Testicular development, histology, and hormone profiles in three yearling angus bulls with spermatogenic arrest

This article discusses the interactions between testis criteria and hormone profiles in Angus bulls with spermatogenic arrest. From 2 to 12 months (mo), testis diameter and hormone concentrations (basal and GnRH-stimulated) were evaluated in 27 bulls. At 12 mo, testes were excised. The z statistical test was used to determine whether parameters in three infertile bulls were different (P < 0.05) from those in 24 bulls with normal spermatouenesis. Bull 1 had Sertoli cell-only syndrome and Bull 2 had 90% of the tubules without germ cells and only A1 spermatogonia in the remaining. In Bull 3, germ cells did not advance beyond the primary spermatocyte stage. At 12 mo, testes of Bull 1 (99 g), Bull 2 (105 g) and Bull 3 (32 g) weighed less than those of normal bulls (251.5 +/- 56 g). Sertoli cell numbers/testis in Bull 1 (3.8 x 10(9)) and Bull 2 (4.3 x 10(9)) were not different from those in normal bulls (4.9 +/- 0.3 x 10(9)), but were reduced in Bull 3 (1.6 x 10(9)). The number of Leydig cells per gram of testis parenchyma was higher in Bull 1 (5.4 x 10(7)), Bull 2 (7.3 x 10(7)) and Bull 3 (19 x 10(7)) than in normal bulls (3.6 +/- 0.2 x 10(7)). In Bulls 1 and 2, basal and GnRH-stimulated LH, FSH, testosterone (T), androstenedione (delta4A) and estradiol 17-beta (E2) were within normal ranges at most ages. However, basal FSH and LH were greater in Bull 3 than in normal bulls, probably the causes for higher Leydig cell density. Also in the same animal, GnRH induced lower responses in LH and FSH, consequence of low basal T and E2 at some ages. Basal and GnRH-stimulated delta4A in Bull 3 were greater than in normal bulls after 6 mo, indicating impairment of Leydig cell differentiation. Deficiency in hormone secretion did not appear to be the cause of infertility, which points toward impaired gonadal responses or secretion of intratesticular factors, or genetic defects. Moreover, infertile animals may not always show pronounced changes in hormone secretion, but evaluation of testis growth around puberty can help identify those animals that do not have proper gonadal development.     

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.     

A comparison of three methods of assessing sex-drive in yearling beef bulls and relationships with testosterone and LH levels

Three methods of assessing sex-drive were compared in 113 yearling beef bulls. These were the serving capacity score (SC), the libido score (L) and reaction time to first service (R). Ovariectomized heifers restrained in service crates were the stimulus for all tests. Bulls were assessed twice by each method. For the first libido and serving capacity tests (L1 and SC1), the heifers were induced to show estrus. For the second tests (L2 and SC2), the heifers were not induced to show estrus. On a non test day, single blood samples were taken from all bulls and assayed for LH and testosterone. Reaction times to first service (R1 and R2) in the two serving capacity tests were not significantly correlated. Although the numbers of services (SC1 and SC2) in both serving capacity tests, were significantly correlated (r = .67), 57% of the bulls did not achieve a service in both the tests with heifers in heat and with heifers not in heat. Libido scores between the test with heifers in heat and heifers not in heat were significantly correlated (r = .67). The libido score method had the advantage that more bulls received a positive score and the test duration was shorter than in the serving capacity test. Of the three scoring procedures compared, libido score appeared to have most advantage in assessing sex-drive in yearling beef bulls. The total number of services achieved in the first serving capacity tests (using 'estrus' heifers) did not differ from that achieved in the second tests (using non-estrus heifers). The total services achieved in 10-min compartments of the 30-min serving capacity tests showed no difference either within or between tests. It is concluded that a 10-min test provides as much comparative information on the sex-drive of yearling beef bulls as longer tests. Further, the use of females in estrus appears unnecessary to satisfactorily assess bull sex-drive, provided proper restraint and presentation of stimulus females is employed. LH and testosterone values were not significantly correlated or were poorly correlated with sex-drive measurements.     

The effect of testosterone on LH and testosterone responses to GnRH in bulls

Clinical and andrological-spermatological investigations in male beagles revealed the status of the gonads before and after fistulating the vas deferens.When semen samples were collected reqularly, no siqnificant differences could be observed in comparison to ejaculates before surgical intervention, as judged by spermatological parameters. Only an increased incidence of immature spermatic cells was found. Changes in the gonads and spermatozoa respectively were found in animals with irregular collection of spermatozoa via fistula which induced irrep-arable occlusion of the fistula and subsequent spermio-stasis.Insemination of beagle bitches with spermatozoa from fistulae led to fertilisation of 3 animals from the group of 4.     

Lack of influence of mounting on plasma FSH,LH and testosterone concentrations in azoospermic and normospermic young bulls

The influence of mounting and ejaculation on the FSH, LH and testosterone secretory patterns was studied in three azoospermic (including one 61 XXY; one Sertoli-cell-only Syndrome and one secretory-excretory azoospermic) and three control normospermic bulls. Sexual activity did not result in any alteration in the release of these three hormones. There was no difference between the secretory patterns before and after ejaculation in the two classes of bulls. One of the main features was the elevated concentrations of FSH in the bull with Klinefelter's Syndrome, but the mounting test did not result in any significant effect on this concentration. The LH and testosterone patterns were similar for all individuals. From these results, it can be concluded that the mounting test applied under these experimental conditions had no effect on the pituitary-gonadal axis in bulls characterized by either impairment of spermatogenesis or normal semen production.     

Temporal relationships among peripheral blood concentrations of corticosteroids, luteinizing hormone and testosterone in bulls

Interrelationships among peripheral blood concentrations of corticosteroids (CS), luteinizing hormone (LH) and testosterone (T) were evaluated over a 24-hr period in four Angus bulls (18 months of age and 450 kg in body weight). Concentrations of LH and T were determined by radioimmunoassay and concentrations of CS by competitive protein binding assay of blood samples collected via jugular cannula at hourly intervals for 24 consecutive hr. A positive temporal relationship was observed between LH and T as significant positive correlations were obtained between concentrations of LH at one hour and concentrations of T at the subsequent hour in 3 of 4 bulls. Although LH peaks preceded T peaks by 1 hr, variation in this temporal relationship was observed as LH peaks occurred which were not accompanied by T peaks in some bulls. LH peaks were usually preceded by basal or declining concentrations of CS and prolonged elevations in concentrations of CS were often coincident with basal concentrations of LH and T. Negative correlations were obtained between concentrations of CS at one hour and concentrations of LH and T at the subsequent hour. These data describe the positive regulatory role of LH in testicular T production in the bull and suggest that alterations in endogenous concentrations of CS may influence peripheral concentrations of LH and T in the bull.     

Alterations in plasma concentrations of testosterone, LH, and prolactin associated with mating in the male rat.

The hormonal response of the male rat to sexual activity was investigated in two studies. In the first, no evidence of a chronic elevation in plasma levels of testosterone (T), LH, or prolactin (PRL) was observed in sexually experienced rats compared to naive controls. Both groups showed an acute increase in plasma levels of all three hormones following mating, but the increases shown by the experienced group were more pronounced. In the second study, plasma levels of T, LH and PRL rose in sexually experienced male rats following exposure to a mating arena whether it contained an estrous female, an anestrous female, or no other animal. However, the increases were considerably larger in the group exposed to estrous females. It is suggested that plasma hormones rise in anticipation of mating, although not to the same extent as following mating, and that the anticipatory rise may function to initiate or facilitate mating behavior.     

Effect of naloxone on plasma concentrations of prolactin and LH in lactating sows

Six lactating sows were injected through an indwelling vena cava cannula with naloxone (2.5 mg/kg body weight) on Day 15 post partum. Blood samples were collected through the cannulas at 10-min intervals for 8 h before and 10 h after naloxone administration. Plasma prolactin and LH concentrations were measured by radioimmunoassay. Naloxone caused a marked suppression of plasma prolactin concentrations lasting 4-6 h. LH concentrations were also affected by naloxone: LH rose to reach maximum values 20-50 min after naloxone treatment. Pretreatment values were recorded 200-300 min after the treatment. These results indicate that endogenous opioids are involved in causing the endocrine patterns occurring during lactation, i.e. high prolactin and low LH concentrations.     

Effects of hemicastration and unilateral vasectomy on the remaining gonad and on the FSH,LH and testosterone blood concentration in bulls

The effects of unilateral castration and vasectomy on the weight and microscopic appearance of the contralateral testis and on the blood levels of testosterone, LH and FSH, were studied in German Fleckvieh bulls. Testicular weights were higher in hemicastrated bulls (P < 0.01) and unilaterally vasectomized bulls (P < 0.05) when compared to controls, 377 ± 45g (x ± s, N = 4 and 281 ± 12g, N = 4 vs 226 ± 38g, N = 3, respectively.Testosterone concentrations were higher during the weeks 14 to 22 after surgery in both treated groups. LH levels were not different from controls, but FSH levels increased significantly (P < 0.01) two weeks after hemicastration and unilateral vasectomy.Different factors appear to regulate the LH and the FSH concentrations in bulls. The increase of FSH after hemicastration may indicate a reduced production of inhibin or an inhibin-like substance from the testes, and a similar increase after unilateral vasectomy suggests that this substance may be resorbed distal to the testes.     

Effect of vascular cannulation on serum concentrations of LH, FSH, prolactin and cortisol in prepubertal gilts

Two experiments were conducted to determine whether cannulation of the jugular vein in gilts alters serum concentrations of LH, FSH, prolactin (PRL) or cortisol (C). In Experiment 1, 12 crossbred prepubertal gilts weighing 95 +/- 1.3 kg were immobilized by snaring, and tygon tubing was threaded into the anterior vena cava through a 12-gauge needle inserted into the jugular vein. Five hours later, blood samples were drawn at 20-min intervals for 4 h (Day 0). Samples were also drawn at 20-min intervals for 4-h periods 24 h (Day 1) and 48 h (Day 2) after cannulation. Serum concentrations of LH were similar (P=0.26) among Day 0 (0.40 ng/ml), Day 1 (0.39 ng/ml) and Day 2 (0.34 ng/ml). Serum PRL was similar (P=0.07) among Day 0 (4.10 ng/ml), Day 1 (3.87 ng/ml) and Day 2 (3.43 ng/ml). Serum concentrations of C were greater (P < 0.001) on Day 0 (8.32 ng/ml) than Day 1 (4.48 ng/ml) or Day 2 (3.54 ng/ml). In Experiment 2, cannulas were placed in 29 prepubertal gilts. Two days after initial cannulation, six blood samples were drawn at 20-min intervals. Gilts were then immobilized by snaring, and a second cannulae was inserted into the contralateral vein. Five blood samples were taken at 2-min intervals during the second cannulation and then six samples were drawn at 20-min intervals. Serum LH and FSH were not altered by cannulation or elevated during the subsequent 2-h sampling period (P>0.05). In contrast, serum concentrations of PRL rose slowly (P<0.05) during cannulation and remained elevated for 60 min before returning to baseline. Serum concentrations of C rose within 6 min of cannulation, remained elevated for 30 min, and then declined over the next 90 min. From these two experiments, it appears that secretory patterns of LH and FSH can be accurately assessed immediately after cannulation in prepubertal gilts. Measurements of serum PRL and C that reflect nonstressed conditions, however, cannot be obtained until at least 2 h or 1 d after cannulation, respectively.     

Circadian variations of plasma LH and testosterone in adult swamp buffalo bulls

Three swamp buffalo bulls aged 1.5, 1.10 and 2 years were submitted to frequent blood sampling every 15 m during a period of 25 h using an indwelling infusion set. Plasma LH and testosterone were quantified by radioimmunoassay technique. The levels of the two hormones in each individual exhibited episodic and nonrhythmic patterns. The number of LH peaks varied according to individval, ranging from no peak in one bull to 2 in the other two bulls. The mean LH concentrations during the period of study for each bull were 0.74, 0.33 and 1.17 ng/ml. Whereas the number of testosterone peaks varied between 1-10 and the average testosterone concentrations were 0.1, 0.33 and 0.55 ng/ml for the younger to the older bulls respectively. The testosterone peaks related to the LH peaks in each individual bull.     

Effect of mouse epidermal growth factor on plasma concentrations of LH, FSH and testosterone in rams

Two experiments were conducted to examine the effects of mouse epidermal growth factor (EGF) on the concentrations of testosterone, LH and FSH in jugular blood plasma and on the pituitary responsiveness to LHRH. In 20 rams treated with subcutaneous doses of EGF at rates of 85, 98 or 113 micrograms/kg fleece-free body weight, mean plasma LH and testosterone concentrations were significantly reduced (P less than 0.05) at 6 h after treatment but not at 24 h. EGF treatment at 130 micrograms/kg fleece-free body weight suppressed the plasma content of these hormones for up to 48 h. Mean plasma FSH concentrations decreased significantly (P less than 0.05) for up to 48 h after EGF treatment, the effect being most pronounced in rams with mean pretreatment FSH values greater than or equal to 0.5 ng/ml. Intravenous injections of 1.0 micrograms LHRH given to each of 5 rams before and at 6 h, 24 h and 72 h after EGF treatment produced LH and testosterone release patterns which paralleled those obtained in 5 control rams similarly treated with LHRH. These results suggest that, in rams, depilatory doses of mouse EGF temporarily impair gonadotrophin and androgen secretion by inhibiting LHRH release from the hypothalamus. Such treatment appears to have no effect on the responsiveness of the pituitary to LHRH.     

Effect of dexamethasone on plasma concentrations of LH, FSH and testosterone in women with hirsutism.

Thirty sexually mature women with hirsutism were treated with 3 x 1.5 mg dexamethasone per day over a period of three days. Before and after treatment, plasma concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), and testosterone were determined. While an effect of dexamethasone on LH plasma levels could not be established statistically, FSH and testosterone plasma concentrations decreased significantly in comparison to their initial values (p less than 0.01). Special attention is directed to the different effects of dexamethasone on LH and FSH plasma concentrations.     

Effect of zearalenone and estradiol benzoate on serum concentrations of LH, FSH and prolactin in ovariectomized gilts

Six ovariectomized gilts were given zearalenone (Z), estradiol benzoate (EB) or vehicle in a replicated 3 x 3 Latin square design. Zearalenone was added to 2.3 kg of a corn-soybean ration at a dose of 1 mg Z/kg body weight; EB was given intramuscularly at 0.1 mg EB/kg body weight. Control gilts received vehicle solvent for both Z and EB. Blood samples were collected from indwelling jugular cannulas at 6-h intervals for 48 h before Z, EB or vehicle was given. After treatment, blood samples were drawn at 6-h intervals for an additional 84 h. Serum concentrations of luteinizing hormone (LH) decreased (P<0.001) from 4.67 ng/ml to 0.29 ng/ml within 6 h of EB. From 54 to 84 h after EB, serum concentrations of LH rose to 15.60 ng/ml (P<0.001). Serum concentrations of LH were reduced (P<0.001) in a similar pattern after Z (3.70 ng/ml to 0.49 ng/ml), but a rise in serum LH was not observed 54 to 84 h after Z (1.30 ng/ml). Serum concentrations of LH remained unchanged (P=0.55) in gilts given vehicle. Serum concentrations of follicle stimulating hormone (FSH) were suppressed (P<0.03) at 6 h in EB (19.10 vs 11.35 ng/ml) and Z gilts (16.16 vs 11.41 ng/ml) but remained unchanged in vehicle gilts. Serum concentrations of FSH did not change in EB or Z gilts during the next 36 h. These data indicate that the suppressive action of Z on serum concentrations of LH and FSH was similar to that of EB, while the biphasic stimulatory effect of EB for LH was not manifested by Z.     

Effect of active immunisation against testosterone-3-BSA on circulating levels of testosterone, LH, prolactin and testosterone antibody titre in the male rat

Male Sprague-Dawley rats were actively immunised against testosterone-3-bovine serum albumin (T-3-BSA) and on appearance of detectable anti-testosterone antibodies, elevated serum testosterone and LH concentrations were observed. These concentrations reached values of >28 μg/100ml testosterone and 16 μg/100ml LH in some animals after 5 months of immunisation. The corresponding prolactin values did not appear to differ significantly from controls. The circulating bound testosterone fraction as determined by equilibrium dialysis, rose from 65.0 ± 2.75% before immunisation to 98.7 ± 0.75% in those animals possessing high titre antisera. This entailed a nett $\text{decrease}$ in the concentration of unbound steroid from 144 ± 49 ng/100 ml to 78 ± 25 ng/100ml.     

Effect of mating stimuli on LH release in bulls

Fourteen crossbred beef bulls were assigned at random to receive one of four sexual stimuli treatments. Treatments consisted of: (1) controls (four bulls), no visual or physical contact with any cows; (2) false mount (two bulls), allowed to mount an estrual cow; (3) mated (four bulls), allowed to mount an estrual cow with intromission and ejaculation; (4) electroejaculated (four bulls), no exposure to cows. Serum from blood samples taken at 15-min intervals from -15 min to 2 hr from sexual stimuli were radioimmunoassayed for luteinizing hormone (LH). Four bulls had elevated levels at the pretreatment bleeding, but none of the stimuli induced or were associated with LH releases. Basal levels of LH were consistent within bulls but varied considerably among bulls. Conclusion is that stimuli associated with mating do not cause a release of LH.     

Plasma concentrations of testosterone and LH in the male dog

Blood samples were withdrawn every 20 min from 3 conscious intact and 2 castrated mature males during non-consecutive periods of 12 h during the light and dark phases of the lighting schedule (intact dogs) and of 11 h during the light period (castrated dogs). In the intact dogs testosterone concentrations ranged from 0.4 to 6.0 ng/ml over the 24-h period. LH concentrations varied from 0.2 to 12.0 ng/ml. In all animals, LH peaks were clearly followed, after about 50 min, by corresponding testosterone peaks, but no diurnal rhythm could be established. LH concentrations in the castrated dogs were high (9.8 +/- 2.7 (s.e.m.) ng/ml), and still showed an episodic pattern in spite of the undetectable plasma testosterone levels.