Ultrastructural study of the regressing prothoracic glands of blattarian insects

Summary The prothoracic glands, source of the molting hormone ecdysone, regress within a few days after the final molt, a process which was analyzed with electron microscopic methods in the cockroaches Leucophaea and Blaberus. This strictly timed event is accompanied by drastic alterations in cellular fine structure. Early signs of breakdown appear in groups of nuclei whose substance becomes segregated into patches of contrasting electron density characteristic of pyknosis.The most conspicuous change in the cytoplasm of parenchymal cells concerns the appearance of large, heterogeneous inclusion bodies in which various cellular elements become segregated. These compartments seem to represent autophagic vacuoles within which the gradual degradation of much of their contents takes place, presumably under the influence of lysosomal enzymes. Undigested swirls of membranous character may remain sequestered within these packets for some time.At advanced stages of cellular atrophy, plasma membranes and nuclear envelopes have gradually disappeared, and masses of protoplasm undergoing autolysis become invaded by a greater number of hemocytes than are present in nymphal glands. These phagocytic elements appear to engulf debris of parenchymal cells as well as some degenerating connective tissue elements. After the completion of the regressive process, the axial band of musculature characteristic of the nymphal gland persists on its own. Whether or not some parenchymal cells (or possibly their precursors) capable of reactivation persist in the proximity of this muscle is unknown.The resorption of the prothoracic gland in the newly emerged insect is the result of physiological autolysis and seems to be aided by the activity of phagocytic hemocytes.Dedicated to Professor W. Bargmann on his 60th birthday in friendship and admiration.This study was supported by Research Grants AM-03984, NB-02145 and NB-05219 from the U.S.P.H.S.I wish to express my thanks to Mrs. S. Wurzelmann, Mrs. C. Jones, Mrs. C. Grubman, and Mr. S. Brown for their excellent technical assistance.     

Photoperiodic and osmotic influences on the ultrastructure of the hypothalamic neurosecretory system of the White-crowned sparrow,Zonotrichia leucophrys gambelii

Summary An attempt was made to correlate functional changes in the neurohypophysis of the White-crowned Sparrow, Zonotrichia leucophrys gambelii, with morphologic features on the light- and electron-microscope levels. The aldehyde-fuchsin-staining anterior median eminence possesses essentially the same ultrastructural features as the non-staining posterior median eminence. The axon terminals are characterized by the presence of a large number of small vesicles (approximately 400 Å in diameter) and occasional electron-dense granules. The more-or-less depleted anterior median eminence occasionally evident in the photosensitive bird showing testicular development is indistinguishable ultrastructurally from the more intensely staining median eminence generally characteristic of the photorefractory bird. In the median eminence, stainability and functional state do not seem to be correlated with changes in the type, size or number of vesicles. A slight increase in the number of granules was noted in the photorefractory bird but this was considered insufficient basis to account for differences in stainability.The pars nervosa, on the other hand, responded to osmotic stimuli (saline drinking water) by loss of stainability and decrease in numbers of elementary neurosecretory granules. Small vesicles are also present in the pars nervosa axon terminals, but are intermingled with neurosecretory granules in normal birds. Acute-osmotic birds, however, had axon terminals almost entirely occupied by small vesicles.It is to be emphasized that the pars nervosa and the median eminence are two structurally very different regions of the neurohypophysis. The basis for aldehyde-fuchsin staining in the median eminence appears to differ from that in the pars nervosa. The implications of these findings are considered in regard to hypothalamic control over gonadotropic activity in the White-crowned Sparrow.Dedicated to Professor Dr. W. Bargmann in honor of his 60th birthday.This investigation was supported by grant GB-2484 from the National Science Foundation to Professor Bern, grant GB-2819 from the National Science Foundation to Professor Mewaldt, and grant NB-01353 from the National Institutes of Health to Professor Farner. The authors wish to express their appreciation of the technical assistance of Mrs. Irene Brown, Mr. John Butchart, Sally S. Kibby, Mrs. Carol Nicoll, and Mr. John Striffler. Mrs. Emily Reid kindly prepared the histograms.     

Scanning electron microscopic studies of cilia

Summary A simple method for the preparation of ciliated epithelia for study with the scanning electron microscope is described. Ciliary groups are well preserved and it is possible to discern individual cilia and work out their numbers and orientation. Following scanning electron microscopical study some of the material was prepared for transmission electron microscopy and the ultrastructure of the tissue was found to be surprisingly well preserved. The tracheal epithelium of the rabbit, the olfactory epithelia of the goldfish and the rabbit, and the sensory epithelia in the statocyst of a cephalopod mollusc were examined with the scanning electron microscope to demonstrate the possibilities of the method. Acknowledgements. We would like to thank Professor J. Z. Young for his continued interest and support. The scanning electron microscope was purchased with a grant provided by the Science Research Council to Dr. Boyde, Mr. R. Willis helped in the initial stages of the study, Mr. G. Savage provided help with the goldfish material, Mr. S. Waterman provided much photographic assistance, and Mrs. N. Finney the secretarial assistance.     

Some supplementary attributes in the classification of chlorococcum species

Summary Seventeen species of the chlorophycean genus Chlorococcum have been studied comparatively in axenic culture with the purpose of exploring the value of certain supplementary attributes in facilitating identification. The algae were cultivated under standard conditions in defined media. Such attributes as colony characteristics, changes in color and cellular morphology with increasing age, inhibition in the light by organic compounds such as acetate, pyruvate and certain pentose sugars, and differential sensitivity to antibiotic agents were studied and found useful in distinguishing the species of the genus Chlorococcum.This paper is dedicated to Professor Dr. E. G. Pringsheim on the occasion of his 80th birthday.The writers acknowledge with gratitude the assistance of Professor R. E. Alston, Mrs. Mildred Austin, Henry Aldrich and T. C. Massey.     

Some effects of electrical stimulation and exogenous metabolites on the contractile activity and the ultrastructure of the radula-muscle of Buccinum undatum

Summary The effect of various metabolites on the contractile pattern of the radulamuscle of Buccinum undatum was recorded. Its ultrastructure was studied after varying periods of spontaneous and electrically induced work.The effect of added substrates on the contractile behaviour of the radula-muscle was most pronounced after 12 hours of work or more. Addition of succinate caused a relaxation of the muscle, and its contractions were inhibited. Addition of glutamate had the opposite effect and caused a marked contraction of the muscle. The contractile pattern was not influenced by -ketoglutarate. These results indicate that factors other than energy supply influenced the response of the radula-muscle.The ultrastructure of the control muscles and of the electrically stimulated muscles showed some changes referable to the isolation of the muscle. Contrary to the controls, those muscles that had been stimulated for 4 and 12 hours respectively showed a more regular organisation. The filaments were more parallel and electron dense dots occurred arranged in transverse or oblique bands. The distance between the bands was around 1 . This organisation of the radula-muscle was discussed and compared with that of banded muscles of various kinds. Attention was drawn to the similarities in appearance between the bands of the radula-muscle and of Z bands of developing striated vertebrate muscles. The mitochondria showed changes partly referred to the isolation of the muscle and partly to the experimental conditions.Our sincere thanks are due to Professor I. Agrell, Head of the Zoophysiological Institute, Lund, for his advice and support during the work.Thanks are also due to Dr. B. Swedmark and his staff of the Zoological Station, Kristineberg for kind help with animal material and to Professor C. G. Ahlström, Pathological Institute, Lund, for electron microscopic facilities. We thank Mrs. M. Carleson and Mrs. I. Hedström for most valuable technical assistance during the electron microscopic work. The work was supported by grants from Swedish Natural Science Research Council and the Royal Physiographic Society of Lund.     

The role of the thyroid in the development of Mauthner's neuron

Summary Although the normal postmetamorphic atrophy of the giant neurons of Mauthner in anuran amphibians has been attributed to the direct or indirect action of thyroid hormone, the administration of exogenous thyroxin to Rana pipiens larvae appears to facilitate growth of the neuron rather than inhibit it. In the present investigation, Rana pipiens embryos were surgically thyroidectomized. In the absence of the thyroid, metamorphosis never occurred, and hind limb development was arrested at larval stages VI–VIII. However, these animals continued their growth and often exceed their controls in overall size. Mauthner cells of thyroidectomized larvae have nuclei of significantly smaller size than their controls. The observations are consonant with the view that thyroid hormones play an important positive role in the growth and maintenance of Mauthner's cell. It is further indicated that the growth of Mauthner's neuron is more dependent upon the titer of thyroid hormone in the blood than it is upon age, brain size, tail motor influences, or the state of the lateral line system.Dedicated to Professor Berta Scharrer on her 60th birthday. This study was supported by Grants NB-04555, IGS-98 and 5 T 1-GM-102 from the U.S.P.H.S.I am pleased to acknowledge the excellent technical assistance of Mrs. Celia Grubman.     

Morphology of microtubules in rabbit platelets

Summary The morphology of the microtubular wall in rabbit platelets fixed in glutaraldehyde and osmic acid solution and stained with uranyl acetate, or lead hydroxide, or doubly stained, is variable. In cross section, the wall may appear as a uniformly dense annulus, an annulus containing nodular densities about 35 Å in diameter or as a series of contiguous subunits with a circular cross sectional profile about 70 Å wide. The authors relate the varying morphology to the intensity of staining and equate the nodular and circular subunits. Rotational analysis suggests that there are 12 ± 2 subunits in the microtubular wall.We thank Professor A. C. Ritchie for his criticism of the paper and Mrs. M. Lorber and Mrs. M. Mezari for technical assistance. — This work was supported by grants from the Medical Research Council of Canada and the Ontario Heart Foundation.     

Cytologic and metabolic parameters of pineal inhibition by continuous light in the rat (Rattus norvegicus)

Summary Adult rats (Rattus norvegicus) were subjected to continuous light or control conditions (14 hours light/day) for six weeks or longer, and quantitative cytological and metabolic studies were made of the pineal organs. After 11 weeks of continuous light, the pineal parenchymal cell's largest nucleolar eosinophilic mass is significantly reduced in diameter, especially in the medulla of the organ. Evidence of metabolic inhibition includes reduction of pineal glycogen content, succinic dehydrogenase activity, and respiration in the absence of exogenous substrates. Pineal ATP content, P³²-phosphate uptake and 5-hydroxy indole acetic acid content did not appear to be affected. Pineal serotonin content and melatonin-forming activity in the continuously lighted animals were measured but could not be interpreted metabolically, due to the diurnal fluctuations of these in control animals. Results provided here and elsewhere suggest that pineal inhibition by continuous light involves primarily the citric acid cycle, the accumulation of metabolites and lipid, and the synthesis of protein.This investigation was supported by grant GM-05219 from the National Institutes of Health, U.S. Public Health Service.I am grateful to Mrs. Virginia Green Bowers, Mrs. Ann Richards, Mr. Peter Charles Baker and Mr. Jorge Antonio Alvarado for laboratory assistance, and to Dr. Richard Strohman and Mr. David Epel, for advice on the determination of ATP.     

Differentiation of the membrane system in the cells of imaginai discs

Summary Imaginal discs from developing larvae of the fly Calliphora erythrocephala fixed in permanganate or osmium tetroxide and embedded in Epon 812 were observed by electron microscopy. When the larval growth ceases, the differentiation manifests itself through an enlargement of the endoplasmic reticulum, by which continuous membrane contact is established between all cell organelles. During the same time mitochondria swell up and transform into lipid granules and the intercellular contacts weaken.I am indebted to Mrs Mariann Carleson and Miss Brita Nilsson for technical aid.     

Studies on the receptors in Ciona intestinalis

Summary A photoreceptor type structure not previously described has been found in the dorsal wall of the cerebral vesicle of the tadpole larva of Ciona intestinalis. The membranes of this receptor are organised as tubules some 60–100 nm in diameter and up to 1.5 m long. The tubules are confined in bundles about 1.5 m in diameter, which extend from the cell surface into the cavity of the cerebral vesicle. These tubules are similar to those in the rhabdomeric type of photoreceptor. However, in the cells from which the tubule processes arise are structures typical of the bases of cilia, and found in ciliary type photoreceptors.I should like to thank Professor J. Z. Young, F. R. S. for his continuing encouragement and help, and Dr. R. Bellairs for the use of electron microscope facilities. Mr. R. Moss and Mrs. J. Hamilton gave excellent technical assistance.     

Continuity of the membrane systems in the cells of imaginal discs

Summary Imaginal discs from mature larvae of the fly Calliphora erythrocephala fixed in permanganate and embedded in Epon 812 were observed by electron microscopy. Irrespective of the type of the disc, the endoplasmic reticulum in the cells was found to be in unbroken contact with the outer nuclear membrane, Golgi areas, the mitochondria and the cell membrane. The imaginai discs are syncytial in that all cells are in cytoplasmic communication through secondary breaks in the cell membranes.The investigation was facilitated by grants from the Nordic Insulin Foundation.I am indebted to Mrs. Mariann Carleson and Miss Brita Nilsson for technical aid.     

The fine structure of blattarian prothoracic glands

Summary The ultrastructural characterization of the prothoracic glands derives interest from the fact that they are the only known source of a steroid hormone (ecdysone) among invertebrates. The present material includes nymphal glands of two blattarian species studied at graded intervals over a period of several intermolt cycles.Among the prominent cytological features of this organ are long and intricately interwoven cellular processes that may be separated from each other by extracellular channels with an average width of 0.5 . The plasma membranes of the cells facing these spaces are coated by an external lamina (boundary membrane) representing extensions of the same extracellular material that provides the connective tissue sheath of this and other insect organs. Micropinocytotic caveolae and vesicles present along this large cellular surface area may facilitate the transport of material between glandular cells and hemolymph, or vice versa.The cytoplasm contains a fairly moderate amount of membranous structures. Golgi units are distinctive but never spectacular; ergastoplasmic elements are sparse in most cells. This sparsity as well as the absence of membrane-bounded secretory granules is in line with the known steroidal nature of the secretory product involved. Some cells show a striking array of microtubules.Certain structural attributes of nucleoli, mitochondria, and lysosome-like bodies seem to vary in conjunction with distinct phases of the intermolt cycle.Dedicated to an inspiring teacher and loyal friend, Professor F. Wassermann on the occasion of his 80th birthday, August 13, 1964. — Supported by Research Grants AM-03984 and NB-02145 from the U.S.P.H.S. — I wish to thank Mrs. Cynthia Jones and Mr. Stanley Brown for their skillful assistance.     

A family outbreak of tinea capitis due to Trichophyton violaceum in Michigan. Its control with griseofulvin

Summary An outbreak of tinea capitis caused byTrichophyton violaceum in five siblings of a family of eight in Michigan and its control with griseofulvin are described.The griseofulvin (Fulvicin) tablets used in this study were provided by Schering Corporation, Bloomfield, New Jersey, through Dr.G. Kenneth Hawkins.We gratefully acknowledge the help extended by Mrs.Matilde Dolen, Mrs.Miriam Harris, Mr.George Miller and Mr.Walter Boehme of the Norris and Jefferson Schools in Nankin, Michigan.     

Cells with microvillous borders in the cerebral ganglion of Leptodora kindtii focke (Crustacea)

Summary Groups of large cells in the cerebral ganglion of Leptodora kindtii join in intricate patterns to enclose lacunar spaces. The cell surfaces bordering on these lacunae are covered by long, densely packed microvilli that all but fill the spaces. Near their brush borders the cells are joined by adhesion plates; for the rest they are separated from each other by glial septa. The possible significance of these structures is discussed.Dedicated to an inspiring teacher and loyal friend, Prof. F. Wassermann on the occasion of his 80th birthday, August 13, 1964.Supported by Grant No. NB-02145 from the United States Public Health Service. The expert assistance of Mrs. Cynthia Jones, Mrs. Sarah Wurzelmann, and Mr. Stanley Brown is gratefully acknowledged.     

Electron microscopic observations of synaptic endings in cultures of mammalian central nervous tissue

Summary Synapses were found in rat cerebellar and brainstem cultures with the electron microscope. Three distinct types of synaptic terminals were described. The similarity between synapses found in vitro and in vivo was emphasized.Supported by USPHS Grants 5 Tl 459-04 and NB 03114-03S1 from the National Institutes of Health, Bethesda, Maryland.The authors wish to express their sincere appreciation to Mrs. Eleanor Morris for her assistance in preparing the cultures and Mr. Earl Pitsinger for his photographic assistance.     

The nerve fibres in the basement membrane and related structures in Saccoglossus horsti (Enteropneusta)

Summary The structure of the basement membrane of Saccoglossus horsti has been examined with the electron microscope. The membrane consists of two lamellae each of two layers. An outer amorphous layer 150 nm across and an inner fibrillar layer 1–3 m across. The fibrils of the fibrillar layer are two sizes, the majority are 5–9 nm in diameter and at least 2 m long. The thicker 30 nm fibrils occur in small patches and have striations with a 30 nm period.Within the lamellae of the basement membrane are blood spaces. The only regularly found structures in these spaces are blood particles some 12–16 nm in diameter.Nerve fibres of varying diameters traverse all the layers of basement membrane. These fibres run longitudinally and obliquely through the basement membrane, and emerge amongst the muscle cells inserted into the coelomic side of the membrane. No motor end plates have been seen. Preliminary observations suggest that many of the nerve fibres have no sheath other than the cell membrane of the fibre itself.The muscle cells are attached to the basement membrane by structures that resemble hemidesmosomes. The blood vessels of Saccoglossus have a basement membrane on the lumenal side of the endothelial cell cytoplasm.I wish to thank Professor J. Z. Young, F. R. S. for continuous encouragement and advice. To Dr. R. Newell I am indebted for the collection and identification of the specimens. I am pleased to acknowledge my debt to Dr. R. Bellairs for the use of electron microscope facilities, and to Mrs. J. Hamilton and Mr. R. Moss for skillful technical assistance.     

The structure of a photoreceptor organelle in the eye of Pterotrachea mutica

Summary The photoreceptor cell of Pterotrachea consists of an elongated cell some 100 m long with recogniseable inner and outer segments. The photoreceptor membranes point towards the light. There are about 300 discs per photoreceptor, a small number of discs arising from a single ciliary base. There are bout 75–100 such bases on each receptor cell. The receptor cells themselves (the inner segments) have four recognisable regions. The vacuolated region, the region of mitochondria, the nuclear region, and the axonal region.The photoreceptor cells are organised in five roughly parallel rows, and separated from one another by pale supporting cells.My thanks are due to Professor J. Z. Young, F. R. S. for his enthusiastic support and help during this work. Dr. R. Bellairs kindly provided electron microscope facilities. Mr. R. Moss, Mrs. J. Hamilton and Mr. A. Aldridge provided excellent technical and photographic assistance.     

Some aspects of the fine structure of the statocysts of the molluscsPecten andPterotrachea

Summary The statocyst ofPecten is composed of hair cells and supporting cells. The hair cells bear kinocilia and microvilli at their distal ends and the supporting cells bear microvilli. The cilia have a 9+2 internal filament content, and arise from basal bodies that have roots, basal feet and microtubular connections. Two different ciliary arrangements are described, one with a small number of cilia arranged in a ring, and another with many more cilia arranged in rows. Below the hair cells are probable synapses. A ciliated duct connects to the lumen of the static sac and passes through the centre of the static nerve. The hair cells in the statocyst ofPterotrachea bear kinocilia and microvilli. The possible importance of cilia and microvilli in the transduction process is discussed.We would like to thank ProfessorJ. Z. Young for bringing specimens ofPterotrachea from Naples and also the staff of the Stazione Zoologica for the provision of specimens, Dr.M. Land for providing specimens ofPecten, the Science Research Council (U.K.) for providing the electron microscope used in much of the study and also for a grant to one of us (V.C.B.), and Mrs.J. Parkers and Mr.R. Moss and Mrs.J. Hamilton for much photographic and technical assistance.     

The fine structure of cephalopod blood vessels II. The vessels of the nervous system

Summary Blood vessels of the perioesophageal nerve ganglia (brain) of Octopus vulgaris and the stellate ganglia of Sepia officinalis are described. The vessels have an incomplete endothelium, a complete basement membrane and a complete investment of pericytes. The pericytes are joined by specialised membrane junctions but these are not tight junctions. The main type of neuron/vessel arrangement is one where there is a collagen-filled space between the pericytes and the surrounding glial cells. Axons or neurons are sometimes applied directly to the vessel pericytes and in the neuropil, pericytes contact glial cells that ensheath bundles of axons. Blood spaces between neurons are also present.We would like thank Professor J. Z. Young and Dr. E. G. Gray for encouragement and advice, Mrs. Jane Astafiev for drawing Fig. 11, Mr. S. Waterman for photographic assistance and Miss Cheryl Martin for secretarial and other assistance.     

The metabolism of p-hydroxybenzoate by Rhodopseudomonas palustris and its regulation

Summary Rhodopseudomonas palustris metabolizes p-hydroxybenzoate aerobically via a series of specific reactions which are partly described. The pathway similar to that described for aerobic pseudomonads. The requisite enzymes are absent in extracts of cells grown anaerobically under photosynthetic conditions with p-hydroxybenzoate, benzoate and malate, or aerobically with benzoate or malate. An intermediate of the reaction sequence presumably induces in a retrograde fashion.Dedicated to Prof. C. B. van Niel on the occasion of his 70th birthday. I wish especially to acknowledge a debt to Prof. C. B. van Niel, who offered an assistant so much encouragement and suggested this and so many other problems in his summer Microbiology Course.The early portion of this work was done in collaboration with Dr. E. R. Leadbetter in the laboratory of R. Y. Stanier at Berkeley. Mrs. C. Feist generously provided a sample of -hydroxymuconic semialdehyde. This investigation was supported by the U.S. Public Health Service, Grant HD-02448.