Evaluation of mathematical models to describe testicular growth in Blackbelly ram lambs.

The primary objective was to compare various mathematical models to describe scrotal circumference (SC) and paired testis volume development in Blackbelly ram lambs. The study was conducted in the state of Querétaro, México (20° 43' N, 100° 15' W). Spring-born Blackbelly ram lambs (n = 41) were housed outdoors and fed alfalfa hay and concentrate. Body weight, SC, and testis length, diameter, and volume were recorded every 2 wk from 24 to 172 d of age (June 18 to November 3). The following mathematical functions were used to model SC-age and testis volume-age relationship: Von Bertalanffy, Brody, Gompertz, Logistic, and Richards. The suitability of the models was evaluated based on parameter values and standard errors, residual mean square, the coefficient of determination (R(2)), and the average prediction error (APE). All models, except for Brody's, had good fit to SC (R(2) > 0.98) and testis volume (R(2) > 0.95), and produced similar growth curves in the range of ages studied. The logistic model predicted SC at maturity quite well, 33.6 ± 0.6 cm as compared with 33.9 ± 0.5 cm observed in adult animals; all models had APE's smaller than ± 7% between 56 and 168 d of age. The Bertalanffy model predicted testis volume at maturity quite well, 513 ± 22 cm(3) as compared with 488 ± 20 cm(3) calculated for adult animals. The logistic model had a good fit to testis volume during the period of study, but underestimated the volume at maturity by 28%. All models, except for Brody's, had APE's smaller than ± 14% between 98 and 168 d of age. The logistic and Bertalanffy models predicted the inflection point for SC at 83 and 59 d of age, and testis volume at 116 and 109 d of age, respectively. In conclusion, all models, except for Brody's, had good fit to actual SC and testis volume data in the range of age evaluated, whereas the logistic and Bertalanffy's models made the best predictions for adult SC and testis volume, respectively.

Stereological study of postnatal testicular development in Blackbelly sheep.

The objective was to characterize testicular development in Blackbelly sheep, focusing primarily on Sertoli cell number. Lambs (n=43) were allotted into eight groups, and surgically castrated at 0, 3, 6, 9, 12, 15,18 or 21 weeks of age (n=4-6 lambs per group). Testes were fixed and paraffin-embedded, cross-sections (5 microm) were stained and evaluated with quantitative morphometry techniques. Testis weight increased at a greater rate between 9 and 15 weeks of age, which was associated with remarkable changes in testicular histology, including increases in tubular tissue volume, and tubule diameter and length. Spermatogenesis started in a period between 9 and 12 weeks, lumen and elongated spermatids were observed for the first time at 12 weeks (78% of the tubules) and 15 weeks (37% of the tubules), respectively. Total number of Sertoli cells (mean+/-S.E.M.) increased steadily from birth (531+/-76 x 10(6)) to 15 weeks (12,008+/-1722 x 10(6)), with no changes afterwards. Sertoli cell number per gram of testicular tissue decreased as lambs were older, with the most remarkable change occurring between Weeks 9 and 12. An early increase in serum LH was observed at 6 weeks of age, with testosterone (T) increasing at Weeks 12 and 21. In conclusion, Sertoli cells maintained the capacity of proliferating from birth to 15 weeks of age in Blackbelly sheep; furthermore, the period of accelerated testis growth was associated with increased serum T concentration and with important changes in testicular morphology, as a consequence of the beginning and establishment of spermatogenesis and Sertoli cell maturation.

Comparative response of rams and bulls to long-term treatment with gonadotropin-releasing hormone analogs.

The objective was to compare the relative response between rams and bulls in characteristics of LH, FSH and testosterone (T) secretion, during and after long-term treatment with GnRH analogs. Animals were treated with GnRH agonist, GnRH antagonist, or vehicle (Control) for 28 days. Serial blood samples were collected on day 21 of treatment, and at several intervals after treatment. Injections of natural sequence GnRH were used to evaluate the capacity of the pituitary to release gonadotropins during and after treatment. Treatment with GnRH agonist increased basal LH and T concentrations in both rams and bulls, with a greater relative increase in bulls. Endogenous LH pulses and LH release after administration of GnRH were suppressed during treatment with GnRH agonist. Treatment with GnRH antagonist decreased mean hormone concentrations, LH and T pulse frequency, and the release of LH and T after exogenous GnRH, with greater relative effects in bulls. Rams previously treated with antagonist had a greater release of LH after administration of GnRH compared with control rams, while rams previously treated with agonist showed a reduced LH response. Bulls previously treated with agonist had reduced FSH concentrations and LH pulse amplitudes compared with control bulls while bulls previously treated with antagonist had greater T concentrations and pulse frequency. The present study was the first direct comparison between domestic species of the response in males to treatment with GnRH analogs. The findings demonstrated that differences do occur between rams and bulls in LH, FSH and testosterone secretion during and after treatment. Also, the consequences of treatment with either GnRH analog can persist for a considerable time after discontinuation of treatment.

Testicular development of Zebu bulls after chronic treatment with a gonadotropin-releasing hormone agonist.

The objective was to compare testis characteristics of Zebu bulls treated with the GnRH agonist, deslorelin, at different times and for different durations during their development. An additional objective was to determine the usefulness of a stain for the transcription factor GATA-binding protein 4 (GATA-4) as a specific marker for Sertoli cell nuclei in cattle. Bulls (54) were allocated to nine groups (n = 6) and received s.c. deslorelin implants as follows: G1 = from birth to 3 mo of age; G2 = from 3 to 6 mo; G3 = from 6 to 9 mo; G4 = from 9 to 12 mo; G5 = from birth to 15 mo; G6 = from 3 to 15 mo; G7 = from 6 to 15 mo; G8 = from 12 to 15 mo; and G9 (control) = no implant. Bulls were castrated at 19 mo of age. Paraffin sections (10 microm) were subjected to quantitative morphometry and GATA-4 immunohistochemistry. At castration, all bulls in the control group (6/6) had attained puberty (scrotal circumference > or = 28 cm), whereas a smaller proportion (P < 0.05) had reached puberty in G2 (2/5) and G6 (1/6). Bulls in G2 and G6 also had a lesser (P < 0.05) testis weight compared with the control group. Total volume of seminiferous epithelium and total daily sperm production in G2 and G6 were only half that observed in the control group. Spermatids were observed in less than 50% of seminiferous tubules in G2, G6, and G7 compared with 82% in the control group (P < 0.05). Staining for GATA-4 was specific for and abundant in the Sertoli cell nucleus in both pre- and postpubertal bulls, and no other cell nucleus inside the seminiferous tubule was positive for GATA-4. Total number of Sertoli cells was not affected by treatment (P = 0.45), but nuclear volume was smaller in G2 and G6 (P < 0.05) compared with the control group. In conclusion, treatment of Zebu bulls with deslorelin had no apparent beneficial effect on testis development and delayed puberty when treatment was initiated at 3 mo of age. Staining for GATA-4 was a useful method for identifying and quantifying Sertoli cell nuclei in both pre- and postpubertal bulls.

Season of the year influences testosterone secretion in bulls administered luteinizing honrmone.

The objective of the present study was to evaluate the secretion of testosterone (T) in bulls in response to the administration of varying doses of bovine LH (bLH) during the four seasons of the year. Five adult bulls (4 yr of age) were treated with an amount of bLH that was estimated to induce a 5 ng/mL amplitude pulse of LH in blood serum on five consecutive days around the spring equinox, summer solstice, fall equinox, and winter solstice. Five hours after this dose, bulls were treated with bLH in amounts that were estimated to induce a 0.25, 0.5, 1, 2, or 4 ng/mL amplitude LH pulse in blood serum in a Latin square design. Blood samples were collected for 5 h after administration of a dose of bLH that was estimated to induce the 5-ng amplitude LH pulse, and for 3 h after administration of the variable doses of bLH, and were then assayed for concentrations of T. Average concentrations and amplitude of T release after doses of bLH that were estimated to induce the 5-ng amplitude LH pulses were greater during the spring and summer than during the winter (P < 0.05). The area under the release curve (AUC) was greater during the spring than during the winter (P < 0.05). During the 3 h after treatment with the variable doses of bLH, T response was affected by dose (P < 0.001) and season (P < 0.001), but there was no dose x season interaction. Testosterone response increased in a dose-dependent fashion for all variables studied. The greatest average concentrations of T and AUC were observed in the spring compared with the fall and winter (P < 0.05). These data support our working hypothesis that testes of bulls are more responsive in releasing T in response to bLH stimulation in the spring and summer compared with the winter; however, there were no changes in sensitivity of the testes to LH during different seasons of the year as indicated by the lack of a dose of bLH x season interaction.

Chronic treatment with an agonist of gonadotropin-releasing hormone enhances luteal function in cattle.

Our hypothesis was that luteal function, as determined by plasma progesterone concentrations, and corpus luteum (CL) size is enhanced in cattle administered an agonist of GnRH when the CL is developing as compared with administration of an agonist when the CL is fully functional. Cattle were chronically administered a GnRH agonist, azagly-nafarelin, from Day 3 to Day 21 (D3) or Day 12 to Day 21 (D12) or served as untreated control females (Day 0 = behavioral estrus). Blood samples were serially collected on Days 7 and 14 to evaluate LH secretory patterns and twice daily to measure plasma progesterone. Ultrasonographic examinations were conducted daily to record the area of the CL. CL size and plasma progesterone concentrations were both enhanced in the D3 group as compared with the control group. Progesterone was increased in the D12 group on Days 16 and 17 as compared with the control females. Treatment with GnRH agonist increased basal and mean LH concentrations in both D3 and D12 groups as compared with the controls. We rejected our hypothesis because chronic administration of a GnRH agonist increased plasma progesterone when administered both when the CL was developing and when it was fully functional. The enhanced luteal function was likely due to increased basal LH.

Effect of chronic treatment with the gonadotrophin-releasing hormone agonist azagly-nafarelin on basal concentrations of LH in prepubertal bulls.

Administration of GnRH agonist for an extended period inhibits pulsatile LH release but enhances testicular function of bulls. The mechanism whereby long-term administration of GnRH agonist enhances testosterone concentration in the blood of bulls has not been determined. The aim of this study was to determine whether chronic treatment with the GnRH agonist, azagly-nafarelin, increases blood concentrations of LH and FSH in prepubertal bulls. Two different doses of the GnRH agonist were administered via Alzet mini-osmotic pumps for 28 days. Blood samples were collected at 20 min intervals for 24 h at days 2, 13 and 25 of treatment. Agonist-treated groups had reduced testosterone pulse frequency (P < 0.05) and increased mean and basal concentrations of testosterone (P < 0.05) compared with untreated control bulls. Basal LH concentrations were higher in agonist-treated bulls during all three periods (P < 0.05) and overall (1 ng ml(-1) higher, compared with control bulls; P < 0.001). Frequency of LH pulses in the agonist-treated groups was reduced to less than one pulse in 24 h. Agonist-treated bulls tended to have (P < 0.10) or had (P < 0.05) a slight but significant increase in blood FSH concentration. In conclusion, the higher blood testosterone concentration in bulls after prolonged treatment with GnRH agonist may result, at least in part, from changes in the testes induced by enhanced basal concentration of LH.