Effects of synchronization of donor cell cycle on embryonic development and DNA synthesis in porcine nuclear transfer embryos

The relationship between donor cell cycle and the developmental ability of somatic cell nuclear transfer (SCNT) embryos has not fully been elucidated. Donor cells that are usually prepared by serum starvation or confluent-cell culture for SCNT represent a heterogeneous population that includes mainly G0 phase cells, other cells in different phases of the cell cycle and apoptotic cells. In this study, we compared the developmental ability of porcine SCNT embryos reconstructed from G0 phase cells (G0-SCNT embryos) and strictly synchronized-G1 phase cells (G1-SCNT embryos), and examined the developmental rates and timing of first DNA synthesis. The G0 phase cells were synchronized by confluent culture, and the G1 phase cells were prepared from actively dividing M phase cells. The G1-SCNT embryos showed a significantly higher (P<0.05) developmental rate to the blastocyst stage per cleaved embryo (59%) than the G0-SCNT embryos (43%). Moreover, initiation of first DNA synthesis and cleavage occurred significantly earlier in the G1-SCNT embryos than in the G0-SCNT embryos. Delay of initiation of first DNA synthesis in the SCNT embryos by aphidicolin resulted in decreased developmental rates to the blastocyst stage without any effect on cleavage rates. Our data demonstrates that synchronized-G1 phase cells can be used as donor cells for SCNT embryos and that earlier initiation of first DNA synthesis may be important for subsequent development of SCNT embryos. The SCNT system using G1-synchronized cells, in terms of their highly uniform and viable cell states, can be useful for studying the reprogramming processes and embryonic development of SCNT embryos.     

New potyvirus isolated from <Emphasis Type="Italic">Cryptotaenia japonica</Emphasis>

 A potyvirus, for which the name Japanese hornwort mosaic virus (JHMV) is proposed, was isolated from Japanese hornwort plants (Cryptotaenia japonica) with mosaic disease symptoms. The virus was used to inoculate mechanically 34 plants belonging to 33 species of 10 families. Of these species seven from two families were infected. Faint chlorotic spots appeared on the inoculated leaves of Chenopodium quinoa and C. amaranticolor, but no systemic infection occurred in these plants. JHMV systemically infected only Umbelliferae plants; they did not infect 26 other species in eight families. JHMV was transmitted in a nonpersistent manner by aphids (Myzus persicae). The virus was a flexuous rod-shaped particle about 750 nm in length. Sequencing the nucleotides in the 3′ terminal region of JHMV revealed that the coat protein contains 280 amino acids with a molecular mass of 32.2 kDa. The nucleotide sequence of the coat protein of JHMV had the highest similarity with that of Zantedeschia mosaic virus (83.3%) compared to those of other potyviruses (57.0%–64.9%). An antiserum against JHMV reacted strongly with JHMV and weakly with Potato virus Y. These results indicate that JHMV is a new potyvirus. Received: September 9, 2002 / Accepted: November 7, 2002 RID="*" ID="*" The nucleotide sequence determined in this work appears in the DDBJ/EMBL/GenBank nucleotide sequence databases with the accession number AB081518     

Age- and sex-related changes in susceptibility of Wistar rats to Strongyloides venezuelensis infection

The effects of host age and sex on susceptibility to Strongyloides venezuelensis in Wistar rats were examined by counting larvae recovered from the lungs of animals 3 days after infection. The susceptibility of female rats to S. venezuelensis rapidly decreased with age and elevated estrogen. Resistance in female rats inoculated at 6 and 10 weeks of age was nine and twenty-fold higher, respectively than that in the youngest group (3 weeks). In contrast, the susceptibility of male animals was lowest in the youngest group, then increased with age and elevated testosterone. Sex differences in susceptibility were not evident in the youngest group, but became apparent with age.     

Androgen- and estrogen-dependent sex differences in host resistance to Strongyloides venezuelensis infection in Wistar rats

The effects of male and female sex hormones on the protective capacity of Wistar rats against infection with Strongyloides venezuelensis were investigated. Male rats were more susceptible than females in terms of worm recovery from the lungs. Orchidectomy of male animals significantly reduced the plasma testosterone concentration and increased host resistance to the migratory stages of S. venezuelensis larvae. In contrast, ovariectomy of female animals significantly decreased host resistance in association with a significant reduction of estrogen levels. To examine the direct effect of sex hormones, exogenous testosterone and estrogen were implanted into animals. Susceptibility significantly increased or decreased in ovariectomized females given testosterone or estrogen, respectively. These results suggest that male and female sex hormones are important in the down- and up-regulation of host resistance against S. venezuelensis in Wistar rats.     

Improvement of rice (Oryza sativa L.) growth in upland conditions with deep tillage and mulch

The productivity of upland rice in Japan as well as in the world is low and unstable owing to scarce and unpredictable rainfall. The objective of this study was to examine whether agronomic methods could enhance grain yield of upland rice. Four field experiments were conducted from 2001 to 2003 in Nishitokyo, Japan, under upland conditions with different water supplies, in order to quantify the effects of deep tillage combined with deep placement of manure (50-cm depth), straw mulch (6 t ha⁻¹), or their combinations on the growth and grain yield of rice. Mulch kept surface soil moisture higher than without mulch even at reproductive stage, and it increased yield to the greatest extent under the most favourable conditions with much rainfall before heading (i.e., 2003). Deep tillage with deep placement of manure induced deep root proliferation and higher nitrogen uptake, increasing biomass production, and panicle number, and consequently grain yield was enhanced under the two lowest yielding environments with less rainfall before heading. Rice plants with deep tillage with deep manure application without mulch tended to have lower leaf water potential and higher diffusion resistance during drought, and negative effects on grain filling and harvest index in some experiments compared with the control. When deep tillage with deep placement of manure was combined with mulching in two experiments in 2002 and 2003, grain yield always enhanced compared with the control (P < 0.10, 6.0 t ha⁻¹ versus 5.4 t ha⁻¹ on average), suggesting their synergetic mechanisms for yield increase and stabilization. The results showed that deep tillage or mulching can improve grain yield of rice under drought-prone rainfed upland conditions in a temperate climate on an Andosol, and their combination had more consistent and greater positive effects.     

The muscle protein Dok-7 is essential for neuromuscular synaptogenesis

The formation of the neuromuscular synapse requires muscle-specific receptor kinase (MuSK) to orchestrate postsynaptic differentiation, including the clustering of receptors for the neurotransmitter acetylcholine. Upon innervation, neural agrin activates MuSK to establish the postsynaptic apparatus, although agrin-independent formation of neuromuscular synapses can also occur experimentally in the absence of neurotransmission. Dok-7, a MuSK-interacting cytoplasmic protein, is essential for MuSK activation in cultured myotubes; in particular, the Dok-7 phosphotyrosine-binding domain and its target in MuSK are indispensable. Mice lacking Dok-7 formed neither acetylcholine receptor clusters nor neuromuscular synapses. Thus, Dok-7 is essential for neuromuscular synaptogenesis through its interaction with MuSK.     

A simple,low‐cost technique for in situ measurement of leaf P concentration in field‐grown rice

Simple tools for diagnosing crop nutritional status are in great demand by agricultural extension staff, particularly in low‐fertility environments. We developed the first practical method for in situ diagnosis of phosphorus (P) nutrition in field‐grown cereal crops by using a handheld colorimeter. The concentration of P in extracts from fresh leaves of lowland rice grown under various P availabilities was closely correlated with that of oven‐dried leaves measured with the standard molybdenum‐blue method in the laboratory, over a range of values from 0.319 to 1.420 mg P g ^?1 DW (r = 0.885^**, n = 14). It takes only 3–5 min to estimate the P concentration of plant samples using the new technique. Although not as accurate as the laboratory method, the new method can easily detect P deficiency of rice in the field (and possibly other cereal crops) without requiring costly, off‐site equipment. Thus, extension staff and agronomists can easily adopt this technique.     

α-Melanocyte-stimulating hormone directly increases the plasma calcitonin level and involves calcium metabolism in goldfish

International Aquatic Research - The effects of α-melanocyte-stimulating hormone (α-MSH) on calcium metabolism were examined with goldfish. The scales on the left side of goldfish bodies...