Cytological Differences Research Articles

Cytological changes associated with alkaloid production in cultured cells of Coptis japonica and Thalictrum minus

Cell structures were compared between alkaloid-producing and non-producing cell cultures of Coptis japonica and Thalictrum minus by electron microscopic observation. In alkaloid-producing cells of C. japonica, prior to the onset of alkaloid synthesis, the vacuoles showed a greater volume than in non-producing cells. These were characterized by a number of large starch grains in the cytoplasm. Furthermore, alkaloid-producing cells contained stacks of rough endoplasmic reticulum. Comparison of different cell lines suggested that there might be a negative correlation between accumulation of alkaloids and starch. Similar cytological differences were observed with T. minus cell cultures that release berberine into the culture medium. Alkaloid producing cells were found to contain an abundance of cytoplasmic vesicles (0.5 - 1 υm in diameter).

Nucleolar organizing region in theApiaceae (Umbelliferae)

Data available for the nucleolus organizing region in the familyApiaceae are reviewed. An attempt has been made to establish the exact number of this region in various subfamilies and tribes through studies on the karyotype and nucleolus. Most of the taxa have a single nucleolar chromosome per haploid complement. The location of the nucleolar organizing region (NOR) on the chromosome varies. Members ofHydrocotyloideae differ drastically from those of subfam.Saniculoideae andApioideae, with respect to the location of NOR. Despite wide geographical distribution, varied ecological preferences and differences in morphology, anatomy and cytology, Umbellifers have attained stability in the number and location of NORs. Characters of NOR offer scope for utilization in understanding phylogenetic relationships at higher levels of taxonomic hierarchy.

Biotypes of the brown planthopper Nilaparvata lugens (Stål) and strategies in deployment of host plant resistance

Resistant rice varieties have proved to be highly effective against the brown planthopper Nilaparvata lugens (Stal), but their long-term stability is threatened because of the evolution of prolific biotypes. At present, identification of biotypes is based principally on the differential reactions of host rice varieties to pest in greenhouse screenings and in field plantings of test nurseries at many locations. So far, three N. lugens biotypes that attack different rice varieties have been identified in the Philippines. Biotype 1 can survive on and damage only those varieties that do not carry any genes for resistance. Biotype 2 can thrive on varieties carrying Bph I resistance gene and on those susceptible to biotype 1. Biotype 3 can infest and destroy varieties having bph 2 resistance gene and those susceptible to biotype 1. However, none of these biotypes is capable of damaging varieties with Bph 3 and bph 4 genes for resistance, and the variety Ptb 33 carrying two unidentified genes is resistant to almost all known biotypes. Recently, a population of N. lugens was found thriving on the common weed grass Leersia hexandra Swartz on the IRRI farm and in some locations in Luzon, Philippines. This biotype does not survive on rice. Several varieties which are resistant in the Philippines are susceptible in India and Sri Lanka because the South Asian biotypes of N. lugens are more virulent than the Southeast Asian biotypes. To monitor the pest biotypes in different geographical regions and to identify new sources of resistance, an International Rice Brown Planthopper Nursery (IRBPHN) has been established in many cooperating Asian countries. Biotypes can also be differentiated by studying their behavioural and physiological responses to plants of known genotypes. Distinctive differences can be observed in insect feeding, growth, longevity, fecundity, oviposition and population increase on differential varieties and relative susceptibility or tolerance for hosts’ allelochemics. Recent investigations of the existence of subtle morphological, cytological, and electrophoretic differences among N. lugens biotypes and results of hybridization experiments strongly complement other methods of biotype identification. The evolution of biotypes is a complex process that is governed by the interactions of the genetic and other biological characteristics of the pest populations and the genetic makeup of cultivated varieties. While the strategy of sequential release of varieties with major resistance genes has been fairly successful, the monogenic nature of those varieties makes them vulnerable to the development of virulent pest biotypes. Therefore, present breeding endeavors envisage use of both major and minor resistance genes for effective pest suppression.

Cuticular cavities: Storage chambers for cyanoglucoside-containing defensive secretions in larvae of a zygaenid moth

As a defensive reaction against predators the larvae of Zygaena trifolii Esper, 1783, release highly viscous fluid droplets out of cuticular cavities. The fluid appears on the cuticular surface upon contraction of the irritated segments, with no specialized muscles being involved. Two morphologically different types of cavities have been found: the larger ones are located beneath pigment spots, the smaller ones occupy the remaining surface except in the ventral region. Both types have complicated cuticular opening structures. The defensive fluid contains the cyanoglucosides linamarin and lotaustralin, the amino acid β-cyano-L-alanine, proteins and water. Although a considerable amount of fluid (3–6 μ1 per sixth-instar larva) is stored in the cuticle, fine structural examinations of the epidermis do not show any specific cells or cell areas with morphological adaptations for secretion. Further, there do not exist any major cytological differences between the cells below the cavities and in the ventral region, where those cavities are absent.

Microcircuitry of bipolar cells in cat retina

We have studied 15 bipolar neurons from a small patch (14 X 120 micron) of adult cat retina located within the area centralis. From electron micrographs of 189 serial ultrathin sections, the axon of each bipolar cell was substantially reconstructed with its synaptic inputs and outputs by means of a computer-controlled reconstruction system. Based on differences in stratification, cytology, and synaptic connections, we identified eight different cell types among the group of 15 neurons: one type of rod bipolar and seven types of cone bipolar neurons. These types correspond to those identified by the Golgi method and by intracellular recording. Those bipolar cell types for which we reconstructed three or four examples were extremely regular in form, size, and cytology, and also in the quantitative details of their synaptic connections. They appeared quite as specific in these respects as invertebrate "identified" neurons. The synaptic patterns observed for each type of bipolar neuron were complex but may be summarized as follows: the rod bipolar axon ended in sublamina b of the inner plexiform layer and provided major input to the AII amacrine cell. The axons of three types of cone bipolar cells also terminated in sublamina b and provided contacts to dendrites of on-beta and other ganglion cells. All three types, but especially the Cb1, received gap junction contacts from the AII amacrine cell. Axons of four types of cone bipolar cells terminated in sublamina a of the inner plexiform layer and contacted dendrites of off-beta and other ganglion cells. One of these cone bipolar cell types, CBa1, made reciprocal chemical contacts with the lobular appendage of the AII amacrine cell. These results show that the pattern of cone bipolar cell input to beta (X) and probably alpha (Y) ganglion cells is substantially more complex than had been suspected. At least two types of cone bipolar contribute to each type of ganglion cell where only a single type had been anticipated. In addition, many of the cone bipolar cell pathways in the inner plexiform layer are available to the rod system, since at least four types of cone bipolar receive electrical or chemical inputs from the AII amacrine cell. This may help to explain why, in a retina where rods far outnumber the cones, there should be so many types of cone bipolar cells.

Influence of photoperiod on the ultrastructure of the hypophysial pars tuberalis of the Djungarian hamster, Phodopus sungorus.

Conspicuous cytological differences are found between specific secretory cells of the hypophysial pars tuberalis of Djungarian hamsters exposed to long and short photoperiods. The cells differ with respect to the shapes of perikarya and nuclei and show diverse amounts of secretory granules, lysosome-like bodies and glycogen.

Fine structural morphology of identified X- and Y-cells in the cat's lateral geniculate nucleus.

Four physiologically identified neurons in the A laminae of the cat's dorsal lateral geniculate nucleus were filled with horseradish peroxidase and studied using the electron microscope. Two were X-cells and two were Y-cells. Each had electrophysiological properties appropriate for its X- Or Y-cell class, and each also had an axon that projected into the optic radiation, indicative of a geniculocortical relay cell. Representative samples from about 10% of each neuron's entire dendritic arbor (proximal and distal) were taken to obtain an estimate of the types and distributions of synapses contacting these arbors. One X-cell had a cytoplasmic laminar body, but there were no other significant cytological differences seen among the neurons. Common to each of the neurons were the following synaptic features: (i) retinal terminals (r.l.p.) were mostly or entirely restricted to proximal dendrites or dendritic appendages (less than 100 microns from the soma). These terminals constituted about 15-25% of the synapses on the proximal dendrites. (ii) Terminals with flattened or pleomorphic synaptic vesicles (f. terminals) were predominant on the proximal dendrites (30-55% of the total synapses for that region) and were mainly located near the retinal terminals. A smaller percentage (10-20%) were also distributed onto the distal dendrites. (iii) Small terminals with round synaptic vesicles (r.s.d.), many presumably having a cortical origin, predominated (60-80%) on distal dendrites (greater than 100 microns), but also formed a large proportion (40-70%) of the synapses on the intermediate (50-150 microns) dendrites. Total synaptic contacts for one X-cell and one Y-cell were estimated at about 4000 and 5000, respectively. The major fine structural differences observed between X- and Y-cells were almost entirely related to the retinal afferents. First, the retinal synapses for X-cells were mostly made on to dendritic appendages (spines, etc.), whereas Y-cells had most of their retinal synapses onto the shafts of primary and proximal secondary dendrites (that is, near branch points. Second, the retinal terminals that contacted X-cell dendrites nearly always formed triadic arrangements that included nearby f. terminals, but those on Y-cells rarely did so. Finally, the main type of f. terminals associated with X-cells were morphologically different from most of those associated with the Y-cells, and this also related directly to the triadic arrangements; that is, f. terminals in the triadic arrangements were morphologically distinguishable from f. terminals that did not participate in triadic arrangements.(ABSTRACT TRUNCATED AT 400 WORDS)

Effects of Metabolic Inhibitors and Ionophores on the Stomatal Apparatus in Spirodela oligorrhiza (Kurz.) Hegelm. and in Lemna minor L.

The ability to control stomatal opening is studied in Spirodela oligorrhiza and Lemna minor after treatments with CMU, FCCP, valinomycin and nigericin. The results showed that Spirodela stomata close to a various extent following such treatments. In Lemna the above substances seem to exert no effects. The different behaviour of Spirodela and Lemna may possibly be dependent on cytological and morphological differences observed in the guard cells of the two genera examined.

In vitro morphogenesis and ultrastructure of a Frankia sp. ArI3 (Actinomycetales) from Alnus rubra and a morphologically similar isolate (AirI2) from Alnus incana subsp. rugosa

An actinomycete (AirI2) with black spores was isolated from nodules of Alnus incana ssp. rugosa. Although this organism is very different in appearance from the typically white strains of Frankia from Alnus, it has all the main microscopic features of members of this genus. However, cytological differences have been observed when compared with a strain with white spores (ArI3).

Somatostatin and vasoactive intestinal polypeptide-like immunoreactive cells and terminals in the retrohippocampal region of the rat brain

The retrohippocampal region of the rat brain was analyzed by using immunohistochemistry with specific antibodies against somatostatin (SOM) and vasoactive intestinal polypeptide (VIP). Specifically immunoreactive neurons and terminal processes were labeled with either the anti-SOM or anti-VIP antiserum and they were referred to as SOM-like immunoreactive (SOM-LI) or VIP-like immunoreactive (VIP-LI) neurons and processes, respectively. The retrohippocampal region was rich in neuronal cell bodies and terminal processes showing immunoreactivity for SOM and VIP. In the entorhinal area SOM-LI neurons were located mainly in layers IV through VI and the VIP-LI neurons were found mainly in layers I through III. Thick (70-120 microns) sections treated with the immunoperoxidase method to achieve a Golgi-like staining pattern showed that cytological differences existed between SOM- and VIP-positive neurons. SOM-LI neurons were usually multipolar, fusiform, or occasionally pyramidal while VIP-LI neurons were usually bipolar, stellate, or fusiform. SOM-LI and VIP-LI axons and preterminal processes were differentially distributed within the laminae of the retrohippocampal region. VIP-LI terminals were found throughout all layers except layer I. SOM-LI terminals were found primarily in the molecular layers of all areas, layer IV of the medical and lateral entorhinal areas, and in the angular bundle. Thus, SOM-LI and VIP-LI neurons are distinguished by their morphology and their different distribution within the cortical layers and areas of the retrohippocampal region.

Intramitochondrial ATP and cell functions: yeast cells depleted of intramitochondrial ATP lose the ability to grow and multiply.

Cells of the yeast Saccharomyces cerevisiae could be depleted of their intramitochondrial ATP bu culturing on glucose in the presence of antimycin A, which prevents production of ATP in mitochondria, along with bongkrekic acid, which prevents transport of ATP from the cytosol into mitochondria. Alternatively, the depletion could be achieved by culturing respiration-deficient mutants in the presence of bongkrekic acid. The depleted cells of the respiration-deficient mutant did not grow on glucose in a synthetic medium and growth for a few generations was made possible by adding peptone, yeast extract or some amino acids into the medium. The depleted cells did not differ from control cells in their content of amino acids, proteins, nucleic acids and major phospholipids and had preserved the ability to carry on protein and nucleic acid syntheses and to mate to other cells. No conspicuous cytological differences were found between the control and depleted cells. After culturing in a semi-synthetic medium in the presence of bongkrekic acid the cells of the respiration-deficient mutant exhibited almost no cytochrome c in their spectra and their azide-sensitive ATPase activity was drastically reduced. The results suggest that intramitochondrial syntheses of some low-molecular compounds as well as import and/or assembly of some cytoplasmically synthesized mitochondrial proteins into mitochondria may be impaired in cells lacking intramitochondrial ATP and this may be responsible for their inability to grow and multiply.

Heterochromatin and karyotypic differentiation of some neotropical cactus-breeding species of the Drosophila repleta species group

Metaphase chromosomes from neuroblasts of strains from both laboratory stocks and natural populations of D. serido, D. meridionalis, D. borborema and D. buzzatii have been studied using colcemid pretreatment, and air-drying followed by Giemsa staining. The two latter species both show a uniform ‘basic’ metaphase karyotype. D. serido and D. meridionalis, on the other hand, both include a number of different, geographically distinct, metaphase karyotypes involving differences in the major blocks of constitutive heterochromatin present on the sex chromosomes and/or the 6th chromosome (microchromosome). These chromosomal differences are largely due to the acquisition of extra heterochromatin though pericentric inversions appear to be responsible for some of the Y-chromosome variants in D. serido. Moreover, the cytological evidence demonstrates that populations of both species are far from continuous in distribution. The extent to which such cytological differences reflect the existence of subspecific or specific complexes with minimal morphological differentiation is under investigation.

Immunohistochemical identification of the oxytocin and vasopressin neurons in the hypothalamus of the monkey (Macaca fuscata).

The hypothalamic oxytocin and vasopressin neurons of the monkey, Macaca fuscata, were demonstrated in Golgi-like images by a modified immunoperoxidase method. The magnocellular oxytocin and vasopressin neurons were distributed mainly in the supraoptic and paraventricular nuclei. In addition to these main nuclei, both types of magnocellular neurons were found in the accessory supraoptic nucleus, the periventricular and perifornical areas, the nucleus of the stria terminalis, the lateral hypothalamic area, and the pars interna of the globus pallidus. Magnocellular oxytocin neurons were seen immediately ventral to the anterior commissure, and parvocellular vasopressin neurons were localized in the medial portion of the suprachiasmatic nucleus. The preferential distribution of the oxytocin and vasopressin neurons was recognized not only in the supraoptic and paraventricular nuclei, but also in other areas. In all areas observed, the cytological difference between the oxytocin and vasopressin neurons could be identified. The area of the perikarya of the vasopressin neurons was determined to be larger than that of the oxytocin neurons. Most of the axons of the oxytocin neurons issued from the perikarya, while the axons of the vasopressin neurons originated in most cases from the thick proximal dendrites. These results show that the oxytocin and vasopressin neurons are distributed in areas much broader than has hitherto been assumed, and that these two peptidergic neurons can be definitely differentiated morphologically as well as functionally.

Relationships between morphology and viral etiology in mammary tumors of BALB/cfC3H and BALB/cfRIII mice.

The morphologic pattern of mammary tumors induced by milk-transmitted C3H and RIII murine mammary tumor viruses (MuMTVs) in BALB/c hosts infected by foster nursing were investigated further an compared taking into account several tumor and host variables. Comparison of BALB/cfC3H and BALB/cfRIII mammary tumors under identical tumor and host conditions showed highly significant differences in subgross morphology and histologic structure. Differences in cytology were also observed. Data suggest that mammary tumor morphology in high cancer strain mice is controlled by the causative MuMTV.

Functional morphology of the midgut of a sandfly as compared to other hematophagous nematocera

The midgut epithelium of female Lutzomyia longipalpis was investigated by means of electron microscopic morphometry before and during blood digestion. Ultrastructure and cytological changes of the stomach cells upon blood feeding were generally similar to the ones described for Phlebotomus longipes (Gemetchu, 1974) and for mosquitoes (Hecker, 1977), In addition, the quantitative composition of the cells resembled the one of mosquitoes in many respects. Despite some morphological differences in the functional gut cytology, it can be admitted that, in general, digestive processes may run similarly in the midguts of sandflies and mosquitoes.

Tracheobronchial Cytologic Changes during Prolonged Cannulation

The tracheobronchial cytology of eighteen patients undergoing prolonged mechanical ventilation were examined for the type of cell population present, together with the morphologic integrity of the ciliated epithelial cells 1, 3, 5,10, and 20 days after tracheal cannulation. Smears obtained from patients recovering from cardiac surgery were different from all others in that: (1) they contained fewer epithelial cells (ciliated and squamous) which disappeared earlier; (2) they initially had more pus cells which appeared earlier; (3) they originally contained more histocytes but these were not found after the 5th day after cannulation; and (4) their ciliated epithelial cells displayed more cytomorphologic changes. Differences in cytology were attributed to impaired cardiac output in these patients. The reduced perfusion pressure of the vasculature of their tracheobronchial tree was presumably unable to sustain the lateral wall pressure of tube cuffs pressure, which further impaired tracheal perfusion. Less reparative processes (squamous metaplasia and early appearance of histiocytes) were also found in patients following cardiac surgery, together with early invasion of smears by pus cells indicating premature lowered resistance to bacterial infection.

Pollen tube development in Petunia hybrida following compatible and incompatible intraspecific matings.

Pollen tubes formed following compatible and incompatible intraspecific matings in Petunia have been examined with light and electron microscopes. Compatible and incompatible tubes develop in an identical fashion on the stigma but, on entry into the top 1 mm of the stylar transmitting tissue changes occur both to the cytology of the tubes and their rates of growth. The early cytological changes are common to tubes of both compatibilities but, although both types of tube accelerate on entry into the style, incompatible tubes grow more slowly than compatible. Cytological differences became apparent between compatible and incompatible tubes following a short period of growth in the style, the latter possessing thicker cell walls and a cytoplasm packed with both organelles and reserves. Incompatible tubes subsequently burst or simply cease growth and die. The characteristic image afforded by this cytoplasm resembles that or burst or dead compatible tubes, except in that proportions of the cell components may differ. These data are discussed in terms of current models proposed to explain pollen tube growth and the operation of the self-incompatibility response in Petunia.

About the Cytological Difference between the Abnormal Cells of Mild Dysplasia and those of benign Atypia

当科において昭和53年の1年間に採取した細胞診検体は19, 412例であり, このうちciass IIIaの判定を得たのは175例であった. これらの組織学的背景は良性から上皮内癌に至るまで種々である. 今回わたくしどもは組織診によりmild dysplasiaと確認された群20例 (MD群とする) と, chronic cervicitis等の良性病変しか確認できなかった群20例 (non-MD群) との標本を再検討し, 細胞分化度, 細胞質所見, 核形, 核内クロマチン像に関して分類し, その形態学的特異点について比較検討した.さらにMD群, non-MD群について各症例ごとに, 数回のclass IIIa判定を得た標本をすべて通して検討し, その細胞形態学的傾向を研究した. その結果は次のごとくである。1. 頻回に“class IIIa”の判定が下される場合には, dysplasiaの存在を強く疑い, 逆に1回きりの判定の場合にはdysplasiaの可能性はうすい.2. 異常細胞が多数出現する場合には核所見に注目し, 特にN/C比大, 円形腫大かつクロマチン顆粒状多染を示す異常細胞の割合が全異常細胞中30%以上を占める場合にはdysplasiaの存在を疑い, 逆に5%以下の場合にはdyspiasiaの可能性はうすい.3. mild dysplasiaは一般に標本上の異常細胞数が少ないため判定困難な場合が多いが, 上記所見 (1., 2.) のほかに, follow up中に出現した異常細胞を動的にとらえ異常細胞数の総和を一連の流れとし, われわれのパターン認識方式としてとらえることにより, MD群とnon-MD群との識別がかなり可能となる.われわれが細胞診においてdysplasiaの存在を確認できるか否かは悩むところであるが, 今回得た結果により, 今後さらに細胞診判定基準において役立つならば幸いである.

Denervation study of synapses of organ of Corti of old world monkeys.

Fine structural characteristics of synapses in the spiral organ of Corti were examined, with reference to differences between inner and outer hair-cell systems, and to location of neurons of origin of efferent axons. Surgical interruption of crossed olivocochlear bundle, of vestibular nerve, of facial nerve, and excision of superior cervical ganglia were used to determine the pathways of different axons. Interruption of the vestibular nerve near the brainstem results in degeneration of all efferent terminals on outer hair cells. Mid-line lesions at, and caudal to, the facial colliculus result in degeneration of about half of these efferent terminals. Efferent synaptic bulbs to the inner hair-cell system are small, of the order of one micron, and form type 2 junctions with afferent dendrites. They tend to have more large dense-core vesicles (about 80 nm) than the large efferent terminals of the outer hair-cell system, and appear to be the terminals of axons in the habenula perforata, which exhibit varicosities laden with large dense core vesicles. The varicosities are unaffected by excision of the superior cervical ganglia. So far as our material can reveal, it appears that the varicosities in the habenula perforata do not survive vestibular root interruption, nor do the efferent processes in the internal spiral bundle or at the base of inner hair cells. Most interestingly, the afferent processes of the inner hair-cell system, as identified for example by their relation to pre-synaptic bodies in the inner hair cells, are subject to a trans-synaptic reaction after severance of the vestibular root. They undergo a dramatic cytological transformation, characterized by increase of volume, engorgement with microtubules, microfilaments, microvesicles of various sizes, and clusters of lysosomes. Thus, both the efferent and afferent terminals of the inner hair-cell system show marked cytological differences from the corresponding terminals of the outer hair cell system.

Fine structure of the unistellate sperm of the shrimp, Sicyonia ingentis (Natantia)

Sperm of the prawn Sicyonia ingentis were studied cytochemically and ultrastructurally. Striking cytological differences were noted between these natantian sperm and previously studied reptantian sperm. In general, the S. ingentis sperm are composed of a spherical main body that is partially encompassed by a morphologically diverse cap region, from which extends a single appendage or spike. The main body houses an uncondensed, Feulgen-positive nuclear region that is partially surrounded by a cytoplasmic band. A single layer of small, 600 Å, vesicles lines the periphery of the cytoplasmic band. Large membranous vesicles extend from the inner surface of the cytoplasmic band into the nuclear region. The nucleus is separated from the cap or acrosomal complex by a dense plate and a highly organized crystalline lattice, which is composed of geometric squares that are approximately 350 Å in dimension. The cap region also contains convoluted membrane pouches; a central granular core; spherical bodies; an electron-dense, saucer-shaped plate; and a large anterior granule. The convoluted membrane pouches and anterior granule are periodic acid-Schiff (PAS) positive. The anterior granule also demonstrates RNAase-stable red fluorescence with acridine orange staining. A spiralled spike, approximately 6 μm long, extends from the anterior end of the cap. The cap and spike are bound by a double membrane, which results from the fusion of the plasma membrane and the convoluted pouch membrane. The sperm's acrosome is thought to be composed of the two PAS-positive cap components and the spike.