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D H Fitch

Publications and source records attributed to D H Fitch.

17 recordsLinked to original sources

Morphogenesis of the Caenorhabditis elegans male tail tip.

Using electron microscopy and immunofluorescent labeling of adherens junctions, we have reconstructed the changes in cell architecture and intercellular associations that occur during morphogenesis of the nematode male tail tip. During late postembryonic development, the Caenorhabditis elegans male tail is reshaped to form a copulatory structure. The most posterior hypodermal cells in the tail define a specialized, sexually dimorphic compartment in which cells fuse and retract in the male, changing their shape from a tapered cone to a blunt dome. Developmental profiles using electron microscopy and immunofluorescent staining suggest that cell fusions are initiated at or adjacent to adherens junctions. Anterior portions of the tail tip cells show the first evidence of retractions and fusions, consistent with our hypothesis that an anterior event triggers these morphogenetic events. Available mutations that interfere with morphogenesis implicate particular regulatory pathways and suggest loci at which evolutionary changes could have produced morphological diversity.

Animals↗

Evolution of male tail development in rhabditid nematodes related to Caenorhabditis elegans.

The evolutionary pathway that has led to male tails of diverse morphology among species of the nematode family Rhabditidae was reconstructed. This family includes the well-studied model species Caenorhabditis elegans. By relating the steps of male tail morphological evolution to the phenotypic changes brought about by developmental mutations induced experimentally in C. elegans, the goal is to identify genes responsible for morphological evolution. The varying morphological characters of the male tails of several rhabiditid species have been described previously (Fitch and Emmons, 1995, Dev. Biol. 170:564-582). The developmental events preceding differentiation of the adult structures have also been analyzed; in many cases the origins of varying adult morphological characters were traced to differences during ontogeny. In the present work, the evolutionary changes producing these differences were reconstructed in the context of the four possible phylogenies supported independently by sequences of 18S ribosomal RNA genes (rDNA). Two or more alternative states were defined for 36 developmental and adult morphological characters. These characters alone do not provide sufficient data to resolve most species relationships; however, when combined with the rDNA characters, they provide stronger support for one of the four rDNA phylogenies. Assuming a model of ordered transformations for multistate developmental characters generally results in greater resolution. Transformations between character states can be assigned unequivocally by parsimony to unambiguous branches for most of the characters. Correlations are thereby revealed for some of the developmental characters, indicating a probability of a shared developmental or genetic regulatory pathway. Four of the unequivocal character state changes on unambiguously supported branches closely resemble the phenotypic changes brought about by known mutations in C. elegans. These mutations define genes that are known to act in genetic regulatory hierarchies controlling pattern formation, differentiation, and morphogenesis. Although these studies are still at an early stage, these results strongly suggest that parallel studies of developmental mutants in C. elegans and of morphological and developmental evolution among related nematodes will help define genetic changes underlying the evolution of form.

Animals↗

Variable cell positions and cell contacts underlie morphological evolution of the rays in the male tails of nematodes related to Caenorhabditis elegans.

As a first step toward understanding their mechanism of morphological evolution, we compare the morphology and development of the male genitalia in 10 species of Rhabditidae, the family of nematodes that includes Caenorhabditis elegans. We describe a number of variable morphological characteristics and focus in particular on the differing arrangements of the caudal papillae or rays within the acellular fan. We analyze the development of the ray cells within the epidermis of the last larval stage and identify changes in cell positions and cell contacts that underlie evolutionary changes in the arrangement of the rays. Epidermal cell positions were determined by means of indirect immunofluorescence staining with a monoclonal antibody directed towards adherens junctions. Similarities between the species in the cellular arrangements during the earliest developmental stages allow us to propose homologies between the rays in different species. Evolutionary changes in the positions and order of homologous rays are correlated with shifts in cell positions during development. The results suggest that genes for cell recognition or adhesion proteins, or pattern formation genes that regulate cell recognition or adhesion proteins, may be important foci of evolutionary change affecting morphology.

Animals↗

18S ribosomal RNA gene phylogeny for some Rhabditidae related to Caenorhabditis.

We have investigated the molecular evolution of the nucleotide sequences of 18S ribosomal RNA genes (18S rDNA) from a set of nematodes in the family Rhabditidae (Nematoda: Secernentea). Our aim was to evaluate the usefulness of this gene for molecular systematics of this family, as well as to establish phylogenetic relationships within a group that has potential for comparative studies of the relationship between development and evolution. We determined the 18S rDNA sequences of nine species of nematodes representing six genera within this family (Caenorhabditis briggsae, C. vulgaris, C. remanei, Rhabditis blumi, Rhabditis sp. br, Rhabditella axei, Pellioditis typica. Teratorhabditis palmarum, and Pelodera strongyloides dermatitica). Using hypothetical models for secondary structure as well as nucleotide similarity, these sequences were aligned with the 18S rDNA sequence published by Ellis et al. for C. elegans and with the partial sequences published by Nadler for eight ascaridoid species. We find that 18S rDNA is likely to be a useful tool to resolve relationships at the intrafamilial level. However, 18S rDNA sequences cannot be used to resolve relationships between taxa as closely related as the Caenorhabditis species. Parsimony, minimum-evolution, and maximum-likelihood methods strongly reject Andrássy's proposed phylogenetic classification based on adult morphological characters but support that of Sudhaus as one alternative of a few possible phylogenies. Distances between genera in this family are about eight times as great as distances between tetrapod classes, suggesting rapid rates of substitution, ancient divergence, or both.

Animals↗

Widespread occurrence of the Tc1 transposon family: Tc1-like transposons from teleost fish.

We characterized five transposable elements from fish: one from zebrafish (Brachydanio rerio), one from rainbow trout (Salmo gairdneri), and three from Atlantic salmon (Salmo salar). All are closely similar in structure to the Tc1 transposon of the nematode Caenorhabditis elegans. A comparison of 17 Tc1-like transposons from species representing three phyla (nematodes, arthropods, and chordates) showed that these elements make up a highly conserved transposon family. Most are close to 1.7 kb in length, have inverted terminal repeats, have conserved terminal nucleotides, and each contains a single gene encoding similar polypeptides. The phylogenetic relationships of the transposons were reconstructed from the amino acid sequences of the conceptual proteins and from DNA sequences. The elements are highly diverged and have evidently inhibited the genomes of these diverse species for a long time. To account for the data, it is not necessary to invoke recent horizontal transmission.

Amino Acid Sequence↗

Low codon bias and high rates of synonymous substitution in Drosophila hydei and D. melanogaster histone genes.

We have evaluated codon usage bias in Drosophila histone genes and have obtained the nucleotide sequence of a 5,161-bp D. hydei histone gene repeat unit. This repeat contains genes for all five histone proteins (H1, H2a, H2b, H3, and H4) and differs from the previously reported one by a second EcoRI site. These D. hydei repeats have been aligned to each other and to the 5.0-kb (i.e., long) and 4.8-kb (i.e., short) histone repeat types from D. melanogaster. In each species, base composition at synonymous sites is similar to the average genomic composition and approaches that in the small intergenic spacers of the histone gene repeats. Accumulation of synonymous changes at synonymous sites after the species diverged is quite high. Both of these features are consistent with the relatively low codon usage bias observed in these genes when compared with other Drosophila genes. Thus, the generalization that abundantly expressed genes in Drosophila have high codon bias and low rates of silent substitution does not hold for the histone genes.

Animals↗

Fetal recruitment of anthropoid gamma-globin genes. Findings from phylogenetic analyses involving the 5'-flanking sequences of the psi gamma 1 globin gene of spider monkey Ateles geoffroyi.

We determined the nucleotide sequence of a 2.5 kb DNA fragment (1 kb is 10(3) base-pairs) that includes exon 1, intron 1 and about 1.4 kb of 5'-flanking DNA of the spider monkey gamma 1-globin pseudogene locus and compared this sequence to its homologous from other primates and rabbit. This region of the gamma 1 locus of spider monkey still retains conserved regulatory elements, suggesting that it became a pseudogene late in New World monkey phylogeny. In the 250 base-pair region immediately 5' from the transcription start site where many known regulatory elements are located, a higher rate of nucleotide substitutions occurred in the ancestral anthropoid (human, ape and monkey) lineage than in the prosimian (galago) lineage, as was also the case for non-synonymous substitutions in the coding region. The opposite pattern was observed for most other non-coding regions and for synonymous substitutions. These substitution patterns correlate with the embryonic-to-fetal transformation of the gamma-globin genes of the ancestral anthropoids. Analysis of the 5'-flanking sequences suggests that 11 gene conversion events have occurred in the anthropoid gamma-gene lineages. In the parts of the 5'-flanking region where no gene conversions have been detected, gamma 2-gene sequences have accumulated more nucleotide changes than gamma 1, which suggests that the gamma 2 gene was the more redundant duplicate that may have accumulated first the nucleotide changes responsible for the anthropoid fetal pattern of gamma-globin gene expression.

Animals↗

Duplication of the gamma-globin gene mediated by L1 long interspersed repetitive elements in an early ancestor of simian primates.

Regions surrounding the single gamma-globin gene of galago and the duplicated gamma 1- and gamma 2-globin genes of gibbon, rhesus monkey, and spider monkey were sequenced and aligned with those from humans. Contrary to previous studies, spider monkey was found to have not one but two gamma-globin genes, only one of which (gamma 2) is functional. The reconstructed evolutionary history of the gamma-globin genes and their flanking sequences traces their origin to a tandem duplication of a DNA segment approximately 5.5 kilobases long that occurred before catarrhine primates (humans, apes, and Old World monkeys) diverged from platyrrhines (New World monkeys), much earlier than previously thought. This reconstructed molecular history also reveals that the duplication resulted from an unequal homologous crossover between two related L1 long interspersed repetitive elements, one upstream and one downstream of the single ancestral gamma-globin gene. Perhaps facilitated by the redundancy resulting from the duplication, the gamma-globin genes escaped the selective constraints of embryonically functioning genes and evolved into fetally functioning genes. This view is supported by the finding that a burst of nonsynonymous substitutions occurred in the gamma-globin genes while they became restructured for fetal expression in the common ancestor of platyrrhines and catarrhines.

Animals↗

Phylogenetic scanning: a computer-assisted algorithm for mapping gene conversions and other recombinational events.

An algorithm, 'phylogenetic scanning', is described for mapping gene conversion events where comparative DNA sequence data are available from different species. In this algorithm, sets of hypothetical phylogenetic trees are constructed that describe possible sequence relationships due to gene conversions in different species lineages; these trees are then evaluated by the principle of parsimony at intervals in the sequence alignment. When used to map gene conversion events that occurred between the pair of gamma-globin genes of higher primates, the algorithm gives results nearly identical to those obtained using a tedious manual approach. Suggestions are also provided for adaptation of this procedure to the analysis of other recombination events.

Algorithms↗

Molecular evolution of the psi eta-globin gene locus: gibbon phylogeny and the hominoid slowdown.

An 8.4-kb genomic region spanning both the psi eta-globin gene locus and flanking DNA was sequenced from the common gibbon (Hylobates lar). In addition, sequencing of the entire orthologous region from galago (Galago crassicaudatus) was completed. The gibbon and galago sequences, along with published orthologous sequences from 10 other species, were aligned. These noncoding nucleotide sequences represented four human alleles, four apes (chimpanzee, gorilla, organgutan, and gibbon), an Old World monkey (rhesus monkey), two New World monkeys (spider and owl monkeys), tarsier, two strepsirhines (galago and lemur), and goat. Divergence and maximum parsimony analyses of the psi eta genomic region first groups humans and chimpanzees and then, at progressively more ancient branch points, successively joins gorillas, orangutans, gibbons, Old World monkeys, New World monkeys, tarsiers, and strepsirhines (the lemuriform-lorisiform branch of primates). This cladistic pattern supports the taxonomic grouping of all extant hominoids into family Hominidae, the division of Hominidae into subfamilies Hylobatinae (gibbons) and Homininae, the division of Homininae into tribes Pongini (orangutans) and Hominini, and the division of Hominini into subtribes Gorillina (gorillas) and Hominina (chimpanzees and humans). The additional gibbon and galago sequence data provide further support for the occurrence of a graded evolutionary-rate slowdown in the descent of simian primates, with the slowing rate being more pronounced in the great-ape and human lineages than in the gibbon or monkey lineages. A comparison of global versus local molecular clocks reveals that local clock predictions, when focused on a specific number of species within a narrow time frame, provide a more accurate estimate of divergence dates than do those of global clocks.

Animals↗

Pattern formation in the nematode epidermis: determination of the arrangement of peripheral sense organs in the C. elegans male tail.

The developmental process that determines the arrangement of ray sensilla in the Caenorhabditis elegans male tail has been studied. It is shown that the adult arrangement of rays is determined by the placement of ray cells at specific sites in the epidermis of the last larval (L4) stage. Placement of ray cells at specific epidermal sites results from the generation of neurons and support cells in the epidermis near to their final positions, and the subsequent refinement of these positions by an active mechanism involving specific cellular associations. Positions of ray cells and adjacent epidermal cells have been studied during ray development by means of indirect immunofluorescence staining with an antibody to a cell junctional antigen. Mutations are described in six genes that alter the adult arrangement of the rays, frequently resulting in fusion of rays. Changes in the adult pattern of rays in mutants appear to result from prior changes in the epidermal positions of ray cells, and for two mutants it is suggested that this may be due to the inappropriate clustering of processes from neurons and support cells of adjacent rays. Development of the wild-type arrangement of rays appears to require the specification of molecular differences between the rays that affect the specificity of their cellular associations.

Animals↗

Molecular history of gene conversions in the primate fetal gamma-globin genes. Nucleotide sequences from the common gibbon, Hylobates lar.

Comparative and phylogenetic analyses of homologous sequences from closely related species reveal genetic events which have happened in the past and thus provide considerable insight into molecular genetic processes. One such process which has been especially important in the evolution of multigene families is gene conversion. The fetal gamma 1 and gamma 2-globin genes of catarrhine primates (humans, apes, and Old World monkeys) underwent numerous gene conversion events after they arose from a gene duplication event 25-35 million years ago. By including the gamma 1- and gamma 2-globin gene sequences from the common gibbon, Hylobates lar, the present work expands the gamma-globin data set to represent all major groups of hominoid primates. A computer-assisted algorithm is introduced which reveals converted DNA segments and provides results very similar to those obtained by site-by-site evolutionary reconstruction. Both methods provide strong evidence for at least 14 different converted stretches in catarrhine primates as well as five conversions in ancestral lineages. Features of gene conversions generalized from this molecular history are 1) conversions are restricted to regions maintaining high degrees of sequence similarity, 2) one gene may dominate in converting another gene, 3) sequences involved in conversions may accumulate changes more rapidly than expected, and 4) certain elements, such as polypurine/polypyrimidine [Y)n) and (TG)n elements, appear to be hotspots for initiating or terminating conversion events.

Amino Acid Sequence↗

On the origins of tandemly repeated genes: does histone gene copy number in Drosophila reflect chromosomal location?

Widely regarded beliefs about Drosophila histone gene copy numbers and developmental requirements have been generalized from fairly limited data since studies on histone gene arrangements and copy numbers have been largely confined to a single species, D. melanogaster. Histone gene copy numbers and chromosomal locations were examined in three species: D. melangaster, D. hydei and D. hawaiiensis. Quantitative whole genome blot analysis of DNA from diploid tissues revealed a tenfold variability in histone gene copy numbers for these three species. In situ hybridization to polytene chromosomes showed that the histone DNA (hDNA) chromosomal location is different in all three species. These observations lead us to propose a relationship between histone gene reiteration and chromosomal position.

Animals↗

Primate evolution at the DNA level and a classification of hominoids.

The genetic distances among primate lineages estimated from orthologous noncoding nucleotide sequences of beta-type globin loci and their flanking and intergenic DNA agree closely with the distances (delta T50H values) estimated by cross hybridization of total genomic single-copy DNAs. These DNA distances and the maximum parsimony tree constructed for the nucleotide sequence orthologues depict a branching pattern of primate lineages that is essentially congruent with the picture from phylogenetic analyses of morphological characters. The molecular evidence, however, resolves ambiguities in the morphological picture and provides an objective view of the cladistic position of humans among the primates. The molecular data group humans with chimpanzees in subtribe Hominina, with gorillas in tribe Hominini, orangutans in subfamily Homininae, gibbons in family Hominidae, Old World monkeys in infraorder Catarrhini, New World monkeys in semisuborder Anthropoidea, tarsiers in suborder Haplorhini, and strepsirhines (lemuriforms and lorisiforms) in order Primates. A seeming incongruency between organismal and molecular levels of evolution, namely that morphological evolution appears to have speeded up in higher primates, especially in the lineage to humans, while molecular evolution has slowed down, may have the trivial explanation that relatively small genetic changes may sometimes result in marked phenotypic changes.

Animals↗

Molecular phylogeny of the family of apes and humans.

The morphological picture of primate phylogeny has not unambiguously identified the nearest outgroup of Anthropoidea and has not resolved the branching pattern within Hominoidea. The molecular picture provides more resolution and clarifies the systematics of Hominoidea. Protein and DNA evidence divides Hominoidea into Hylobatidae (gibbons) and Hominidae, family Hominidae into Ponginae (orangutan) and Homininae, and subfamily Homininae into two tribes, one for Gorilla, and the other for Pan (chimpanzee) and Homo. Parsimony and maximum likelihood analyses, carried out on orthologous noncoding nucleotide sequences from primate beta-globin gene clusters, provide significant evidence for the human-chimpanzee tribe and overwhelming evidence for the human-chimpanzee-gorilla clade. These analyses also indicate that the rate of molecular evolution became slower in hominoids than in other primates and mammals.

Animals↗

The spider monkey psi eta-globin gene and surrounding sequences: recent or ancient insertions of LINEs and SINEs?

A phylogenetic comparison of the psi eta-globin DNA sequence of the spider monkey (Ateles geoffroyi) to orthologous sequences from other primates provides evidence for an evolutionarily recent (less than 17 million years ago (MYA] insertion of a truncated L1 LINE (long interspersed repetitive element). An Alu SINE (short interspersed element), found 3 kb downstream of the poly(A) addition site, arose less than 40 MYA subsequent to the divergence of platyrrhinies (New World monkeys) from catarrhines (humans, apes, and Old World monkeys). Another Alu element, which appears in the catarrhines 588 bp upstream from the psi eta gene, is absent in Ateles, thus placing its possible time of insertion between 20 and 40 MYA. Alignment of the Ateles psi eta sequence with orthologous hominoid sequences (M. M. Miyamoto, J. L. Slightom, and M. Goodman, 1987, Science 238: 369-373) provides unequivocal evidence against the view (J. H. Schwartz, 1987, "The Red Ape: Orang-utans and Human Origins," Houghton Mifflin, Boston, MA) that the orangutan forms a monophyletic group with human. Furthermore, a critical analysis of potential sources of homoplasy (i.e., parallel evolution) reaffirms a Homo-Pan monophyletic clade (Miyamoto et al., 1987). Reconstruction of primate psi eta sequence phylogeny suggests Ateles and Aotus shared a common ancestor only 16 +/- 1.5 MYA. Finally, the results show a mutation rate of 1.7 +/- 0.2 x 10(-9) mutations site-1 year-1 during the evolution of the Ateles psi eta-globin region since the divergence of catarrhines from platyrrhinies.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

External review in quality assurance.

This paper describes the application of external review to a physical therapy practice setting. The roots of the approach are examined in terms of consultation, accountability, and recognition of professionalization in physical therapy. The use of the structured interview as a data-gathering instrument is presented and the estimating of validity, reliability, and practicality is discussed. Implications for the use of external review as a tool in quality assurance are explored.

Clinical Competence↗