Opioidergic control of gonadotrophin secretion in the prepubertal gilt during restricted feeding and realimentation

Prepubertal gilts, having undergone a 7-day period of feed restriction to a maintenance ration, were allocated to one of 4 treatments; restricted feeding at 09:00 and 17:00 h for an 8th day both with (Group RN) and without (Group R) administration of the opioid antagonist naloxone hydrochloride (1 mg.kg-1 at 09:30 h followed by 0.5 mg.kg-1 at hourly intervals for 7 h), or feed to appetite with (Group ALN) and without (Group AL) naloxone administration. Gilts were bled at 10-min intervals on Day 8 from morning to evening feed and plasma LH, FSH and prolactin concentrations were measured by radioimmunoassay. Compared with Group R gilts, Group AL gilts exhibited significantly (P less than or equal to 0.05) higher mean and maximum LH concentrations and pulsatility, lower prolactin concentrations (P less than 0.05) but no significant difference in FSH secretion. Naloxone significantly depressed the increase in LH after re-feeding (Group ALN) (P less than 0.05). Once again there were no significant effects on FSH secretion. Naloxone also significantly depressed prolactin secretion in feed-restricted gilts (P less than 0.05). These results confirm that re-feeding of feed-restricted prepubertal gilts stimulates an immediate increase in LH secretion and that this elevation is not mediated via a suppression of inhibitory endogenous opioidergic tone. Rather, naloxone treatment appeared to expose a latent inhibition of LH secretion. The control of LH secretion is distinct from that of FSH in this model.     

Response of sows to oestradiol benzoate treatment after weaning at two stages of lactation

The timing and dosage of oestradiol benzoate injected after weaning was critical with respect to the pattern of behavioural oestrus and the ovarian stimulation achieved; treatment on the day of weaning (Day 0) and Day 1 with 60 micrograms oestradiol benzoate/kg body wt produced poor ovulatory responses and abnormal oestrous behaviour. Treatment on Day 2 with 30 micrograms oestradiol benzoate/kg resulted in consistent oestrus and ovulatory responses although the ovulation rates (10 . 6 +/- 1 . 1 in 3-week and 12 . 2 +/- 1 . 7 in 5-week weaned sows) were below those expected in fertile untreated sows weaned at these times. The timing of the preovulatory LH surge (53 . 6 +/- 2 h after oestradiol benzoate) was closely synchronized in all sows and a similar synchronous rise in plasma progesterone concentrations 100 +/- 4 h after oestradiol benzoate suggests a similar synchrony of ovulation. The magnitude of the LH and FSH responses to oestradiol benzoate were similar to those that occur in untreated sows and similar differences also existed in gonadotrophin secretion related to the length of lactation.     

Differences in follicular morphology, steroidogenesis and oocyte maturation in naturally cyclic and PMSG/hCG-treated prepubertal gilts

Ovaries were obtained from naturally cyclic pigs on Days 16-17, 18, 19, 20 and 21 of the oestrous cycle and on the basis of observed follicular characteristics were assigned as representative of the early (Group 1), mid- (Groups 2 and 3) or late (after LH; Group 4) follicular phase. Follicular development in cyclic gilts was compared with that in ovaries obtained from late prepubertal gilts 36 (Group 5) or 72 (Group 6) h after treatment with 750 i.u. PMSG alone, or with a combination of 500 i.u. hCG 72 h after PMSG and slaughter 30-40 h later (Group 7). After dissection of all follicles greater than 2 mm diameter, follicular diameter, follicular fluid volume, follicular fluid concentrations of progesterone, oestradiol and testosterone, and the stage of oocyte maturation were determined. Combined PMSG/hCG treatment of immature gilts resulted in a pattern of follicular development different from that in naturally cyclic gilts during the follicular phase. Overall exogenous gonadotrophin treatment also increased (P less than 0.001) the variability in follicular diameter and fluid volume. Comparisons between appropriate groups also established differences in the variability of both morphological (diameter and volume, Group 1 vs Group 5; P less than 0.05) and biochemical development (follicular fluid oestradiol, Group 3 vs Group 6 and Group 4 vs Group 7; both P less than 0.05). Such differences in both morphological and biochemical characteristics between cyclic and PMSG/hCG-treated gilts were particularly evident in the population of larger (greater than 6 mm) follicles. These results indicate that the pattern of follicular development in naturally cyclic and in PMSG/hCG-treated gilts is dissimilar and suggests that the ovaries of gonadotrophin-treated prepubertal gilts are functionally different from the ovaries of mature females.     

In vitro fertilization of in vitro matured pig oocytes: effects of boar and ejaculate fraction

Ejaculates from 3 young boars were collected on 4 occasions as a series of separate 15-ml fractions. The contribution of different fractions of these ejaculates to observed variability in the quality of the semen when used for IVF was then determined. On the basis of sperm concentration, 3 fractions representing the first peak concentration (Fraction 1), the lowest sperm concentration after Fraction 1 (Fraction 2), and the second peak concentration (Fraction 3) were selected for analysis in vitro. Oocyte-cumulus-granulosa cell complexes were obtained by dissection from slaughterhouse ovaries. In vitro matured oocytes were randomly assigned for fertilization by the 3 semen samples from each boar. Sperm concentration was the same in all the samples during prefertilization incubation, while the final concentration for fertilization was 5 x 10(5) sperm/ml. Data were analysed using ANOVA for a split-plot design. In the presence of fraction effects, Student-Newman-Keuls (SNK) test was used for multiple comparison of treatment means. Oocyte penetration rates differed among fractions (P = 0.001) and varied from 69 to 100% (mean 95.7%) for Fraction 1, from 0 to 100% (mean 53.3%) for Fraction 2, and from 50to 100% (mean 89.9%) for Fraction 3. There were also differences in male pronuclear formation rate (P = 0.028; mean 27.6, 9.3 and 16.4% for Fractions 1, 2 and 3, respectively); in the rate of polyspermy (P = 0.0001; mean 92.3, 31.9 and 76.3% for Fractions 1, 2 and 3, respectively); and in the number of penetrated spermatozoa per oocyte P = 0.002; mean 5.58, 1.94 and 4.07 for Fractions 1, 2, and 3, respectively). The first peak concentration of semen (Fraction 1) showed superiority in fertilizing ability and less variability in penetration rate from replicate to replicate compared with the other 2 fractions. By multiple comparison, Boar 1 showed higher rates of penetration (P < 0.05), male pronuclear formation (P < 0.05) and polyspermy (P < 0.05) than the other 2 boars. There was no fraction-by-boar interaction. The IVM-IVF system adopted proved to be a promising method for boar semen evaluation.     

Role of the tubulin-microtubule system in lymphocyte activation

The role of the tubulin-microtubule system was examined in human peripheral blood leukocytes after activation with phytohemagglutinin (PHA). Soluble tubulin and microtubules were measured with a [(3)H]colchicine-binding assay. It was found that the tubulin content of PHA-activated lymphocytes was consistently increased relative to total protein content after 36 h of culture. There was no increase in the proportion of total tubulin synthesis which was present as microtubules at 36 h. Nevertheless, as a result of increased tubulin synthesis, there was a two-to three-fold increase in total microtubular mass. Colchicine, which disrupts microtubles, was used to assess the role of microtubule assembly in the sequence of events which follow lymphocyte activation, namely lymphokine release, protein synthesis, RNA synthesis, and DNA synthesis. Colchicine consistently inhibited DNA synthesis but did not inhibit release of the lymphokine, osteoclast activating factor (OAF). Protein and RNA syntheses were inhibited much less than DNA synthesis. The fact that some effects of PHA on lymphocytes appear to require intact microtubules and at least one does not suggest that the microtubule dependent step in PHA-stimulated lymphocyte activation occurs at a stage after propagation of the signal from the membrane to the cell interior.     

Time of the preovulatory LH surge in the gilt and sow relative to the onset of behavioral estrus

Two experiments were conducted to study the time of occurrence of the preovulatory LH surge in pigs. Sampling every ten minutes in six cycling gilts before and after onset of standing estrus revealed the preovulatory surge began from 8 hr before to 12 hr after the lordosis reflex was elicited. Three of six gilts initiated the preovulatory LH release coincident with the onset of estrus. Data from 28 postpartum sows, with samples drawn every six hours commencing with the onset of estrus, indicated maximum LH levels were present at the first observance of estrus. Six of the 28 sows had an LH peak 18-24 hr after the onset of estrus.     

Ontogeny of the opioidergic regulation of LH and prolactin secretion in lactating sows. II: Interaction between suckling and morphine administration

Administration of morphine to ten suckled and nine zero-weaned (piglets removed immediately after farrowing) sows was used to investigate the apparent absence of opioid regulation of LH and prolactin secretion in early lactation. Blood samples were collected at 10 min intervals at 24-30, 48-54, 72-78 h post partum, and for a 12 h period from 08:00 to 20:00 on day 10 after farrowing. Morphine (0.1 mg kg-1) was administered as three i.v. bolus injections at intervals of 1 h during the last 3 h of each of the 6 h sampling periods, and at 6, 7 and 8 h after the beginning of sampling on day 10. There were significant (P < 0.001) group (zero-weaned versus suckled), time and morphine effects on LH secretion. Plasma LH concentrations increased (P < 0.001) within 48 h of farrowing in zero-weaned sows. Long-term trends of an increase in mean plasma LH in the sampling periods before treatment were attenuated in both groups by morphine treatment. Morphine also significantly inhibited (P < 0.05) prolactin secretion in suckled sows. In zero-weaned sows, plasma prolactin was already low at the start of sampling and did not change with time or in response to morphine treatment. Therefore, the inability to demonstrate an opioidergic involvement in the suckling-induced inhibition of LH secretion during the early post-partum period in sows is not due to a lack of opioid receptors. Furthermore, in suckled sows, morphine is stimulatory to systems that have an inhibitory effect on prolactin secretion.     

The time of ovulation in relation to estrus duration in gilts

The objective of this study was to determine time of ovulation, monitored by transcutaneous ultrasonography, relative to the duration of estrus in gilts. We exposed 92 cyclic gilts, Camborough x Canabrid terminal line, at Day 19 of their third estrous cycle to vasectomized boars every 6 h for the detection estrus. Transcutaneous ultrasonography was performed every 6 h, starting 24 h after the onset estrus, to determine time of ovulation. Estrus duration was, on average, 52.6 h (range: 30 to 72 h), and ovulation occurred between 30 and 60 h after the onset of estrus (mean: 44 h), about 85 % of the way through the estrus period. The time of ovulation during estrus was dependent on the duration of estrus (Time of ovulation = (duration of estrus) x 0.409 + 22.7; r = 0.57, P = 0.0001). Prediction of the time of ovulation in relation to duration of estrus is important for determining the optimal time for inseminating gilts. This knowledge would contribute to an improvement in the fertilization rate and in reproductive efficiency of the breeding herd.     

Simulated brain tumor growth dynamics using a three-dimensional cellular automaton

We have developed a novel and versatile three-dimensional cellular automaton model of brain tumor growth. We show that macroscopic tumor behavior can be realistically modeled using microscopic parameters. Using only four parameters, this model simulates Gompertzian growth for a tumor growing over nearly three orders of magnitude in radius. It also predicts the composition and dynamics of the tumor at selected time points in agreement with medical literature. We also demonstrate the flexibility of the model by showing the emergence, and eventual dominance, of a second tumor clone with a different genotype. The model incorporates several important and novel features, both in the rules governing the model and in the underlying structure of the model. Among these are a new definition of how to model proliferative and non-proliferative cells, an isotropic lattice, and an adaptive grid lattice.