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PubMed2000

Births of kittens produced by intracytoplasmic sperm injection of domestic cat oocytes matured in vitro.

In Experiment 1, cleavage frequency and in vitro development of domestic cat embryos produced after in vitro maturation of oocytes obtained from ovaries after ovariohysterectomy (in vitro) with that of oocytes retrieved from follicle-stimulating hormone-treated donors at 24 h after administration of luteinizing hormone (in vivo) and fertilization by intracytoplasmic sperm injection (ICSI) or IVF were compared. In each group presumptive zygotes were assessed for cleavage on IVC Days 1 and 4 and for development to blastocysts on IVC Day 7. In vitro matured oocytes had lower frequencies of meiotic maturation (59.2% v. 66.5%), cleavage at Day 1 (41.4% v. 64.9%) and development to the morula stage at Day 4 (65.8% v. 87.9%) than did in vivo matured oocytes, after ICSI and IVF. Development to the blastocyst stage was lower in in vitro matured oocytes (19.0%) than in vivo matured oocytes (29.5%) after ICSI. In Experiment 2, we evaluated the capacity of sperm injected oocytes without a visible polar body to undergo cleavage and in vitro development. More in vivo matured than in vitro matured oocytes underwent cleavage at Day 1 (46.6% v. 12.6%) and developed to the morula stage by Day 4 (66.7% v. 46.1%), but no blastocysts were obtained at Day 7 in either group. In Experiment 3, we evaluated the in vivo viability of domestic cat embryos derived from ICSI of in vitro matured oocytes. Morula stage embryos were transferred to 18 domestic cat recipients either on Day 4 or 5 after oocyte recovery. A total of 3 domestic cat recipients were pregnant after transfer to recipients on Day 5. Two pregnant cats delivered two normal and healthy live male kittens on Day 68 of gestation and the remaining cat delivered a male kitten on Day 62 that died during the last two days of gestation. These results demonstrate that: (1) inadequate cytoplasmic maturation of in vitro matured domestic cat oocytes is the main cause of deficient oocyte activation; (2) the injection of oocytes without a visible polar body is a useful technique to evaluate oocyte cytoplasmic maturation; and (3) blastocysts obtained after ICSI of in vitro matured oocytes are viable and not a result of parthenogenesis.

Animals↗
PubMed2003

Worker policing without genetic conflicts in a clonal ant.

In group-living animals, mutual policing to suppress reproduction is an important mechanism in the resolution of conflict between selfish group members and the group as a whole. In societies of bees, ants, and wasps, policing against the production of males by other workers is expected when egg laying by workers decreases the average inclusive fitness of individual group members. This may result (i) from the relatedness of workers being lower to worker than to queen-derived males or (ii) from a lowered overall colony efficiency. Whereas good evidence exists for policing behavior caused by genetic conflicts, policing caused by efficiency factors has not been demonstrated. We investigated the regulation of reproduction in the ant Platythyrea punctata, a species in which colonies are clones because workers are capable of producing female offspring by thelytokous parthenogenesis. Reproductive conflicts resulting from differences in genetic relatedness are therefore not expected, but uncontrolled reproduction by all workers could lead to the destruction of sociality. Here we show that worker policing by aggressive attacks against additionally reproducing workers keeps the number of reproducing workers low. Furthermore, through experimental manipulation of the number of brood items per colony, we show that worker policing can enhance group efficiency.

Aggression↗
PubMedDate not supplied

Rescuing human embryonic stem cell research: the Blastocyst Transfer Method.

Despite the therapeutic potential of human embryonic stem (HES) cells, many people believe that HES cell research should be banned. The reason is that the present method of extracting HES cells involves the destruction of the embryo, which for many is the beginning of a person. This paper examines a number of compromise solutions such as parthenogenesis, the use of defective embryos, genetically creating a "pseudo embryo" that can never form a placenta, and determining embryo death, and argues that none of these proposals are likely to satisfy embryoists, that is, those who regard the embryo as a person. This paper then proposes a method of extracting HES cells, what might be called the Blastocyst Transfer Method, that meets the ethical requirements of embryoists, and it considers some possible concerns regarding this method. It concludes by encouraging future HES cell research to investigate this method.

Blastocyst↗
PubMed1974

Enzyme variability in natural populations of Daphnia magna. 3. Genotypic frequencies in intermittent populations.

In temporary habitats populations of the cyclical parthenogen, Daphnia magna, are re-established each year from sexual eggs and reproduce parthenogenetically for two or at most three generations. The genetic effects of this breeding system have been investigated by analyzing allozyme frequencies in nineteen intermittent populations.-Genotypic frequencies at polymorphic loci were ordinarily found to be in good agreement with Hardy-Weinberg proportions and disequilibria between loci were not observed. Although significant changes in gene frequencies were observed both during and between successive cycles, there was no evidence of the marked instability of genotype frequencies characteristic of permanent populations. The recombinational degradation of genotypes at the end of each annual cycle in temporary habitats effectively prevents the genotypic structuring which develops when continued parthenogenesis is possible.

Animals↗
PubMed1976

Polymorphism in a cyclic parthenogenetic species: Simocephalus serrulatus.

A survey of sixteen isozyme loci using electrophoretic techniques was conducted for three isolated natural populations and one laboratory population of the cyclic parthenogenetic species, Simocephalus serrulatus. The proportion of polymorphic loci (33%-60%) and the average number of heterozygous loci per individual (6%-23%) in the three natural populations were found to be comparable to those found in most sexually reproducing organisms. Detailed analyses were made for one of these populations using five polymorphic loci. The results indicated that (1) seasonal changes in genotypic frequencies took place, (2) apomicitic parthenogenesis does not lead to genetic homogeneity, and (3) marked gametic disequilibrium at these five loci was present in the population, indicating that selection acted on coadapted groups of genes.

Alkaline Phosphatase↗
PubMed1981

The chromosomes of turkey embryos during early stages of parthenogenetic development.

In the early stages of parthenogenetic development in turkey eggs, many blastoderms are mosaics of haploid, diploid and polyploid cells. The genome composition of these blastoderms can be identified by C-banding. They may be generally described as either A-Z/2A-ZZ/nA-nZ or A-W/2A-WW/nA-nW and are found in a nearly 1:1 ratio. The blastoderms showing the W body (W+) become lethal within two days of incubation. The haploid cell proportion decreases rapidly during the early stage of development, and, as haploid cells decrease, the proportion of polyploid cells appears to increase. At six days of incubation, various kinds of parthenogenetic development can be observed. Their genome compositions are either diploid (2A-ZZ) or mosaic (A-Z/2A-ZZ). These findings suggest that diploid parthenogenesis occurs by either suppression of meiosis II or chromosome doubling some time after the first cleavage division. The frequent occurrence of mosaic blastoderms indicates that the majority, if not all, of the parthenogenetic embryos initiate their development in haploid ova.

Animals↗
PubMed1989

Cytogenetic analysis and developmental capacity of normal and abnormal embryos after IVF.

Chromosome abnormalities represent the major cause of pre- and post-implantation embryo wastage. Indeed, 29% of embryos produced by in-vitro fertilization have an abnormal karyotype. Parthenogenesis (1.6% of the cases) and triploidy (6.4%) are the only abnormalities detectable 17 h after insemination. A total of 23% of activated oocytes with a single pronucleus (1 PN) and 18% of fertilized eggs with three pronuclei (three PN) remained uncleaved, compared with 4% for diploid eggs. The rate of cleavage for parthenogenes is not different from normal eggs. On the contrary, we observed that 29% of three PN-eggs reached the 5- to 8-cell stage 42 h after insemination when compared to 15% for diploids mainly due to a direct division of 56% of three PN-eggs in three cells. A model of development for triploid eggs is proposed, taking into account physiological and cytogenetic observations. The quality of embryos expressed in terms of morphology did not show any difference between embryos proceeding from parthenogenes or diploid eggs. On the contrary, the rate of fragmentation is decreased for triploid (3%) compared with diploid (13%) or activated (17%) embryos. Embryos resulting from one, two, three or four pronucleate ova display different developmental capacities during 5 days' culture in vitro.

Chromosome Aberrations↗
PubMed2026

The twofold cost of sex reconsidered: meiotic mechanisms protect anisogamous populations from invasion by thelytoky.

Most multicellular animals practice anisogamy (fertilization between eggs and sperm). When mothers produce sons and daughters at a 1:1 ratio, the "twofold cost of males" arises because males do not directly contribute to population growth. If thelytokous parthenogens producing only daughters invade a population, they should spread rapidly. Although thelytoky has repeatedly evolved across invertebrate and vertebrate taxa, it remains a minority. Why? The evolutionary transition from anisogamy to thelytoky requires eggs to initiate embryonic development without fertilization. However, in metazoan animals, meiotic metaphase (MM) arrest halts oogenesis midway and normally resumes only after stimulation by sperm penetration. Empirical and experimental evidences indicate that release of MM arrest without fertilization is extremely difficult, providing a strong mechanistic barrier against parthenogenesis. Even if MM arrest were released, oogenesis would proceed to produce either a haploid embryo or a diploid embryo through refusion with the second polar body (terminal fusion automixis). Outbred species typically accumulate more than one lethal equivalent of recessive deleterious alleles per genome as heterozygotes. Upon transition to haploid or automictic development, these recessive lethals normally masked in outbred diploids would be exposed simultaneously, causing embryonic death and creating the next barrier. Thus, thelytoky cannot be achieved simply by modification of the existing meiotic system; instead, other mechanisms, such as apomixis, that bypass meiosis are required. Mathematical models and simulations support this "meiotic constraint" hypothesis. Combined with recently proposed immediate benefits of anisogamy and traditional genetic benefits (e.g., Red Queen), it may largely explain the maintenance of costly anisogamy.

Animals↗
PubMed2006

Human factors facilitating the spread of a parasitic honey bee in South Africa.

Workers of the honey bee subspecies Apis mellifera capensis (Eschscholtz) produce female offspring by thelytokous parthenogenesis and can parasitize colonies of other subspecies. In 1990, translocation of 400 colonies of A. m. capensis into the distribution area of A. m. scutellata by a commercial beekeeper triggered a dramatic parasitic phenomenon. Parasitized colonies died within a few months of infestation, and this resulted in the loss of tens of thousands of colonies by commercial beekeepers in the A. m. scutellata range in South Africa. To deal with the problem and to identify methods that would limit the impact of the social parasite, we investigated the link between beekeeping management and severity of parasitic infestations in terms of colony mortality and productivity. We demonstrate that colonies from apiaries subjected to migrations are very susceptible to infestation and consequently show dramatic mortality. Their productivity is also inferior to sedentary colonies and those in isolated apiaries in terms of honey yield and brood quantity. Furthermore, by concentrating hives in small areas and often in the vicinity of other beekeepers, cross-infestations can easily occur. This can undermine previously parasite-free beekeeping businesses. As a result of our surveys, we propose beekeeping practices based on locally trapped bees, reduced migration, and better control of parasite spread, thus promoting the conservation of these pollinators. If followed by all the South African beekeepers, these measures should limit the spread of the parasite until it is eliminated within a few years, after which full migratory beekeeping practices could resume.

Animal Husbandry↗
PubMed2006

Multiple parthenoforms of Empoasca leafhoppers from Madeira Island: where are these unisexual forms coming from?

There are controversial opinions on whether asexual reproduction is more common on islands than on the mainland. Although some authors consider that the evidences of geographical parthenogenesis support the view that asexual reproduction is more common on islands, comparative data on the modes of reproduction of insular and continental taxa confirming this statement are very limited. In this work, we report the presence of three unisexual forms and three bisexual species of the genus Empoasca (Cicadelloidea, Hemiptera, Insecta) from Madeira Island. Experimentally, the unisexual forms reproduced in the absence of males for several generations. The chromosome analysis has shown that the bisexual species differ from one another in chromosome number, and unisexual forms are apomictic and also each have different chromosome numbers. Of parthenoforms, one is triploid and two are of obscure level of ploidy, 2n or 3n. The results obtained show that for this genus unisexual forms are more common on Madeira Island than in the nearby continental areas. It is suggested that unisexual forms may be more plentiful on islands than on the mainland because if an asexual reproduction event occurs, the relaxing competition in these underexploited and enemy-free habitats may favor the establishment of new parthenogenetic lineages.

Animals↗
PubMedDate not supplied

Turkey-chicken hybrids: a cytological study of early development.

Reciprocal hybridization experiments were performed between the line of Beltsville Small White turkeys that has been selected for high incidence of parthenogenesis and two strains of chickens. The eggs were examined cytologically at early stages of development. In the cross of turkey females X chicken males, it was found that both male (2A-ZZ) and female (2A-ZW) zygotes are formed, but the latter are much less viable and usually die in a very early stage of development. Besides the hybrid embryos, parthenogenetic-type embryos also were found. In the cross of chicken females X turkey males, no evidence of successful hybridization was obtained. However, cell division was initiated in 100 percent of the chicken eggs. This fact suggests that turkey sperm had some enhancing effect on parthenogenetic cell division in chicken eggs.

Animals↗
PubMed1998

Observations on ploidy of cells and on reproductive performance in parthenogenetic turkeys.

Using flow cytometry, the ploidy levels of parthenogenetic turkeys were quantified from blastodisc stage to adulthood. Eggs were collected from noninseminated hens of the Beltsville Small White flock, known for their high degree of parthenogenesis, and the blastodermal cells from developing embryos were compared with those of embryos produced by hens inseminated with semen from males of the same flock. Erythrocytes of parthenogens from Day 10 of incubation to 27 mo of age were also used for ploidy determination. Sperm and erythrocyte preparations from normal males of the above flock served as haploid and diploid standards, respectively. In parthenogenetically developing blastoderms, 40.3 +/- 14.5% of the cells were haploid and 48.9 +/- 11.9% diploid; blastoderms from fertilized eggs had no haploid cells. The haploid cell content of parthenogens declined from the blastodermal stage to adult life, with 1.9 +/- 2.3% at 10 to 20 d of embryonic development, 1.5 +/- 1.4% at 21 to 29 d of development, 1.4 +/- 2.6% at 4 wk posthatch, and 1.3 +/- 1.9% in adulthood, although changes between the 1st mo after hatch and adult stage were not significant. It is possible, therefore, that parthenogenetic embryos with a low proportion of haploid cells could be the ones that survive to Day 10 of development and beyond, whereas those with a higher proportion of haploid cells fail to develop. The semen volume of male parthenogens was significantly lower than that of normal males, although the concentration of spermatozoa and their fertilizing capacity did not vary significantly between groups, suggesting that the germ cells of these parthenogens are capable of normal meiosis and sperm maturation leading to a normal fertility.

Animals↗
PubMed1998

Differentiating between parthenogenetic and "positive development" embryos in turkeys by molecular sexing.

In mated or inseminated turkeys, 5 to 15% of eggs set for incubation show only rudimentary development. Most of these embryos die during the first 24 to 48 h of incubation and contain only unorganized sheets of tissue. This abnormal development is termed "positive development" (PD). Turkey eggs also show incidence of parthenogenesis and the resulting progeny is believed to be always male. As both types of embryos are morphologically similar at the early stage of incubation, it has been speculated that PD embryos may in fact be parthenogens. By identifying the sex at the blastodermal stage with the help of DNA markers, we have differentiated between the PD embryos and parthenogens. Parthenogenetic embryos were obtained from eggs laid by uninseminated or virgin Beltsville Small White (BSW) hens, and the PD embryos were obtained from eggs of inseminated Nicholas and British United Turkeys of America (BUTA) hens. DNA was extracted from blastoderms of parthenogenetic and PD embryos. Turkey W-chromosome specific DNA probe and primers were used to detect females in all samples by Southern blot and polymerase chain reaction (PCR), respectively. No female was detected among the 35 parthenogens examined, whereas there were 3 females among the 11 PD embryos. The presence of both males and females among PD embryos suggests that they are products of fertilization, and that at least these 3 female embryos, if not all the 11 PD embryos, are not of parthenogenetic origin. It is concluded, therefore, that PD embryos result from errors in fertilization or from early embryonic mortality following successful fertilization, and that they are unlikely to be of parthenogenetic origin.

Animals↗
PubMed2004

Apoptosis in parthenogenetic preimplantation porcine embryos.

Parthenogenesis (PA) of the oocyte is essential to a number of oocyte- or embryo-related technologies such as intracytoplasmic sperm injection and cloning by nuclear transfer. This study investigated the onset and frequency of apoptosis in PA- porcine embryos and the morphological changes that conform to the general criteria of apoptotic cell death by using a terminal deoxynucleatidyl transferase-mediated deoxyuridine 5-triphosphate nick-end labeling (TUNEL) assay. PA embryos had a higher degree of apoptotic cell death during in vitro culture, a lower cleavage rate (45% vs. 71%), and a lower development rate to the blastocyst stage (16% vs. 29%), relative to in vitro fertilization (IVF). The earliest positive TUNEL signal in the PA embryos was detected on Day 6, 1 day later than that in IVF embryos. Apoptosis in PA embryos increased from 15% of the embryos on Day 6 to 29% on Day 8. The mean level of apoptosis of the PA embryos was statistically higher than that of IVF embryos, except on Day 5. In particular, apoptosis in PA embryos was twice that of IVF embryos on Day 6 (15% vs. 6.7%) and Day 8 (29% vs. 13%). The mean cell number in PA blastocysts was significantly lower than that of IVF blastocysts, whereas the percentage of apoptosis in PA blastocysts was significantly higher than that of IVF blastocysts. There was a high percentage of haploid (62.5%) PA blastocysts. The ploidy may contribute to a high level of apoptosis. These results may help to explain the mechanism of parthenogenetic developmental failure and may lead to methods that will improve parthenogenetic development.

Animals↗
PubMed1998

Development of parthenogenetic and cloned ovine embryos: effect of activation protocols.

Preliminary experiments carried out on ovine oocytes were designed to establish correlations between activation protocols and subsequent rates of embryonic development. The best activation protocols were thereafter used in studies on ovine parthenogenesis and cloning. The first study established that chemical activators induce pronuclear development at a slightly higher rate than physical activation (ionomycin, 96%; ethanol, 95%; electro activation, 80%). Inhibition of second polar body extrusion and one single pronucleus were observed in the majority of the oocytes (approximately 90%) treated for 3 h with 6-dimethylaminopurine (6-DMAP) following either ionomycin or ethanol activation. While over 80% of these oocytes cleaved after transfer to the oviducts of recipients, progression to the blastocyst stage was higher after ionomycin as compared with ethanol activation (58% vs. 19%). The ionomycin plus 6-DMAP activation protocol was used to produce parthenogenetic blastocysts whose subsequent development was monitored both by ultrasonography and by direct fetal examination. Over 70% of parthenogenotes were viable on Day 21 of pregnancy but dead by Day 25. The effects of 6-DMAP on nuclear remodeling and fetal development of cloned embryos was then investigated. Control cloned embryos underwent nuclear envelope breakdown (NEBD), premature chromatin condensation (PCC), and inhibition of DNA synthesis. By contrast, reconstructed embryos treated with 6-DMAP exhibited intact nuclear membranes, interphase chromatin, and no interference on DNA synthesis. Moreover, cloned embryos developed to blastocyst stage in higher percentage after 6-DMAP treatment (83% vs. 25%). We conclude that ionomycin followed by 6-DMAP incubation yields high percentages of diploid parthenogenetic embryos that develop to Day 25 before dying. Cloned embryos activated by the ionomycin-6-DMAP protocol develop readily to term.

Adenine↗
PubMed1998

Parthenogenetic activation of marmoset (Callithrix jacchus) oocytes and the development of marmoset parthenogenones in vitro and in vivo.

Mammalian oocytes can be induced to resume meiosis without fertilization, and the resulting parthenogenetic embryos carry only maternal chromosomes. Human oocytes can be activated by many chemical and physical stimuli, but postimplantation studies of human parthenogenetic embryos are not ethically acceptable. The common marmoset monkey (Callithrix jacchus) is a good model for studying primate parthenogenetic development postimplantation, since follicular aspiration, embryo transfer, and early postimplantation development of biparental embryos have already been described. Marmoset oocytes were either subjected to two series of six electrical pulses (DC; 2 kV/cm and 70 microsec) or were incubated in 7% ethanol in PBS. Ninety-two percent (68 of 74) and 20% (8 of 40) of marmoset oocytes were activated by electrical stimulus or ethanol, respectively. Parthenogenetic (n = 3) or in vitro-fertilized (n = 2) embryos were transferred at the 4-cell stage to synchronized recipient female marmosets (n = 5). Progesterone, chorionic gonadotropin, and inhibin in the peripheral plasma of recipient animals were measured. After 33 days of gestation, recipient animals were perfused and the uteri were collected. The 2 females that had received biparental embryos and 2 of the 3 females that had received parthenogenetic embryos displayed biochemical and histological evidence of implantation. This is the first report that a primate embryo comprising only parthenogenetic cells is capable of implantation. This highlights the need to scrutinize levels of parthenogenesis associated with human assisted reproductive technologies. Marmoset parthenogenones also provide a unique model for elucidating the roles of parental genomes in primate development.

Animals↗
PubMed1992

Clonal structure of the introduced freshwater snail Potamopyrgus antipodarum (Prosobranchia: Hydrobiidae), as revealed by DNA fingerprinting.

Multi-locus DNA fingerprints were obtained from individuals of the hydrobiid snail, Potamopyrgus antipodarum (= P. jenkinsi), by using an RNA derivative (pSPT 18.15) of Jeffrey's 33.15 minisatellite core sequence. Whole-body homogenization of snails yielded 3.21 +/- 0.09 micrograms DNA per individual, producing complex profiles comprising 12-22 fragments within the 1.0-20.0 kilobase (kb) size range. Fingerprints from natural and experimental populations identified three distinct clonal genotypes corresponding to morphological strains A, B and C, with only rare mutational variants. Mother-offspring comparisons of genetic fingerprints revealed genetic stability during apomictic parthenogenesis. Data support the notion that British populations of P. antipodarum comprise three widespread obligate parthenogenetic clones resulting from a mid-19th Century introduction from Australasia. The present-day low levels of genotypic diversity are discussed in relation to the typical occurrence of P. antipodarum in man-made or immature habitats.

Animals↗
PubMed1996

Molecular characterization of cyclic and obligate parthenogens in the aphid Rhopalosiphum padi (L.).

Holocyclic clones of the aphid Rhopalosiphum padi (L.) reproduce by cyclic parthenogenesis, whereas anholocyclic individuals are obligate parthenogens. Mitochondrial DNA and (mtDNA) and random amplified polymorphic DNA markers in R. padi as well as plasmid DNA markers of its bacterial endosymbiont, Buchnera aphidicola, were examined to determine the extent of genetic divergence between clones with these differing breeding systems. These analyses revealed that cyclically parthenogenetic lineages possessed differing mtDNA and plasmid haplotypes than most obligately asexual clones. The extent of sequence divergence between the maternally inherited molecules suggest a relatively ancient origin of asexuality. Our work also identified a random amplified polymorphic DNA marker linked to the life-cycle variation in R. padi. This marker not only permits the rapid diagnosis of breeding system but sets the stage for studies to identify the gene(s) controlling this variation in mode of reproduction.

Animals↗