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Biomedical subjects

M Himes

Publications and source records attributed to M Himes.

9 recordsLinked to original sources

Crypthecodinium and Tetrahymena: an exercise in comparative evolution.

Nucleotide sequences have been determined for the highly variable D2 region of the large rRNA molecule for over 60 strains of dinoflagellates. These strains were selected from a worldwide collection that represents all the known sibling species (compatibility groups, Mendelian species) in the sibling swarm referred to as Crypthecodinium cohnii. A phylogenetic tree has been constructed from an analysis of the variations in a length of about 180 bases, using PHYLOGEN string analysis programs. The Crypthecodinium tree is compared with the previously published but here augmented tree constructed upon the same rRNA region for the sibling species of a worldwide collection of ciliated protozoa related to the genus Tetrahymena. The first reported sequence of Lambornella clarki, the parasite of tree-hole mosquitoes, is included. The dinoflagellate species complex is much more homogeneous with respect to ribosomal variation. The mean number of differences among sequences from different Crypthecodinium species is about 7, in comparison with 22 differences among the ciliate species examined. Moreover, all the diversity in the dinoflagellates can be explained by base substitutions, whereas insertions and deletions are common in the ciliates. The dinoflagellates are also much more uniform with respect to nutritional and genetic economies. The two complexes differ also in the relationship between molecular variations and breeding compatibility. All tetrahymenine sibling species thus far examined are monomorphic in the D2 region, but several dinoflagellate species are polymorphic. Several different dinoflagellate species, moreover, have identical D2 regions. This kind of ribosomal identity of incompatible strains is found in these ciliates only in one tight cluster of species--Group C. The tetrahymenine swarm is apparently much older than the Crypthecodinium swarm, and the dinoflagellate species produce incompatible progeny species much more readily than do the ciliates, perhaps by the acquisition of mutations that potentiate incompatibility in sympatric populations.

Animals↗

Genetic analysis in the dinoflagellate (Crypthecodinium (Gyrodinium) cohnii: evidence for unusual meiosis.

The atypical structure and behavior of dinoflagellate chromosomes suggests that the genetics of these organisms might show comparable peculiarities. We have begun genetic analysis of the neritic, marine heterotroph Crypthecodinium (Gyrodinium) cohnii by means of motility mutants that show complementation shortly after zygote formation, permitting identification of heterozygotes. Six mutations, conferring four microscopically distinguishable phenotypes, have been isolated and investigated. These "genes" were found to complement in double heterozygotes in all pairwise combinations, indicating that the lesions are recessive and non-allelic. Clones of all possible combinations of these factors have been established and each complements only those combinations expected on the assumption that there are six independent recessive mutant "genes." Tetrad analysis following isolation of over 200 complementing zygotes showed: (1) regular segregation with recovery of parental phenotypes and genotypes; (2) independent assortment, with one possible exception; (3) segregations that were always 1:1, that is, in all tetrads showing recombination, only the two reciprocal recombinant genotypes were found; there were no tetratypes. This behavior could result from centromere linkage or the absence of crossing over in an otherwise conventional meiosis, or it could result from an unusual one-division "meiosis." Some evidence is provided that favors the latter hypothesis.

Animals↗

DNA-protein binding in interphase chromosomes.

The metachromatic dye, azure B, was analyzed by microspectrophotometry when bound to DNA fibers and DNA in nuclei with condensed and dispersed chromatin. The interaction of DNA and protein was inferred from the amount of metachromasy (increased beta/alpha-peak) of azure B that resulted after specific removal of various protein fractions. Dye bound to DNA-histone fibers and frog liver nuclei fixed by freeze-methanol substitution shows orthochromatic, blue-green staining under specific staining conditions, while metachromasy (blue or purple color) results from staining DNA fibers without histone or tissue nuclei after protein removal. The dispersed chromatin of hepatocytes was compared to the condensed chromatin of erythrocytes to see whether there were differences in DNA-protein binding in "active" and "inactive" nuclei. Extraction of histones with 0.02 N HCl, acidified alcohol, perchloric acid, and trypsin digestion all resulted in increased dye binding. The amount of metachromasy varied, however; removal of "lysine-rich" histone (extractable with 0.02 N HCl) caused a blue color, and a purplish-red color (micro-peak absorption) resulted from prolonged trypsin digestion. In all cases, the condensed and the dispersed chromatin behaved in the same way, indicating the similarity of protein bound to DNA in condensed and dispersed chromatin. The results appear to indicate that "lysine-rich" histone is bound to adjacent anionic sites of a DNA molecule and that nonhistone protein is located between adjacent DNA molecules in both condensed and dispersed chromatin.

Animals↗

An analysis of heterochromatin in maize root tips.

The B chromosomes of maize are condensed in appearance during interphase and are relatively inert genetically; therefore they fulfill the definition of heterochromatin. This heterochromatin was studied in root meristem cells by radioautography following administration of tritiated thymidine and cytidine, and was found to behave in a characteristic way, i.e. it showed asynchronous DNA synthesis and very low, if any, RNA synthesis. A cytochemical comparison of normal maize nuclei with nuclei from isogenic maize stock containing approximately 15-20 B-chromosomes in addition to the normal complement has revealed the following: (a) the DNA and histone contents are greater in nuclei with B chromosomes; (b) the proportion of DNA to histone is identical with that of nuclei containing only normal chromosomes; (c) the amount of nonhistone protein in proportion to DNA in interphase is less in nuclei with B chromosomes than in normal nuclei. In condensed B chromosomes the ratio of nonhistone protein to DNA is similar to that in other condensed chromatin, such as metaphase chromosomes and degenerating nuclei. The B chromosomes appear to have no effect on nucleolar RNA and protein. Replication of B chromosomes is precisely controlled and is comparable to that of the ordinary chromosomes not only in synthesis for mitosis but also in formation of polyploid nuclei of root cap and protoxylem cells.

Autoradiography↗

Ribosomal RNA sequencing of members of the Crypthecodinium cohnii (Dinophyceae) species complex; comparison with soluble enzyme studies.

Sixty-five members of the Crypthecodinium cohnii species complex were analyzed for sequence differences within the D2 region of the 23S ribosomal RNA molecule. On the basis of 46 sequence differences the strains fell into 19 distinct ribosets (strains of identical sequence), some with many members. Members of four of the seven major sibling species (widespread breeding groups) were each found within single ribosets. Members of three other major sibling species were each, however, divided into two ribosets by a single sequence difference correlated with geographic separation and with previously reported electrophoretic polymorphisms of soluble enzymes within the sibling species. In addition to members of major sibling species, some ribosets include many minor sibling species (each represented by only one strain). Of 38 minor sibling species, 22 shared sequence with a major sibling species. Of these 22, 14 were identical in soluble enzymes to their related major sibling species or differed by only one of three enzymes. Other minor sibling species appear to have diverged extensively from any others in both rRNA sequence and electrophoretic profile. As a group, major sibling species differ markedly in the number of minor sibling species associated with them, suggesting differences in frequency of sexually isolating events in their past histories. These findings are discussed in the context of the previously proposed model of sympatric speciation.

Animals↗