prevailed but diverged. In the 3-month photoperiodic cycle, these relationships were out of synchrony. In the second 4-month cycle, two bucks "missed" shedding and subsequent casting, but commenced antler growth in the following cycle with an in-time shedding. The possibility of desynchronisation of physiological conditions and the question of an endogenous circannual mechanism interacting with daylight are discussed. At the end of the 3-month cycle, experimental indoor and natural outdoor casting was coincident so the group was transferred to outside conditions for re-synchronisation. After spending altogether 36 months in frequency altered photoperiods, the represented values were neither synchronized nor revealed their typical relationships to the antler cycle, except of AP and neck volume. In the second cycle of re-synchronisation, all parameters, except of creatinine, appeared to be resynchronized.     

Photoperiodic control of antler cycles in deer. VI. Circannual rhythms on altered day lengths

Groups of sika deer were exposed to light and dark periods of equal lengths but different from 12 hr. Light cycles were 4.94L/4.94D, 6L/6D, 8L/8D, and 21L/21D. In all experiments, deer underwent circannual cycles of antler replacement, testis size, molting, and coat color. The results indicate that the previously reported abolition of circannual cycles on 12L/12D was due to the 12-hr duration of the light or dark periods, not their equivalence. They also eliminate the possibility that the circannual cycle might be the sum of 365 circadian cycles. Circannual antler cycles appear to be expressed under artificial light cycles to which the deer cannot entrain.     

Stability of spermatogenic synchronization achieved by depletion and restoration of vitamin A in rats

Studies of synchronization of spermatogenesis following vitamin A deficiency have suggested that this may provide an in vivo model for the study of stage-dependent changes in hormonal action and protein secretion within the seminiferous epithelium. However, until now, no information on the stability or durability of this condition has been available. In this study, 200 seminiferous tubules from each of 40 rats (including controls) were classified according to their spermatogenic stage after withdrawal and replenishment of vitamin A. Following 15 wk withdrawal and subsequent replenishment of vitamin A, spermatogenesis was initiated in a synchronous fashion. This synchrony remained stable for more than 10 cycles of the seminiferous epithelium (2.5 spermatogenic cycles). In association with the extended period of vitamin A deficiency, a proportion of tubules (30%) showed morphological characteristics of either Sertoli cells only or Sertoli cells plus spermatogonia with occasional pachytene spermatocytes. During the 11-wk period of observation in this study, no significant change in proportions of damaged tubules were observed. Testicular testosterone concentrations, although elevated with respect to controls, showed no correlation with the stage of the cycle of the seminiferous epithelium observed, whereas pituitary and serum follicle-stimulating hormone levels were elevated, probably due to the number of damaged tubules observed. The persistence of synchrony in spermatogenesis following vitamin A treatment suggests that this model is applicable for studies of paracrine actions within the testis. However, the decreased ratio of synchrony observed with time may provide evidence that duration of the individual stages of the cycle of the seminiferous epithelium might be subject to temporal variation, leading to a progressive desynchronization of spermatogenesis in this model system.