Deletion of the trichodiene synthase gene of Fusarium venenatum: two systems for repeated gene deletions.

The trichodiene synthase (tri5) gene of Fusarium venenatum was cloned from a genomic library. Vectors were created in which the tri5 coding sequence was replaced with the Neurospora crassa nitrate reductase (nit3) gene and with the Aspergillus nidulans acetamidase (amdS) gene flanked by direct repeats. The first vector was utilized to transform a nitrate reductase (niaD) mutant of F. venenatum to prototrophy, and the second vector was utilized to confer acetamide utilization to the wild-type strain. Several of the transformants lost the capacity to produce the trichothecene diacetoxyscirpenol and were shown by hybridization analysis to have gene replacements at the tri5 locus. The nit3 gene was removed by retransformation with a tri5 deletion fragment and selection on chlorate. The amdS gene was shown to excise spontaneously via the flanking direct repeats when spores were plated onto fluoroacetamide.     

Development of the choroid plexus anlage and supraependymal structures in the fourth ventricular roof plate of human embryos: scanning electron microscopic observations

The developing anlage of the choroid plexus and supraependymal structures in the fourth ventricular roof plates of nine normal human embryos ranging from Carnegie stages 14 to 19 were investigated with scanning electron microscopy. In the human embryos at stage 18, the first semimacroscopic choroidal anlage developed in the form of bilateral evaginations that ran dorsomedially and caudally from the bilateral corners of the rhombencephalon. The anlage became evident with even smaller and parallel ridges in the embryo at stage 19. Embryos at earlier stages exhibited surface membrane modifications such as convexity, microvilli, cilia, and spherical protrusions at the middle one-third of the rhombencephalon, which corresponded to the future choroidal anlage region. Two morphologically different groups of supraependymal cells (SE cells) were elucidated throughout the stages examined. Type 1 SE cells has spindle or tear-drop-like bodies, frequently with one or more long cytoplasmic processes. Type 2 SE cells were globular, with numerous fine pseudopodial processes. Type 1 SE cells were distributed mainly at the future choroidal anlage regions or on the anlage itself and were less frequently located at the rostral end of the roof. We found no general pattern in the distribution of type 2 SE cells. Supraependymal fibers (SE fibers) were seen as fine processes that were distributed similarly to type 1 SE cells and extended transversely for a long distance.     

Thymus-independent anti-DNP antibody responses to DNP-casein and DNP-gelatin.

Although studies on the molecular nature of thymus-independent antigens suggested that the polymeric structure with repeated antigenic determinants and slow metabolism are responsible for thymus-independence, we found that anti-DNP antibody responses to DNP-casein and DNP-gelatin were thymus-independent as well as macrophage-independent. These antibody responses were not affected by in vivo treatment with carrageenan or anti-thymocyte serum. In addition, responses of athymic nude mice to both antigens did not show any significant differences when compared with heterologous nu/+ mice. The findings were confirmed by in vitro experiments; non-adherent spleen cells or T cell-depleted spleen cells responded well to both antigens to the same extent as normal spleen cells. Since both casein and gelatin are polyclonal B cell activators and are not presumed to be high polymer or slow-metabolizing substances, we suggest that thymus-independence in many kinds of antibody response should be reconsidered.     

Genetic study on the mandibular molar size of rats

Tooth size is determined by genetic and environmental factors like other quantitative characters such as body weight and body height. However, the degree of the relative contribution of both factors to the determination of tooth size has not been well clarified. In order to study the genetic and environmental factors affecting tooth size, we carried out a diallel cross mating by the cohabitation of pairs of males and females among 10 strains of rats. The bucco-lingual widths of the first, second, and third molars of the right mandible were measured in each offspring of F1 population. The body weight was also measured as a parameter that might indicate systemic growth factor in connection with tooth development. The quantitative genetic analysis was performed based on Wearden's model (Heredity 19:669-680, 1964). As a result, the size of the first and the second molars was more significantly controlled by genetic effect than maternal effect, while maternal effect could not be ignored for the size of the third molar in addition to genetic effect. The genetic effect on body weight became greater with age, while the maternal effect showed its maximum influence upon the body weight around the weaning. It is concluded that the size of the molar teeth beginning to develop in the uterus and to be calcified just after birth was mainly controlled by genetic factor, and that the size of the molar teeth beginning to develop approximately after birth was mainly controlled by maternal effect affecting body weight at the same period.