A new role of H89: Reduces capacitation-like changes through inhibition of cholesterol efflux, calcium influx, and proteins tyrosine phosphorylation during sperm cryopreservation in buffalo.

It is a known fact that cryopreservation initiates premature capacitation in spermatozoa during the cryopreservation process. Protein tyrosine phosphorylation is a landmark of cascade reaction accountable for capacitation or capacitation-like changes in spermatozoa. Therefore, our hypothesis was to test an inhibitor (H89) that reversibly inhibits the cascade reaction responsible for capacitation during the cryopreservation process but does not hamper normal capacitation and fertilizing ability of sperm. For this, sixteen ejaculates were collected from Murrah buffalo bulls (n = 4). Each ejaculate was divided into four equal aliquots and diluted in an egg yolk-based semen dilutor supplemented with 0, 2, 10, and 30 µM concentrations of H89 and cryopreserved. Interestingly, H89 reduces cholesterol efflux from spermatozoa and protects spermatozoa from membrane damage during the cryopreservation process. H89 did not prevent lipid peroxidation of the sperm membrane. H89 reduced intracellular calcium concentration in spermatozoa in a dose-dependent manner, but tyrosine phosphorylation reduction was observed in the 2 and 10 µM H89 groups. The CTC assay revealed that the percentage of uncapacitated spermatozoa in different treatment groups increases in a dose-dependent manner. In the in vitro capacitation medium, the effect of H89 is abolished and spermatozoa underwent normal capacitation, but H89-treated spermatozoa attached to zona pellucida in large numbers compared to untreated spermatozoa. In conclusion, H89 does not only inhibit tyrosine phosphorylation of spermatozoa but it reduces cholesterol efflux and calcium influx, and ultimately reduces capacitation-like changes during the cryopreservation process.

A new role for RU486 (mifepristone): it protects sperm from premature capacitation during cryopreservation in buffalo.

The objective of this study was to determine the mechanism by which RU 486 (mifepristone) protects sperm to undergo premature capacitation during cryopreservation. For this, semen ejaculate (n = 20) was divided into four equal fractions and diluted using egg yolk-based extender supplemented with different concentrations of RU 486 (0, 5, 10 and 20 µM) and cryopreserved. We found that RU 486 did not impair the post-thaw sperm kinetics and motility but prevented cholesterol efflux, calcium influx, and protected CatSper channels during cryopreservation. The RU 486 protected sperm from premature capacitation which was confirmed by intracellular calcium level, expression of tyrosine phosphorylated proteins (75 and 80 kDa) and CTC (chlortetracycline) assay. Furthermore, antioxidant ability of RU 486 was reflected by the ferric reducing ability, lower production of sperm malondialdehyde and intracellular reactive oxygen species. Also, we demonstrated that RU 486 treated sperm underwent normal capacitation, zona pellucida binding and zygote cleavage indicating normal fertilizing ability of sperm. In conclusion, we report a new role of RU 486 in protecting buffalo sperm from premature capacitation during cryopreservation.

Diagnosis of pathological conditions of kidney by two-dimensional and three-dimensional ultrasonographic imaging in dogs.

AIM: The objective of the study was to obtain and compare two-dimensional (2D) and three-dimensional (3D) ultrasonographic images of the kidney in different disease conditions. MATERIALS AND METHODS: In this study, 11 clinical cases of different age groups of dogs suffering from kidney diseases were diagnosed by 2D and 3D ultrasonography at Teaching Veterinary Clinical Complex, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar. The ultrasound (US) machine used for this study was 3D US machine (Nemio-XG: Toshiba, Japan) having four-dimensional (4D) volumetric probe. The images were acquired with 3-6 MHz 2D curvilinear transducer and 4.2-6 MHz 4D volumetric curvilinear transducer. RESULTS: Nephritis was diagnosed in four dogs aged between 5 months and 6 years. In all the cases of nephritis diffuse increase in echogenicity of kidney, parenchyma was observed. Two dogs with end-stage kidney disease were also diagnosed. In both 2D and 3D ultrasonography, the kidney size was decreased and architectural details were also lost in them. The cases of regional renal diseases diagnosed were hydronephrosis and nephrolithiasis. Dilated renal pelvis was the common finding in all the three cases of hydronephrosis in both 2D and 3D ultrasonogram. Nephroliths were observed in one case with the history of hematuria and oliguria. The multifocal renal disease diagnosed in this study was dysplastic polycystic kidney. In 2D ultrasonogram, six anechoic cavities appeared with thin strip of renal parenchyma. In 3D ultrasonogram, the cysts appeared as black anechoic areas. CONCLUSION: The result of the current study showed that the clinical conditions of kidney such as nephritis, end-stage kidney, hydronephrosis, polycystic kidney, and nephrolithiasis can be diagnosed easily using 2D and 3D ultrasonography. Visualization of renal structures was clear in 2D ultrasonography in the conditions of nephritis and end-stage kidney. However, the conditions such as hydronephrosis, polycystic kidney, and nephrolithiasis were visualized clearly in both 2D and 3D ultrasonography equally.

Study on development of accessory sex glands in prepubertal kids using two-dimensional ultrasonography.

AIM: The present study was undertaken to study growth pattern of accessory sex glands in prepubertal kids from 2 weeks to 6 months of age using two-dimensional ultrasonography. MATERIALS AND METHODS: The study was conducted on six Beetal kids. The scanning of accessory sex glands was done in standing position using rectal probe and measurements were recorded. Data collected were statistically analyzed using one-way ANOVA followed by Duncan multiple range test was performed using the SPSS (16.0) system for windows. RESULTS: With the advancement of age all the dimensions of glands increased. Both the lobes of prostate gland showed an increase in width with advancement of age. Width of prostate above the urethra (W1) showed a significant increase at 2, 10, and 20 weeks of age, whereas non-significant increase from 2 to 8, 10 to 19, and 20 to 24 weeks of age was recorded. Width of prostate below the urethra (W2) showed a significant increase at 20 weeks of age, whereas non-significant increase was recorded during rest of period of growth. Left and right bulbourethral gland showed a similar pattern of growth with the advancement of age. The circumference dimensions increased significantly at 2, 16, 20, and 21 weeks of age for both glands. The increase was non-significant from 4 to 14, 16 to 19, and 20 to 23 weeks of age. The same pattern was observed for left and right seminal vesicular gland. CONCLUSION: Significant growth in three accessory sex glands in prepubertal kids was not observed at the same age. The trend observed was that the prostate was the first gland to show significant growth at 10 weeks of age followed by a significant increase in seminal vesicles and bulbourethral gland at 14 and 16 weeks of age, respectively.

A comparative study of two-dimensional and three-dimensional ultrasonography in evaluation of gastric affections in dogs.

AIM: The objective of the study was to obtain and compare the two-dimensional (2D) and three-dimensional (3D) ultrasonographic images of pathological conditions of the stomach in dogs in clinical cases. MATERIALS AND METHODS: In our study, 12 clinical conditions of the stomach were recorded using ultrasonography. The ultrasound machine used for this study was 3D ultrasound machine (Nemio-XG: Toshiba, Japan) having four-dimensional volumetric transducer. RESULTS: Present study was done to compare 2D and 3D ultrasonographic images in different gastric affections in dogs. In case of uremic gastropathy due to inflammatory response, the wall of the stomach was 0.6 cm thick and hyperechoic and gastric folds were also hyperechoic indicative of gastritis. In second, third, and fourth case of gastritis the wall of the stomach was 0.7, 0.6, and 0.55 cm, respectively thick and hyperechoic. In fifth and sixth case of gastritis, inflammatory response due to ingestion of polythene and sand led to gastritis and ultrasonographically, the wall of the stomach was 0.6 cm and 0.7 cm thick, respectively, and hyperechoic. In case of gastric ulcer, ultrasonographically, there was a disruption of gastric mucosal layer. In cases of gastric dilatation, anechoic content indicating fluid was seen in stomach area and due to dilatation boundary of the stomach was not clear and the increase in the lumen of the stomach was observed. In case of foreign body, ultrasonographically the wall of the stomach was 0.55 cm thick and hyperechoic. In the middle of the stomach, multiple hyperechoic shadows of the foreign bodies i.e. leather and bunch of straw of grass were observed. In case of pyloric stenosis ultrasonographically, anechoic lumen of the pylorus surrounded by 0.5 cm hypoechoic thickened muscle. In some cases, 3D ultrasonography was not diagnostic i.e. gastric foreign bodies and gastric dilatation. These conditions were better visualized on the 2D sonogram. CONCLUSION: The appearance of clinical conditions of the stomach such as gastritis and pyloric stenosis were more distinct on 3D ultrasonogram than 2D ultrasonogram. The 3D ultrasonography was not diagnostic in cases of gastric foreign bodies and gastric dilatation.

Diagnosis of urinary bladder diseases in dogs by using two-dimensional and three-dimensional ultrasonography.

AIM: The objective of this study was to obtain and compare two-dimensional (2D) and three-dimensional (3D) ultrasonographic images of the urinary bladder in different disease conditions. MATERIALS AND METHODS: The present study was conducting on total 10clinical cases of the urinary bladder in dogs. The ultrasound (US) machine used for this study was 3D US machine (Nemio-XG: Toshiba, Japan) having a four-dimensional volumetric probe. RESULTS: In the present study, the inflamed thickened wall was clearly visible with the distinction of different layers of the urinary bladder wall in some of the cases of cystitis using 2D ultrasonography. In 3D sonogram, the urinary bladder was visualized as a large anechoic structure with no distinction of different layers of the bladder wall. The cystoliths were clearly visible as hyperechoic structures with distal acoustic shadow in 2D sonogram and appeared as a bright echogenic area in 3D sonogram. In case of urinary bladder neoplasia in 2D ultrasonogram, the bladder lumen was found to be occluded with a large growth imaged as focal anechoic areas in the tissue of mixed echogenicity with small hyperechoic dots in this tissue parenchyma. In 3D ultrasonogram, a tissue of mixed echogenicity of pus was also observed. CONCLUSION: From the present study it was concluded that 2D and 3D ultrasonography is very helpful for diagnosis of different clinical conditions of the urinary bladder such as cystitis, cystoliths, and urinary bladder neoplasia. The cavity of urinary bladder was more clearly visualized in 3D ultrasonography, but the distinction of different layers of the bladder wall was visualized only in 2D ultrasonography. The distinct shadow of pus and cystoliths were visible in 2D ultrasonogram. The visualization of pus in 3D ultrasonography was done for the first time in present study.

Sexual maturation in the bull.

In this review, we describe the process of sexual maturation in the bull calf. The testes of the bull grow relatively slowly until approximately 25 weeks of age and then a rapid phase of growth occurs until puberty, at 37-50 weeks of age. During the early postnatal phase of slower growth of the testis pre-spermatogonia and some spermatogonia are established, adult Leydig cells appear and undifferentiated Sertoli cells are produced. The rapid testicular growth, after 25 weeks of age, consists of marked increases in the diameter and length of the seminiferous tubules, dramatic proliferation and differentiation of germ cells, with mature spermatozoa occurring between 32 and 40 weeks of age. The adult Leydig cell population is largely in place by 30 weeks of age and that of Sertoli cells by 30-40 weeks of age. Serum concentrations of LH increase from 4 to 5 weeks of age, to an early postnatal peak at 12-16 weeks of age, followed by a decline to 25 weeks of age. Serum FSH concentrations are high postnatally, declining to approximately 25 weeks of age. Serum testosterone concentrations increase during the phase of rapid testicular growth. Hypothalamic opioidergic inhibition may abate transiently to allow the early postnatal increase in LH secretion, while testicular androgenic negative feedback probably contributes to the decline in gonadotropin secretion to 25 weeks of age. Several lines of study have led us to suggest that early postnatal gonadotropin secretion is pivotal in initiating the process of sexual maturation in the bull calf.

Blockage of urine by intravesical ejaculate in cynomolgus monkeys.

BACKGROUND: The present communication reports intravesical semen coagulation and formation of a larger precipitate in two Cynomolgus monkeys. METHODS: Ultrasound of the urinary bladder and light microscopy of intravesical coagulates. RESULTS: These monkeys suffered from complete blockage of urine output and surgery was required to remove the sperm mass. Microscopic examination of the urine revealed millions of sperm as a cause of the mass and the blockage of urine output. CONCLUSIONS: Retrograde ejaculation of sperm may cause coagulation of ejaculates in the bladder of the cynomolgus monkey Macaca fascicularis. However, involvement of sperm mass in blockage of urine passage has not been described in this species.

Opioidergic, dopaminergic and adrenergic regulation of LH secretion in prepubertal heifers.

Studies have shown inhibitory effects of endogenous opioids on LH secretion in early post-natal heifers. However, it is not clear whether these effects change during the rest of the prepubertal period or whether the inhibitory influences on the GnRH neurones are direct or by way of other neuronal systems. Two experiments were performed in heifer calves to study the developmental patterns of opioidergic, dopaminergic and adrenergic regulation of LH and the possible interactions between opioids and dopaminergic and adrenergic neuronal systems, in the regulation of LH secretion. In Expt 1 four groups each of five heifer calves were used. Blood samples were taken every 15 min for 10 h and each calf received one of the following treatments as a single injection at 4, 14, 24, 36 and 48 weeks of age: (i) naloxone (opioid antagonist, 1 mg kg(-1), i. v.); (ii) sulpiride (dopamine D2 antagonist, 0.59 mg kg(-1), s.c.); (iii) naloxone and sulpiride combined; or (iv) vehicle (control group). Treatments began after the first blood sample was taken. The design of Expt 2 was similar; a separate group of heifer calves was assigned to receive one of the following treatments as a single injection at 4, 14, 24, 36 and 48 weeks of age: (i) naloxone; (ii) phenoxybenzamine (an alpha-adrenoreceptor blocker, 0.8 mg kg(-1), i. v.); (iii) naloxone and phenoxybenzamine; (iv) or vehicle. Results from Expt 1 showed that the maximum concentration of LH and the number of calves responding to treatments with an LH pulse was higher in the first hour after treatments at 36 and 48 weeks of age in the naloxone group compared with the control or sulpiride groups (P < 0.05). These values in the naloxone group also increased over time and were greatest at 48 weeks of age (P < 0.05). In heifers given naloxone + sulpiride treatment at 36 and 48 weeks of age, maximum concentrations of LH in the first hour after treatment did not differ from the naloxone and control groups. In Expt 2, at 36 and 48 weeks of age, treatment with naloxone with or without phenoxybenzamine resulted in higher concentrations of LH than in the controls (P < 0.05). No pulses were seen over the first hour of treatment at 36 and 48 weeks of age in heifers treated with phenoxybenzamine. The 10 h periods of blood sampling at 48 weeks of age revealed that phenoxybenzamine alone suppressed LH pulse frequency and mean serum concentrations of LH compared with the control group (P < 0.05). It was concluded that a strong or more acute inhibition of LH secretion by endogenous opioids developed in mid- to late prepubertal heifers, or alternatively, that removal of opioidergic inhibition at the GnRH neurone unmasked stimulatory inputs that were greater in heifers close to first ovulation. Since sulpiride appeared to negate in part the effects of naloxone on LH release, the suppressive effects of opioids could be exerted in part through the inhibition or blocking of a stimulatory dopaminergic system. alpha-Adrenergic neuronal systems have stimulatory effects on LH release, especially during the late prepubertal period, but do not appear to mediate opioidergic inhibition of LH secretion in prepubertal heifer calves.

Transcriptional control of development, protein synthesis, and heat-induced heat shock protein 70 synthesis in 2-cell bovine embryos.

Experiments were performed to evaluate the role of transcription in early development of bovine embryos. Two transcription inhibitors-5, 6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DRB) and actinomycin D-were used to test whether 1) the inhibitors alter the rate of early embryonic development and protein synthesis, 2) heat shock increases the steady-state amounts of mRNA for the inducible form of heat shock protein 70 (HSP70) in embryos, and 3) this latter effect is blocked by transcription inhibitors. Addition of either DRB or actinomycin D to culture medium beginning 8 h postinsemination (hpi) reduced the proportion of oocytes that had undergone cleavage by 32-34 hpi. Both transcription inhibitors also reduced the proportion of cleaved embryos that reached the 4-cell stage by 32-34 hpi. Incorporation of (35)S-labeled amino acids into de novo synthesized protein by bovine 2-cell embryos was lower for embryos cultured with DRB. Using reverse transcription-polymerase chain reaction, HSP70 mRNA in 2- and 4-cell embryos was increased by exposure to 42 degrees C. Both inhibitors reduced amounts of HSP70 mRNA at 42 degrees C. Results indicate that bovine embryos can undergo transcription in response to heat shock as early as the 2-cell stage. Moreover, the observations that transcription inhibitors reduce rates of cleavage and early development point out the importance of transcription for development from the earliest period of embryonic life.

Reproductive and endocrine function in rams exposed to the organochlorine pesticides lindane and pentachlorophenol from conception.

There is controversy over the potential endocrine modulating influence of pesticides, particularly during sensitive phases of development. In this study, ram lambs were exposed to lindane and pentachlorophenol from conception to necropsy at 28 weeks of age. The rams (and their mothers) were given untreated feed (n = 7) or feed treated with 1 mg kg-1 body weight per day of lindane (n = 12) or pentachlorophenol (n = 5). Semen was collected from 19 weeks onwards and reproductive behaviour was tested at 26 weeks. Serum was collected every 2 weeks and at 27 weeks every 15 min for 6 h during both day and night, and for 1 h before and 5 h after stimulation with GnRH, adrenocorticotrophic hormone and thyroid-stimulating hormone. The pesticides did not affect body weight and ejaculate characteristics, or cause overt toxicity. In pentachlorophenol-treated rams, scrotal circumference was increased. However, seminiferous tubule atrophy was more severe and epididymal sperm density was reduced in comparison with untreated rams at necropsy (P < 0.05). Thyroxine concentrations were lower in pentachlorophenol-treated rams than in untreated rams (P < 0.05). However, after thyroid-stimulating hormone treatment, the thyroxine response was unaltered. Reproductive behaviour was reduced in lindane-treated rams compared with control rams (P < 0.05). Serum LH and oestradiol concentrations during reproductive development, LH pulse frequency at 27 weeks and testosterone secretion after GnRH treatment were lower in lindane-treated rams than in untreated rams (P < 0.05). In summary, the effects of pentachlorophenol on the testis may be linked to a decrease in thyroxine concentrations, and reduced reproductive behaviour in lindane-treated rams may be related to decreased LH, oestradiol and testosterone concentrations.

Ovarian antral follicular dynamics and their relationships with endocrine variables throughout the oestrous cycle in breeds of sheep differing in prolificacy.

Transrectal ultrasonography of ovaries was performed each day in non-prolific Western white-faced (n = 12) and prolific Finn ewes (n = 7), during one oestrous cycle in the middle portion of the breeding season (October-December), to record the number and size of all follicles > or = 3 mm in diameter. Blood samples collected once a day were analysed by radioimmunoassay for concentrations of LH, FSH and oestradiol. A cycle-detection computer program was used to identify transient increases in concentrations of FSH and oestradiol in individual ewes. Follicular and hormonal data were then analysed for associations between different stages of the lifespan of the largest follicles of follicular waves, and detected fluctuations in serum concentrations of FSH and oestradiol. A follicular wave was defined as a follicle or a group of follicles that began to grow from 3 to > or = 5 mm in diameter within a 48 h period. An average of four follicular waves per ewe emerged during the interovulatory interval in both breeds of sheep studied. The last follicular wave of the oestrous cycle contained ovulatory follicles in all ewes, and the penultimate wave contained ovulatory follicles in 10% of white-faced ewes but in 57% of Finn ewes. Transient increases in serum concentrations of FSH were detected in all animals and concentrations reached peak values on days that approximated to follicle wave emergence. Follicular wave emergence was associated with the onset of transient increases in serum concentrations of oestradiol, and the end of the growth phase of the largest follicles (> or = 5 mm in diameter) was associated with peak serum concentrations of oestradiol. Serum FSH concentrations were higher in Finn than in Western white-faced ewes during the follicular phase of the cycle (P < 0.05). There were no significant differences in serum concentrations of LH between Western white-faced and Finn ewes (P > 0.05). Mean serum concentrations of oestradiol were higher in Finn compared with Western white-faced ewes (P < 0.01). It was concluded that follicular waves (follicles growing from 3 to > or = 5 mm in diameter) occurred in both prolific and non-prolific genotypes of ewes and were closely associated with increased secretion of FSH and oestradiol. The increased ovulation rate in prolific Finn ewes appeared to be due primarily to an extended period of ovulatory follicle recruitment.

Short communication: lack of breed differences in responses of bovine spermatozoa to heat shock.

An experiment was conducted to test whether the magnitude of effects of heat shock on spermatozoal function were less for thermotolerant breeds (Brahman and other breeds with Brahman influence) than for breeds that evolved in northern Europe (Angus and Holstein). Frozen spermatozoa were thawed, purified by Percoll gradient centrifugation and incubated at 38.5, 41, or 42 degrees C for 4 h. Sperm motility was then analyzed with a Hamilton Thorn Motility Analyzer. Heat shock reduced the percentage of sperm that were motile, mean track speed, and mean path velocity. There were no significant breed x temperature interactions for these traits. The mean frequency of tail beat tended to be reduced by heat shock in bulls of Brahman-influenced breeds and, to a lesser extent, in Brahman bulls, but it was not affected by heat shock in Angus or Holstein bulls. For no traits were there significant temperature x bull within breed interactions. Overall, results indicate that 1) heat shock reduces motility of bovine spermatozoa and 2) genetic effects are unlikely to be an important determinant of the function of ejaculated sperm following heat shock.

Effects of season of birth on the prepubertal pattern of gonadotropin secretion and age at puberty in beef heifers.

There is an early transient rise in gonadotropin secretion in spring-born prepubertal heifers and there is an indication that this pattern is different in autumn-born heifers. The effect of season of birth on age and weight at puberty is equivocal. This study was designed to compare the temporal patterns of LH and FSH secretion between spring- and autumn-born heifers and to determine the effects of season of birth on age and weight at puberty. Blood samples from 2 groups of heifer calves born in spring (last week of March, n = 5) or autumn (last week of October, n = 5) were collected every other week from birth to puberty and every 15 min for 10 h at 6, 12, 18, 24 and 32 wk of age. Timing of puberty was determined by measuring progesterone in plasma samples collected every 2 to 3 d starting at 42 wk of age. Age and weight at onset of puberty did not differ between the 2 groups of heifers (P > 0.05); however, the autumn-born heifers tended to mature in a wider range of ages and weights. Based on the 10-h sampling periods, mean serum concentrations of LH and LH pulse frequency and amplitude were higher in spring-born heifers at 18 wk of age than in autumn-born heifers (P < 0.05). In spring-born heifers, LH pulse frequency increased over time to 32 wk of age, and LH pulse amplitude was higher at 12 and 18 wk than at 32 wk of age (P < 0.05). Autumn-born heifers had higher LH pulse frequency at 6 wk and showed a decrease in mean concentrations of LH at 12 and 18 wk of age (P < 0.05). The FSH pulse frequency of spring-born heifers was higher at 12 wk of age than in autumn-born heifers (P < 0.05), FSH pulse amplitude in autumn-born heifers decreased from 6 to 32 wk of age. It was concluded that although the mean age and weight at puberty did not differ between spring- and autumn-born heifers, the range in age and weight at puberty was wider in the autumn-born heifers. The patterns of LH secretion differed between spring- and autumn-born prepubertal heifers, with spring-born calves exhibiting an early rise in LH secretion, while mean serum concentrations of LH decreased during this period in autumn-born heifers.

Excitatory amino acid regulation of gonadotropin secretion in prepubertal heifer calves.

The mechanisms controlling the pulsatile release of gonadotropins in prepubertal heifers are not completely understood. We examined the role of excitatory amino acid neurotransmitters, via activation of N-methyl-D-aspartate (NMDA) receptors, in the control of pulsatile LH and FSH release during prepubertal development in heifers. Hereford heifer calves received 4.7 mg/kg of N-methyl-D,L-aspartic acid (NMA), a potent NMDA receptor agonist (n = 5, i.v.), or saline (n = 5, i.v.), as single doses, at 4, 8, 12, 24, 36, and 48 wk of age. Blood samples were collected every 15 min, for 1 h before and 9 h after injection, on the days of treatment. Injection of NMA resulted in an acute release of LH (p < 0.001) in 0, 3, 3, 4, 5, and 5 calves (p < 0.01) and of FSH (p < 0.001) in 0, 1, 2, 4, 3, and 2 calves at 4, 8, 12, 24, 36, and 48 wk of age, respectively. The peak response of LH and FSH release to NMA was at 15 min posttreatment, and these peak responses were highest at 36 wk of age (p < 0.05). We suggest that neuroexcitatory amino acids, through NMDA receptors, are involved in prepubertal development of LH and FSH secretion in heifer calves.

Effects of treatment with LH releasing hormone before the early increase in LH secretion on endocrine and reproductive development in bull calves.

Between 6 and 20 weeks of age an early increase in LH secretion has been reported in Hereford bull calves. Delaying this early increase in LH secretion delays testicular development. This study was designed to determine whether a premature increase in LH secretion during the early postnatal period enhances testicular development. Ten age- and body weight-matched Hereford bull calves were divided into two groups. One group (n = 5) received 200 ng LH releasing hormone (LHRH) i.v. every 2 h for 14 days, between 4 and 6 weeks of age. On the basis of blood samples taken every 15 min for 10 h, mean serum LH and testosterone concentrations and LH pulse frequency were increased by LHRH treatment (P < 0.05). Serum concentrations of FSH were not significantly influenced by treatment (P > 0.05). In treated animals at 24 weeks of age, mean serum testosterone concentrations and LH pulse amplitude were increased (P < 0.05). The concentrations of spermatozoa in electroejeculates collected at 52 weeks of age were greater in LHRH-treated compared with control calves. Testicular growth was enhanced by LHRH treatment and histological evaluation of the testis at 54 weeks of age showed increased spermatogenesis and also larger numbers of Sertoli cells per tubule cross-section as a result of LHRH treatment. We conclude that treatment with LHRH before the early increase in LH secretion altered testicular development and suggest that the early increase in LH secretion in bull calves may be critical for initiating and regulating the progression of reproductive maturation.

Ultrasonography of the developing reproductive tract in ram lambs: effects of a GnRH agonist.

In spring-born ram lambs, the testes (from 2 wk), prostate and vesicular glands (from 4 wk) were examined by ultrasonography every 2 wk up to 26 wk of age. Image analysis was done (numerical pixel values). Ram lambs were treated with a long acting formulation of a GnRH superagonist (Leuprolide acetate; 1.5 mg/kg) at 3 and 7 wk of age. In blood samples taken every 15 min for 8 h, mean serum LH, LH pulse amplitude, and basal and mean serum FSH concentrations were lower at 5 wk of age, and LH pulse frequency was lower at 15 wk of age in animals given Leuprolide acetate compared with those of the controls. There were no differences (P>0.05) in testis, prostate or vesicular gland development between treated and control animals. Testicular diameter of the left and right testes in transverse and longitudinal planes increased slowly to 8 wk of age, more rapidly to 18 wk of age, then more slowly to 26 wk of age (P<0.05). Numerical pixel values of testicular images decreased from 2 to 8 wk of age, increased to 22 wk of age and then plateaued. Width of the prostate increased from 4 to 26 wk of age, but length and width of the vesicular glands increased slowly to 8 wk of age, more rapidly to 18 wk of age and then plateaued (P<0.05). Numerical pixel values for the prostate declined from 4 to 8 wk and for the vesicular glands, declined from 4 to 10 wk of age; numerical pixel values increased to 12 wk and then decreased to a nadir at 18 wk, followed by a steady increase to 26 wk of age (P<0.05). We concluded that developmental patterns of numerical pixel values of the testes, prostate and vesicular glands in ram lambs reflect stages of development, but treatment with a GnRH superagonist at 3 and 7 weeks of age did not affect growth of testes, vesicular or prostate glands.

Assessment of development of the testes and accessory glands by ultrasonography in bull calves and associated endocrine changes.

The testes, prostate and vesicular glands of 10 bull calves were examined by ultrasonography every 2 wk from 2 to 46 wk of age, at which time the scrotal circumference (SC) of all the calves had reached pubertal size (28 cms). Computer-assisted image intensity analysis (numerical pixel values) was conducted. Blood samples were collected every other week from 2 to 46 wk of age. Testicular diameter increased in a linear manner from 2 to 46 wk of age, but the diameter measured in a transverse plane (caudal) was greater between 10 and 34 wk of age than when measured in a longitudinal (lateral) plane (P<0.05). Growth of the prostate and vesicular glands, based on dimensions, was linear, but vesicular gland length increased more rapidly after 32 wk of age (P<0.05). Image intensity of the vesicular glands and prostate declined from birth or 8 wk of age, respectively, to 14 wk of age, increased to 18 wk and then declined to a nadir at 30 wk, followed by a rapid increase to 34 wk of age for the vesicular glands and to 46 wk of age for the prostate (P<0.05). Image intensity of the testes showed an early increase to 6 to 8 wk of age and a subsequent increase from about 20 wk of age to 46 wk of age, with an inflection at 30 wk of age (P<0.05). There was a transient increase in mean serum concentrations of LH between 6 and 20 wk of age (P<0.05), and LH concentrations appeared to increase again after 36 wk of age (P>0.05). Mean serum concentrations of FSH declined with age (P<0.05). Mean serum concentrations of testosterone increased after 32 wk of age (P<0.05) In summary, numerical pixel values comprising the ultrasound images of the developing testes, prostate and vesicular glands revealed a complex development pattern that may reflect important details of developmental stages.

The involvement of dopaminergic and opioidergic neuronal systems in the control of the early rise in LH secretion in bull calves.

There is an early rise in secretion of luteinizing hormone (LH) in bull calves between 6 and 20 weeks of age. This study was designed to examine the involvement of opioidergic and dopaminergic neuronal systems in the control of this early rise in gonadotrophin secretion. Four groups of five bull calves were bled every 15 min for 10 h and administered one of the following treatments i.v. at 6, 14 and 24 weeks of age: a) saline; b) 1 mg naloxone/kg body weight every hour for 10 h; c) 0.8 mg pimozide/kg body weight as a single injection, after the first blood sample; d) 1 mg naloxone + 0.8 mg Pimozide/kg body weight. The early rise in LH secretion had started by 6 weeks of age, was at a peak at 14 weeks of age and was complete by 24 weeks of age (P<0.05). Naloxone (an opioid antagonist) treatment resulted in increased basal and mean serum concentrations of LH and LH pulse frequency at 14 weeks of age, increased mean serum LH concentrations of LH pulse frequency at 24 weeks of age, decreased FSH pulse amplitude at 14 weeks of age and increased mean serum concentrations of FSH at 24 weeks of age compared to control calves (P<0.05). Pimozide (a dopamine antagonist) decreased LH pulse amplitude at 24 weeks of age compared to all other groups of calves (P<0.05). Naloxone and pimozide in combination resulted in LH secretory patterns similar to naloxone treated calves or intermediate to naloxone and control calves at 14 weeks of age but similar to control calves at 24 weeks of age. In calves given the combination treatment, parameters of FSH secretion resembled those in the other groups of calves except for basal and mean serum concentrations of FSH at 24 weeks of age, which were similar to control or pimozide treated calves, but lower than in naloxone treated calves (P<0.05). At 14 weeks of age mean serum concentrations of testosterone were greater in calves given naloxone and pimozide in combination compared to controls, but at 24 weeks of age they were greater in the naloxone treated calves and lower in pimozide treated calves compared to the control calves (P<0.05). We concluded that an opioidergic inhibition of LH secretion was seen from the middle of the early rise in LH secretion (14 weeks of age) onwards but inhibition of FSH secretion was only seen at 24 weeks of age. A dopaminergic drive for LH and FSH secretion developed after the completion of the early rise in gonadotrophin secretion (24 weeks of age). Opioidergic inhibition of gonadotrophin secretion largely involved inhibition of a dopaminergic drive at the end of the early rise in LH secretion (24 weeks of age) but not at earlier ages.