Electron microscopic observations of lung alveolar epithelial cells of normal young mice,with special reference to formation and secretion of osmiophilic lamellar bodies

Summary Using the electron microscopy and electron microscopic histochemistry the authors studied the lung alveolar epithelial cell of normal young mice.Type II cell of the alveolar epithelium has characteristically numerous osmiophilic lamellar bodies. The lamellar boies are formed in the cytoplasmic vesicle, and never originate from the mitochondrion. These bodies have abundant acid phosphatase activity in their limiting membrane therefore it is considered to be lysosomal origin, but the mitochondria have no such enzyme activity.The body which is newly formed in the cytoplasmic vesicle grows up to the large lamellar body as a result of an accumulation of the fibrous dense substance, migrates to the free margin of the type II cell of alveolar epithelium, and then is discharged into the alveolar lumen as a merocrine type secretion.Acknowledgement is given to Professor Dr. Y. Sano and Professor Dr. H. Fujita, Department of Anatomy, and Assistant Professor Dr. S. Fujita, Department of Pathology, for their kind advice and criticism.     

The chromomere map of the pachytene spermatocyte of the Turkish hamster (Mesocricetus brandti).

The chromomere map of the early to mid pachytene spermatocyte of the Turkish hamster (Mesocricetus brandti) is described. Each autosomal bivalent was identified and a total of 304 chromomeres was found. A sex bivalent with a despiralized Xq protruding from the sex vesicle and a small number of the polymorphic 16q bivalents were observed.     

Trivalent behavior during prophase I in male mice heterozygous for three Robertsonian translocations: an electron-microscopic study

A synaptonemal complex (SC) analysis was carried out in male mice heterozygous (CHT/+) for three Robertsonian translocations. All pachytene preparations studied showed the presence of three trivalents. At early pachytene, the nonhomologous centromeric regions of the acrocentric chromosomes were unpaired. Heterosynapsis subsequently took place with complete pairing of the trivalents. Association between one of the three trivalents and the sex vesicle was observed in 30.4% of the nuclei. Association between the unpaired regions of two trivalents was present in 14.4% of the cells, suggesting that the relationship between unpaired regions of structural rearrangements and the X-Y bivalent may simply reflect the tendency of unpaired regions to establish end-to-end associations or heterosynapses among them, which are usually resolved during the pachytene stage of prophase I. Since the sex bivalent always has unpaired regions, these associations often affect the sex chromosomes.     

Pachytene chromosomes in male and female mice heterozygous for the Is(7;1)40H insertion

Pairing of pachytene chromosomes was studied in oocytes and spermatocytes of mice heterozygous for the male-sterile Is(7;1)40H insertion using light and electron microscopy for synaptonemal complex analysis in surface-spread, silver-stained preparations. The data comprised four males and four female embryos. The insertion/deletion configurations appeared as either two bivalents or one quadrivalent in both sexes, but the proportion of bivalents was higher in oocytes. Some insertion and deletion bivalents showed synaptic adjustment. The insertion/deletion configurations were associated with, or adjacent to, the XY bivalent in the majority of spermatocytes. End-to-end association of different bivalents was more frequent in oocytes than in spermatocytes. It is suggested that physiological differences between male and female gametocytes may lead to the difference in their reproductive potential.The authors warmly dedicate this paper to the Founder and Senior Editor of Chromosoma, Professor Hans Bauer, on the occasion of his 80th birthday.     

XYY spermatogenesis in XO/XY/XYY mosaic mice

The relative frequencies of XYY and XY cells in XO/XY/XYY mosaic mice were compared between somatic cells (bone marrow) and spermatogonia, and between spermatogonia and pachytene or MI spermatocytes. The results indicated there was no selection either for or against XYY spermatogonia. There was, however, a strong selection against XYY spermatocytes during pachytene, with their almost total elimination by the first meiotic metaphase. At pachytene, most XYY cells had trivalent or X univalent/YY bivalent configurations. These findings are contrasted with previous studies of XYY spermatogenesis in mice and are discussed with respect to a model that invokes sex-chromosome univalence as the cause of XYY spermatogenic failure.     

The fine structure of sensory receptor processes in the auricular epithelium of the planarian,Dugesia tigrina

Summary The marginal epithelium of the lateral auricles of the planarian, Dugesia tigrina, includes a cell type with surface cilia and microvilli, a basal nucleus, and dense cytoplasm containing secretory vacuoles, Golgi elements, mitochondria and ribosomes. Through channels within the epithelial cytoplasm, cellular processes, interpreted as extensions of neurosensory receptor cells located in the subepidermis, project to the surface. The receptor processes, containing microtubules, mitochondria, vesicles and an agranular tubular reticulum, project beyond the epithelial cell surface; one or two cilia each emerge from a basal body in the apex of the projection. Close to the point of emergence to the epithelial surface, each cylindrical receptor process is surrounded by a collar-like septate junction between adjacent plasma membranes. The cilia of the projections differ from those of the epithelial cells in diameter, density of matrix and in the banding patterns of the rootlets. A few projections appear with the apex and basal body retracted below the epithelial surface. The possible function of these ciliated processes in sensory reception is discussed.This work was supported by Grant No. SO 1 FR 5369 from the U.S. Public Health Service to the University of Illinois at the Medical Center.I thank Dr. J. P. Marbarger, Director of the Research Resources Laboratory, for use of the electron microscope facilities, Miss Irena Kairys for technical help, Miss Marie Jaeger for assistance with photography, and Mr. Robert Parshall for the drawing.To Professor Arthur Wagg Pollister, I respectfully dedicate this article on the occasion of his retirement from Columbia University.     

The fine structure of photoreceptors in a marine flatworm

Summary The ocelli or eyes of the marine polyclad turbellarian Notoplana acticola are clustered on the paired dorsal nuchal tentacles and in two longitudinal bands lateral to the cerebral ganglion. The ocelli, studied by electron microscopy, were characterized as rhabdomeric and non-ciliary in origin. There are 60 to 80 ocelli per animal each enclosed in a fibrous capsule to which muscle fibers may attach. An ocellus consists of a pigmented eyecup into which 30 to 50 photoreceptor cells send dendritic processes through interruptions in or among pigment cell projections across the eyecup opening. The dendritic processes terminate in numerous long intertwined microvilli which fill the eyecup. The nucleated cell body of each photoreceptor cell lies outside the eyecup and projects an axonal process to the cerebral mass. Within the dendritic processes are observed mitochondria, ribosomes, neurotubules, multivesicular bodies, vesicles and vacuoles. The cell body contains smaller mitochondria, endoplasmic reticulum, ribosomes, vesicles and prominent Golgi complexes.After dark adaptation, there are some structural alterations in terms of swelling of microvilli, increased numbers of vacuoles associated with the microvilli and dendritic processes, and changes in the pigment cell projections.This work was supported by Grant No. GM 10292 from the U.S. Public Health Service to Professor Richard M. Eakin, Department of Zoology at the University of California, Berkeley, U.S.A., where this investigation was conducted during the author's sabbatical leave of absence from the University of Illinois, and by Grant No. 1 SO 1 FR 5369 from the U.S. Public Health Service to the University of Illinois at the Medical Center.I express appreciation to Professor Eakin for interesting discussions and generous hospitality to me as a guest in his laboratory, and to the John Simon Guggenheim Memorial Foundation for the Fellowship which I held during 1964–65. I thank Dr. John P. Marbarger, Director of the Aeromedical Laboratory for the electron microscope facilities used at the University of Illinois.     

M31, a murine homolog of Drosophila HP1, is concentrated in the XY body during spermatogenesis.

The formation of the sex vesicle, or XY body, during male meiosis and pairing of the sex chromosomes are thought to be essential for successful spermatogenesis. Despite its cytological discovery a century ago, the mechanism of XY body formation, particularly heterochromatinization of the sex chromosomes, has remained unclear. The HP1 class of chromobox genes are thought to encode proteins involved in the packaging of chromosomal DNA into repressive heterochromatin domains, as seen, for example, in position-effect variegation. Study of the distribution of a murine HP1-like chromodomain protein, M31, during spermatogenesis revealed spreading from the tip of the XY body in mid-stage pachytene spermatocytes to include the whole of the XY body in late-pachytene spermatocytes. We also demonstrate that the formation of the XY body during spermatogenic progression in neonatal mice coincides with the expression of a novel nuclear isoform of M31, M31(p21). These results support the view that a common mechanistic basis exists for heterochromatin-induced repression, homeotic gene silencing, and sex-chromosome inactivation during mammalian spermatogenesis.     

The elimination and differentiation of chromosomes in the germ line of sciara

The germ line chromosomes of S. coprophila have been followed from the time of origin of the germ cells up to the time of meiosis in the male and up to first larval molt in the female. The mechanism which prevents the accumulation of L (limited) chromosomes in the germ line is a unique process of chromosome elimination: it occurs in male and female embryos after the germ cells have migrated from the pole plasm to the definitive gonad site, and it involves the movement of whole L chromosomes through the nuclear membrane into the cytoplasm. The extra paternal X chromosome is eliminated from the germ cells at the same time and in the same manner. Following this elimination there is a cytological differentiation of the chromosomes remaining inside the nucleus. First, the 4 paternal homologues of the regular complement undergo a loosening of coils and become light-staining whereas the maternal homologues remain condensed like the L's. Next, the L chromosomes undergo a process of extreme attenuation and dispersion following which they return to the condensed state. H³-thymidine autoradiography on gonial and premeiotic cells in the testis reveals that the L chromosomes undergo DNA replication at the end of the S period, also that there are asynchronies in DNA synthesis among the regular chromosomes. The phenomena of differential chromosome staining and asynchronous DNA replication are considered in the light of current theory regarding heterochromatization and gene inactivation, also in relation to the phenomenon of chromosome imprinting encountered in this genus.The studies reported here were supported by the National Science Foundation grants GB-42 and GB-2857, and in part by Contract No. AT-(40-1)-2690 under the Division of Biology and Medicine, U.S. Atomic Energy Commission.Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy, in the Faculty of Pure Science, Department of Botany, Columbia University. This work was carried out in the laboratory of Professor J. Herbert Taylor and has been supported in part by U.S. Public Health Training Grant No. 2 T 1-GM-216-05. Grateful acknowledgement is made to Professor Spencer W. Brown, Department of Genetics, University of California, Berkeley, in whose laboratory the final studies were completed.     

EM autoradiographic evidence that DNA synthesis occurs at recombination nodules during meiosis in Drosophila melanogaster females

Serial section electron microscopic autoradiography was used to examine the relationship between recombination nodules and ³H-thymidine incorporation during pachytene in Drosophila melanogaster females. For both ellipsoidal and spherical recombination nodules, the number of nodules that are associated with an autoradiographic grain is higher than that expected by chance; this observation is consistent with the hypotheses that recombination involves DNA synthesis and that recombination nodules are the sites of meiotic recombination. Moreover, general DNA replication (S-phase) and synapsis (synaptonemal complex formation) were found to be temporally distinct events, contrary to previous reports; Drosophila females therefore are not exceptional in this regard.This paper is dedicated to Herschel Roman on the occasion of his 65th birthday in fond appreciation for his many and varied contributions to Genetics as Professor and Chairman of the Department of Genetics, University of Washington, Seattle Washington, U.S.A.     

An analysis of meiotic impairment and of sex chromosome associations throughout meiosis in XYY mice

The existing XYY meiotic data for mice present a very heterogeneous picture with respect to the relative frequencies of different sex chromosome associations, both at pachytene and diakinesis/metaphase I. Furthermore, where both pachytene and diakinesis/MI data are available for the same males, the frequencies of the different configurations at the two stages are very different. In the present paper we utilise "XYY" and "XY/XYY" mosaic mice with cytologically distinguishable Y chromosomes to investigate the factors responsible for this heterogeneity between different males and between the two meiotic stages. It is concluded (1) that the initial pattern of synapsis is driven by the relatedness of the three pseudoautosomal regions (PARs); (2) that the order and extent of PAR synapsis within radial trivalents are also affected by PAR relatedness and that this leads to chiasmata being preferentially formed between closely related PARs; (3) that trivalents with a single chiasma resolve into a bivalent + univalent by the diakinesis stage; (4) that although many spermatocytes with asynapsed sex chromosomes are eliminated between pachytene and diakinesis, those that survive this phase of elimination progress to the first meiotic metaphase (MI) and accumulate in large numbers, leading to an over-representation of those with univalents as compared to radial trivalents; and (5) that the arrested MI cells are eventually eliminated, so that very few "XYY" cells contribute products to MII.     

Formation of giant nuclei in continuously irradiated tumors

Summary Transplantable malignant tumors of mice and rats have been irradiated continuously for several days by means of embedded glass beads containing Sr⁹⁰-Y⁹⁰. Concomitant enlargement of tumor cell nuclei occurs in areas where mitosis is inhibited, i.e. associated with doses of 1200–2000 rads per day and above. Some enlarged cells survive at least a week in areas where the dose rate is much higher than this. Intense incorporation of thymidine into enlarged cells can be seen at least a week after beginning irradiation.Work supported by the U.S. Atomic Energy Commission.Dedicated to Professor Friedrich Wassermann with admiration and affection on the occasion of his eightieth birthday.     

Q & A. [interview

George Oster is Professor of Biophysics, University of California, Berkeley. He received his B.S. at the U.S. Merchant Marine Academy and his Ph.D. at Columbia University. He began his career in biophysics as a postdoc at the Weizmann Institute under Aharon Katchalsky, where his research involved membrane biophysics and irreversible thermodynamics. His concern for environmental issues led him into population biology, which shaded into evolutionary biology and thence to developmental biology, cell biology and, most recently, protein motors and bacterial motility and pattern formation. His tools are mathematics, physics and computer simulation. He is currently a faculty member in the Departments of Molecular and Cellular Biology and the College of Natural Resources at Berkeley.     

Analysis of male meiotic "sex body" proteins during XY female meiosis provides new insights into their functions

During male meiosis in mammals the X and Y chromosomes become condensed to form the sex body (XY body), which is the morphological manifestation of the process of meiotic sex chromosome inactivation (MSCI). An increasing number of sex body located proteins are being identified, but their functions in relation to MSCI are unclear. Here we demonstrate that assaying male sex body located proteins during XY female mouse meiosis, where MSCI does not take place, is one way in which to begin to discriminate between potential functions. We show that a newly identified protein, "Asynaptin" (ASY), detected in male meiosis exclusively in association with the X and Y chromatin of the sex body, is also expressed in pachytene oocytes of XY females where it coats the chromatin of the asynapsed X in the absence of MSCI. Furthermore, in pachytene oocytes of females carrying a reciprocal autosomal translocation, ASY associates with asynapsed autosomal chromatin. Thus the location of ASY to the sex body during male meiosis is likely to be a response to the asynapsis of the non-homologous regions [outside the pseudoautosomal region (PAR)] of the heteromorphic X-Y bivalent, rather than being related to MSCI. In contrast to ASY, the previously described sex body protein XY77 proved to be male sex body specific. Potential functions for MSCI and the sex body are discussed together with the possible roles of these two proteins.     

Sex chromosome configurations in pachytene spermatocytes of an XYY mouse

Karyotypic investigation of a phenotypically normal but sterile male mouse showed the presence of an XYY sex chromosome constitution. The synaptic behaviour of the three sex chromosomes was examined in 65 pachytene cells. The sex chromosomes formed a variety of synaptic configurations: an XYY trivalent (40%); an XY bivalent and Y univalent (38.5%); an X univalent and YY bivalent (13.8%); or X, Y, Y univalence (7.7%). There was considerable variation in the extent of synapsis and some of the associations clearly involved nonhomologous pairing. These observations have been compared with previously published information on chromosome configurations at metaphase I from other XYY males.