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Biomedical subjects

M G Hunter

Publications and source records attributed to M G Hunter.

At least 55 records · Page 3Linked to original sources

Effects of testosterone and oestradiol on [3H]-thymidine incorporation by porcine granulosa and theca cells.

Experiments were carried out to investigate the effects of varying physiological concentrations (0, 10, 100, and 1000 ng ml-1) of oestradiol or testosterone on [3H]-thymidine incorporation by porcine granulosa and theca cells in vitro. Granulosa cells only were recovered from small (1-3-mm) follicles and both granulosa and theca cells recovered from large (4-8-mm) porcine follicles. Cells were cultured for 72 h in medium containing 10% foetal calf serum, 24 h in serum-free medium, and finally 40 h in serum-free medium containing [3H]-thymidine and appropriate steroid treatment. Although DNA per well was significantly higher (P < 0.05) at the end of culture in the theca cells than in the granulosa cells, neither steroid treatment had a significant (P > 0.1) effect on DNA concentration in either cell type. Overall, cells from small follicles incorporated significantly (P < 0.01) more [3H]-thymidine than those from medium follicles. Both oestradiol and testosterone significantly (P < 0.01) inhibited thymidine incorporation by cells from both follicle size categories, with a significant (P < 0.05) hormone X dose interaction. Finally, there was a highly significant (P < 0.001) interaction between the response of cells to different hormone concentrations and the follicle size from which they were recovered. These results indicate that both oestradiol and testosterone may act in an autocrine/paracrine manner to inhibit proliferation and encourage differentiation in follicular cells and thus are likely regulators of the later stage of antral follicle development in the pig.

Animals↗

Dissociation of the Jak kinase pathway from G-CSF receptor signaling in neutrophils.

Activation of the granulocyte colony-stimulating factor receptor (G-CSFR) induces rapid tyrosine phosphorylation of multiple intracellular substrates in proliferating cells and nonproliferating, terminally differentiated neutrophils. The kinases that couple ligand binding to tyrosine phosphorylation of cellular substrates by the G-CSFR with activation of specific functional programs are largely unknown. In this study, we examined early signaling events in proliferating and terminally differentiated cells following G-CSF stimulation to determine whether identical signaling cascades are activated. In murine Ba/F3 cells transfected with the human G-CSFR and NFS-60 cells constitutively expressing the murine G-CSFR, G-CSF induced tyrosine phosphorylation and activation of Jak1, Jak2, and Tyk2. Tyrosine phosphorylation of Stat3 and, to a lesser extent, Stat1 was also detected following G-CSF stimulation. Using a mitogenically incompetent human G-CSFR mutant in which Pro639 and Pro641 were substituted by alanine, the box 1 PDP motif was found to be required for activation of Jak kinases, tyrosine phosphorylation of the G-CSFR, and recruitment of Stat proteins. Notably, no activation of Jak1, Jak2, Tyk2, Stat1, or Stat3 was observed in neutrophils following G-CSF stimulation. In addition, there was no detectable activation in neutrophils of the recently cloned Jak3 kinase, which has been reported to be expressed at high levels as myeloid cells undergo terminal neutrophilic maturation. These results indicate a lack of involvement of Jak kinases in signaling by the G-CSFR in neutrophils, and suggest utilization of alternative signal transduction pathways distinct from those in proliferating cells. Activation of the Jak-Stat pathway correlates with proliferative signaling by the G-CSFR and requires the membrane-proximal box 1 PXP motif, which is conserved in members of the cytokine receptor superfamily.

Animals↗

Ovum pick up in swine: the influence of aspiration vacuum pressure on oocyte recovery from preovulatory follicles.

Endoscopical ovum pick up (OPU) in swine is a minimal invasive and reliable technique to aspirate oocytes from preovulatory follicles for studying intrafollicular development and oocyte maturation as well as for IVM/IVF-programs. The aim of the present study was to determine the effect of different aspiration vacuum pressures on oocyte recovery and on the morphology of cumulus oocyte complexes (COCs). Oestrus of crossbred Landrace gilts (n = 33) was synchronized by feeding altrenogest and follicular growth was stimulated with 1,000 IU PMSG 24 h after the last altrenogest application. On day 4 after PMSG application preovulatory healthy follicles of > 5 mm diameter were aspirated laparoscopically. Aspiration was carried out using a two-way cannula and an electronic aspiration pump. Five different vacuum pressures were used: 10, 17, 32, 47 and 66 ml water/min, corresponding to 30, 60, 125, 250 and 375 mm Hg, respectively. Fluids from different follicles were pooled per ovary and the morphology of cumulus oocyte complexes (COCs) was determined microscopically immediately after aspiration. COCs were classified into oocytes with intact cumulus (i.e. compact or slightly expanded COC), oocytes with corona radiata and denuded oocytes. A total of 695 follicles were aspirated and 501 COCs recovered. Increasing the aspiration pressure stepwise from 10 ml water/min up to 66 ml water/min resulted in a decrease in oocyte recovery rate. A significant higher (P < 0.05) rate of oocyte recovery (77.4% v.s. 59.8%) was achieved using a vacuum pressure of 17 ml water/min compared to 66 ml water/min, respectively. There was a tendency to reduce the portion of COCs with intact cumulus from 82 to 88% to 77% if the vacuum pressure increased to more than 47 ml water/min. A higher aspiration pressure provoked an increase (P < 0.05) in the number of denuded oocytes: 0 to 3% at 10 to 32 ml water/min, respectively, compared to 10% at 47 ml water/min to 17% at 66 ml water/min. These results demonstrate that variation in aspiration pressure affects oocyte recovery rate and COC quality. Aspiration vacuum pressure of 17 to 32 ml water/min was found to be optimal in swine both for oocyte recovery and COC quality.

Animals↗

BB-10010/MIP-1 alpha in vivo maintains haemopoietic recovery following repeated cycles of sublethal irradiation.

Macrophage inflammatory protein-1 alpha (MIP-1 alpha) is an inhibitor of stem cell proliferation affording protection against damage from agents that express their cytotoxicity specifically in the DNA synthesis phase of the cell cycle. Its ability also to modify the self-renewal capacity of the regenerating cells is now shown to improve and maintain haemopoietic recovery following therapy (sublethal irradiation) whose cytotoxic damage is not limited solely to the DNA-S phase of this cycle. Such non-cell cycle-active cytotoxic agents are used clinically in repeated treatment regimens, which are often limited or terminated because of accumulating haemopoietic damage. BB-10010, a non-aggregating variant of MIP-1 alpha, was administered as a continuous dose (1600 micrograms kg-1 24 h-1) via a subcutaneously implanted pump over a period of 7 days. A dose of 4.5 Gy total-body gamma-rays was given 3-4 h after implantation. Day 8 and 12 spleen colony-forming units (CFU-S) were assayed on days 1, 7 and 14 after irradiation. This cycle of treatment was repeated four times (total 56 days), and on day 14 of the last two cycles the marrow-repopulating ability (MRA) was also measured. In the control bone marrow (no BB-10010) CFU-S fell to < 1% of normal within 1 day of irradiation and recovered to 40% at 14 days. Repeated treatments increased the level of damage, and after four cycles CFU-S recovered to only 10% of normal. BB-10010 afforded little benefit in the first treatment cycle, but by the end of the fourth cycle CFU-S still recovered to 35% of normal. MRA was reduced to 7% of normal by the irradiation protocol-about half that maintained by BB-10010 protection. We conclude that BB-10010 (MIP-1 alpha) reduces the degree of accumulated haemopoietic stem cell damage following repeated non-cell cycle-specific cytotoxic insults-a principle which should be valuable in repeated clinical cytotoxic therapy regimens.

Animals↗

Role of estrogen and prostaglandin F2 alpha in premature luteal regression in monovulatory and superovulated red deer (Cervus elaphus).

The superovulation of red deer hinds with eCG is commonly associated with premature luteal regression. This study was an investigation of the endocrine mechanisms regulating luteal function after superovulation. Four groups of hinds (n = 7-8 per group) were treated with progesterone-impregnated intravaginal controlled internal drug-releasing (CIDR) devices for 12 days to synchronize estrus (CIDR device withdrawal = Day 0). Group 1 served as controls; group 2 received an i.m. injection of 0.25 mg estradiol benzoate (EDB) at 72, 84, 96, and 108 h after removal of the device; group 3 received an i.m. injection of 1200 IU eCG at -72 h; group 4 received both EDB and eCG treatments. Oxytocin-induced prostaglandin F2 alpha (PGF2 alpha) release was assessed on Day 4 by oxytocin challenge. Ovarian response was determined by laparoscopy on Day 14. Plasma steroid profiles were determined from thrice-weekly plasma samples collected from Day -13 to Day 35 (progesterone) and Days 0 to 14 (estradiol). EDB increased the incidence of premature luteal regression in monovulatory and eCG-treated animals (p < 0.05) and reduced the number of CL (p < 0.05) in eCG-treated animals. EDB and eCG each elevated plasma concentrations of estradiol and increased the incidence of significant oxytocin-induced PGF2 alpha release. These results support the hypothesis that eCG causes premature luteal regression by inducing prolonged estrogen secretion that sensitizes the endometrium to oxytocin, thereby eliciting PGF2 alpha release during the early luteal phase.

Animals↗

Effects of exogenous oxytocin and progesterone on GnRH-induced short luteal phases in anoestrous ewes.

Two experiments investigate the effects of oxytocin and progesterone on premature luteolysis in ewes. In Expt 1, 20 anoestrous ewes were induced to ovulate by multiple injections of GnRH (250 ng i.v. every 2 h for 24 h) followed by a bolus injection of GnRH (125 micrograms, i.v.). Ten ewes received a continuous infusion of oxytocin from the day after the GnRH bolus injection and the other ten ewes were infused with saline. Oxytocin infusion had no significant effect on the proportion of ewes with short luteal phases (P > 0.05). All ewes that had luteal phases of normal duration from either group (n = 9) exhibited a transient increase in plasma concentrations of progesterone 2 h after insertion of the pump. In Expt 2, 25 anoestrous ewes were treated with GnRH as in Expt 1. Five ewes were pretreated with progestagen for 11 days and ten ewes received progesterone (12 mg, i.m.) 24 h after the bolus injection of GnRH. All animals received an oxytocin injection (1 microgram, i.v.) on day 4 after the GnRH bolus. All five ewes that were pretreated with progestagen had normal luteal function and none exhibited a 13, 14-dihydro-15-keto PGF2 alpha (PGFM) response to oxytocin. None of the ten ewes injected with progesterone had a normal luteal phase and six ewes exhibited a PGFM response to oxytocin. Four ewes in the control group had normal luteal function and three had short luteal phases. It is concluded that (1) administration of oxytocin from about the time of ovulation does not prevent premature luteal regression; (2) a transient increase in progesterone at about the time of ovulation is associated with luteal phases of normal duration; (3) a more extended exposure to progesterone at about the time of ovulation prevents normal luteal function and may inhibit luteinization and (4) pretreatment with progesterone prevents luteolysis by reducing the uterine response to oxytocin early in the luteal phase.

Anestrus↗

Periovulatory endocrinology in high ovulating Meishan sows.

The present study was designed to investigate the hormone profiles (oestradiol, LH, FSH, inhibin, progesterone) in high ovulating Meishan sows (MS; n = 9) and in contemporary Large-White hybrid control sows (LW; n = 9) during the follicular phase, the periovulatory period and the early luteal phase. Ovulation rate was higher in MS than LW animals (23.7 and 16.6 respectively; P < 0.001) and overall was correlated with the area of the oestradiol peak (P < 0.05) and inhibin concentrations (P < 0.05). Both the duration of and the area of the oestradiol peak were greater in MS than LW (P < 0.01; P < 0.02), as were inhibin concentrations both before and after the LH surge (P < 0.05). Neither basal nor peak concentrations of LH or FSH differed between the breeds (P > 0.05), although FSH concentrations were correlated with the area under the oestradiol peak (P < 0.05). Finally, the time-interval from the onset of the LH surge until the rise in plasma progesterone was shorter in MS than LW (54.5 and 74.3 h respectively; P < 0.01). In conclusion, these results show for the first time that the higher ovulation rate in MS is associated with enhanced oestradiol and inhibin secretion with no significant difference in LH or FSH concentrations. The more rapid luteinization response to the LH surge by MS in terms of plasma progesterone concentrations may be important in ensuring the high level of embryo survival in this breed.

Animals↗

BB-10010: an active variant of human macrophage inflammatory protein-1 alpha with improved pharmaceutical properties.

The stem cell inhibitor, macrophage inflammatory protein-1 alpha (MIP-1 alpha) or LD78, protects multipotent hematopoietic progenitors in murine models from the cytotoxic effects of chemotherapy. Clinical use of human MIP-1 alpha during chemotherapy could therefore lead to faster hematologic recovery and may allow dose intensification. We have also shown that human MIP-1 alpha causes the rapid mobilization of hematopoietic cells, suggesting an additional clinical use in peripheral blood stem cell transplantation. However, the clinical evaluation of human MIP-1 alpha is complicated by its tendency to associate and form high molecular weight polymers. We have produced a variant of rhMIP-1 alpha, BB-10010, carrying a single amino acid substitution of Asp26 > Ala, with a reduced tendency to form large polymers at physiologic pH and ionic strength. This greatly increases its solubility, facilitating its production and clinical formulation. We confirmed the potency of BB-10010 as a human MIP-1 alpha-like agonist in receptor binding, calcium mobilization, inhibition of colony formation, and thymidine suicide assays. The myeloprotective activity of BB-10010 was shown in a murine model of repeated chemotherapy using hydroxyurea. BB-10010 is therefore an ideal variant with which to evaluate the therapeutic potential of recombinant human MIP-1 alpha.

Amino Acid Sequence↗

Mobilization of early hematopoietic progenitor cells with BB-10010: a genetically engineered variant of human macrophage inflammatory protein-1 alpha.

BB-10010 is a genetically engineered variant of human macrophage inflammatory protein-1 alpha with improved solution properties. We show here that it mobilizes stem cells into the peripheral blood. We investigated the mobilizing effects of BB-10010 on the numbers of circulating 8-day spleen colony-forming units (CFU-S8), CFU-S12, and progenitors with marrow repopulating ability (MRA). A single subcutaneous dose of BB-10010 caused a twofold increase in circulating numbers of CFU-S8, CFU-S12, and MRA 30 minutes after dosing. We also investigated the effects of granulocyte colony-stimulating factor (G-CSF) and the combination of G-CSF with BB-10010 on progenitor mobilization. Two days of G-CSF treatment increased circulating CFU-S8, CFU-S12, and MRA progenitors by 25.7-, 19.8-, and 27.7-fold. A single administration of BB-10010 after 2 days of G-CSF treatment increased circulating CFU-S8, CFU-S12, and MRA even further to 38-, 33-, and 100-fold. Splenectomy resulted in increased circulating progenitor numbers but did not change the pattern of mobilization. Two days of treatment with G-CSF then increased circulating CFU-S8, CFU-S12, and MRA by 64-, 69-, and 32-fold. A single BB-10010 administration after G-CSF treatment further increased them to 85-, 117-, and 140-fold, respectively, compared with control. We conclude that BB-10010 causes a rapid increase in the number of circulating hematopoietic progenitors and further enhances the numbers induced by pretreatment with G-CSF. BB-10010 preferentially mobilized the more primitive progenitors with marrow repopulating activity, releasing four times the number achieved with G-CSF alone. Translated into a clinical setting, this improvement in progenitor cell mobilization may enhance the efficiency of harvest and the quality of grafts for peripheral blood stem cell transplantation.

Animals↗

Point mutations in the conserved box 1 region inactivate the human granulocyte colony-stimulating factor receptor for growth signal transduction and tyrosine phosphorylation of p75c-rel.

The human granulocyte colony-stimulating factor receptor (hG-CSFR) belongs to the cytokine receptor superfamily. As with other members of this family, the cytoplasmic domain of hG-CSFR lacks intrinsic tyrosine kinase activity. To identify critical regions mediating growth signal transduction by hG-CSFR, deletions or site-directed amino acid substitutions were introduced into the cytoplasmic domain of hG-CSFR, and the mutant cDNAs were transfected into the murine interleukin-3 (IL-3)-dependent Ba/F3 and FDCP cell lines. Truncation of the carboxy-terminal end of the receptor to the membrane-proximal 53 amino acids of the cytoplasmic domain, which retained the conserved Box 1 and Box 2 sequence motifs, decreased the ability of hG-CSFR to transduce G-CSF-mediated growth signals without an associated loss in receptor binding affinity. Substitution of proline by alanine at amino acid positions 639 and 641 within Box 1 completely abolished the G-CSF-mediated growth signal. Rapid induction of tyrosine phosphorylation of several cellular proteins, including a 75-kD protein (p75) identified as c-rel, was an early event associated with transduction of proliferative signals by hG-CSFR in Ba/F3 transfectants. Mutant receptors containing Pro-to-Ala substitutions that inactivated the receptor for mitogenic activity also inactivated the receptor for tyrosine-specific phosphorylation of p75. These results show that the conserved Box 1 sequence motif (amino acids 634 to 641) is critical for mitogenesis and activation of cellular tyrosine kinases by hG-CSFR.

Amino Acid Sequence↗

Plasminogen mutants activated by thrombin. Potential thrombus-selective thrombolytic agents.

Plasmin, the enzyme responsible for degradation of fibrin in blood clots and thus thrombolysis, is normally formed when its zymogen plasminogen is activated by cleavage of the Arg561-Val562 bond by specific plasminogen activators. We have altered the activation characteristics of plasminogen by substituting the P3, P2, and P1' cleavage site residues with sequences from thrombin-cleavable proteins to produce a novel thrombolytic agent which instead is activated by the blood clotting system. Plasminogen variants with thrombin cleavage sites from fibrinogen, the thrombin receptor, factor XIII, and factor XI were cleaved by thrombin with times to 50% cleavage of 28 h, 2.5 h, 5.7 min, and 3 min, respectively. In vitro clot lysis studies have shown that a variant in which the P3-P1' residues of plasminogen were substituted by the P7-P1' residues (Thr363-Ile370) from factor XI (T51) was sufficiently rapidly cleaved by thrombin to be activated by the endogenous thrombin produced by the coagulation cascade, resulting in rapid clot dissolution. Thrombin-activatable plasminogen therefore has the capacity to short circuit the physiological hemostatic mechanisms and produce fibrinolytic activity localized to the site of thrombin formation, that is, at the thrombus itself. The novel activation mechanism combined with the natural long circulating half-life of plasminogen gives this type of thrombolytic agent the potential for thrombus-selective plasmin generation and an extended duration of action.

Amino Acid Sequence↗

Intrauterine and peripheral steroid concentrations and conceptus development in Meishan and Large White hybrid gilts.

Six Meishan and five Large White hybrid gilts were naturally mated to boars of the same breed during their tenth or third oestrous cycle respectively. Maternal serum progesterone and total oestrone were monitored throughout the pregnancy period. On Day 30 of gestation, all gilts were slaughtered and ovulation rate, embryonic survival, conceptus development and intrauterine steroidogenesis were evaluated. The results of the study confirm previous reports that Meishan pigs have a higher number of live conceptuses (P < 0.03), a higher rate of embryonic survival (92.1% v. 78.6% for Large White hybrids) and a higher ovulation rate (P < 0.02) than Large White hybrid gilts. Embryos from Large White hybrid gilts were heavier (P < 0.001) than Meishan embryos and placental lengths (P < 0.001) and weights (P < 0.001) were greater. The volume of allantoic fluid per conceptus was greater (P < 0.03) in Large White hybrid gilts. The oestradiol concentration in the allantoic fluid was greater in Large White hybrid gilts (P < 0.002), but the progesterone concentration in allantoic fluid did not differ (P > 0.15) between the breeds. More oestradiol was synthesized in vitro on a wet weight basis from placental tissue in Large White hybrid gilts than in Meishan gilts (P < 0.001); however, a positive linear relationship existed in both breeds between oestradiol synthesis and placental length (P < 0.005). Progesterone concentrations in maternal serum tended to be higher overall (P < 0.1) in Meishan gilts than in Large White hybrid gilts throughout the 30-day period of study and were significantly higher (P < 0.02) from Day 13 to Day 30.(ABSTRACT TRUNCATED AT 250 WORDS)

Allantois↗

Gonadotropin secretion in ovariectomized Chinese Meishan and hybrid large white gilts; responses to challenges with estradiol benzoate, gonadotropin-releasing hormone, or porcine follicular fluid.

Castrated Meishan (MS) and hybrid Large White (hLW) gilts were used in a series of challenge experiments to establish differences in the control of gonadotropin secretion. Acutely ovariectomized gilts (72 h) were injected i.m. with 10 micrograms/kg estradiol benzoate (EB) and serially bled for evaluation of estradiol clearance from the blood and patterns of LH and FSH release. Estradiol was more rapidly metabolized from the circulatory system of the acutely ovariectomized MS gilts than from the hLW gilts; as a consequence, hLW gilts had higher concentrations of estradiol than MS gilts at 24-32 h (p < 0.05) and 48-60 h (p < 0.01) post-EB treatment. The hLW but not the MS gilts exhibited a classic negative/positive LH response to EB; MS gilts responded with a random release of large episodes of LH. Plasma FSH concentrations were consistently greater (p < 0.05) in acutely ovariectomized MS gilts than in hLW gilts. Both breeds demonstrated a slight negative FSH response to EB and then exhibited increased variability in plasma FSH with time after treatment. Mean plasma LH was immediately elevated in chronically ovariectomized (> 14 days) hLW and MS gilts given 1 microgram/kg of GnRH i.m., but the magnitude of the LH response was greater (p < 0.01) in the hLW gilts. Comparable differences (p < 0.01) in FSH secretion were also observed, indicating the presence of greater pools of releasable FSH and LH in the hLW gilts, or a greater sensitivity to GnRH.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Quantitative control of oxytocin-induced PGF2 alpha release by progesterone and oestradiol in ewes.

The effect of oestradiol and progesterone concentrations on the uterine PGF2 alpha response to oxytocin was investigated by measuring 13,14-dihydro-15-keto PGF2 alpha (PGFM) secretion. One week after ovariectomy, 27 ewes were administered progestagen for 10 days followed by oestradiol for 2 days. Day 0 was designated as the time of the last 'oestrous' oestradiol injection. Six groups of ewes (n = 4) were then treated for 12 days with a high or low dose of progesterone (36 or 12 mg day-1) either alone or with a high or low dose of oestradiol (36 or 12 micrograms day-1) administered (in 1 ml of corn oil by i.m. injection, at intervals of 8 h) in a pattern designed to simulate a natural oestrous cycle profile. A control group (n = 3) was given corn oil alone. Ewes were treated with 1 microgram oxytocin (i.v.) on days 4, 8 and 12 of the simulated cycle and plasma was collected for assay of PGFM. An oxytocin-induced PGFM response occurred only on day 12, when the response was suppressed by high doses of progesterone and stimulated by high oestradiol doses. There was a significant effect of progesterone (P < 0.05) and a highly significant effect of oestradiol (P < 0.01) on the pattern of PGFM release in response to oxytocin. Low progesterone/high oestradiol stimulated the largest and most sustained increase in PGFM following oxytocin. There was a significant relationship between the oestradiol:progesterone ratio and the mean PGFM response on day 12 (P < 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Effects of bovine follicular fluid and exogenous oestradiol on the GnRH-induced short luteal phase in anoestrous ewes.

The role of oestradiol in the control of premature luteolysis (previously shown to occur by the normal luteolytic mechanism involving PGF2 alpha and oxytocin) was investigated in anoestrous ewes induced to ovulate using GnRH (250 ng every 2 h for 24 h followed by 125 micrograms on day 0) without progesterone pretreatment. Seven ewes were administered charcoal stripped bovine follicular fluid (bFF) on days 1-5 (2 ml by s.c. injection every 8 h) together with oestradiol on days 2-4 (4 micrograms in 1 ml of corn oil by i.m. injection every 8 h). Ten ewes were treated with bFF and corn oil (as above), and ten ewes received saline and corn oil (control group). All ewes were treated with 1 microgram oxytocin (i.v.) on day 4 and plasma was collected for measurement of 13,14-dihydro-15-keto PGF2 alpha (PGFM). Blood samples were collected for measurement of progesterone and oestradiol (day--2 to 15). The ewes in the control group that responded to GnRH formed either normal (50% of ewes) or short-lived (50% of ewes) corpora lutea identified by progesterone profiles. The proportion of ewes that displayed premature luteolysis was reduced (P < 0.05) by bFF treatment alone (to 11% of ewes), and increased (P < 0.001) by bFF plus oestradiol treatment (to 100%). bFF treatment suppressed oestradiol concentrations (P < 0.01), whereas bFF plus oestradiol treatment increased oestradiol concentrations (P < 0.001) on days 1-5.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Factors affecting the age at onset of puberty, ovulation rate and time of ovulation in Chinese Meishan gilts.

Experiments were carried out to study: the effects of season on age at puberty, the influence of reproductive age on ovulation rate, and the time interval from the onset of oestrus to ovulation in Chinese Meishan gilts. Gilts approaching puberty either in the spring (n = 88) or in the autumn (n = 40) were housed indoors under natural daylight conditions and observed daily for oestrous behaviour. Gilts approaching puberty in the spring were younger (P < 0.001) and more likely to reach puberty by 100 days of age (P < 0.01) than were those approaching puberty in the autumn. Ovulation rate was estimated in gilts at second (n = 22), third (n = 24), fourth (n = 18), fifth to ninth (n = 9) and tenth to twenty-first (n = 17) oestrous cycle and in primiparous Meishan sows (n = 12) by counting the number of corpora lutea or corpora albicantia at laparoscopy, laparotomy or at ovarian recovery following slaughter. Ovulation rate increased (P < 0.001) with reproductive age and approached that of primiparous sows only when gilts had experienced > or = 10 oestrous cycles (19.2 versus 21.0). The time interval between the onset of oestrus and ovulation was studied in six naturally cyclic Meishan gilts and nine Meishan gilts administered hCG at the onset of oestrus. All glits were observed six times a day for the commencement of oestrous behaviour and were subsequently examined by laparoscopy at 32 h following onset of oestrous and every 8 h till ovulation, which was a maximum of 56 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗

Differences in follicular aromatase activity between Meishan and large-white hybrid gilts.

Two experiments were carried out in which follicular aromatase activity was compared between Meishan and Large-White hybrid gilts. In Expt 1, preovulatory follicles (n = 10 largest per animal) were recovered from Meishan and Large-White hybrid gilts (n = 5 per breed) on the day before predicted onset of behavioural oestrus, and the granulosa cells and theca tissue incubated to determine aromatase activity. Follicles recovered from Meishan pigs were smaller (P < 0.01) and contained fewer granulosa cells (P < 0.05), but follicular oestradiol content of the breeds was similar (P > 0.1). Aromatase activity was higher in the theca tissue (P < 0.05) and tended to be higher in the granulosa cells recovered from Meishan follicles (P = 0.065). In Expt 2, granulosa cell aromatase activity was investigated during the early follicular phase (estimated day 16 of cycle) in Meishan and Large-White hybrid gilts (n = 6 and 5, respectively). The number of follicles > or = 1 mm diameter recovered per animal was 171 for both breeds (P > 0.1), whereas the number of follicles > or = 2 mm diameter was 65 and 101 (P < 0.05) from Meishan and Large-White hybrid gilts, respectively. The mean diameter of all follicles recovered was smaller in the Meishan gilts (P < 0.001). Overall, neither the number of granulosa cells per follicle, as indicated by DNA estimation, nor the oestradiol content differed between the breeds at this time (P > 0.1).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Follicle development and oocyte maturation during the immediate preovulatory period in Meishan and white hybrid gilts.

A study was carried out to compare preovulatory ovarian events in prolific Meishan gilts (which have high levels of embryo survival) with contemporary white hybrid control gilts. Gilts of similar reproductive ages (second or third oestrous cycle) were observed three times a day for oestrous behaviour and ovaries recovered at a time estimated as within 7 h of ovulation (white hybrid, n = 13; Meishan, n = 16). Preovulatory follicles were recovered (n = 195, white hybrid; n = 252, Meishan), and oocytes were aspirated and fixed for later identification of meiotic stage, and follicular fluid was frozen for subsequent determination of progesterone. The number of presumed ovulatory follicles recovered per animal was similar to the expected ovulation rate (15.0, white hybrid; 15.75, Meishan; P > 0.1); however, follicles from Meishan gilts were smaller (8.1 versus 9.0 mm, P < 0.001) and contained less follicular fluid (139.9 versus 187.6 microliters, P < 0.02) than did white hybrid gilts. Total follicular progesterone (88.7 ng per follicle, white hybrid; 77.4 ng per follicle, Meishan) and concentration of progesterone in follicular fluid (526 ng ml-1, white hybrid; 640 ng ml-1, Meishan) did not differ between the two breeds (P > 0.1). There were no breed differences in the extent of heterogeneity in follicular diameter, volume of follicular fluid, and total follicular progesterone and progesterone concentration (P > 0.1). However, classification of recovered oocytes into seven discrete meiotic stages revealed that more oocytes recovered from Meishan preovulatory follicles were in the more advanced stages of meiosis than were those recovered from white hybrid gilts (P < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗