bluuebunny/crossref_metadata_2025 huggingface dataset:
Actions of oxytocin and vasopressin on oestrogen-induced electromyographic activity recorded from the uterus and oviduct of anoestrous ewes
Authors: Ayad VJ , Gilbert CL , McGoff SA , Matthews EL , Wathes DC
Published: October 2005

Oxytocin and the related peptide [Arg8]vasopressin (AVP) have previously been shown to bind with equally high affinity to oxytocin binding-sites (presumed oxytocin receptors) present within the uterus and oviduct of oestrous ewes. There is a possibility, therefore, that AVP mediates oxytocic actions through these binding sites. For the present study, ewes in seasonal anoestrus were treated with oestradiol-17 beta (50 micrograms subcutaneously, daily for 2-4 days). It was shown initially that this treatment stimulated the development of high-affinity oxytocin binding-sites (Kd 4.4 +/- 0.8 nmol L-1) which had similar affinity for AVP (Kd 4.2 +/- 0.9 nmol L-1) in the myometrium. The efficacy of oxytocin and AVP in vivo were compared by recording electromyographic (EMG) activity from the ampullary-isthmic junction of the left oviduct and the left uterine horn of four conscious ewes. Before oestradiol treatment there was no EMG response to oxytocin even at supraphysiological (1000 mU) doses. During oestradiol treatment, EMG activity was consistently increased in response to injections of 25 mU and 100 mU oxytocin via the jugular vein, but not to saline or 100 mU AVP. Higher doses of AVP were not investigated because of the possibility of cardiovascular side effects. A subsequent blood sampling experiment showed that maximal concentrations of oxytocin and AVP (achieved in peripheral plasma during the first 2 min following injection into the jugular vein) were of a similar order of magnitude after injection of equivalent doses of the two peptides. It is concluded that AVP probably does not mediate biological activity through the oxytocin receptor in non-pregnant ewes.(ABSTRACT TRUNCATED AT 250 WORDS)
Porcine conceptus and endometrial retinoid-binding proteins
Authors: Harney JP , Ali M , Vedeckis WV , Bazer FW
Published: October 2005

Porcine conceptus secretory proteins were obtained from medium in which pig conceptuses, collected on Day 15 of pregnancy, were cultured for 30 h. Culture medium was pooled, dialysed and concentrated by Amicon ultrafiltration for retinol and retinoic acid (RA) binding studies. Proteins in the 20-kDa range, conceptus-secreted retinol-binding protein (RBP), bound both [3H]retinol and [3H]RA specifically. Cross-competition experiments indicate that [3H]RA was completely displaced with excess cold retinol; however, excess cold RA did not completely displace [3H]retinol, suggesting that conceptus RBP has greater affinity for retinol than RA. Cellular RBP and retinoic acid receptor (RAR)-alpha and RAR-gamma mRNA transcripts (0.7 kb; 3.8 and 2.8 kb; 3.4 kb respectively) were detected in poly (A)+ RNA isolated from Day-15 conceptus, Day-15 pregnant endometrium, late pregnant myometrium and late pregnant fetal tissues of pigs by Northern blot analysis. RAR-alpha and RAR-gamma immunoreactive proteins were detected in extracts of Day-15 conceptus, Day-15 pregnant endometrium and late pregnant fetal tissues by Western blot analysis. Collectively, results indicate that biochemical molecules required for retinoid transport, metabolism and regulatory effects are present in porcine conceptus and endometrial tissues during early pregnancy in swine.
Silastic implants for delivering physiological concentrations of progesterone to mice
Authors: Milligan SR , Cohen PE
Published: October 2005

Silastic implants filled with progesterone in arachis oil were designed to provide a convenient and reliable method for the delivery of physiological concentrations of progesterone to mice. Placement of the implants in ovariectomized mice resulted in a rapid increase in circulating progesterone within 6 h; stable levels could be maintained for many days. Removal of the implants resulted in a very rapid fall in progesterone concentrations. The delivery of progesterone from the implants could be controlled by varying both the length of the implants and the concentration of progesterone internally. This allowed plasma progesterone concentrations to be maintained and controlled over the entire physiological range.
Contraceptive vaccine development
Authors: Alexander NJ , Bialy G
Published: October 2005

Recent advances in antigen definition and production have made the development of a contraceptive vaccine more attainable. Such a vaccine must evoke an immune response that blocks an indispensable step in the reproductive process. Vaccine research involves many approaches to fertility prevention. Vaccines are being developed that could interrupt fertility by inhibition of gonadotrophin release, the function of follicle-stimulating hormone or the effects of human chorionic gonadotrophin (hCG); alternatively, they may prevent fertilization by interfering with the transport of spermatozoa or with sperm-zona pellucida binding. The most advanced prototype is a vaccine based on antibodies to beta hCG. Such vaccines are being studied for clinical efficacy. Many hurdles remain in contraceptive vaccine development. Since the antigens are peptides or small proteins, the resultant immune response is usually moderate, and better adjuvants and delivery systems must be developed to enhance and maintain the immune response. Improvement of the mucosal immune response may be necessary for vaccines incorporating sperm antigens. Research on vaccines that control fertility has resulted in a fascinating base of scientific knowledge that, it is hoped, can be converted into products that will allow another option for individuals who wish to control their fertility.
Prefertilization gamete maturation events in marsupials
Authors: Rodger JC
Published: October 2005

Despite many fundamental similarities between the gametes of marsupials and placental mammals, the regulation and timing of prefertilization gamete maturation are quite different. The marsupial acrosome is remarkably stable and an acrosome reaction (AR) is not induced by reagent effective for the sperm of placental mammals. The ultrastructure of the marsupial sperm AR is essentially similar to that of placental mammals, however, whether an equatorial segment (ES) persists to serve as the site of sperm-egg membrane fusion is unclear. Diacylglycerol induction of the AR suggests that the sperm of Australian species lack an ES, yet an ES-like region appears to be involved in fertilization in the opossum Monodelphis. The marsupial oocyte, unlike those of placentals, continues to grow throughout follicular life and major cytoplasmic maturation events occur late in oocyte development. Cortical granules only become evident shortly before ovulation and mature dark granules may only appear after ovulation. Further, the zona pellucida (ZP) changes in character and function during the peri-ovulatory period. In vitro fertilization has been achieved for an opossum but not for any Australian marsupial, owing to failure of sperm-ZP binding. Requirement for a sperm maturation process is likely, but capacitation treatments used for placental sperm in vitro have been ineffective. Since it is now feasible to experimentally manipulate marsupial gametes in vitro major advances in our understanding of their function can be expected.
Studies of in vitro differentiation with embryonic stem cells
Authors: Pedersen RA
Published: October 2005

Embryonic stem (ES) cells were first cultured from mouse embryos little more than a decade ago, yet they are now widely used in transgenic studies which are revolutionizing mammalian genetics. Although drawing less attention, in vitro studies of mouse ES cells have also contributed widely to the understanding of mechanisms of embryonic cell differentiation and proliferation. This review focusses on the application of ES cells as in vitro models for cellular and molecular events in the early mammalian embryo. Future studies with cultured ES cells of mouse and other species should provide insights into the factors regulating the differentiation of intermediate stem cells and terminal cells for the various embryonic lineages, thus contributing profoundly to the understanding of mammalian embryogenesis as well as providing cells for therapeutic applications.
Systems for production of calves from cultured bovine embryonic cells
Authors: First NL , Sims MM , Park SP , Kent-First MJ
Published: October 2005

The development of totipotent bovine embryonic cell cultures has great value in cattle breeding. They provide: (1) a mechanism for making large numbers of clonal offspring by nuclear transfer; (2) an efficient gene transfer system through the use of selectable markers to select transgenic cells; and (3) a mechanism for site-specific gene transfer or deletion by homologous DNA sequence recombination. Bovine embryonic cell cultures have been established from blastocyst inner cell mass (ICM) cells, morulae and the precompaction 16-20-cell stage. All have exhibited similar morphology to mouse embryonic stem (ES) cells, pluripotency on differentiation and proliferation in culture. Culture systems have consisted of microdrop loose suspension short-term cultures or long-term cultures on bovine or murine fibroblast feeder layers, in either a microdrop or a culture dish. The relative merit of culture systems or media requirements for mitosis and prevention of differentiation have not been determined. At present, totipotency is also unknown for cultured cells of the 16-20-cell stage. For cultured ICM cells, totipotency was demonstrated by the birth of four calves from ICM cells cultured 27 days or less in a loose suspension microdrop. Advanced pluripotency and perhaps totipotency was demonstrated in one fetus in a recently reported study where morulae cells cultured in vitro were chimaerized with non-cultured cells. DNA fingerprinting to associate cell lines with offspring and karyotyping to ascertain chromatin normalcy is important in ES cell research. Data pertaining to the use of each are presented.
Development and validation of swine embryonic stem cells: a review
Authors: Wheeler MB
Published: October 2005

The establishment of embryonic cell lines from swine should be useful for studies of cell differentiation, developmental gene regulation and the production of transgenics. This paper summarizes the establishment of porcine (Sus scrofa) embryonic stem (ES) cell lines from preimplantation blastocysts and their ability to develop into normal chimaeras. ES cells can spontaneously differentiate into cystic embryoid bodies with ectodermal, endodermal, and mesodermal cell types. Further, culture of ES cells to confluence or induction of differentiation with retinoic acid or dimethylsulfoxide results in morphological differentiation into fibroblasts, adipocytes, and epithelial, neuronal, and muscle cells. These ES cells have a normal diploid complement of 38 chromosomes. Scanning electron microscopy of the ES cells reveals a rounded or polygonal, epithelial-like cell with numerous microvilli. The differentiation of these embryonic cell lines into several cell types indicates a pluripotent cell. Furthermore, chimaeric swine have been successfully produced using such ES cells.
Strategies for the isolation and characterization of bovine embryonic stem cells
Authors: Cherny RA , Stokes TM , Merei J , Lom L , Brandon MR , Williams RL
Published: October 2005

The practical application of advanced breeding technologies and genetic manipulation of domestic animals is dependent on the efficient and routine isolation of embryonic stem (ES) cell lines from these species. ES cell lines of proven totipotency have thus far been isolated only from the mouse. Murine ES cells can be identified by a number of criteria including morphology and characteristics in culture, the presence of specific markers, differentiative capacity and contribution to chimaeras. Reported cell lines derived from ruminant preimplantation embryos do not stably exhibit these characteristics. As demonstrated for the mouse, primordial germ cells may provide an alternative source for pluripotential cell lines. The isolation, culture and preliminary characterization of bovine primordial germ cell-derived (PGCd) cells are described in this paper. The PGCd cells are capable of differentiation in vitro and display murine ES cell markers including alkaline phosphatase. With farm animals, long generation intervals and small numbers of offspring make it important to develop techniques for evaluating chimaeric embryos in vitro before embarking on expensive in vivo programmes. A method for labelling putative pluripotential cells with a fluorochrome marker to follow the fate of such cells was developed. Labelled PGCd cells were injected into blastocysts and the chimaeric embryos were monitored in vitro. Preliminary results demonstrate that the labelled PGCd cells incorporate preferentially within the inner cell mass of the host blastocyst.(ABSTRACT TRUNCATED AT 250 WORDS)
Germ line transmission of yeast artificial chromosomes in transgenic mice
Authors: Montoliu L , Schedl A , Kelsey G , Zentgraf H , Lichter P , Schutz G
Published: October 2005

Several groups have recently reported the successful generation of transgenic mice harbouring yeast artificial chromosomes (YACs). Different methodological approaches have been shown to produce similar results, namely, the faithful expression of the transgenes carried on YAC DNA. In this paper, we compare the reported techniques for obtaining transgenic mice carrying YACs using a 250-kb YAC bearing the mouse tyrosinase gene. These methods include: microinjection of gel-purified YAC DNA into pronuclei of fertilized mouse oocytes, yeast spheroblast fusion with embryonic stem (ES) cells and lipofection of YAC DNA into ES cells. Taken together, these reports show that the delivery of large genomic regions covering a gene of interest (such as those cloned in YAC vectors) is feasible, and will ensure appropriate temporal and spatial expression of the transgene at a level comparable to that of the endogenous counterpart.
Chromosomal position effects and the modulation of transgene expression
Authors: Clark AJ , Bissinger P , Bullock DW , Damak S , Wallace R , Whitelaw CB , Yull F
Published: October 2005

Chromosomal position effects can influence strongly the transcription of foreign genes in transgenic animals. This results in low frequencies and levels of gene expression and, in some cases, in aberrant patterns of expression. Strategies for overcoming these effects are described with particular reference to their application in embryonic stem cells.
Cloning and characterization of a fox sperm protein FSA-1
Authors: Beaton S , Cleary A , ten Have J , Bradley MP
Published: October 2005

A monoclonal antibody was raised to a fox sperm protein (FSA-1) which was found to be localized to the inner acrosomal compartment of sperm fixed in methanol. Western blots of testicular germ cell membrane extracts probed with this antibody identified a major protein band with a molecular weight of 36,000. Immunofluorescent studies on fox testis sections showed that the antigen is expressed on round and elongating spermatids on a crescent-shaped structure, which probably represents the developing acrosome. An antibody specific for FSA-1 was used to screen a fox testis cDNA library for its cognate gene. An 875-bp cDNA clone was isolated and sequenced revealing an open reading frame. Searches of the GenBank and EMBL databases with the nucleic acid sequence revealed significant homology (86%) of FSA-1 with 406 bases of an unidentified RNA transcript from human fetal brain (EST02625). Northern blot analysis of fox testis RNA samples identified an RNA transcript of approximately 0.9 kb during the months when spermatogenesis is active. Zoo Northern blots (at high stringency) reveal an RNA transcript of a similar size present in testis RNA from dogs and mice. Zoo Southern analysis (high stringency) reveal genomic sequences present in dogs, mice, cattle and sheep. At present, the function of the FSA-1 gene product remains unknown, but it may play a role as a structural protein component of the acrosome.
Timing of seasonal breeding in birds, with particular reference to New Zealand birds
Authors: Cockrem JF
Published: October 2005

A model to explain the timing of seasonal breeding in birds is presented. It is assumed that, despite the wide range in egg-laying seasons, there are common physiological mechanisms which underlie seasonality in birds and that most, if not all, birds are photoperiodic. Birds are considered to possess an internal rhythm of reproduction which is synchronized with seasonal changes in the environment by external factors, particularly the annual cycle of daylength. The rhythm consists, at least in part, of regular changes in the photoperiodic response between states of photosensitivity and photorefractoriness. Avian breeding seasons effectively start in autumn when birds become photosensitive, regardless of when egg-laying occurs. The timing of breeding is then influenced by the rate of increase of hypothalamic 'drive' and by the sensitivity of the hypothalamus and pituitary gland to inhibitory feedback from gonadal steroids. If sensitivity is high, gonadal growth will not occur until the threshold daylength for photostimulation is exceeded after the winter solstice. Egg-laying then starts in late winter, spring or summer. Alternatively, steroid feedback may be relatively low and gonadal growth may be sufficiently rapid once the birds become photosensitive that breeding occurs in late autumn or winter. The time of egg-laying in birds may also be strongly influenced by supplementary information, such as social cues, food availability, temperature and rainfall and, in some species, this information is more important than daylength in determining the timing of breeding. The review also includes the first summary of the breeding seasons of New Zealand birds. The pattern of egg-laying is exactly the same in native birds, in birds introduced to New Zealand and in other Southern hemisphere birds from similar latitudes, with a broad peak of egg-laying occurring from September to December. In addition, annual cycles of steroid hormone concentrations in the North Island brown kiwi, the yellow-eyed penguin and the kakapo are consistent with results from many studies on Northern hemisphere birds. This model for the timing of breeding in birds can be applied to New Zealand birds and it is concluded that the physiological control mechanisms for the timing of seasonal breeding in New Zealand birds are similar to those of other birds.
Inhibitory potency of isoprenaline on guinea-pig and gravid human myometrium following extraneuronal uptake blockade
Authors: Kousides M , Story ME , Pennefather JN , Ziccone SP , Ross AW
Published: October 2005

The hypothesis that inhibitory effects of isoprenaline on myometrial contractility may be constrained by activation of putative intracellular beta-adrenoceptors negatively-coupled to adenylate cyclase was examined. Field-stimulated preparations of guinea-pig and human myometrium were used to examine the influence of the catecholamine extraneuronal uptake2 inhibitors, corticosterone and beta-oestradiol, on the inhibitory effects of the beta-adrenoceptor agonist, isoprenaline, on uterine contraction. Longitudinal and circular myometrial layers were obtained from guinea-pigs in dioestrus, primed with oestrogen before progesterone, or pregnant (Days 62-65). In the guinea-pig myometrium, corticosterone (30 microM) did not affect responses to isoprenaline. beta-oestradiol (10 microM) induced a small potentiation of the effects of isoprenaline on longitudinal myometrium from dioestrus guinea-pigs. Myometrial preparations were obtained from pregnant women (36-40 weeks gestation) undergoing caesarean section. Isoprenaline inhibited stimulation-evoked contractions in 7 of 10 preparations of the inner myometrial layer and in 5 of 8 preparations of outer myometrial layer. Corticosterone (30 microM) reduced the effects of isoprenaline on the inner layer and did not affect the outer layer. These results do not support the existence of mechanism involving isoprenaline-sensitive intracellular receptors which constrain responses to beta-adrenoceptor agonists.
Distribution of variance associated with measurement of post-thaw function in ram sperm
Authors: Edward AY , Windsor DP , Purvis IW , Sanchez-Partida LG , Maxwell WM
Published: October 2005

Post-thaw characteristics of ram semen frozen as pellets were assessed using biochemically (amidase activity) or motility-based (Hamilton Thorn Motility Analyzer) techniques. The total variation associated with each semen characteristic measured was partitioned between rams (5), ejaculates within rams (5), pellets within ejaculates (5) and within pellets (2). A variety of variance distributions were observed for the characteristics measured. Of the 18 post-thaw characteristics examined, 10 had > 50% of variance distributed between within-ejaculate components. This has important implications for the way in which such measurements may be used in post-thaw semen analysis.
A comparative study of the corpus luteum
Authors: Gemmell RT
Published: October 2005

The corpus luteum (CL) is a transitory organ which has a regulatory role in reproduction. Sharks, amphibians and reptiles have corpora lutea that produce progesterone which influences the rate of embryonic development. The egg-laying monotremes and the two major mammalian groups, eutherian and marsupial, have a CL that secretes progesterone. Most eutherians have allowed for the uterine development of their young by extending the length of the oestrous cycle and the CL or placenta actively secretes progesterone until birth. Gestation in the marsupial does not extend beyond the length of an oestrous cycle and the major part of fetal development takes place in the pouch. Where the extension of the post-luteal phase in the eutherian has allowed for the uterine development of young, the marsupial has extended the pre-luteal phase of the oestrous cycle and has evolved an alternative reproductive strategy, embryonic diapause. The mechanism for the secretion of hormones from the CL has been controversial for many years. Densely-staining secretory granules have been observed in the CL of sharks, marsupials and eutherians. These granules have been reported to contain relaxin, oxytocin or mesotocin, and progesterone. A hypothesis to suit all available data is that all hormones secreted by the CL are transported within such granules. In conclusion, although there are obvious differences in the mode of reproduction in the two main mammalian groups, it is apparent that there is a great deal of similarity in the hormonal control of regression of the CL and parturition.
Implication of inhibin and related proteins in fetal development
Authors: Jenkin G , McFarlane JR , de Kretser DM
Published: October 2005

Initial studies on inhibin, activin and follistatin focussed on their role as regulators of adult gonadal function via feedback regulation of anterior pituitary hormones and via intragonadal control of steroid hormones. The discovery of isoforms of follistatin which are either secreted or retained on the cell surface and which bind activin and, to a lesser extent, inhibin adds a further dimension to the regulation of these peptides. More recently, the cloning of inhibin and activin, and the observation of their close homology to the transforming growth factor-beta family of peptides, has led to an interest in their possible role as growth and differentiation factors. Activin, inhibin and follistatin are expressed in embryonic and fetal tissues, as well as in the placenta. However, although activin is a potent regulator of growth and differentiation in a number of cell types, their role in embryonic and fetal development has yet to be established. High concentrations of inhibin have been observed in the fetal gonads, particularly the testes, and in the fetal adrenals of a number of species and a sex difference in fetal plasma concentrations has also been observed. Although the stimulus for high concentrations of inhibin in the fetus is not know, they are associated with decreased testicular testosterone and a decrease in the concentration of circulating follicle-stimulating hormone (FSH); this suggests that, as in the adult, inhibin may be involved in the regulation of fetal testicular androgen and pituitary FSH secretion during late gestation. The recent reports of elevated concentrations of inhibin and, particularly, activin in amniotic fluid during late gestation and its ability to stimulate the production of prostaglandin E2 by fetal membranes provides yet another potential role for this hormone in the regulation of events leading to parturition.
Functional development of fetal limb muscles: a review of the roles of activity, nerves and hormones
Authors: Walker DW , Luff AR
Published: October 2005

Animals that are immature at birth with respect to postural and locomotor control (e.g. cats, rats) possess incompletely differentiated 'fast-twitch' and 'slow-twitch' muscles at birth; full development proceeds slowly in the postnatal period and involves myogenic, hormonal, neural and behavioural factors. The gradual emergence of specific motor patterns and the exercise of individual muscle groups is thought to play a major role in the final development of each muscle and the fibre types which comprise them. In contrast, precocial species such as the sheep are born with skeletal muscles, especially those of the limbs, which are fully differentiated at birth. The relative importance of neural and hormonal factors in allowing this functional specialization to occur in the presumed absence of significant load-bearing exercise in the intrauterine environment is unclear. In this brief review, the changes which occur in contractile function and fibre type differentiation during the last one-third of gestation in fetal sheep are described, and some of the factors which influence this development are considered.
Use of epididymal sperm for assisted reproduction in men with acquired, irreparable obstructive azoospermia
Authors: Patrizio P , Ord T , Balmaceda JP , Asch RH
Published: October 2005

Microsurgical epididymal sperm aspiration (MESA) and in vitro fertilization (IVF) is primarily offered to men with congenital absence of the vas deferens (CAVD). However, the IVF capacity of these epididymal sperm is low ( < 15%) and unpredictable. In this study, IVF and intracytoplasmic sperm injection (ICSI) results in patients with non-congenital, irreparable obstructive azoospermia were analysed. Thirty-three patients were evaluated for a total of 37 cycles of MESA and IVF. Most had obstruction secondary to failed vasectomy reversal and to epididymal blockage. The overall fertilization rate was 30% with regular IVF and 26% with ICSI, and six clinical pregnancies were obtained. Both rates are significantly higher than the IVF rate previously reported with sperm from men with CAVD (13%, P < 0.00001). In men with non-congenital obstructive azoospermia, a significant difference was found in the average sperm count (56.9 x 10(6) v. 12.3 x 10(6), P < 0.04) and total motile count (16.6 x 10(6) v. 1.6 x 10(6), P < 0.01) respectively for patients who achieved IVF and those who did not. It is concluded that (a) the real IVF capacity of human epididymal sperm is 30%, or 42%, if calculated only for patients who achieved fertilization, (b) this higher rate is an indirect support of the hypothesis that sperm from men with CAVD have intrinsic biochemical defects, related to cystic fibrosis mutations, responsible for their low and unpredictable IVF rate, and (c) MESA and IVF can be offered at the same time or as an alternative to patients requesting vasectomy reversal.
Molecular aspects of the ontogeny of the pituitary-gonadal axis
Authors: Huhtaniemi I
Published: October 2005

The endocrine function of the mammalian pituitary-gonadal axis begins in utero. This is important particularly for the ontogeny and function of the male reproductive organs, the induction of which is critically dependent on the two fetal testicular hormones, testosterone and anti-mullerian hormone. In contrast, ovarian endocrine activity begins only after birth. The earliest phases of testicular hormone production are probably under autocrine or paracrine regulation, but the dependence on gonadotrophins starts in fetal life. During maturation of the hypothalamic-pituitary-testicular axis, the target organs acquire their responsiveness (viz receptors) before the onset of secretion of the tropic hormonal stimulus. The last link to develop is the feedback regulation, and the whole axis is functional in the developing male rat during the last days of gestation. Although gonadotrophin secretion starts in both sexes simultaneously, the fetal ovary is endocrinologically quiescent--its gonadotrophin responsiveness and endocrine activity begin only after birth. The fetal and postnatal periods of testicular activity have crucial effects on male sexual differentiation, whereas in the female, early sexual development occurs autonomously without influence of ovarian function. The purpose of this review is to elucidate some of the recent findings on the molecular mechanisms involved in the perinatal maturation of the rat hypothalamic-pituitary-gonadal axis.
Effects of gonadotrophins, growth hormone and prolactin on developmental competence of domestic cat oocytes matured in vitro
Authors: Schramm RD , Bavister BD
Published: October 2005

Specific aims were to (1) examine the developmental capacity of felid oocytes matured in vitro and (2) determine the effects of gonadotrophins, growth hormone and prolactin on nuclear and cytoplasmic maturation oocytes in vitro. Oocytes were obtained from excised ovaries of 21 cats, and were matured for 45-46 h in modified CMRL-1066 culture medium (1 mM glutamine, 1 mM pyruvate and 20% bovine calf serum), with one of the following: (1) gonadotrophins (1.0 micrograms mL-1 hFSH+10 micrograms mL-1 hLH), (2) gonadotrophins+10 micrograms mL-1 growth hormone, (3) gonadotrophins+10 micrograms mL-1 prolactin, or (4) no hormones. Oocytes were inseminated with ejaculated cat sperm capacitated in TALP medium. Embryos were cultured in modified CMRL-1066 medium until developmental arrest, then stained with Hoechst 33342 to assess nuclear status or cell number. Gonadotrophins enhanced (P < or = 0.05) the incidence of nuclear maturation, but neither gonadotrophins, growth hormone nor prolactin improved fertilization or developmental potential of oocytes matured in vitro. Mean percentages of mature oocytes that were fertilized and cleaved to or beyond the 2, 4, 8 and 16-cell stages were 80, 77, 66, 42 and 24%, respectively. Three embryos progressed to 40-60 cells, but none developed a blastocoel. Thus, although gonadotrophins enhance nuclear maturation of oocytes in vitro, and mature oocytes are capable of fertilization and development to the morula stage, culture with growth hormone, prolactin or gonadotrophins during maturation in vitro does not enhance developmental competence or overcome the morula-to-blastocyst-stage block in development of domestic-cat oocytes matured in vitro.
Development in vitro and in vivo of aggregated parthenogenetic bovine embryos
Authors: Boediono A , Saha S , Sumantri C , Suzuki T
Published: October 2005

Mature bovine oocytes were activated with 7% ethanol followed by cytochalasin B or D treatment. Most oocytes extruded a second polar body and formed one pronucleus when treated with 7% ethanol alone [35/43 (81%)]. With ethanol followed by cytochalasin B or D, overall activation frequency was 70% (309/441), with activated oocytes containing two pronuclei. The cleavage rate was not significantly different between treatment with ethanol alone and ethanol followed by 5 micrograms mL-1 cytochalasin B, but it was significantly lower than in fertilized oocytes (P < 0.01). However, the blastocyst production rate was significantly different (P < 0.01) among the treatments. The incidence of parthenogenetic embryos with normal (diploid) complements and with chromosome anomalies (2N/4N) was 68% (17/25) and 32% (8/25) respectively, and this was not affected by cryopreservation treatment. The longitudinal diameter of aggregated-four embryos cultured in vitro was greater (P < 0.01) than aggregated-two or single embryos. One of the aggregated-four parthenogenetic embryos was further cultured in vitro and developed up to Day 27 after activation, with a diameter of 2980 microns. The aggregated-four parthenogenetic embryos were transferred to five recipients. The oestrus was prolonged in three recipients and they returned to oestrus on Day 57, 62 and 67 after the previous oestrus. These results indicate that aggregating parthenogenetic embryos can prolong their survival in vitro and in vivo.
Motility, acrosome integrity and fertility of frozen ram spermatozoa treated with caffeine, pentoxifylline, cAMP, 2-deoxyadenosine and kallikrein
Authors: Maxwell WM , Robinson SJ , Roca J , Molinia FC , Sanchez-Partida LG , Evans G
Published: October 2005

The motility, acrosome integrity and fertility of ram spermatozoa were examined after treatment with five compounds (caffeine, pentoxifylline, cAMP, 2-deoxyadenosine and kallikrein). Semen was extended with a Tris-based medium and frozen in pellet form. The compounds were added at various concentrations to the semen before freezing or after thawing. Motility and acrosome characteristics were assessed over a 6-h post-thaw period of incubation at 37 degrees C. Of the five compounds examined, only pentoxifylline, when added after thawing, stimulated motility but had not effect on the acrosome integrity of spermatozoa. Pregnancy rates for ewes inseminated in the uterus with semen treated after thawing with Dulbecco's phosphate-buffered saline (PBS, control) or PBS containing pentoxifylline (15 mM in the thawed semen), were 16/39 (41%) and 22/42 (52%) respectively. Motility characteristics of spermatozoa assessed by image analysis (Hamilton-Thorne analyser) were initially better after treatment of thawed semen with pentoxifylline than with PBS ('percent motility', 'percent progressive motility', 'percent rapid', 'percent moderate' and 'percent static', P < 0.01 or P < 0.001), but there was a greater deterioration in these characteristics during post-thaw incubation in semen treated with pentoxifylline than in the controls. The deterioration in motility characteristics occurred within 4 h of thawing and this may have been responsible for the modest improvement in fertility of spermatozoa treated with pentoxifylline.
Nucleotide sequence of cDNA encoding bonnet monkey (Macaca radiata) zona pellucida glycoprotein-ZP3
Authors: Kolluri SK , Kaul R , Banerjee K , Gupta SK
Published: October 2005

The cDNA encoding bonnet monkey zona pellucida ZP3 from bonnet ovary has been amplified by polymerase chain reaction. The ZP3 gene has an open reading frame of 1272 nucleotides encoding a polypeptide of 424 amino acid residues which shares 93.9% overall identity with human ZP3. Bonnet ZP3 has four potential attachment sites for N-linked sugar chains which are also conserved in human ZP3. Bonnet ZP3 has 14 cysteine residues compared with 15 in human ZP3. The highest disparity between these molecules was restricted to a domain represented by amino acid residues 370-398. These results have important implications for the use of bonnet monkey as an animal model for evaluation and development of contraceptive vaccine based on ZP3 for human use.
Concentrations of cytochrome P450 cholesterol side-chain cleavage enzyme and 3 beta-hydroxysteroid dehydrogenase during prostaglandin F2 alpha-induced luteal regression in cattle
Authors: Rodgers RJ , Vella CA , Young FM , Tian XC , Fortune JE
Published: October 2005

Prostaglandin F2 alpha (PGF2 alpha)-induced regression of the corpus luteum causes both plasma progesterone concentrations and luteal concentrations of mRNA encoding the steroidogenic enzyme 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) to fall in parallel. To investigate the hypothesis that a decline in the concentrations of mRNA encoding steroidogenic enzymes causes plasma progesterone to fall, the luteal concentrations of the enzymes 3 beta-HSD and cytochrome P450 cholesterol side-chain cleavage were measured during induced luteolysis. Holstein heifers were treated with PGF2 alpha (25 mg Lutalyse) on Day 6 or Day 7 of the oestrous cycle and corpora lutea were collected 0 h, 2 h, 12 h, and 24 h later (n = 6, 4, 4, and 4 respectively). Analyses of the steroidogenic enzymes were carried out by Western immunoblotting. The luteal concentrations of both steroidogenic enzymes did not decrease over the 24-h period. It is concluded that, although the concentrations of mRNA encoding steroidogenic enzymes may decline in response to PGF2 alpha, this does not lead to a sufficiently rapid reduction in the concentrations of the enzymes to precede, and thus cause, the decline in plasma progesterone concentrations. Thus, the mechanism for the initial decline in plasma progesterone concentrations during luteolysis is still not known.
Nuclear transplantation using bovine primordial germ cells from male fetuses
Authors: Delhaise F , Ectors FJ , de Roover R , Ectors F , Dessy F
Published: October 2005

The developmental potential of nuclei of bovine gonial cells was investigated by nuclear transfer. Gonial cells were collected from male fetuses at about 175 days post coitum (p.c.). They were fused with enucleated oocytes; reconstituted embryos were cultured in vitro for 7 days. Embryos reaching the compacted morula or blastocyst stage were either fixed for cell counting or transferred into recipients. Out of 115 oocyte-gonia fusions, 101 (87.8%) gave rise to cleaved embryos at Day 3 and 26 (22.6%) had reached the 8-cell stage. At Day 7, 1 (1%) developed to the morula stage and 5 (4%) reached the blastocyst stage. Three blastocysts were fixed and showed normal cell numbers (135; 90; 76 cells). Three blastocysts and one morula were transferred in four recipients; two recipients were pregnant at Day 21 but only one was positive at Day 35 p.c.; this last one aborted around Day 40 p.c. No conceptus was collected. These results indicate that gonial cell nuclei can be partially reprogrammed; they are able to develop into blastocysts and to initiate gestation. However, more experiments will be necessary to prove the nuclear totipotency of bovine gonial cells.
Effect of neonatal allylestrenol treatment (hormonal imprinting) on the serum testosterone and progesterone concentration in adult rat
Authors: Pap E , Csaba G
Published: October 2005

Serum testosterone concentration was significantly elevated in adult male rats by a single perinatal allylestrenol administration. One week after a second allylestrenol treatment in adulthood the hormone concentration dropped below the control values. Serum progesterone concentration was significantly lowered in adult female rats by a single perinatal allylestrenol administration. Following a second allylestrenol treatment in adulthood the hormone concentration reached the control values. These experiments demonstrate that the hormonal imprinting caused by allylestrenol (a steroid used in the treatment of endangered pregnancy) not only acts at receptor level and produces changes in sexual behaviour, but also induces modifications in serum hormone concentrations.
Testosterone concentrations in chicken gonads and plasma during the peri-etching period
Authors: Motelica-Heino I
Published: October 2005

In chick embryos of the black sex-link strain, the concentration of testosterone was much lower on Days 17 and 18 of the incubation period in both gonads (tests and left ovary) than on the next two days. Then, at hatch, and especially at 1 h after hatching, the concentration decreased in both gonads. Between 2 and 4 h after hatching, the concentration of T reached the highest value in each sex and then decreased progressively within the next 18 h. Concentrations on Days 19 and 20 (in both gonads) differed significantly from those on Days 17 and 18 of the incubation period, but no significant sex differences were found at these four sampling times. In newly hatched chickens, significant differences were found only between the values obtained at 1 h after hatching (in both sexes) and those from 2-4 h after hatching; no significant sex differences in T concentrations were found in gonads on Day 1 after hatch. The concentration of T in plasma from females was a little higher than in males on Day 19 of incubation, but no significant differences were found. At hatch, the concentrations were of the same order of magnitude in both sexes. At 1 and 2 h after hatching, increased values were found only in males; pooled values of these two sampling times were significantly different from those obtained either in embryos or at two periods after hatching (8-20 h and 48-68 h respectively). No significant changes occurred in T concentration in plasma from females over all these sampling times. By 3-6 h after hatching, the concentrations of T returned to near hatching concentrations in both sexes, and then decreased progressively between 8 and 68 h after hatching. No significant differences in plasma T concentrations were found between males and females, at hatch, or within the next 68 h. In red sex-link strain chickens, the concentrations of T in plasma were about the same in both sexes, at hatch and on Day 1 after hatching, but the concentrations were much lower than those recorded in black sex-link strain chickens at the same sampling times. A peak of T was found only in plasma from newly hatched male chickens of the black sex-link strain at 1-2 h after hatching but not in females, and not in red sex-link strain (of both sexes) either.
Fetal and maternal plasma lipids in chronically catheterized mares in late gestation: effects of different nutritional states
Authors: Stammers JP , Hull D , Silver M , Fowden AL
Published: October 2005

The effects of different nutritional states on plasma lipid concentrations have been examined in pregnant mares and their fetuses. Maternal and fetal arterial catheters were inserted into 12 pony mares between 244-303 days' gestation (term 320-360 days) and observations made from 5 days following the insertion of catheters. After recovery from surgery maternal and fetal arterial samples were withdrawn from 7 mares with normal feeding patterns (Group IA), from four of these mares at the end of a 30 h fast and 3 h later following refeeding (Group IB) and six mares who failed to re-establish normal feeding patterns (Group II). The fatty acid concentrations and composition of the plasma free fatty acid (FFA), triacylglycerol and phospholipid fractions were analysed. Maternal FFA, triacylglycerol and phospholipid concentrations were significantly raised in the fasted (Group IB) and under-fed (Group II) mares. Fetal concentrations of FFA and phospholipid increased significantly in the group of under-fed (Group II) mares but not in the fasted (Group IB) mares. In the fetal plasma the proportions of polyunsaturated fatty acids derived from essential fatty acids in the FFA and phospholipid fractions were much higher than those in the mare. In the fasted (IB) and under-fed (II) groups the relative amounts of the polyunsaturated fatty acids in each fraction remained unchanged (P > 0.05). These results show a short fast or prolonged undernutrition result in raised maternal plasma lipid concentrations which in turn can effect the total amount of lipid in the fetal circulation. However any increases in polyunsaturated fatty acids in the fetus (e.g. in Group II) are unlikely to come from the maternal circulation; likely sources of these fatty acids are the placenta or fetal tissues.
Flow and velocity waveform indices in the ovine fetal abdominal aorta with changes in behavioural state
Authors: Parkinson S , Belleau C , Homan J , Richardson B
Published: October 2005

The purpose of this study was to determine the extent to which flow and velocity waveform indices are comparable as measured from the ovine fetal abdominal aorta and reflect downstream vascular resistance with changes in electrocortical activity. Nine chronically catheterized fetal sheep were studied near term with continuous measurement of electrocortical activity, perfusion pressure, and fetal heart rate. A cuffed Doppler crystal and a Transonic flow probe on the descending abdominal aorta were used for the simultaneous measurement of velocity waveforms and flow waveforms, with the Resistance Index (RI), calculated for each. Mean blood flow was also obtained from the Transonic flow probe. Whereas the change in cardiovascular parameters between the low- and high-voltage electrocortical activity (ECOG) states was small, with little relationship to the change in RI measurements, across all animals there was an inverse correlation between the RI measurements and fetal heart rate, perfusion pressure and aortic blood flow during both low- and high-voltage ECOG states. The RI as measured from the velocity waveforms showed only a modest correlation to those measured from the flow waveforms. It is concluded that over the range of circulatory change generated with spontaneous change in behavioural state, the RI as measured in the abdominal aorta of the ovine fetus is primarily determined by pulsatile cardiovascular variables rather than lower body vascular resistance.
Postnatal changes in size and actomyosin content of rabbit gastric myocytes
Authors: Tomomasa T , Xie Y , Morikawa A , Kuroume T , Hyman PE
Published: October 2005

Maximal tension generated by gastric muscle is three to four times greater in weanlings than in newborn rabbits. To determine if this functional maturation is accompanied by structural changes, we compared length-tension relationships, myocyte number and size, and actomyosin content in muscle from the gastric body of newborn (1 day) and weanling (12 weeks) rabbits. Passive tension at optimal length (Lo) was six times greater in circular smooth muscle strips from weanling rabbits than from newborn rabbits. Active tension at Lo in weanling rabbits was three times greater than in newborn rabbits. For morphometry, muscle cross-sections stretched to Lo in the circular axis were photographed with electron microscopy (5300x). Cell number/unit area was counted in circular muscle layers from newborns and weanlings. Cross-sectional area of each cell was measured by computerized planimetry. There were 2.5 times more cells per unit area in newborn than in weanling tissue, P < 0.001. However, the mean cell area in newborns (5.4 +/- 4.6 microns2) was less than that in weanlings (13.5 +/- 11.7 microns2). Consequently, the muscle cells occupied similar total areas in newborns and weanlings. We measured actin and myosin heavy chain in full-thickness muscle homogenates using SDS gel electrophoresis and densitometric scanning. Actin and myosin concentrations were lower in newborns (9.6 +/- 1.3 micrograms g-1 wet weight and 5.6 +/- 0.7 micrograms g-1 wet wt, respectively) than in weanlings (17.7 +/- 3.0 micrograms g-1 wet wt and 8.2 +/- 1.6 micrograms g-1 wet wt respectively), each P < 0.01. The proportion of myosin heavy chain isozymes did not change with age. We conclude that there are postnatal increases in cell size and the quantity of actin and myosin in rabbit gastric muscle. The increase in quantity of contractile protein may be in part responsible for age-dependent increases in maximal tension.