Spindle imaging in human oocytes: the impact of the meiotic cell cycle.

Recent studies using polarized light microscopy revealed a correlation between the presence of a spindle in human metaphase II meiotic oocytes and the fertilization rate following intracytoplasmic sperm injection (ICSI). Using a new spindle imaging system, it was possible to visualize the spindle image and the conventional light microscopic view of the oocyte simultaneously. Using this system, time-lapse studies of the meiotic cycle of human oocytes were performed. The video sequences showed that during the transition from metaphase I to metaphase II, the spindle completely disappears for approximately 40-60 min. These data support the idea that at least in some oocytes, the absence of the spindle is more likely an indicator for physiological progression through an important developmental stage of meiosis rather than a cellular disturbance. In view of the low fertilization rates of oocytes with absence of spindles as reported in the literature, the underlying problem could simply be the incorrect timing of ICSI.

Calystegines as chemotaxonomic markers in the Convolvulaceae.

An extended GC-MS study of 129 convolvulaceous species belonging to 29 genera (all 12 tribes) including the results of a previous survey (65 spp.) revealed the occurrence of one to six polyhydroxy alkaloids of the nortropane type (calystegines) in 62 species belonging to 22 genera of all tribes except the unique parasitic Cuscuteae. The large genus Ipomoea turned out to comprise calystegine-positive species in at least eight out of ten sections checked. The number of the calystegines used as reference compounds has been increased from seven (previous survey) to 11 (present study). Furthermore, the results concerning these additional four alkaloids could also be completed for all species of the previous survey. The plant material (epigeal vegetative parts and/or roots, flowers, fruits/seeds) was obtained from collections in the wild from a wide range of tropical, subtropical, and temperate locations of all continents as well as from cultivation in the greenhouse. All plant organs turned out to be potential locations for the occurrence of these metabolites though they are detectable often only in certain organs of a given species. Three genera (Cuscuta, Operculina, Polymeria) might have lost the ability to synthesize these plesiomorphic characters in the course of the evolution since the examination of several different organs and/or provenances of five species each failed to show calystegines as constituents. Nevertheless, the present data clearly demonstrate that the occurrence of calystegines is an almost consistent trait in the Convolvulaceae in principle, from basal to most advanced tribes.

Ipangulines and minalobines, chemotaxonomic markers of the infrageneric Ipomoea taxon subgenus Quamoclit, section Mina.

A comprehensive GC-MS analysis of 8 Ipomoea species belonging to the subgenus Quamoclit, section Mina revealed that the members of this taxon form combinations of two necine bases with rare necic acids resulting in unique pyrrolizidine alkaloids. The occurrence and diversity of these metabolites show remarkable variations: Some species, especially Ipomoea hederifolia and Ipomoea lobata, are able to synthesize a large number of alkaloids whereas others, especially Ipomoea coccinea and Ipomoea quamoclit, are poor synthesizers with only a few compounds. However, these metabolites are apparently chemotaxonomic markers of this infrageneric taxon in general. They represent either esters of (-)-platynecine (altogether 48 ipangulines and 4 further esters including results of a previous study) or esters of (-)-trachelanthamidine, an additional novel structural type called minalobines (altogether 21 alkaloids). Both types are characterized by section-specific rare necic acids, e.g., ipangulinic/isoipangulinic acid, phenylacetic acid. The alkaloids of Ipomoea cholulensis, I. coccinea, I. hederifolia, Ipomoea neei, and Ipomoea quamoclit were mono and diesters of platynecine. Minalobines turned out to be metabolites of I. lobata (Cerv.) Thell. (syn.: Mina lobata Cerv.) lacking ipangulines. The major alkaloid of this species, minalobine R, has been isolated and identified as 9-O-(threo-2-hydroxy-2-methyl-3-phenylacetoxy-butyryl)-(-)-trachelanthamidine on the basis of spectral data. Apparently only two of the species included in this study, Ipomoea cristulata and Ipomoea sloteri, are able to synthesize both, ipangulines as well as minalobines. Minalobine O could be isolated as a major alkaloid of I. cristulata, its structure has been established as 9-O-(erythro-2-hydroxy-2-methyl-3-tigloyloxy-butyryl)-(-)-trachelanthamidine on the basis of spectral data.