Joseph Hall Bodine

Factors Influencing the Rate of Respiratory Metabolism of a Developing Egg (Orthoptera)

DESPITE DESPITE the fact of their widespread occurrence and extensive use as biological experimental materials, little if any information of a quantitative nature seems to exist concerning the factors involved in the embryological development of insects. Reference to the literature on the question of fundamental changes in single developing insect eggs shows that practically nothing quantitative concerning these interesting phenomena is recorded. Such knowledge as does exist has usually been obtained from experiments carried out on large numbers of eggs and without regard to environmental or other history (Von Fiirth, 1903; Krogh, 1916; Winterstein, 1924; Sayle, 1928; and others). In a previous article some facts concerning the effects of temperature on the embryonic development of grasshoppers were presented (Bodine, 1925). The present article is a continuation of this study in which the rates of respiratory exchange and other related phenomena during the development of the eggs have been followed under normal as well as experimental conditions. It is the specific purpose of these investigations to make as complete a correlation as is possible between the morphology and physiology during the development of these forms so that a more or less rigid standard can be established as a basis for extensive experimental cellular investigations already under way in this laboratory. The material seems of such a unique nature that it will doubtless not be out of place here to point out again some of its outstanding

Hibernation and Diapause in Certain Orthoptera. III. Diapause: A Theory of Its Mechanism

XISTING data on the effects of temperature, etc., on the embryonic development of grasshoppers are of much interest; but when considered in view of more recent knowledge concerning embryonic diapause, they can be shown to be of little value except as starting-points for more fundamental investigations (Bodine, 1925; Uvarov, 1928; Parker, 1930.) To satisfactorily analyze problems on the physiology of development in these forms, it becomes more and more evident that studies must be carried out on single eggs, the morphological, physiological, and genetical histories of which are accurately known. The closest possible correlations between the morphology and physiology of the egg must be made before an appreciation of either of the two can be fully understood. From existing and recent data, however, it is possible to show something of the fundamental working of diapause and other factors during embryonic development. The following discussion, an interpretation based upon existing data and that more recently acquired, is an attempt to give a possible explanation of various phenomena connected with diapause occurring during the embryonic development of certain grasshoppers. Existing theories as to causes of diapause will not be discussed at this time, and the reader is referred to the recent monograph of Uvarov (1931) for complete bibliography of same. As pointed out in the preceding article (Bodine, 1932), two main classes of eggs in Melanoplus differentialis seem to be produced: (i) a non-diapause type, in which no embryonic diapause can be demonstrated, the embryo developing at a uniform rate at constant high temperature (above developmental zero) from laying until hatching; and (2) a diapause type, in which a marked cessation of embryonic

The Action of Certain Stimulating and Inhibiting Substances on the Respiration of Active and Blocked Eggs and Isolated Embryos

IN A recent publication (Bodine and Boell, 1936a) attention was directed to the importance of considering the reactions of the egg as a whole, as well as those of the contained embryo, in studies dealing with the various problems in the field of developmental and cellular physiology. Such precautions seem especially applicable when definite and concrete information concerning the responses of the embryo and cells is desired. The present study deals with results of further investigations into the responses of the entire egg, as well as those of the contained embryo, and, in addition, treats of the reactions to various stimulating and inhibiting reagents of normal, developing, and active embryos as well as those blocked or in a state of diapause. The eggs of the grasshopper, Melanoplus diferentialis, have been used throughout; and the reader is referred to former publications for details as to procedures followed, etc., in work with such forms (Bodine and Boell, 1936a).

A Study of the Mechanism of Cyanide Inhibition: I. Effect of Concentration on the Egg of Melanoplus differentialis

R RESULTS obtained from studies of the depressing action of cyanide on the respiration of animal cells and tissues have teen used by Warburg (1928) in support of a theory that respiration is due primarily to an activation of oxygen by an iron-containing enzyme. A complete discussion of this subject is found in Warburgs original works (1928) and in reviews by Dixon (1929) and Meldrum (1934) to which the reader is referred.

Hibernation and Diapause in Certain Orthoptera. II. Response to Temperature during Hibernation and Diapause

O UESTIONS of hibernation and diapause among animals, in addition to their inherent interest as such, are of much concern to the physiologist, since their analyses may give information or throw light upon the fundamental actions and reactions of the cells of an organism as well as the relations between the organism and its environment. Such fundamental problems as the causes of cessation of development and differentiation, of inhibition of mitosis, and other cellular activities have occupied the minds of biologists for some time. Recent extensive literature, reviews, and discussions by Uvarov (1931) and others contain rather complete surveys on hibernation, diapause, etc., in lower animals, and particularly inI insects, so that no attempt will be made here to duplicate such effort. The reader is referred to these monographs, where rather complete bibliographies and detailed data may be found. In the present paper some phases of the physiology of hibernation and diapause in certain Orthoptera (grasshoppers) will be discussed. In a previous communication (Bodine, 1923) certain aspects of this problem have been presented. The present data are concerned almost exclusively with questions of hibernation and diapause in the nymphs of Chortophaga viridifasciata and in the eggs of Melanoplus differentialis. The life-cycles of these two species are sufficiently different to justify a brief review of them before going farther. Chortophaga viridifasciata, in the vicinity of Iowa City, lays its eggs in the soil in late spring or early summer; and these hatch in late summer and early fall. The nymphs, reaching the second or third instars, hibernate, and in spring grow and become mature in early summer. Melanoplus differentialis, on the other hand, normally lays its eggs in late fall, the eggs remaining in the ground during the winter and

Oxygen Consumption and Rates of Dehydration of Grasshopper Eggs (Orthoptera)

I INVESTIGATIONS regarding the effect of the removal of water from living materials may be classified into three general categories: first, the effect of desiccation on behavior of animals; second, the vital limit of desiccation and the rate of water loss; third, the effect of dehydration on physiological processes, such as respiration, etc. In the first of these, results have, in general, shown an increased irritability as the first response of animals to desiccation (Shelford,

Changes in riboflavin during embryonic development as functions of the embryo.

tion of the substance, especially in the pharyngeal and anal regions. In the case of a worm with fission, the two adjacent sides of the fission zone appear to develop new gradients. These disappear following starvation of the worm. A piece that is cut from any body level of the worm reveals the same reaction as that region would show in an intact worm. If it is allowed to regenerate for 2 or 3 days, the regenerating tissue will develop a new gradient pattern, decreasing from the cut end or ends, the intensity of the color reaction depending on the amount of tissues regenerated. A possible relationship between the sulphydryl gradient and the regeneration potency gradient is discussed.

Riboflavin and other fluorescent compounds in a developing egg (Orthoptera).

inorganic phosphate at all stages investigated. The relative amount of ATP breakdown at different stages in response to nitrogen was found to parallel the amount of inhibition of development. Cleavage proceeds normally in nitrogen with relatively little ATP breakdown, while gastrulation is arrested in response to a nitrogen atmosphere, with correspondingly greater ATP breakdown. The release of inorganic phosphate from ATP appears to occur rapidly after exclusion of air and to be readily reversible. Sodium azide inhibition of gastrulation was found to be accompanied by increased inorganic phosphate at the expense of the ATP content of gastrulae. Evidence that phosphate-bond energy is coupled to gastrulation came also from the study of arrested hybrid gastrulae, whose lower rate of respiration and glycolysis was found to be accompanied by a decreased capacity to keep ATP (as well as an unidentified organic phosphate ester) in the phosphorylated condition.

Structure and endogenous oxygen uptake of embryonic cells.

BODINE, J. H., and BOELL, E. J. 1936. Respiration of the embryo versus egg (Orthoptera). Jour. Cell. and Comp. Physiol., 8:357-66. BODINE, J. H., and Lu, K. H. i95oa. Oxygen uptake of intact embryos, their homogenates, and intracellular constituents. Physiol. Zo6l., 23:301-8. . 1950b. Methylene blue, 2,4-dinitrophenol, and oxygen uptake of intact and homogenized embryos. Proc. Soc. Exper. Biol. and Med.,

Succinic Dehydrogenase in Mitotically Active and Blocked Embryonic Cells

AT succinic dehydrogenase is widely distributed in organisms AT seems well established (Lardy, 1949). The degree of activity of this enzyme has been shown to vary for different tissues and even for the same tissue under varied physiological conditions. Most succinoxidase (succinic dehydrogenase plus cytochrome oxidase) is thought to be located for the most part in vertebrates in the large granule fraction of the cytoplasm-the mitochondria; and in the case of rat liver some 70-80 per cent of the activity of the original homogenate has been found in these structures (Hogeboom, Schneider, and Pallade, 1947). The importance of the cytochrome system and the part played by it in the normal reactions of succinic dehydrogenase have been repeatedly pointed out (Lardy, 1949). Preliminary results of experiments with the colorimetric triphenyltetrazolium chloride method for estimating succinic dehydrogenase with the embryo of the grasshopper failed to show the presence of the enzyme in this organism (Bodine and Fitzgerald, 1949b). Further investigations, however, using manometric as well as colorimetric tech-