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Biomedical subjects

M Anniko

Publications and source records attributed to M Anniko.

At least 127 records · Page 7Linked to original sources

Developmental morphology of the mouse inner ear. A scanning electron microscopic observation.

The developing inner ears of mice (CBA/CBA), ages ranging from gestational day 12 through postnatal day 21, were examined using scanning electron microscopy following the epoxy-embedding/freeze-fracture technique. This technique provides unique three-dimensional views of surface and fractured structures of the developing inner ear, thus allowing excellent preservation of the relationships between the developing sensory epithelium and the overlying membranes (i.e. the tectorial membrane and cupula) during their development. The tectorial membrane is formed of two distinct parts: the major (medial) and the minor (distal). The major portion is produced by the cells of the greater epithelial ridge prior to the formation of the minor part, which is produced largely by the primordial supporting cells of the lesser epithelial ridge. The developing tectorial membrane has two types of fibers: radial (found mainly in the major part) and slanted (found mainly in the minor part). The slanted fibers become the cover net, which fuses with the marginal band. The marginal zone of the developing tectorial membrane is completely sealed during development by the third row of Deiters' cells. The surfaces of cells that produce the tectorial membrane are characterized by numerous long microvilli which are largely lost when the tectorial membrane completely forms and separates from the supporting cells. The surface of developing auditory sensory cells is initially covered with numerous microvilli, some of which become future stereocilia. Stereocilia form stepped rows in the shape of a "W", with the tallest row located at the periphery of the cell. As the sensory cell matures, the short transitional stereocilia gradually disappear. Kinocilia on hair cells are still seen in the 14-day-old mouse (even though the organ of Corti is morphologically mature) but not in the 21-day-old mouse, indicating that complete maturation of the sensory cells in all turns is attained by 21 days of age. The mouse has upper radial tunnel fibers and basal tunnel fibers. Neural contacts of the upper radial tunnel fibers with the outer hair cells at the apical portions are frequent in the developing organ of Corti. The external sulcus cells undergo drastic changes during development, forming numerous pits that are often covered with mucus-like droplets or grape-like spherical structures of varying sizes. This phenomenon was observed only during postnatal days 6 and 14. The developing cupula starts as a thin amorphous membrane, which later becomes compact and fibrotic-like as the mass increases. By the 6th postnatal day well-developed cupular canals occur.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Applications of scanning electron microscopy and X-ray microanalysis in inner ear pathology.

Surface pathology of inner ear structures so far described in detail concern cochlear and vestibular hair cells and the stria vascularis. In man, surgical intervention into the inner ear is very uncommon and when performed is in general with the primary objective of destroying the diseased peripheral end organs. The vast majority of inner ear tissue available for use with scanning electron microscopy (SEM) is therefore obtained from animals. The present paper reviews the progression of surface pathology caused by aminoglycoside antibiotics, acoustic overstimulation and in a guinea pig strain with genetic inner ear disease. The primary site of onset of surface pathology differs, depending on the underlying cause. Advanced surface pathology shows a similar type of morphological degeneration independent of cause. The combination of SEM and energy dispersive X-ray microanalysis (XRMA) of inner ear pathology has as yet been reported in only three studies, all concerning inner ear fluids or otoconia.

Aminoglycosides↗

Freeze-drying and related preparation techniques for biological microprobe analysis.

An X-ray microanalytical and morphological investigation has been carried out on rapidly frozen, freeze-dried or freeze-substituted tissues. A comparison was made between different embedding and polymerisation procedures following freeze-substitution and freeze-drying. The investigation also included an analysis of specimens infiltrated, embedded and polymerised by ultraviolet irradiation at low temperatures with Lowicryl HM20. The morphological preservation of Lowicryl embedded tissue was adequate for the identification of different cell structures like nuclei, mitochondria, lysosomes and different types of endoplasmic reticulum. X-ray microanalytical investigation of low temperature embedded material displayed an elemental composition of cells and organelles similar to that found in freeze-dried cyosections. Compared with freeze-dried cryosections, low temperature embedded material could be sectioned for light microscopy and area of interest chosen for further thin sectioning. This is of great importance in work with tissues with complicated morphology and heterogenous cell populations.

Animals↗

DNA ploidy and cell phase in human pituitary tumors.

The flow-cytofluorometric technique for nuclear DNA analysis was used in the study of 47 consecutive cases with pituitary tumors. The material consisted of 18 tumors secreting growth hormone (GH), 10 secreting GH and prolactin (PRL), 10 prolactinomas, 2 secreting thyroid-stimulating hormone (TSH), 2 secreting ACTH, and 5 were nonsecreting. An aneuploid DNA pattern occurred in 23 of 47 (49%) cases ranging from 1.4 to 3.6 N thus deflecting from the normal diploid 2N chromosome content of somatic nonreproductive cells. In four tumors two cell lines occurred, but one of them was always dominant, comprising 80% to 90% of all cells analyzed which totally amounted to 20,000 to 30,000 cells. The highest percentage of aneuploid tumors occurred in those with a concomitant secretion of GH and PRL (8 of 10, 80%). The percentage of diploid cells in tumors with an aneuploid nuclear DNA content was very low, 2.3 +/- 2.9% (+/- 0.65% SE). In 11 cases with aneuploidy, diploid cells were not detected. The mean age in patients with diploid and aneuploid tumors was rather similar in the different endocrinologic types of tumors.

Acromegaly↗

Early development of cochlear hair cell stereociliary surface morphology.

The early development of the surface structures of differentiating cochlear hair cells (guinea-pig) was analysed by scanning electron microscopy (SEM). A basal-to-apical gradient was evident in hair cell maturation. Inner hair cells developed before outer hair cells at the same level in the cochlea. The first sign of the onset of hair cell differentiation was a regularization of the pattern of microvilli on the future hair cell. Later, the cluster of regularized microvilli was rebuilt to form the stereociliary bundle, with a stepwise increase in the length of those stereocilia facing the stria vascularis.

Animals↗

A unique striated muscle: further morphological and x-ray microanalytical investigation of the stapedius muscle of the guinea pig using thin and thick cryosections.

Thin (130-200 nm) and thick (10-16 micrometers) cryosections of stapedius muscle of guinea pig were examined by X-ray microanalysis in the scanning- and scanning-transmission mode of electron microscopy. To further explore the unusual elemental composition of stapedius muscle and to evaluate previously used method of analysis of thick cryosections cut at -30 degrees C (Wróblewski et al., 1981), thin cryosections cut at -100 degrees C were investigated. The ultrastructural organisation of stapedius muscle with the centrally located nuclei and the mitochondria could be easily recognized in unstained thin cryosections. X-ray microanalytical investigations of thin sections revealed nearly the same elemental composition of single organelles as results obtained on defined areas in thick cryosections. Analysis of mitochondrial matrix. The elements appeared, however, at the same ratio (1:1). The P/Ca ratio was approximately 6:1 in the nuclei. Myofibrils presented low K and higher Ca concentrations. The levels of other elements (P. S. Cl) were close to those found in the quadriceps muscle of the guinea pig, rat, or human.

Animals↗

[The in vitro system in internal ear research].

The knowledge about in vitro culture of the embryonic inner ear dates back to the 1920's. During the following decades it was mainly the avian embryonic labyrinth which was used for organ culture. Emphasis was put on histological differentiation of individual cells and tissues. A poor morphogenesis/organogenesis was accepted. In the 1950's a very important field of research emerged which concerned the morphogenetic relationship between cells and tissues of different types. A great progress in basic understanding of in vitro conditions for normal and tumour tissues from mammalian species including man occurred during the 1960's. In the early 1970's this was applied to research on the cytodifferentiation and morphogenesis of the embryonic mammalian inner ear anlage. The organ culture techniques have been applied to studies mainly on the embryonic development of the mouse inner ear anlage but also to some extent on early postnatal development. In vitro studies on other species are few. The in vitro technique for inner ear studies has been evaluated with regard to morphology and to some extent biochemical parameters as phospholipid composition, the transport enzyme adenylate cyclase, etc. With the introduction of the organ culture technique for the inner ear of the mouse and guinea pig (higher vertebrates) many of the experiments earlier restricted to lower vertebrates performed in vivo could now be performed with great accuracy in vitro using higher vertebrates and under considerably better controlled conditions: neural induction, epithelio-mesenchymal interactions, neurotrophic interaction, "fate mapping", tissue interactions for normal morphogenesis and studies on especially vulnerable stages during inner ear development and applied research as e.g. ototoxicity. It must be emphasised that applications of the organ culture technique cannot give information on the systemic effects of an agent, only about its direct action on a given tissue.

Animals↗

The freeze fracture technique in inner ear research.

Freeze-fracture studies on the inner ear have been focused mainly on the normal structure of junctions sealing the endolymphatic compartment, the compartmentalization of the stria vascularis and the junctional stability of the hair cells towards adjacent supporting cells. The hair cells have a very tight type of zonulae occludenetes as compared with other non-sensory epithelia in the inner ear. In contrast to other epithelial cells, the mature hair cells are in most species lacking gap junctions. During embryonic development a loss of gap junctions is an early and significant feature of cells differentiating into future hair cells. The tight junctions in the secretory epithelia (stria vascularis in the cochlea and dark cells around vestibular organs) are morphologically mature before the onset of the ionic maturation of endolymph. Freeze-fracture studies on inner ear pathology are few. The structural alterations of tight junctions in the diseased inner ear are minimal. The functional significance of such small morphological derangements is not known.

Animals↗

Inductive tissue interactions during inner ear development.

Inductive tissue interactions were studied in the otocyst of the CBA/CBA mouse. Otocysts with surrounding mesenchyme explanted at the 12.5-13th gestational day and cultured in vitro for 4 days underwent morphogenesis with formation of semicircular canals, vestibular organs, and some cochlear coiling. Without their surrounding mesenchyme only little, if any, development was seen. However, otocysts without mesenchyme but grown in a medium precultured with mesenchyme did develop normally. A soluble and diffusible induction factor is apparently produced by mesenchyme and also by other fetal organs. Cell-cell contact may not be needed for induction.

Animals↗

Organ culture of the crista ampullaris of the embryonic guinea pig.

The embryonic inner ear of the guinea pig was cultured one to ten days in vitro. The explantation occurred on the 30th and 40th gestational days, respectively. Hair cells were found in various stages of cytodifferentiation. Both hair cells and supporting cells were ultrastructurally preserved at the end of the period in vitro. The gross morphologic features of the crista ampullaris were still immature. The embryonic inner ear of the guinea pig can be maintained in organ culture until at least as late as the 46th gestational day.

Aging↗

Postnatal maturation of cochlear sensory hairs in the mouse.

The surface morphology of inner hair cells in the cochlea of the mouse is fully developed at birth. At this time a regular pattern of elongated microvilli covers the surfaces of outer hair cells at all levels of the cochlea. The sensory hair on outer hair cells undergo a morphological maturation during the first days postnatally. A mature surface morphology is reached 5-7 days after birth. There are two gradients in hair cell maturation in the cochlea: (1) basal to apical, and (2) from inner hair cells to the 3rd row of outer hair cells. Polarization of sensory hairs on an outer hair cell, i.e. polar differentiation of the sensory hairs in the radial direction, occurs as a stepwise increase in length of those facing the stria vascularis.

Animals↗

Aggressive pituitary tumor growth.

Three cases with extrasellar extension from our material of 132 pituitary tumors are reported. One tumor secreted growth hormone, one secreted prolactin, and one secreted adrenocorticotropic hormone (ACTH). All three patients had a short case history. The tumors recurred rapidly after surgery. Light microscopy showed pleomorphic cells but no signs of morphologic malignancy. Ultrastructural analysis revealed intracellular filamentlike inclusions, with a cross-banded substructure in two cases. The DNA analysis of two of the cases showed aneuploidy. Our three cases of pituitary tumors are likely to represent a particularly aggressive type of tumor growth.

Adenoma, Chromophobe↗

Gamma irradiation effects on human ACTH-producing pituitary tumors in organ culture.

Morphology (studied by light and electron microscopy) and hormone secretion were followed in organ cultures of adrenocorticotropic hormone (ACTH)-secreting hormones after in vitro irradiation with a single dose of 70 Gy. The increase is ACTH secretion immediately after exposure was interpreted as an irradiation-induced direct effect on cell membrane permeability. The low secretional capacity for 120 h after irradiation is probably caused by an impairment of cell metabolism. Cell morphology slightly deteriorated 24-48 h after exposure. Later, cell morphology was restored and specimens investigated 5 days after irradiation showed normal morphology. It is concluded, that the fluctuations in serum ACTH in patients with Cushing's disease treated by stereotactic radiosurgery are caused by changes in the tumor cells proper rather than being due to hypothalamic influence.

Adrenocorticotropic Hormone↗

Human pituitary tumors with two cell lines.

From the total material of 47 pituitary tumors, 42 had one cell line and five had two cell lines. The latter were found in tumors causing acromegaly either with secretion of growth hormone (GH) alone (n = 2) or in combination with prolactin (PRL) (n = 3). In tumors secreting only GH, biclonal cell lines occurred in four of 20 (20%) of the total number of cell lines. In tumors with concomitant secretion of GH and PRL, the corresponding proportion was six of 13 (46%). In only one case was it possible to identify two separate cell populations morphologically.

Adult↗

TSH-secreting pituitary tumor. A case report.

A pituitary tumor secreting thyroid-stimulating hormone (TSH) only was diagnosed in a 43-year-old man who originally showed signs and symptoms of thyrotoxicosis which recurred twice. Despite normal serum levels of T3 and T4, increased levels of TSH were diagnosed. The pituitary tumor was operated by the transsphenoidal approach. Morphological analysis of tumor tissue showed cellular pleomorphism and regressive changes. Nuclear DNA analysis showed that the tumor had only one cell line and an aneuploid DNA pattern. In vitro culture of tumor tissue confirmed that the tumor secreted TSH only.

Adult↗

Oestrogen and testosterone effects on hormone secretion and cell morphology of human pituitary tumours.

Estradurin (a polymer of 17-beta-oestradiol) has an inhibitory effect on the synthesis/secretion of PRL. This effect is assumed to be caused by the strong inhibitory effect of the drug on phosphatases. With increasing time of low doses of Estradurin (0.001 micrograms/ml) in vitro, the inhibitory effect on PRL secretion was overcome and the synthesis/secretion of PRL increased. The secretion of GH was unaffected in all concentrations (0.001-0.1 micrograms/ml) except 1 microgram/ml. A stimulatory effect of 17-beta-oestradiol (0.01 micrograms/ml) on the synthesis/secretion of PRL was suggested by the tissue from one post-pregnancy pituitary. Short time organ cultures from prolactinomas or tumors with a concomitant secretion of GH and PRL show no changes of GH or PRL secretion following incubation with (0.001-0.1 micrograms/ml) of 17-beta-oestradiol. However, electron microscopy of 17-beta-oestradiol incubated specimens revealed an increased number of lysosomes and inclusion bodies in the cell cytoplasm. Incubation with testosterone (0.001-1 microgram/ml) cause inhibition of PRL synthesis/secretion in two of three prolactinomas. In GH secreting tumours testosterone did not effect PRL or GH secretion in vitro when compared with controls. The ultrastructural analysis of specimens in which a decrease of PRL synthesis/secretion had occurred showed myelin figures in the cell cytoplasm and accumulations of amorphous electron dense inclusion bodies.

Adult↗

Cellular localization of adenylate cyclase in the developing and mature inner ear of the mouse.

The localization of adenylate cyclase in the developing and mature inner ear of the CBA/CBA mouse' in in Shaker-1 and Shaker-2 mutants and in organ culture was determined by a histochemical procedure with adenylyl imidodiphosphate as substrate and Sr2+ as capture ion. Enzymatic activity was associated with cell membranes in stria vascularis and Reissner's membrane but could not be demonstrated in other cochlear structures. In stria vascularis, adenylate cyclase was found on the contraluminal infoldings of marginal cells but not on their luminal surface, while Reissner's membrane showed activity on its endolymphatic surface. Activity was also associated with utricular dark cells. Essentially the same localization but differing in quantitative aspects was observed during development, in organ culture and in the mutant strains. The speculative role of adenylate cyclase in cochlear ion and fluid balance is discussed.

Adenylyl Cyclases↗