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C L Littlefield

Publications and source records attributed to C L Littlefield.

13 recordsLinked to original sources
PubMed1997

Embryogenesis in hydra.

Embryogenesis in hydra includes a variable period of dormancy; and this period, as well as subsequent stages through hatching, takes place within a thick cuticle that hinders observation. Thus, although the early stages of development have been well-characterized qualitatively, the middle and later stages are only poorly understood. Here, we provide a detailed description of the stages of embryogenesis, including the time required to traverse each of the stages, and the changes that occur in the type and number of cells throughout the stages. The events of cleavage and gastrulation occur within the first 48 h. Cleavage is holoblastic and unipolar and leads to a single-layered coeloblastula. Gastrulation occurs by ingression and is followed by the deposition of the thick cuticle. Thereafter, during the variable period of dormancy ranging from 2-24 weeks, little occurs; the important events are the conversion of the outer layer into an ectoderm and the appearance of the interstitial cell lineage. During the last 2 days before hatching, the endoderm and gastric cavity form, while stem cells of the interstitial cell lineage proliferate and differentiate into neurons, nematocytes, and secretory cells. Finally, the cuticle cracks, and the hatchling enlarges and emerges from the cuticle as a functional animal. The formation of the gastric cavity and the hatching of the embryo are both explicable in terms of the osmotic behavior of the animal and the hydrostatic forces generated by this behavior. Characteristics of development that are common to hydra and triploblastic phyla are presented.

Animals↗
PubMed1991

Spermatogenesis in Hydra oligactis. II. How temperature controls the reciprocity of sexual and asexual reproduction.

Hydra oligactis undergo two mutually exclusive modes of reproduction: at warm temperatures (18-22 degrees C) animals reproduce asexually by budding, while at cold temperatures (10-12 degrees C) gamete differentiation occurs. Using a monoclonal antibody which is specific for cells of the sperm lineage, it was discovered that under conditions where sperm differentiation does not occur (18-22 degrees C), cells continually enter the sperm pathway but progression down the pathway is prematurely halted, effectively blocking the production of sperm. To elucidate the mechanism by which completion of sperm differentiation is controlled, the cell cycle times of interstitial cells entering the sperm pathway at both the restrictive (18 degrees C) and permissive (10 degrees C) temperatures were examined. It was envisaged that at the restrictive temperature the cell cycle times of committed cells would lengthen as they proceeded down the pathway, leading to dilution and eventual loss of cells at later stages of sperm differentiation. This did not occur. Although cells of the sperm lineage were found overall to divide more slowly at 18 degrees C than at 10 degrees C, at both temperatures the cell cycle times shortened as cells proceeded further down the pathway, making a dilution mechanism untenable. The effect of high temperature on the survival of cells was then tested by subjecting animals to a heat shock. Within 12 hr of the increase in temperature, the total number of sperm lineage interstitial cells dropped 10-fold while the total numbers of epithelial and somatic interstitial cells remained virtually unchanged. A distinct consequence of this cell loss was the disappearance of cells furthest down the sperm pathway. It is proposed that as cells move down the sperm pathway, they become increasingly sensitive to high temperature which adversely affects their survival; the higher the temperature, the earlier in the pathway cells die. The lethal effect is abolished by lowering the temperature, allowing sperm differentiation to continue to completion. The possible adaptive advantages of temperature controlling gametogenesis are discussed.

Adaptation, Physiological↗
PubMed1986

Germ cells in Hydra oligactis males. II. Evidence for a subpopulation of interstitial stem cells whose differentiation is limited to sperm production.

Animals containing germline-restricted interstitial cells were obtained by treating males from a clone of Hydra oligactis with hydroxyurea (HU) to lower the interstitial population to 1 or 2 cells per animal. A 3-day HU treatment produced animals whose interstitial cells did not form somatic cells, but did produce sperm. The isolation of these cells in HU-treated animals has lead us to propose that the interstitial cell population may contain subpopulations which possess different growth dynamics and developmental potentials. Through asexual propagation, we have cloned several animals containing only sperm precursor interstitial cells and have examined the growth and differentiation behavior of these cells in offspring propagated over a 2-year period. Evidence has been obtained which demonstrates (1) the extensive self-renewal capacity of the sperm precursor interstitial cells, and (2) the restricted differentiation capacity of these interstitial stem cells. Factors which affect cells entering and traversing the spermatogenic pathway are also presented.

Animals↗
PubMed1986

Sex determination in hydra: control by a subpopulation of interstitial cells in Hydra oligactis males.

The stability of sexual phenotype was examined in a single clone of Hydra oligactis males maintained at two culture temperatures, 18 and 22 degrees C. At these temperatures animals of this species do not reproduce sexually, but reproduce asexually by budding, and males and females are morphologically indistinguishable. When the temperature is lowered to 10 degrees C gametogenesis is induced and sexual phenotype can be assayed. Males cultured for several years at 18 degrees C expressed a stable sexual phenotype when induced to undergo gametogenesis; males remained male. Those cultured at 22 degrees C for 1 year, however, expressed a low frequency of sex reversal from male to female; males ceased sperm differentiation and began producing eggs. Male sex reversal in cultures maintained at the higher temperature was correlated with the loss of a specific subpopulation of interstitial cells, those that bind the monoclonal antibody, AC2, which labels cells specific to the spermatogenic pathway in H. oligactis males. When interstitial cells restricted to this pathway were reintroduced into sex-reversed males (phenotypic females), the male phenotype was reestablished and animals reverted to sperm production. To further investigate the role of AC2+ cells in the masculinization of females, normal males (containing AC2+ cells) and sex-reversed males (lacking AC2+ cells) were grafted to females. In grafts between normal males and females, egg production ceased and sperm differentiation ensued, whereas those between sex-reversed males and females continued to produce eggs. Thus, the presence of AC2+ interstitial cells is strictly correlated with male sexual phenotypes and it is only in their absence that the female phenotype is expressed.

Animals↗
PubMed1985

Spermatogenesis in Hydra oligactis. I. Morphological description and characterization using a monoclonal antibody specific for cells of the spermatogenic pathway.

A morphological description of cells participating in sperm formation in Hydra oligactis males using a maceration procedure is presented. These descriptions are corroborated by the use of a monoclonal antibody, AC2, that binds to both a subpopulation of interstitial cells that appears to participate exclusively in gamete formation, and to all the gamete-differentiation products, including sperm intermediate cells, spermatids, and sperm. Use of the antibody as an interstitial cell marker has allowed an analysis of the behavior of the gamete-precursor (AC2+) subpopulation of interstitial cells during the asexual state and the early stages of gamete formation, when no differentiating sperm intermediates are present. The results indicate there is a gamete-producing subpopulation of interstitial cells which is present in low numbers in asexual males and undergoes extensive growth following the onset of spermatogenesis to give rise to sperm intermediate cells, and, eventually, the sperm. No input from the AC2- interstitial cells is required to account for this growth or subsequent sperm production. We speculate that the AC2+ interstitial cells may represent a unique subpopulation which is developmentally restricted to sperm production.

Animals↗
PubMed1985

Germ cells in Hydra oligactis males. I. Isolation of a subpopulation of interstitial cells that is developmentally restricted to sperm production.

Single clones of interstitial cells were generated and analyzed to determine if one interstitial cell has the capacity to differentiate both somatic and germ cells. Such clones were produced by using hydroxyurea to selectively eliminate interstitial cells from normal Hydra oligactis males. The number of animals devoid of interstitial cells within the population was determined by staining whole animals with toluidine blue which renders the interstitial cells visible. The number of animals containing single clones of interstitial cells was then estimated using single hit Poisson statistics. In treatments which rendered 60-80% of the population devoid of interstitial cells, the majority of the animals containing interstitial cells lost the ability to produce somatic cells, including nerves and nematocytes, but retained the capacity to produce sperm. This result strongly suggests the presence of a separate germ line in hydra.

Animals↗
PubMed1984

The interstitial cells control the sexual phenotype of heterosexual chimeras of hydra.

The three stem cell populations in hydra, the epithelial cells of the ectoderm and endoderm, which make up the body of the hydra, and the interstitial cells, which give rise to nerve cells, nematocytes, and gametes, were tested for their effects on determining the sexual phenotype of individuals. This was done by creating epithelial hydra, which are devoid of interstitial cells and their derivatives, of one sexual type and repopulating them with interstitial cells from individuals of the other sexual type. The resulting heterosexual chimeras were found in all cases to display the same sexual phenotype as that of the interstitial cell donor, indicating this cell type is responsible for the sex of the animal. The epithelial tissue had no influence in determining which gamete type was produced.

Animals↗
PubMed1984

Regulative interactions between cells of wing discs from different dipteran species.

The mechanism by which patterns are produced appears to be repeated in each segment of an animal, and it has been proposed that it may even have been conserved in evolution so that different species would have the same system of positional information. This idea has been tested by mixing cells of a defined fragment of the wing disc of Drosophila melanogaster with wing disc fragments of five other dipteran species to assay the ability of these disc fragments to stimulate intercalary regeneration of the D. melanogaster cells. The genetically marked (y; mwh) D. melanogaster fragment was mechanically mixed with wing discs or wing disc fragments of four drosophilids (D. melanogaster as a control, D. virilis, D. hydei, Zaprionus vittiger), of Musca domestica, and of Piophila casei. The mixed aggregates were cultured in vivo for 7 days, then metamorphosed in D. melanogaster larval hosts. The D. melanogaster fragments were only stimulated to regenerate when combined with complementary fragments from D. melanogaster or D. virilis wing discs. In the combination between D. melanogaster and D. hydei, the tissue formed integrated mosaic patterns, but no regeneration ensued. The one positive result (D. melanogaster mixed with D. virilis) shows that positional cues can be exchanged and correctly interpreted between cells of different species. The negative results do not prove that the mechanism for establishing patterns is different in the tested species, but may be due to incompatibilities that are not related to pattern formation.

Animals↗
PubMed1978

Regulative interactions between cells from different imaginal disks of Drosophila melanogaster.

The regulative behavior of cells from the imaginal wing disk of Drosophila melanogaster can be modified by interaction with cells from different disk types. Both thoracic and nonthoracic disks are able to interact, but there are major differences in the effectiveness of interaction. The finding lends experimental support to the idea that cells in different fields within the same organism use the same mechanism for specifying positional information. A similar conclusion has been reached by Wilcox and Smith based on studies of the mutation wingless.

Animals↗