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Biomedical subjects

R Saxod

Publications and source records attributed to R Saxod.

At least 37 records · Page 2Linked to original sources
PubMed1991

Morphometry of human nerve biopsies by means of automated cytometry: assessment with reference to ultrastructural analysis.

The morphometric analysis of myelinated fibres is a tool for diagnosis of neuropathies and for assessing nerve regeneration after reconstructive surgery. In order to obtain reliable information, sampling techniques should be avoided and the measurement of all the fibres within a nerve fascicle requiring the use of automated cytometry is necessary. We have developed a programme on the SAMBA cytological image analyser for the automated measurement of myelinated fibres. Different techniques of segmentation were tested and adaptive grey level thresholding gave the more reliable result. Accuracy and reproducibility were tested using a set of five human superficial peroneal nerve biopsies that were previously analysed by means of semi-automatic ultrastructural morphometry. A maximum difference of 9% in the number of fibres counted was obtained when comparing the automatic and the semi-automatic methods. In all cases the histograms of the morphometrical variables (fibre and axon diameters, myelin sheath thickness) were found to be identical to the reference histograms obtained by the semi-automatic method.

Evaluation Studies as Topic↗
PubMed1990

Effects of tunicamycin on the avoidance reaction of epidermis by sensory neurites in co-cultures.

In 7-day chick embryo dorsal root ganglia and epidermis or dermis co-cultures, nerve fibres establish contacts with dermis while avoiding epidermis. Previous results have indicated that factor(s) secreted by epidermis could be involved in this avoidance reaction. The present study demonstrates that the avoidance reaction is abolished when epidermal cells are treated by the N-linked glycoproteins synthesis inhibitor, tunicamycin. The same result is obtained after monensin treatment. The epidermal cell viability, development and total protein secretion are not significantly affected by tunicamycin, as demonstrated by trypan blue exclusion, electron microscopy and SDS-PAGE electrophoresis after 35S-methionine labelling. It has thus been concluded that the avoidance factor is glycoproteic in nature. It is also suggested that this factor possibly contains chondroitin-6-sulphate moieties.

Animals↗
PubMed1989

There is no simple adequate sampling scheme for estimating the myelinated fibre size distribution in human peripheral nerve: a statistical ultrastructural study.

Morphometric studies of peripheral nerves (PN) usually involve some sampling of the myelinated fibres (MF). In order to scrutinize the statistical properties of the sampling processes in common use and the reliability of the resulting estimates, a quantitative analysis of human superficial peroneal nerves from 8 different normal subjects was undertaken at the ultrastructural level, both in terms of MF spatial distribution and of their size distribution. This study used sampling rates involving more than 10% of the whole myelinated fibre population observed in each nerve fascicle. However, in nearly all the fascicles evaluated, the sampling fluctuations are so high that neither the number of axons nor their diameter distribution can be assessed with enough accuracy. A systematic study of the myelinated fibres shows that the spatial distribution of their size is not uniform. This marked heterogeneity in the MF size distribution imposes measurement of large enough samples (500 or 600 MFs usually represent about one-half or two-thirds of the whole MF population) in a way to secure a reliable enough estimate of the density and size distributions. However, the practical usefulness of sampling schemes requiring more than one-half of the whole MF population in a nerve fascicle, is questionable.

Adult↗
PubMed1989

Influence of glycosaminoglycans on neurite morphology and outgrowth patterns in vitro.

The neuritic growth patterns obtained on substrates made of several glycosaminoglycans (GAGs) bound to type I collagen were analysed and compared in primary cultures of chick embryo dorsal root ganglion grown in serum-free supplemented medium. In 2-day cultures grown on type I collagen or heparan sulphate (HS)-collagen surfaces, ganglionic explants exhibit a dense, symmetrical network of long, parallel neuritic processes and very few flat migrating non-neuronal cells. In contrast, on either dermatan sulphate (DS), chondroitin-6-sulphate (C6S) or hyaluronic acid (HA)-bound collagen substrates, neurons form irregular nerve fibre patterns; indeed, neurites follow convoluted paths and often, after abrupt turns, totally reverse their direction of extension. Experiments were carried out in which a choice was given to growing neural processes between collagen or GAG-collagen substrates. While growth cones elongating over type I collagen easily cross the border with HS-bound collagen surface and indiscriminately extend on this substrate, in contrast, neurites generally avoid surfaces coated with DS, C6S or HA and change their direction of growth in order to stay on collagen. The binding of DS, C6S or HA, but not HS, to type I collagen thus decreases its ability to promote neurite elongation. The interaction of neuronal cells with these extracellular matrix components by restricting neurites in their paths of extension may, therefore, play a role in the patterning of the nervous circuitry.

Animals↗
PubMed1989

Automated morphometric study of human peripheral nerves by image analysis.

In this paper we describe a program using the image analyzer SAMBA, which allows an automatic analysis of silver stained semithin nerve sections. The operator can interactively delimit the contour of the fascicle to be analysed by means of a digitizing tablet connected to the system which generates a mask of the region. Segmentation of the fibre images is conducted as a function of brightness threshold defined by the operator. Fibre clusters are automatically separated using morphological procedures like dilatation. Morphometric parameters such as the external and axonal diameters, myelin sheath thickness and circularity are measured. We are now testing this method on normal and pathological human superficial peroneal nerves. Preliminary results are promising and the development of adequate statistical analysis of morphometric data will provide us with a new tool for the diagnosis of peripheral neuropathies.

Humans↗
PubMed1989

Differentiation of pure chick embryo epidermis grown in primary serum-free culture.

The differentiation of precocious embryonic epidermis in serum-free primary culture was analyzed by light and electron microscopic methods. Explants of 7-day chick embryo epidermis were grown on collagen or poly-L-lysine substrates in the absence of dermal mesenchyme. The serum substitute consisted of a mixture of insulin, transferrin, putrescine and seleneous acid together with (or without) Nerve Growth Factor. These culture conditions were shown to support proliferation, growth and development (evaluated using morphological criteria) of the epidermal explants up to 4-5 days; during this period, the epidermis underwent stratification; well-developed desmosomes as well as tonofilaments were formed and the epidermis achieved a morphology close to that of 10-11 day epidermis in ovo. However long-term survival of the explants was not obtained as cellular death, starting on day 5, progressively led to the necrosis of most parts of the explant. This morphological study demonstrates that the early phases of epidermal growth and maturation can occur to some extent in the virtual absence of dermal elements and serum factors. Chick embryo epidermal cells may thus possess the intrinsic ability to go through, at least for short periods in vitro, their differentiation programme. Then, at the onset of epidermal keratinization (12 days in ovo), they require specific exogenous factors to fully differentiate in vitro.

Animals↗
PubMed1988

Schwann cell proliferation in the sciatic nerve of hypothyroid chick embryos studied by autoradiography and image analysis.

The proliferation of Schwann cells in the sciatic nerve of chick was studied from day 11 to day 27 of development in control and thyroid-deficient embryos. Hypothyroidism was induced by tetramethylthiourea injection on days 8 and 19 of incubation. The parameters of the cell cycle were determined using autoradiographs (tritiated thymidine) and by image analysis of Feulgen-stained nuclear smears. The duration of the cell cycle was lengthened and the growth fraction was reduced in hypothyroid animals, at 11 and 15 days of incubation. At later stages (days 21 and 27), these parameters were not significantly different from the controls as if the sensitivity of Schwann cells to thyroid hormones was scheduled to occur during a limited period of development. The total number of axons was the same in control and hypothyroid animals suggesting that the slowing down of Schwann cell proliferation is not a consequence of neuronal cell death. The consequence of that slowing down is a delay in the isolation of promyelin axons and a reduction in the proportion of myelinated axons at all the stages studied.

Animals↗
PubMed1988

Substrate effects on the dynamics of neurite growth in vitro: a quantitative multi-parametric analysis.

Embryonic chick dorsal root ganglia were cultured in serum-free medium on natural (collagen, fibronectin and hyaluronic acid) and artificial (polylysine and polyornithine) substrata. The movement of individual growth cones was quantified by measuring five parameters using time-lapse cinematography combined with a digitizing-computer system, and the neurite behaviour was compared between the different substrata with multivariate statistical methods. For each substratum, the morphometry of the growth cone was quantified by measuring six morphological parameters. The most discriminative parameters proved to be mean velocity and straightness index for neurite extension, and projected area and cumulated length of filopodia for growth cone morphometry. A good correlation was obtained between behavioural and morphological parameters and the larger the cone area and the filopodia length, the faster and the straighter the neuritic growth. Both quantitative analyses showed highest values for polyornithine and the lowest for hyaluronic acid, and divided the substrata into two opposite groups, artificial and natural. It is concluded that growth cone behaviour and conformation is modulated by substratum properties.

Animals↗
PubMed1986

Differential growth of sensory neurons in vitro in presence of dermis and epidermis. A quantitative time-lapse analysis.

The influence of dermal and epidermal cells on the growth of nerve fibres from chick embryo sensory neurons was investigated in vitro. A previous quantitative analysis showed that the growth of nerve fibres is profoundly modified in the close vicinity of epidermis. This change is mainly characterized by erratic trajectories of nerve fibres resulting from numerous lateral displacements of the growth cones. In contrast, no such behaviour is observed far away from the epidermis or in the presence of dermis. In this latter case, neurites exhibit a straighter direction of extension. These observations suggest that the epidermis exerts some kind of control on the establishment of nerve fibre pattern in the dermis.

Animals↗
PubMed1985

The density of myelinated fibres is related to the fascicle diameter in human superficial peroneal nerve. Statistical study of 41 normal samples.

The density of myelinated fibres in the superficial peroneal nerve was studied in 41 samples from 24 control human subjects. Photographic montages of the whole nerve fascicle were made from semithin and ultrathin transverse sections and used for a statistical analysis of sampling procedures, range of variations and relations between density and other variables. The results indicate that the spatial distribution of myelinated fibres within a nerve is often non-uniform. Therefore, it was not possible to define a statistically valid sampling system. The study of relations between variables shows the lack of any correlation between density and age and a considerable variation in the density. In contrast, there is a strong positive linear correlation between the surface area of the nerve fascicle and its content of myelinated fibres. That is, the fibre density of a given normal nerve is related to its diameter and can be predicted within a narrow range of error. We propose the term "derived density" for this value, and its application as a tool in the diagnosis of peripheral neuropathies is now being studied.

Adolescent↗
PubMed1983

[Quantitative microcinematographic analysis of interactions between sensory nerve cells and cutaneous tissues in vitro: demonstration of a difference in the behavior of nerve fibers with respect to dermal and epidermal cells].

Dynamic and morphometric analysis of interactions between sensory nerve cells and cutaneous tissue of 7 day Chick embryos in co-culture show that nerve endings behave differently when interacting with dermis of epidermis. In particular, nerve fibers display a deviation reaction in the proximity of epidermis.

Animals↗
PubMed1979

[Development of cutaneous innervation in the chick: ultrastructural and quantitative analysis (author's transl)].

In the chick, at the thoracic level, the dorsal branches of spinal nerves form at 4 days of incubation (stage 25) and reach the skin between 5 1/2 and 6 days (stages 28-29). At 6 days, the growing nervous peripheral processes ("axons") form large bundles (200-1,000 fibers). At 10 days, young Schwann cells divide the bundles into groups of axons. The perineurium and endoneurium differentiate between 10 and 14 days (but epineurium is formed after hatching). At 14 days of incubation, the adult pattern of cutaneous innervation is established. At this same stage, myelogenesis begins but develops mainly after hatching : 1% of the axons is myelinated at 16 days of incubation, 4% at hatching, 40% in 6-week old chickens and 60% in adults. Thus, less than 10% of myelinated axons of the adult are already myelinated at hatching. Two modes of myelogenesis were observed: 1) early myelination, starting in the embryo around axons measuring about 1 micrometer in diameter: 2) delayed myelination, occurring in the older chickens after an increase in axon diameter. These observations suggest that there is, in the development of chick skin innervation, a critical stage (14-15 days of incubation) apparently corresponding to the stabilization of cutaneous nerve supply.

Animals↗
PubMed1978

[Formation of cutaneous nerve trunks in the chick. Ultrastructural and quantitative analysis].

In chick skin structuration of nervous trunks takes place at 14-15 days of incubation, at the time of formation of the adult pattern of cutaneous innervation. At this same stage, myelogenesis begins, but develops mainly after hatching: only 4% of axons are myelinated at hatching, 40% in 6-week old Chickens and 60% in adults. This "critical" stage (14-15 days of incubation) apparently corresponds to the stabilization of cutaneous nerve supply.

Aging↗
PubMed1978

Ultrastructure of Merkel corpuscles and so-called "transitional" cells in the white Leghorn chicken.

In the chicken Merkel corpuscles are located in the dermis and consist of specialized Merkel cells, discoid nerve endings and lamellar cells. Merkel cells contain characteristic membrane-bound dense-core granules and bundles of microfilaments. Asymmetric junctions, synapse like, with thickened membranes and clusters of dense-core vesicles were observed between the Merkel cells and the nerve endings. The nerve ending is derived from myelinated nerves and sometimes contains clusters of clear vesicles. A laminar system formed by lamellar cells of the Schwann cell type encloses the Merkel cells and the nerve endings. So called "transitional" cells, showing some of the morphological features of both keratinocytes and Merkel cells, were observed in the basal layer of the epidermis. One was located partly in the epidermis and partly in the dermis. The structure of Merkel corpuscles is compared with that of Merkel cells in other tetrapods. The developmental significance of "transitional" cells and the origin of Merkel cells are discussed.

Animals↗
PubMed1978

Combination of cholinesterase staining of nerves and stereoscopic viewing for three-dimensional study of skin innervation on whole mounts.

A histochemical staining technique for cholinesterases is reported for the visualization of skin innervation on whole mounts in the chick. A method for realization of stereoscopic pictures showing the nerve fiber pattern in the full thickness of preparations is described. The combination of these 2 techniques allows an excellent three-dimensional demonstration of skin innervation.

Animals↗