Is the egg size determining gene,Esd, on the W chromosome identical with the sex-linked giant egg gene,Ge, in the silkworm?

The presence of the egg size-determining (Esd) gene, which acts as a quantitative gene, on the W chromosome of the silkworm was revealed in a previous study by using two types of triploid females, ZZW and ZWW, (Kawamura, Genetica 76: 195–201). The females with the sex-linked giant egg (Ge) gene deposit eggs as large as those laid by tetraploids. If the Ge mutant is induced by translocation of a fragment of the W carrying Esd onto the Z by chance, the egg size increase in the Ge strain and in tetraploids may be easily explained by the double dose of Esd. The measurement of the length of the Z-W bivalent in oocytes showed that the Z of the Ge strain was much longer than that of the other strains which do not carry the Ge gene. The result suggests that the Ge gene is identical with the Esd on the W chromosome of the silkworm.     

Phorbol ester facilitates apoptosis in murine fibroblasts pretreated by mild ultraviolet radiation.

Although phorbol 12-myristate 13-acetate (PMA) inhibits apoptosis and promotes the growth of some types of cells, it induces apoptosis in other cells. We evaluated the apoptotic effects of PMA on murine fibroblasts (L-929) that had been exposed to ultraviolet-B (UV-B) radiation at 312 nm, which promotes tumor cell growth. Exposure to PMA alone did not induce Fas, Fas-L, or apoptosis. Cells exposed to mild UV-B irradiation (80 J/m(2)) alone exhibited a slight expression of Fas and Fas-L 36 to 48 h after the exposure, and exhibited apoptosis as evidenced by DNA fragmentation 72 h after exposure. The addition of PMA (0.8 x 10(-5) to 3.2 x 10(-5) M) to the medium 24 h after the UV-B exposure markedly and dose-dependently enhanced these cell responses. Confluent untreated cells, cells cocultured with PMA, and cells cocultured with PMA for 24 h after the UV-B exposure consistently expressed mRNAs for wild-type p53, bcl-2, and ICE. Expression of c-myc mRNA was initially observed, but became undetectable in the cells cocultured for 24 h with a high concentration of PMA (3.2 x 10(-5) M) following UV-B exposure. Such cells subsequently exhibited the maximal apoptotic response. We conclude that mild exposure to UV-B altered murine fibroblast cells in such a way as to facilitate their death by apoptosis upon addition of PMA.     

Visual inputs to the dorsocaudal fastigial nucleus of the cat cerebellum. An experimental study using single unit recordings and horseradish peroxidase labelling

Visual afferents to the cat fastigial nucleus (FN) have been studied with single unit recordings and horseradish peroxidase techniques. A total of 158 cells responding to electrical stimulation of the optic chiasm (OCh) were extracellularly recorded from the dorsocaudal part of the FN. They were classified into three groups: type-1 cells (48%) which showed early suppression and late facilitation; type-2 cells (38%) which showed early facilitation and late suppression; type-3 cells (14%) which exhibited long lasting suppression with no signs of facilitation. Eight of 32 cells tested showed the same response patterns to photic stimulation as to electrical stimulation of the OCh. None of the cells responding to electrical and photic stimulation, however, were found in the rostral and ventral parts of the FN. Furthermore, 29 cells which responded to electrical stimulation of the OCh were tested for responses to moving pattern stimulation. Seven (4 type-2 cells and 3 type-3 cells) of the 29 cells showed clear modulation, reflecting the velocity component of a horizontally moving pattern. However, none of 13 type-1 cells tested exhibited apparent modulation in relation to movement of the pattern. In order to trace the possible pathways mediating visual signals to this part of the FN, the horseradish peroxidase (HRP) method was used. Injection of HRP into the caudal FN resulted in the labelling of many cells, predominantly in the medial (M) and the descending (D) vestibular nuclei and in lobules VI and VII of the cerebellar vermis. A series of experiments further indicated the presence of possible pathways propagating visual signals to the caudal FN. The main routes are: 1) via the nucleus of the optic tract (NOT)--the dorsal part of the medullary reticular formation--the M and the D vestibular nuclei--to the FN, and 2) via the superior colliculus--the pontine nuclei--vermal lobules VI and VII--to the FN. The different physiological response patterns of FN cells may indicate that several types of visual signals involved with visually guided movements impinge upon the dorsocaudal FN.     

Initial phase of infection with Tyzzer's organism in cultured mouse hepatocytes

The initial phase of infection with Tyzzer's organisms in cultured mouse hepatocytes was observed using indirect immunofluorescence (IF) and a plaque assay. The organisms adhered poorly to aldehyde- or acetone-fixed cells, but once adhered to host cells, whether methanol-fixed or unfixed, they were not removed by methanol or acetone. By the IF as well as the plaque assay, both of which discriminated intra- and extracellularly located organisms, the number of cell-associated organisms increased linearly up to 3 h post-inoculation. The number of intra-cellular organisms increased rapidly in the first 1 h, followed by linear increase at a much lower rate. Similar results were obtained by the plaque assay.     

Introduction of amino acid residues at the N-terminus of the zeocin-resistance protein increases its expression in Saccharomyces cerevisiae

Nucleotide sequences proximal to the initiation codon of a gene are known to affect the expression efficiency of that gene. We screened 10-bp random sequences upstream of the initiation codon of the zeocin-resistance gene to identify sequences that could enhance its expression in Saccharomyces cerevisiae. Of the isolated sequences, 20 sequences exhibited a common feature, i.e. ATG at the position −9 through −7, which resulted in the incorporation of three amino acids at the N-terminus of the protein. The introduction of these sequences upstream of the initiation codon increased the expression levels of zeocin-resistance protein by 2.2–6.5-fold. One of these sequences increased the expression levels of three out of four human proteins, thereby suggesting that this sequence may also enhance the expression efficiency of mammalian proteins in yeast.     

The early embryonic mitosis in normal and cooled eggs of the silkworm,Bombyx mori

Early embryonic mitosis of the silkworm, Bombyx mori, was morphologically studied in the normal eggs and in the eggs treated by low temperature (?10°C). The first embryonic mitosis is observed in the eggs at 120 to 150 minutes after deposition at 26°C. After egg and sperm pronuclei unite, a spindle is formed in each of the pronuclei independently. At metaphase and anaphase paternal and maternal chromosomes are in separate groups on a spindle (gonomeric) and karyogamy takes place at telophase when they reach the poles. The second embryonic mitosis is shown in the eggs at 180 to 210 minutes after deposition. The division of two nuclei is not synchronous in the silkworm, and the mitosis is not gonomeric. In the eggs treated by low temperature, spindle fibers are not observed at all at ?10°C, and chromosomes, which form two deeply stained masses of irregular shape, are seen in the less stained area of spindle shape. When the eggs are returned to 26°C, some eggs go into normal gonomeric division, while some form two small and compact spindles, which seem to be derived from each of the pronuclei. It was observed that these compact spindles are able to continue mitosis.