Progesterone and testosterone concentrations during oestrous cycle and pregnancy in the common vole (Microtus arvalis Pallas)

Serum progesterone and testosterone concentrations were measured during different stages of oestrous and pregnancy in paired and unpaired female common voles (Microtus arvalis). Hormone concentrations were measured by ELISA, and cycle stages were determined by vaginal smears. Paired females usually had serum progesterone concentrations of more than 10 ng/ml in the oestrous cycle. A significant maximum was detected in prooestrous (51.70 +/- 7.84 ng/ml, mean +/- S.D.). Serum progesterone concentrations increased from about 40 ng/ml at the beginning of pregnancy to about 70 ng/ml on days 15 and 16. The last 2 days before parturition (days 19 and 20) were characterised by a decrease of progesterone concentrations to ca. 30 ng/ml. The maximum concentration of testosterone was found in prooestrous (1.58 +/- 0.31 ng/ml). Concentrations during pregnancy varied between 1.5 and 2.1 ng/ml. In two of three cases unpaired females exhibited progesterone values below 10 ng/ml, but with varying vaginal smear patterns. The combination of progesterone concentrations and vaginal smear patterns was found to be regular in only 23.8% of the cases. The most frequent cycle stage found was the oestrous (44.2%). Mean concentrations of progesterone (10.43 +/- 13.81 ng/ml) and testosterone (0.85 +/- 1.11 ng/ml) in unpaired females were significantly lower than in paired females, thereby denoting reproductive inactivity in the former. The study presents basic data for several parameters of the reproductive biology in the common vole and confirms the importance of combining hormone assays and vaginal smear monitoring in reproductive research.     

Intratesticular distribution of testosterone in rats and the relationship to the concentrations of a peptide that stimulates testosterone secretion

Methods have been established and validated for quantitative assessment of the distribution of testosterone in the testis, by measurement of testosterone concentrations in whole testis, in isolated seminiferous tubules and in testicular interstitial fluid. These measurements were made in individual rats injected 2-40 h previously with saline (0.9% NaCl) or a potent antiserum to ovine LH. Testosterone concentrations in interstitial fluid and seminiferous tubules were closely correlated (r = +0.98; n = 60) and their relationship was log linear over a 200-fold range. However, although the concentrations of testosterone in interstitial fluid and seminiferous tubules decreased progressively with time after LH antiserum injection, this decrease was far more pronounced for interstitial fluid. In association with this change there was a significant increase in the amounts of a locally-produced factor in interstitial fluid which stimulates basal and hCG-stimulated testosterone production by isolated purified Leydig cells. This increase was reversed by injection of hCG but not by peripheral injection of a dose (20 mg) of testosterone propionate which restored normal intratesticular concentrations of testosterone. It is concluded that the tubular 'conservation' of testosterone, which occurs as interstitial fluid levels of this steroid decrease, may be a consequence of restricted diffusion of testosterone out of the tubules, but is also associated with increased amounts of a peptide stimulator of testosterone production.     

Ratios of serum concentrations of testosterone and progesterone from yearling bulls with small testes

Thirty crossbred bulls, 12 to 13 mo of age, were used to examine the relationship of testosterone and progesterone concentrations and testosterone: progesterone ratio to measurements of testicular function. Bulls were allotted to 1 of 2 groups based on scrotal circumferences (SC) as follows: the Small SC (n=20) group had scrotal circumference less than 28 cm while the Large SC (n=10) group had scrotal circumference greater than 28 cm. All bulls were administered GnRH (100 mug, im), and blood was obtained immediately prior to injection (t=0), 30 min after injection (t=30) and 2 to 3 h after injection (t=150). Serum was assayed for concentrations of testosterone and progesterone. Semen was evaluated for the percentage of morphologically normal spermatozoa. Testicular parenchyma was sectioned and stained, and 300 cross sections per testis of seminiferous tubules were examined under a light microscope and classified as either active (spermatocytes and spermatids present) or inactive (no spermatocytes or spermatids present). Although progesterone concentrations varied widely (range: 21 pg/ml to 1070 pg/ml), repeated measurements from individual bulls were highly correlated (r(2)=0.74) and did not change significantly (P > 0.1) in response to GnRH treatment. Small SC bulls had a higher percentage of inactive seminiferous tubules (P < 0.001) and a lower percentage morphologically normal spermatozoa (P < 0.001) than Large SC bulls, but no differences in testosterone or progesterone concentrations or in the ratio of testosterone: progesterone were detected. Mean serum testosterone concentration increased (P < 0.0001) by 30 min after GnRH treatment and continued to increase (P < 0.0001) through t=150 but did not differ (p > 0.1) between groups. Normal testosterone secretion in response to GnRH injection suggested that no biochemical lesions in the testosterone production pathway were present in bulls with very small scrotal circumference.     

Testicular development and maturation of the hypothalamic-pituitary-testicular axis in the male tammar, Macropus eugenii

Testicular growth and maturation of the hypothalamic-pituitary-testicular axis were assessed in male tammars from 12 to 25 months of age to establish the time of sexual maturity. The testicular dimensions and body weights of 20 male tammars, approximately 12 months of age at the beginning of the study, were measured monthly for 1 year. Groups of 3 animals were castrated at 13, 19 and 25 months of age and their testes sectioned for histological examination. Testicular volume increased between 12 and 24 months of age and was highly correlated with body weight (r = 0.91). In the 13-month group the seminiferous tubules were closed with few mitotic figures. Spermatogenesis had begun in 2 of the 19-month animals. All stages of spermatogenesis were present in the other 19-month male, and in all of the 25-month males. Basal FSH concentrations increased with the age of the animal (21.0 +/- 32.48, 94.40 +/- 55.18 and 193.05 +/- 40.21 ng/ml (mean +/- s.d.) at 19, 20 and 25 months respectively) while basal LH concentrations were similar at 20 months and 25 months (0.43 +/- 0.18 and 0.58 +/- 0.25 ng/ml respectively). Basal testosterone concentrations were also similar 0.11 +/- 0.04, 0.35 +/- 0.16 and 0.22 +/- 0.10 ng/ml in 13-, 19- and 25-month-old animals. LHRH injection in tammars at 13, 19 and 25 months of age induced release of both LH and testosterone 10-30 min after injection. The hormone concentrations increased in both magnitude and duration with increasing age.(ABSTRACT TRUNCATED AT 250 WORDS)     

Intercellular transport of steroids in the infused rabbit testis

Tritiated-pregnenolone and tritiated-testosterone were infused via the testicular artery into the rabbit testis in situ, in order to determine if steroids can pass the "blood-testis barrier". After various periods of infusion (5-60 minutes) the testis were frozen cryostat sections were cut and freeze-dried. Interstitium and tubules were isolated by micro dissection and radioactivity per mcg dry weight was measured in both tissue compartments. Radioactive steroids could be isolated from the interstitial tissue as well as from the seminiferous tubules. Levels of radioactivity in the testes after infusion of tritiated-pregnenolone varied from 4 to 50% of the infused dose and were found to be dependent on the type of perfusion medium and the duration of the perfusion. Separation and identification of the radioactive steroids after infusing tritiated prognenolone showed that pregnenolone and testosterone were the major radioactive steroids in both interstitium and seminiferous tubules. After infusion with tritiated-testosterone, both tritiated-testosterone (77%) and tritiated-androstenedione (23%) were dound in the seminiferous tubules. It is concluded that steroids can be transported from the Leydig cells to the seminiferous tubules.     

Intratesticular androgen levels, androgen receptor localization, and androgen receptor expression in adult rat Sertoli cells

In the rat, quantitatively normal spermatogenesis is maintained only when intratesticular testosterone (ITT) levels greatly exceed the peripheral T concentration. When ITT concentrations fall below a threshold, germ cells are lost at specific stages of the seminiferous cycle. Germ cells can be restored by high doses of T that binds to androgen receptors (AR) in Sertoli cells. However, the relationships between germ cell dynamics, AR-mediated molecular events, and ITT concentrations are not established. ITT levels may regulate germ cell life and death through an effect on AR localization and AR mRNA or protein levels within Sertoli cells at specific stages of the cycle. We determined AR localization and mRNA and protein expression in adult rat Sertoli cells in relation to reduced and then restored ITT concentrations in vivo. ITT levels were reduced by implanting rats with T- and estradiol (E)-filled capsules for 7-28 days and subsequently restored with large T-filled capsules. AR is normally localized within Sertoli cell nuclei at stages VII-VIII of the seminiferous epithelium. After T/E treatment, AR immunostaining in Sertoli cell nuclei became nondetectable by 14-28 days but was restored 6 h following T restoration. The loss of Sertoli cell nuclear AR localization correlated with increasing numbers of apoptotic germ cells. AR mRNA levels in isolated Sertoli cells did not change through 14 days of T/E treatment, increased significantly by Day 28, and remained elevated 24 h after T restoration. AR mRNA levels in microdissected tubules at stages II-IV, VI-VIII, and IX-XII did not decrease through 14 days of T/E treatment. In contrast, AR protein levels were reduced in seminiferous tubules by Day 14 and in testes at Day 28 post-T/E treatment but were restored within 24 h by T repletion. Therefore, the reduction of ITT concentration results in a time-dependent redistribution of AR and reduced AR protein but not AR mRNA levels in Sertoli cells. Repletion of T restored AR protein and it relocated to Sertoli cell nuclei. By an unknown mechanism, T regulates AR localization within Sertoli cells to determine germ cell life or death.     

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.     

Nuclear testicular 1,25-dihydroxyvitamin D3 receptors in Sertoli cells and seminiferous tubules of adult rodents

1,25(OH)2D3 receptors were studied in whole testes, Sertoli cells, seminiferous tubules, Leydig cells and spermatogonia of adult NMRI mice and SD rats. Specific reversible high affinity binding (KD 1.4 x 10(-10)M; Nmax 72 fmol/mg protein) by a 3.5 S macromolecule was demonstrated in whole testes, Sertoli cells and seminiferous tubules. With identical techniques, no receptors were found in Leydig cells despite previous reports of 1,25(OH)2D3 actions on Leydig cell function.     

Stage dependent and androgen inductive expression of orphan receptor TR4 in rat testis

In this study, we investigated the expression of TR4 in different stages of seminiferous tubules and the relationship between TR4 and androgen in rat testis. We found that TR4 was stage-dependently expressed in rat seminiferous tubules, T withdrawal induced by high doses of testosterone undecanoate and ethane dimethane sulfonate inhibit TR4 expression in rat testis, and testosterone induced TR4 expression in co-cultured primary germ/Sertoli cells. Furthermore, we demonstrated that androgen receptor could enhance TR4-mediated transactivation activity in testis cells in the presence of testosterone. Together, these data indicate that the expression of TR4 in rat testis is stage dependent and androgen inductive, and suggest the important role of orphan receptor TR4 in spermatogenesis.     

Spatial arrangement of the stages of the cycle of the seminiferous epithelium in the Japanese quail, Coturnix coturnix japonica

The spatial arrangement of the stages of the cycle of the seminiferous epithelium of the Japanese quail was investigated by preparing three-dimensional reconstructions of a seminiferous tubule from each of 3 quails. It was found that the stages were not distributed at random, but were arranged in a wave which spiralled helically along a seminiferous tubule. Adjacent stages in space were always adjacent numbers in the cycle of the seminiferous epithelium. Complete spermatogenetic waves were found in which all 10 stages of the cycle were in sequential order. However, in most waves the sequential order of stages was disturbed by the occurrence of modulations. The area of a cellular association varied from 4600 to 41,600 microns 2 with a mean +/- s.e.m. (3 animals) of 17,902 +/- 2614 microns 2. The number of Sertoli cells involved in an association ranged from 4 to 35, with a mean +/- s.e.m. (3 animals) of 13.5 +/- 2.8. The findings support our earlier suggestion that the kinetics of spermatogenesis in the quail are fundamentally similar to the pattern which has been described for mammals.     

Isolation and staging of horse seminiferous tubules by transillumination

Stages of the spermatogenic cycle in the horse were determined by trans-illumination of enzymically isolated, seminiferous tubules and were verified by whole-mounted tubules observed by Nomarski optics and by conventional histology. Isolated tubules were obtained from young (less than 2 years) and adult (4-10 years) horses by enzymic digestion. Dispersed tubules were separated into three different groups based on the presence, size, and intensity of a dark region in the centre of the tubules: (1) pale--homogeneously light, (2) spotty--light on the periphery with a wide spotty region in the central two-thirds, or (3) dark--an intensely dark, narrow region through the central one-third. Seminiferous tubules from young stallions separated easily, but were only of the homogeneously light pattern as they lacked mature spermatids. After observation by Nomarski optics and bright-field microscopy, pale tubules under transillumination largely contained Stages I and II, spotty tubules contained Stages V and VI, and dark tubules contained Stages VII and VIII of the spermatogenic cycle. In-vitro incorporation of [3H]thymidine in spermatogonia and preleptotene/leptotene primary spermatocytes of these tubules confirmed the viability of germ cells in isolated tubules, and ultrastructural analysis confirmed excellent preservation of normal structure of seminiferous epithelium in isolated tubules. Hence, segments of seminiferous tubules in specific stages of the spermatogenic cycle can be obtained from enzymically digested horse testes when viewed by transillumination.     

Stages and duration of the cycle of the seminiferous epithelium in oncilla (Leopardus tigrinus, Schreber, 1775)

Six adult Leopardus tigrinus (oncilla) were studied to characterize stages of the seminiferous epithelium cycle and its relative frequency and duration, as well as morphometric parameters of the testes. Testicular fragments were obtained (incisional biopsy), embedded (glycol methacrylate), and histologic sections examined with light microscopy. The cycle of the seminiferous epithelium was categorized into eight stages (based on the tubular morphology method). The duration of one seminiferous epithelium cycle was 9.19 d, and approximately 41.37 d were required for development of sperm from spermatogonia. On average, diameter of the seminiferous tubules was 228.29 μm, epithelium height was 78.86 μm, and there were 16.99 m of testicular tubules per gram of testis. Body weight averaged 2.589 kg, of which 0.06 and 0.04% were attributed to the testis and seminiferous tubules, respectively. In conclusion, there were eight distinct stages in the seminiferous epithelium, the length of the seminiferous epithelium cycle was close to that in domestic cats and cougars, and testicular and somatic indexes were similar to those of other carnivores of similar size.     

Germ cells with nuclear DNA fragmentation related to apoptotic cells in rat testis in experimental hyperprolactinemia induced by metoclopramide

The cells with nuclear DNA fragmentation related to apoptosis were detected by TUNEL technique in the seminiferous epithelium of control rats and of rats with experimental hyperprolactinemia induced by metoclopramide. The percentage of convoluted tubules with apoptotic cells and the number of apoptotic cells (predominantly spermatogonia and spermatocytes) was increased in the experimental group. The results indicated stage-specific germ cell apoptosis. In the experimental group, apoptotic cells were most evident at early (I-IV), middle (VII-VIII) and late (XII-XIV) stages of the seminiferous epithelium cycle, as revealed by light and electron microscopy. We suggest that a decreased concentration of testosterone and an increased concentration of prolactin could disturb spermatogenesis and contribute to the intensive apoptosis of germ cells in rats with hyperprolactinemia. Sertoli cells which have receptors for testosterone and prolactin and play an important role in spermatogenesis and in the initiation of apoptosis in seminiferous epithelium, could mediate such an influence of both hormones.     

Impact of season on seminal characteristics and endocrine status of adult free-ranging African buffalo (Syncerus caffer)

Pituitary, gonadal and adrenal activity were compared in free-living, adult African buffalo bulls during the breeding and nonbreeding seasons. Frequent blood samples were collected for 2 h from anaesthetized bulls treated intravenously with saline, gonadotrophin-releasing hormone (GnRH, 200 micrograms), human chorionic gonadotrophin (hCG, 10,000 i.u.) or adrenocorticotrophic hormone (ACTH, 1.5 mg). Electroejaculates also were collected from anaesthetized bulls during the breeding and nonbreeding seasons. Pretreatment testosterone concentrations among bulls varied more during the breeding (0.17-23.0 ng/ml) than the nonbreeding (0.15-2.21 ng/ml) season. The variation within the breeding season was attributed to 8 of 25 bulls producing higher (P less than 0.05) serum testosterone (High-T; 16.28 +/- 2.03 ng/ml) and testicular LH receptor (1.53 +/- 0.22 fmol/mg testis) concentrations compared with their seasonal counterparts (Low-T; 0.95 +/- 0.26 ng/ml; 0.38 +/- 0.04 fmol/mg) or with all bulls during the nonbreeding season (0.90 +/- 0.27 ng/ml; 0.31 +/- 0.04 fmol/mg). The magnitude of GnRH- and hCG-induced increases in serum testosterone was similar (P greater than 0.05) between Low-T bulls and bulls during the nonbreeding season. In the High-T animals treated with GnRH or hCG, serum testosterone did not increase, suggesting that secretion was already maximal. Peak serum LH concentrations after GnRH were greater (P less than 0.05) in bulls during the nonbreeding than the breeding season; FSH responses were similar (P greater than 0.05). ACTH treatment did not increase serum cortisol concentrations above the 2-fold increase measured in bulls treated with saline, hCG and GnRH (P greater than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)     

Disruption of seminiferous epithelial function in the rat by ovarian protein

The granulosa cell produces an inhibitor of aromatase activity, which recently was purified to homogeneity (follicle-regulatory protein: FRP). Since extracts of testicular homogenates also contain factor(s) with biological properties similar to FRP, including inhibition of granulosa cell aromatase, we examined the effects of ovarian FRP on testicular function. Forty-five-day-old rats received daily FRP injections (100 micrograms or 300 micrograms). After 15, 30, 45, and 70 days of therapy, (n = 5 each group), trunk serum was measured for testosterone, androstenedione, estradiol, and FSH levels by established radioimmunoassays (RIA). One testis from each rat was homogenized, centrifuged, and evaluated for sperm head counts; the other testis was dissected by transillumination, and the length of seminiferous epithelial stages determined. After 15 (control: 4.8 +/- 0.2 mm; 100 micrograms: 6.0 +/- 0.3 mm; 300 micrograms: 6.6 +/- 0.3 mm) and 30 days (control: 4.6 +/- 0.2 mm; 100 micrograms: 6.3 +/- 0.2 mm; 300 micrograms: 5.9 +/- 0.2 mm) of treatment the length of the "strong" seminiferous tubule segment in FRP-treated rats was greater than in control rats (p less than 0.05). Serum levels of steroids and FSH were similar in all groups. After 30, 45, and 70 days of treatment, the sperm head counts for the 100-micrograms and 300-micrograms dosages were 26%, 29%, 30% and 20%, 34%, and 24% of control values, respectively. By 70 days of treatment, cycle Stage VII was markedly reduced or absent in FRP-treated rats, and their round spermatids contained ring chromatin; both conditions indicate degeneration. FRP (50 micrograms/ml) was added to rat Sertoli cell cultures for 4 days after which transferrin and androgen-binding protein (ABP) were measured. FRP enhanced Sertoli cell secretion of ABP (58 vs. 138 +/- 7 microliters eq/culture) and transferrin (2.1 vs. 4.7 +/- 0.6 microgram/culture). In conclusion, systemic injection of FRP alters seminiferous epithelial function by reducing maturation of mature sperm forms. Adding FRP to Sertoli cells in culture enhances secretion of transferrin and ABP; this suggests that maturation of the germinal elements may be linked to the secretory function of seminiferous tubules.     

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.     

Altered concentrations of gonadotrophin, prolactin and GnRH receptors, and endogenous steroids in the abdominal testes of adult unilaterally cryptorchid rats

Testicular descent was prevented unilaterally in newborn rats by cutting the gubernaculum testis. At 100 days of age, the number of Leydig and Sertoli cells per testis, the concentration of receptors for LH, FSH, prolactin and GnRH, and endogenous concentrations of progesterone and testosterone were determined. The weight of the abdominal testes was reduced by 80%, but in spite of this they contained as many Sertoli (32.8 +/- 1.3 X 10(6), mean +/- s.e.m., n = 6) and Leydig (28.2 +/- 1.7 X 10(6) cells as did scrotal testes (32.1 +/- 2.5 X 10(6) and 24.3 +/- 1.2 X 10(6) respectively). The numbers of receptors for LH (3.2 +/- 0.2 and 1.0 +/- 0.2 pmol/testis, mean +/- s.e.m., n = 11), FSH (358 +/- 11.0 and 96.3 +/- 12.6 fmol/testis) and prolactin (535 +/- 32.7 and 92.4 +/- 13.2 fmol/testis) were reduced (P less than 0.001) in abdominal testes, but the number of GnRH receptors was unaffected (8.9 +/- 1.4 and 12.1 +/- 1.8 fmol/testis, n = 6). Testicular testosterone concentration (30.9 +/- 4.4 vs 15.4 +/- 3.2 ng/g, n = 11, P less than 0.001), but not that of progesterone (0.87 +/- 0.10 vs 1.01 +/- 0.21 ng/g), was decreased in abdominal testes. The decreased receptor and androgen values reflect functional disturbances in the abdominal testes. The changed local milieu within abdominal testes may reduce hormone receptor concentrations which are then involved in the observed Leydig cell dysfunction.     

The cycle of the seminiferous epithelium in Landrace boars

The present study describes the morphological features of the eight stages of the seminiferous epithelium in Landrace boars according to the tubular morphology method, as well as their relative frequency, length, and duration. In Landrace boars the pre-meiotic stages occupied the 31.9 +/- 19.9% of the spermatogenic cycle and had a total length of 1788.8 +/- 1153.0 microm and a duration of 2.78 days; they were mainly characterised by the presence of leptotene and pachytene spermatocytes and round spermatids. Meiotic stages, with a relative frequency of 16.4 +/- 6.8%, a length of 787.1 +/- 603.1 microm and a duration of 1.41 days, contained spermatocytes in advanced meiosis I and/or in meiosis II and elongating spermatids grouped in bundles. Post-meiotic stages occupied the 50.6 +/- 20.4% of the spermatogenic cycle and had a length of 2096.8 +/- 1175.0 microm and a duration of 4.37 days; the most important event of these stages was the spermiation, which included the complete remodelling of sperm head and tail and the releasing of spermatozoa into the lumen, as well as the formation of residual bodies. From data obtained we concluded that both germ cell associations of the stages maintain constant among Landrace boars, and that the relative frequency, length and duration of the stages were directly dependent of the cytological transformations on the seminiferous tubules.     

Relationship of testosterone pulses to androgen binding in the pituitary of rams

The effects of testosterone on cytosol and nuclear androgen receptors of ram pituitary were examined in two experiments. In Exp. I, 500 micrograms testosterone were injected intravenously and groups of 4 rams were slaughtered at 0, 15, 30, 45, 90 and 360 min after injection. Cytosolic receptor concentration decreased from 21 +/- 0.9 to 6 +/- 0.9 fmol/mg protein 30 min after the testosterone injection (P less than 0.001), and then returned towards the preinjection level after 90 min. The pattern of nuclear receptor concentration was the opposite; a maximal increase (12 +/- 3.5 to 32 +/- 5.7 fmol/mg protein) was observed 30 min after injection (P less than 0.001), followed by a progressive but incomplete decrease by 360 min. In Exp. II, blood was collected every 20 min for 17 h in three successive series, each of 12 rams, which were then slaughtered. Plasma LH and testosterone concentrations were measured by radioimmunoassay. No changes were observed in cytosol receptor concentration, but nuclear receptor concentration was negatively correlated with the interval elapsed since the beginning of the last testosterone pulse (r = -0.62; P less than 0.001). The highest values for nuclear receptor concentrations were observed at an interval equal to or less than 120 min. These results indicate that natural pulses are associated with androgen binding particularly in the pituitary nuclei.     

Characterization of male killer whale (Orcinus orca) sexual maturation and reproductive seasonality

Longitudinal serum testosterone concentrations (n=10 males) and semen production (n=2 males) in killer whales were evaluated to: (1) characterize fluctuations in serum testosterone concentrations with respect to reproductive maturity and season; (2) compare morphologic changes to estimated age of sexual maturity, based on changes in serum testosterone concentrations; and (3) evaluate seasonal changes in sperm production. Classification of reproductive status and age class was based on differences (P < 0.05) in serum testosterone concentrations according to age; juvenile males ranged from 1 to 7 years (mean+/-S.D. testosterone, 0.13+/-0.20 ng/mL), pubertal males from 8 to 12 years (2.88+/-3.20 ng/mL), and sexually mature animals were 13 years and older (5.57+/-2.90 ng/mL). For captive-born males, serum testosterone concentrations, total body length and height to width ratio of the dorsal fin were 0.7+/-0.7 ng/mL, 495.6+/-17.5 cm and 1.14+/-0.13c m, respectively, at puberty; at sexual maturity, these end points were 6.0+/-3.3 ng/mL, 548+/-20 cm and 1.36+/-0.1cm. Serum testosterone concentrations were higher (P<0.05) from March to June than from December to February in pubertal animals (4.2+/-3.4 ng/mL versus 1.4+/-2.6 ng/mL) and than from September to December in sexually mature animals (7.2+/-3.3 ng/mL versus 4.0+/-2.0 ng/mL). Ejaculates (n = 90) collected from two males had similar (P > 0.05) sperm concentrations across all months. These data represent the first comprehensive study on male testosterone concentrations during and after sexual maturation, and on reproductive seasonality in the killer whale.