Differential expression of allelic carboxylesterase-2 genes in oocytes of loach (Misgurnus fossilis L.) and heterogeneity of loach oocytes and eggs for the expression of allelic carboxylesterase-2 genes

Tissue and cell differences have been found in the expression of allelic carboxylesterase-2 (E-2) (carboxylic ester hydrolase, E.C. 3.1. 1.1) genes of the loach. The relative activity of two allozymes in the brain and muscles of heterozygotes is similar, whereas in oocytes and eggs of the same fish the activity of one of the allozymes is considerably greater than that of the other. Oocytes and eggs of some heterozygotes were shown to be heterogeneous for the expression of E-2 alleles. The phenomenon implies that roe produced by a heterozygous female can contain two or more egg types: in some the alleles are equally expressed, while in others one of the alleles is predominantly expressed.     

Electrophoretic analysis of newly synthesized and stored maternal RNA during oogenesis ofCalliphora erythrocephala (Dipt.)

Summary Ovaries ofC. erythrocephala synthesize large amounts of poly(A)⁺ and poly(A)^– RNA during early and middle stages of oogenesis as shown by labelling with³H-uridine in vivo. After incubation for 1 h, a striking difference in the electrophoretic pattern of newly synthesized labelled poly(A)⁺ RNA and the poly(A)⁺ RNA present in sufficient amounts for optical density measurements (steady state poly(A)⁺ RNA) was observed. During early and mid-oogenesis, in the poly(A)^– RNA fraction, 4S predominantly mature rRNA, 5S RNA and tRNA were labelled. These fractions were no longer synthesized during late oogenesis, whereas poly(A)⁺ RNA was labelled continously During oogenesis stage specific differences in the size distribution of newly synthesized and steady state poly(A)⁺ RNA were not obvious. However, different sizes of labelled poly(A)⁺ RNA species were detected in 0–2h old preblastoderm embryos, after injection of³H-uridine into females either 3–4 days (stage 3–4 of oogenesis) or 24 h before oviposition (stage 5–6 of oogenesis). This difference in RNA synthesis was related to the presence of active nurse cell nuclei. The poly(A)⁺ RNA fraction represents about 2–3% of the total RNA in both ovaries and freshly laid eggs as judged by measurements of optical density and radioactivity bound to oligo(dT). The length of poly(A)-segments in ovarian poly(A)⁺ RNA varied from about 30 to 200 nucleotides.     

Application of cyanide hydrolase from Klebsiella sp. in a biosensor system for the detection of low-level cyanide

Abstract A partially purified preparation of cyanide hydrolase (cyanidase) from a bacterium, Klebsiella sp., was applied as a biocatalyst in a biosensor system for low-level cyanide detection. In the biosensor system cyanide hydrolase converts cyanide into formate and ammonia. The formate produced in the cyanide degradation was detected with a formate biosensor, in which formate dehydrogenase (FDH; E.C. 1.2.1.2) was co-immobilized with salicylate hydroxylase (SHL; E.C. 1.14.13.1) on a Clark electrode. The principle of the formate sensor is that FDH converts formate into carbon dioxide using -nicotinamide adenine dinucleotide hydrate (NAD⁺). The corresponding NADH produced is then oxidized to NAD⁺ by SHL using salicylate and oxygen. The oxygen consumption is monitored with the Clark electrode. The optimum buffer pH and temperature for the enzymatic hydrolysis of potassium cyanide were studied. The preliminary experiments including the pretreatment of cyanide with cyanide hydrolase and then detection by the formate sensor gave a detection limit at 7.3 mol l^–1 cyanide. The linear range of the calibration curve was between 30 mol l^–1 and 300 mol l^–1 cyanide.     

Transformation of grape (Vitis vinifera L.) zygotic-derived somatic embryos and regeneration of transgenic plants

Summary Transgenic grape plants were regenerated from somatic embryos derived from immature zygotic embryos of seedless grape (Vitis vinifera L.) selections. Somatic embryos were bombarded twice with 1 m gold particles using the Biolistic PDS-1000/He device (Bio-Rad Laboratories) and then exposed to Agrobacterium tumefaciens strain C58/Z707 containing the binary plasmid pGA482GG or pCGN7314. Following cocultivation, secondary embryos were allowed to proliferate on Emershad/Ramming proliferation (ERP) medium for 6 weeks before selection on ERP medium containing 20–40 g/ml kanamycin (kan). Transgenic embryos were identified after 3–5 months under selection and allowed to germinate and develop into rooted plants on Woody Plant Medium containing 1 M 6-benzylaminopurine (BAP), 1.5% sucrose, 0.3% activated charcoal and 0.75% agar. Integration of the foreign genes into these grapevines was verified by growth in the presence of kan, positive GUS and PCR assays, and Southern analysis.     

Expression inEscherichia coli K12 strains of two synthetic human calcitonin genes differing in codon composition

Two genes coding for a Val⁸-variant of the human calcitonin (hCT) are synthesized on two different codon biases: the native codons for the hCT gene and the codons preferential forEscherichia coli. Both genes are fused to a synthetic human interferon-gamma (IF) gene [6] and expressed in various strains ofE. coli K12. It is found that, in all host strains used, the level of expression of both genes is similar and much lower (1/50–1/100) than that of the IF gene alone.     

Distribution and biology of marine cladocerans in the coastal waters of southern China

Geographical and seasonal distributions of marine cladocerans in the coastal waters of southern China were studied. Penilia avirostris was the most common species, followed by Evadne tergestina and Podon schmackeri. P. avirostris and E. tergestina were most common during summer. P. schmackeri, found only in a small bay northeast of Hong Kong, showed no clear seasonal pattern of occurrence. P. avirostris and E. tergestina were found at temperatures ranging from 16–32°C and salinity ranging from 7.3–37.2. P. schmackeri was restricted to a temperature range of 17–29°C and a salinity range of 31.0–37.2. No significant relationships between marine cladoceran abundance and chlorophyll a concentration were found in samples taken from Tolo Harbour. Parthenogenetic brood size of P. avirostris and E. tergestina ranged from 1 to 14, while P. schmackeri was found to carry up to 19 embryos per brood. No geographical trend in fecundity patterns was observed. No correlation was found between body length and brood size. The occurrence of females with resting eggs was rare.     

E-1020, a water soluble imidazopyridine,has direct effects on Ca2+-dependent force and ATP hydrolysis of canine and bovine cardiac myofilaments

E-1020 is a cardiotonic agent that acts as a cyclic-AMP phosphodiesterase inhibitor but also may have actions which alter myofilament response to Ca²⁺. To identify direct actions of E-1020 on cardiac contractile proteins, effects of E-1020 on myofibrillar Ca²⁺ dependent MgATPase and force generation in chemically skinned fiber bundles were measured. In bovine cardiac myofibrils, E-1020 (100 M) significantly increased myofilament Ca²⁺ sensitivity and Ca²⁺-dependent ATPase activity at submaximal pCa values. At pCa 6.75, E-1020 significantly increased ATPase activity in bovine (10–100 pM) and canine (1–100 pM) cardiac myofibrils but had no effect on rat cardiac myofibrils. Moreover, in one population of canine ventricular fiber bundles, E-1020 (0.0–10 M) significantly increased isometric tension at pCa 6.5 and 6.0, whereas in another population of bundles E-1020 had no effect on tension. In no case was resting (pCa 8.0) or maximal tension (pCa 4.5) increased by E-1020. Measurements of Ca²⁺ binding to canine ventricular skinned fiber preparations demonstrated that E-1020 does not alter the affinity of myofilament troponin C for Ca²⁺. We conclude that part of the mechanism by which E-1020 acts as an inotropic agent may involve alterations in the responsiveness of contractile proteins to Ca²⁺. The lack of effect of E-1020 on some preparations may be dependent on isoform populations of myofilament proteins.     

Statistical and Frequency-Amplitude Characteristics of Ultraweak Emissions of the Loach Eggs and Embryos under the Normal Conditions and upon Their Optic Interactions. 2. Changes in Characteristics of Ultraweak Emissions upon Optic Interaction of Groups of Embryos of Different Ages

We compared the characteristics of ultraweak emissions from groups of loach embryos of different ages in the presence or absence of optic interaction. The percentage of zero values of emission gradually increased during the first hour of optic interaction. The number and height of rare big pulses estimated by the value of kurtosis increased in parallel. In addition, the correlation between the Fourier spectra of optically interacting samples decreased at a higher rate than in the absence of optical contact. Just after the 1-hour optic interaction was terminated, the number of high pulses decreased in a younger interacting group and increased in the older one and the farther away the partner groups were in developmental stages, the more pronounced these differences were. Measurements of the Fourier spectra after long-term (12–22-hour) optic interactions have shown that an exchange of autocorrelation characteristics of the spectra took place among the samples: the sums of autocorrelation coefficients were inverted in the vast majority of cases, often with an overshoot or, at least, were smoothed over with reference to the control samples. We conclude that the previously described effects of optic interactions between groups of loach embryos of different ages could be due to changes in the frequency spectra of their ultraweak emissions.     

The relative content of oocyte and somatic 5S rRNA at different stages of embryonic development as an index of the replacement of maternal ribosomes by ribosomes of the embryo in the loach

Relative content of the oocyte and somatic 5S rRNA in loach Misgurnus fossilis L. during development was determined electrophoretically. Embryos before hatching contain 70% and swimming larvae no less than 50% of the oocyte 5S rRNA. We assume that the relative content of 5S rRNA fractions reflects the proportion between ribosomes synthesized during oogenesis and those synthesized in embryos and larvae. We calculated using previous data (Timofeeva, Kafiani, 1964) the rates of maternal ribosome decay and ribosome synthesis in the embryo. During organogenesis these rates appear to be 1.17-1.09 x 10(6) and 1.7 x 10(6) molecules/sec per embryo, respectively.     

Effect of chronic ingestion of the cysteine proteinase inhibitor, E-64, on Colorado potato beetle gut proteinases

Chronic ingestion of the highly active, specific cysteine proteinase inhibitor, E-64, has a profound effect on Colorado potato beetle (CPB) larval growth, development and survival, as well as on adult fecundity. However, the number of insects surviving to the adult stage did not decrease below 26% with increasing E-64 concentration above 1.5 g E-64 cm^–2 leaf surface. The development time to the pupal stage was increased from 13 days, when larvae were reared on control leaves, to 21 days at a concentration of 1.5 g E-64 cm^–2 . The most significant effect of dietary E-64 was on adult fecundity, with mated females reared on untreated leaves laying an average 62 ± 5.7 eggs daily in the first 10 days, and those maintained on 0.5 g E-64 cm^–2, laying only 16 ± 2.4 eggs day^–1. Females given 1 g E-64 cm^–2 laid few if any eggs, but started producing egg masses as large as control insects about 5 days after being switched to control leaves. These effects on the insect life cycle were directly related to the degree of inhibition of cysteine proteinase activity in gut extracts. The general proteinase activity in control extracts was 6.5 ± 0.16 units min^–1 mg gut^–1, which decreased to 1.9 ± 0.16 in guts of insects reared on 1 g E-64 cm^–2. The proportion of proteinase activity inhibitable by E-64 decreased from 66% in control guts to 10-15% in guts from larvae reared on 1 g E-64 cm^–2. The aspartate proteinase inhibitor, pepstatin, decreased proteinase activity by 35% in control guts. There was no induction of pepstatin-inhibitable proteinases in response to inhibition by E-64, and no inhibition of gut enzyme activity by soybean trypsin inhibitor from larvae fed any of the E-64 concentrations. This study demonstrates that proteinase levels must be significantly reduced to have a pronounced effect on larval growth and survival, while fecundity of mated females is affected by lower concentrations of inhibitor. It also suggests that the CPB may be a difficult pest to control using a more specific, plant-derived cysteine proteinase inhibitor, such as oryzacystatin.     

Isozyme patterns in zygotic and somatic embryogenesis of carrot

Isozyme patterns of carrot (Daucus carota L.) zygotic embryos between the torpedo stage up to 5-day-old seedlings have been compared with those of the similar stages from the embryogenic cell suspension culture to the late somatic plantlet. Somatic embryos blocked at the torpedo stage by -cyclodextrine have also been analyzed. All these stages have been analyzed with respect to seven different enzyme systems: arylesterase, glucosephosphate isomerase, phosphogluconate dehydrogenase, alcohol dehydrogenase, isocitrate dehydrogenase, aspartate aminotransferase and phosphoglucomutase (EC 2.7.5.1, PGM). The relationships between the different stages of both types of embryogenesis have been visualized using an unrooted tree. Generally, profiles of somatic embryos were different from those of zygotic embryos. Interestingly however, a typical zygotic embryo pattern was found in the cyclodextrine-blocked somatic embryos. Only aspartate aminotransferase patterns revealed a similarity between zygotic and somatic torpedo embryos. Both plantlet types showed close patterns with common isozymes. Moreover, similarities were evident between somatic plantlets and cell suspensions. A few isozymes appeared to be stage specific markers: esterase 10–11 were specific to achenes and early germination, phosphogluconate dehydrogenase 8 was specific to 4–5 day-old seedlings and phosphoglucomutase 1 and 7 and alcohol dehydrogenase 4 were markers for zygotic embryos. No somatic embryogenesis specific isozyme could be found. We show that patterns can be associated with particular tissue formation: mainly, aspartate aminotransferase 2 and 1, phosphoglucomutase 8 and 9 and phosphogluconate dehydrogenase 7 coincided with apical meristem initiation and phosphoglucomutase 4 and 5, zones b and d of esterase and zone b of phosphogluconate dehydrogenase coincided with vascular bundle formation.Abbreviations ADH alcohol dehydrogenase - CD -cyclodextrine - CS cell suspension culture - 2,4-D 2,4-dichlorophenoxyacetic acid - EDTA ethylenediaminetetraaeetie acid - LiBo lithium hydroxide/boric acid - PEG polyethylene glycol - PVP polyvinylpyrrolidone - SEg somatic embryo at the globular stage - SEh heart stage - SEte early torpedo stage - SEtl late torpedo stage - SEce early cotyledonary stage - SEcl late cotyledonary stage - SECD somatic embryo blocked at the torpedo stage with -cyclodextrine - EST esterase - GOT aspartate aminotransferase - IDH isocitrate dehydrogenase - MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide) - PMS phenazonium methosulfate - PGD phosphogluconate dehydrogenase - PGI glucosephosphate isomerase - PGM phosphoglucomutase - SO dry seed - S1–3 seed after 1–3 days of germination - SP1–2 young and old somatic plantlets - ZE zygotic embryo - ZP4–5 4–5 day-old seedlings     

Somatic embryogenesis in the woody legume Calliandra tweedii

In vitro somatic embryogenesis was established from 1–1.2 cm long internodal and petiolar segments excised from 20-year-old woody legume, Calliandra tweedii Benth trees. Within 6–7 weeks, globular, heart, torpedo-shaped and dicot embryos differentiated directly. Among the cytokinins and auxins tested, 1 M 2iP in Murashige and Skoog's (MS) medium induced well-organized normal embryos in 71% of the internodal segments. Petiolar explants responded best on 0.5 M NAA wherein 30% of the cultures developed normal embryos. Somatic embryos developed roots and shoots when they were retained for 90 days in old desiccated MS medium. Plants ready for transfer to soil were regenerated through somatic embryogenesis in 4–5 months. These grew well after transfer to soil.     

Effects of paper mill effluents on the fish fauna of stony shores of Lake Päijänne

The fish fauna of the stony littoral in the central parts of L. Päijänne was studied by electric fishing on four occasions during 1988–1989. Ten fish species and 1681 individuals (14.5 kg) were caught in the 15 fishing sites (4096 m²) which gives a mean density of 0.41 ind. m^–2 and biomass of 3.5 gm^–2. Effluent from two large paper mills causes a clear zonation of the fish fauna in the area. In the most polluted shores, only burbot and perch occurred regularly and the reproduction of other species was inhibited. In the semipolluted area (5–15 km from the Kaipola paper mill), burbot and stone loach occurred regularly, but owing to low numbers of perch and bullhead the total densities were usually low.The clean shores were characterized by an abundance of stone loach and bullhead and locally also of minnow, which seemed to be very sensitive to pollution. The strong floods during summer 1988 had positive effects on the reproduction of perch and pike, but the densities of other species were highest during the normal water level in autumn 1989.     

Identification of Potential Pleiotropic Genes for Immune and Skeletal Diseases Using Multivariate MetaCCA Analysis

BackgroundImmune and skeletal systems physiologically and pathologically interact with each other. Immune and skeletal diseases may share potential pleiotropic genetics factors, but the shared specific genes are largely unknown.ObjectiveThis study aimed to investigate the overlapping genetic factors between multiple diseases (including rheumatoid arthritis (RA), psoriasis, osteoporosis, osteoarthritis, sarcopenia, and fracture).MethodsThe canonical correlation analysis (metaCCA) approach was used to identify the shared genes for six diseases by integrating genome-wide association study (GWAS)-derived summary statistics. The versatile Gene-based Association Study (VEGAS2) method was further applied to refine and validate the putative pleiotropic genes identified by metaCCA.ResultsAbout 157 (p<8.19E-6), 319 (p<3.90E-6), and 77 (p<9.72E-6) potential pleiotropic genes were identified shared by two immune diseases, four skeletal diseases, and all of the six diseases, respectively. The top three significant putative pleiotropic genes shared by both immune and skeletal diseases, including HLA-B, TSBP1, and TSBP1-AS1 (p<E-300), were located in the major histocompatibility complex (MHC) region. Nineteen of 77 putative pleiotropic genes identified by metaCCA analysis were associated with at least one disease in the VEGAS2 analysis. Specifically, the majority (18) of these 19 putative validated pleiotropic genes were associated with RA.ConclusionThe metaCCA method identified some pleiotropic genes shared by the immune and skeletal diseases. These findings help to improve our understanding of the shared genetic mechanisms and signaling pathways underlying immune and skeletal diseases.     

DNA polymorphisms and linkage relationship of the human complement componentC6,C7, andC9 genes

In this report we describe the linkage between genes encoding human complement componentsC6,C7, andC9. Polymorphisms have been described at the DNA level for theC7 andC9 genes. We have studied 20 individuals by Southern blot analysis with fourC6 cDNA subclones to detect restriction fragment length polymorphisms (RFLPs). We have found a Taq I polymorphism defined by two alleles of 8.0 (C6 H) and 6.0 (C6 L) kilobases (kb). RFLP segregation for theC6, C7, andC9 loci in informative families allowed us to estimate the maximum Lod scores at a recombination fraction of =0.0 (C6–C7), =0.0 (C7–C9), and =0.0 (C6–C9). Significant linkage disequilibrium was found betweenC6 andC7 and betweenC7 andC9 loci in directly determined haplotypes of unrelated parents. Data from this study show that the genes encoding the human terminal complement componentsC6, C7, andC9 define a cluster in the short arm of chromosome 5. We propose that the clusters involving theC8A andC8B and theC6, C7, andC9 genes be referred to as MACI and MACH, respectively.     

Analysis of the genetic stability of <Emphasis Type="Italic">Eucalyptus globulus</Emphasis> Labill. somatic embryos by flow cytometry

Flow cytometry was used to measure the nuclear DNA content of Eucalyptus globulus Labill. somatic and zygotic embryos and leaves in order to determine if somatic embryogenesis induces DNA content and ploidy changes in this species. Mature zygotic embryos derived from open-pollination orchard families were collected from a location in the centre of Portugal. One group was kept for nuclear DNA content and ploidy analysis, and the other group was used for establishing embryogenic cultures. Mature zygotic embryos were grown on Murashige and Skoog (MS) medium supplemented with 3% (w/v) sucrose and 3 mg l^–1 -naphthaleneacetic acid (NAA) for 3 weeks and then transferred to MS medium without growth regulators. Globular somatic embryos from approximately 8-month-old embryogenic cultures were used in the assay. DNA ploidy levels and the nuclear DNA content of mature zygotic embryos, somatic embryos and leaves from the mother field tree were determined using flow cytometry combined with propidium iodide staining. Zygotic embryos had a nuclear DNA content of 1.32 pg/2C, somatic embryos had a nuclear DNA content of 1.39 pg/2C and leaves from the field tree had a nuclear DNA content of 1.40 pg/2C. The values estimated for the somatic embryos and mother plant did not differ statistically from each other (P0.05), but both differed from those of the zygotic embryos (P0.05). These results clearly indicate that no changes were induced during the embryogenic process. However, the differences found between the field plants and zygotic embryos did suggest that some aspects must be evaluated carefully, as propidium iodide fluorescence may potentially be influenced by the presence of secondary compounds (e.g. anthocyanins, tannins) in E. globulus somatic embryos and mature leaves. Therefore we believe that the somatic embryogenesis methodology used did not induce major genetic changes in the somatic embryos and that our primary goal of true-to-type propagation was assured.     

Plant regeneration from cultured protoplasts of the cooking banana cv. Bluggoe (Musa spp., ABB group)

Summary Suspensions of embryogenic cells of a triploid banana (Musa spp., cv. Bluggoe) were initiated from the uppermost part of meristematic buds, and used as protoplast source. After 20 weeks in culture, the suspension contained a mixture of globular structures or globules and embryogenic cell clusters, as well as single cells. Two types of protoplasts were obtained from embryogenic suspension culture: small (20–30 m) and larger (30–50 m) protoplasts with a dense cytoplasm and large starch grains respectively. The small protoplasts probably originated from embryogenic cell clusters, and also from pseudocambial cells of globules, while larger protoplasts were probably released from oval starchy cells and those of the globule peripheral area. In co-culture with a suitable feeder, consisting of suspensions of diploid banana cells, the protoplasts of triploid banana reformed the cell wall within 24 h and underwent sustained divisions leading to the formation of small clusters of 2–3 cells within 7 days. The latter developed directly into embryos without passing through an apparent callus phase. 10% of such embryos gave rise to plantlets when subcultured in 2.2 M 6-benzylaminopurine and 2 M 4 amino-3,5,6-trichloropicolinic acid for 1 week, before transfer to MS medium containing 10 M 6-benzylaminopurine. The rest of the embryos underwent intensive direct secondary embryogenesis which could lead to the formation of plantlets with a frequency of up to 50% upon further transfer to hormone-free medium.Abbreviations BAP 6-benzylaminopurine - MS Murashige and Skoog (1962) medium - 2,4-D dichlorophenoxyacetic acid - UV ultraviolet light - FDA fluorescein diacetate - MES 2-(N-morpholino)ethanesulfonic acid - Picloram 4 amino-3,5,6-trichloropicolinic acid     

Somatic embryogenesis and plant regeneration in Gymnema sylvestre

Somatic embryogenesis and whole plant regeneration were achieved in callus cultures derived from hypocotyl, cotyledon and leaf explants excised from seedlings of Gymnema sylvestre. Embryogenic callus was induced on Murashige and Skoog (MS) medium containing 2,4-D (0.5–5.0 M) +BA (0.5–2.0 M) and 2% (w/v) sucrose in 6–8 weeks of culture. Globular/heart stage embryos developed on induction medium. These embryos produced torpedo and cotyledon stage embryos upon sub-culturing on embryo maturation medium EM8 (medium containing MS salts, B5 vitamins, 0.5 M BA and 2% sucrose). Embryo germination and plantlet formation was achieved by sub-culturing mature embryos on fresh EM8 medium. The plantlets were acclimatized in the greenhouse.     

Hemopoietic sites and development of eosinophil granulocytes in the loach,Misgurnus anguillicaudatus

Summary Unique eosinophils, each of which contained only one eosinophilic granule, have been found in the peripheral blood of the loach (itMisgurnus anguillicaudatus). Several loach organs have been studied by light and electron microscopy to determine the hemopoietic site of this cell type. Eosinophils are produced mainly in the spleen and to a small extent in the kidney, but not in other organs.Presumed myeloblasts are identified as large lymphoid cells containing a number of small-dense granules (diameter, 0.12–0.16 m) in the cytoplasm. These granules have been observed throughout eosinophilopoiesis but they are most abundant in the promyelocyte stage. The largest cells have been identified as myelocytes which contain a number of large granules (diameter, 0.7–1.4 m) with electron-dense crystalline cores. These large granules are present from the myelocyte to metamyelocyte stage. Metamyelocytes differ from myelocytes in having more large granules. Mature eosinophils are morphologically similar to metamyelocytes but are characterized by the presence of only one very large electron-dense granule (diameter, 2.5–2.8 m) with a crystalline core.The nature of these granules has been studied by enzyme digestion using pepsin and trypsin. The results indicate that the crystalline cores are almost pure protein.     

Transforming Growth Factor α (TGFα) Modulates the Effects of Genomic Imprinting and Prolongs Development of Parthenogenetic Mouse Embryos

The effect of transforming growth factor (TGF) on the development of diploid parthenogenetic mouse embryos (CBA × C57BL/6)F₁was studied. The embryos were in vitro treated with the TGF at the morula stage. Upon reaching the blastocyst stage, each embryo was implanted into uterus of a pseudopregnant female. At a dose of 5 ng/ml, the TGF was found to improve development of parthenogenetic embryos before implantation, increase significantly the number of developing blastocysts, and promote embryo implantation into uterus. After treatment with TGF at a dose of 10 ng/ml, 4% of parthenogenetic embryos reached the stage of 30–45 somites and had forelimb and hindlimb buds; the crown rump length of the embryo size from vertex to sacrum was 2.0 to 3.8 mm. A well-developed placenta was observed in 6% of TGF-treated parthenogenetic embryos that reached the somite stages. In the parthenogenetic embryos with the most prominent development (40–45 somites) treated with 10 ng/ml of TGF, the placental diameter was 4.0 to 4.2 mm on day 12 of gestation, which is close to the placental size of the normal (fertilized) 11-day-old mouse embryos. Our results suggest that exogenous TGF can modulate the effects of genomic imprinting significantly improving formation of trophoblast derivatives and promoting longer postimplantation development of parthenogenetic embryos.