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Biomedical subjects

N Cuenca

Publications and source records attributed to N Cuenca.

17 recordsLinked to original sources
PubMed1996

Formation and dissolution of spinules and changes in nematosome size require optic nerve integrity in black bass (Micropterus salmoides) retina.

Teleost retinas adapted to light show numerous spinules invaginated in the cone pedicles and small nematosomes in the distal horizontal cells. Darkness induces the dissolution of spinules and the presence of large and numerous nematosomes. The aim of this work is to study the influence of optic nerve integrity on spinule formation/dissolution and changes in nematosome size during light or dark adaptation of black bass (Micropterus salmoides) retinas. Eyes from fish, dark- or light-adapted, were removed and the eyecups placed in oxygenated Ringer's solution and immediately exposed to light or dark, respectively, for 1 h. The number of spinules per pedicle and the nematosome diameter were measured on electron micrographs. Isolation of eyecups in the dark, impaired both spinule formation and nematosome size reduction when they were superfused in light. In the same way, isolation of eyecups in the light, impaired both spinule dissolution and nematosome size increase when they were superfused in dark. No significant differences in spinule number and nematosome size, following dopamine superfusion, were found in comparison to retinas superfused with Ringer's solution only. Our results suggest: (1) optic nerve integrity is necessary to yield spinule formation/disruption and changes in nematosome size during light or dark adaptation. (2) dopamine does not appear to be the primary agent responsible for spinule formation.

Adaptation, Ocular↗
PubMed1995

Substance P-immunoreactive neurons in the human retina.

Substance P (SP) is a neuropeptide that acts as a neurotransmitter or a neuromodulator in the retina. The aim of this study was to identify the type(s) and the distribution of the SP-immunoreactive (SP-IR) cells in the human retina. We have used an antiserum to SP to immunostain neurons in postmortem human retinae. Immunostained retinae were processed with the avidin-biotin complex (ABC) to visualize the cells either whole mounted in glycerol or embedded in plastic. Some retinae were also sectioned at 20 microns in order to obtain radial views of stained cells. SP-IR amacrine cells stain intensely and appear to be of a single type in the human retina. They are large-field cells with large cell bodies (16 microns diameter) lying in normal or displaced positions on either side of the inner plexiform layer (IPL). Their sturdy, spiny, and appendage-bearing dendrites stratify in stratum 3 (S3) of the IPL, where many overlapping, fine dendrites intermingle to form a plexus of stained processes. Either cell bodies or primary dendrites emit an "axon-like" process that, typically, divides into two long, fine processes, which run in opposite directions for hundreds of micrometers in S5 and S3 before disappearing as distinct entities in the stained plexus in S3. Long, fine dendrites also pass from the dendritic plexus to run in S5 and down to the nerve fiber layer to end as large varicosities at blood vessel walls. In addition, fine processes are emitted from the dendritic plexus that runs in S1, and some pass up to the outer plexiform layer (OPL) to run therein for short distances. The SP-IR amacrine cell has many similarities to the thorny, type 2 amacrine cells described from Golgi studies. In addition to the SP-IR amacrine cells, a presumed ganglion cell type is faintly immunoreactive. Its 20-22 microns cell body gives rise to a radiate, sparsely branched, wide-spreading dendritic tree running in S3. Its dendrites and cell body become enveloped by the more intensely SP-IR processes and boutons from the SP-IR amacrine cell type. The SP-IR ganglion cell type most resembles G21 from a Golgi study.

Cell Count↗
PubMed1995

Two types of mitochondria are evidenced by protein kinase C immunoreactivity in the Müller cells of the carp retina.

The localization of protein kinase C (PKC) was studied immunocytochemically in the Müller cells of the carp retina. Electron microscope immunocytochemistry (using a monoclonal antibody to the alpha and beta isoenzymes of PKC) showed PKC-immunoreactivity mainly inside some mitochondria, especially along the mitochondrial cristae whereas other mitochondria in the same Müller cells showed no staining. Despite a detailed analysis we did not find any significant morphological difference between labeled and unlabeled mitochondria. These results demonstrate, for the first time, the presence of PKC immunoreactivity inside mitochondria and suggest that individual mitochondria may differ in signal transduction pathway.

Adenosine Triphosphate↗
PubMed1995

Substance P: a neurotransmitter of amacrine and ganglion cells in the vertebrate retina.

A short history and summary of the occurrence of substance P in the vertebrate body is presented. Substance P is now generally accepted to be a neurotransmitter and can be visualized by immunocytochemistry to occur in various nerve cells in the CNS. In the retina, substance P-immunoreactivity (SP-IR) occurs in amacrine cell populations in all the species so far studied. In some vertebrates retinas SP is also apparent in one or more ganglion cell types. Anatomical investigations have revealed the morphology and connectivity of SP-IR amacrine cells: they branch in several strata of the inner plexiform layer receiving input from bipolar and amacrine cells and making synapses upon bipolar and ganglion cells. Most commonly SP-IR amacrines emit axon-like process that pass to both the outer plexiform layer and the ganglion cell and nerve fiber layers. These processes often end upon the retinal vasculature. SP-IR ganglion cells have been described in turtle, rabbit and human retinas. In turtle, intracellular dye injection has revealed the morphology of one type of SP-IR ganglion cell as being a large-field monostratified cell with a branches in the outer stratum of the inner plexiform layer. It may correspond to a "Dogiel cell" type. Intracellular investigation of SP-IR amacrine cells in turtle reveal their physiological responses to be ON-OFF in nature with some color-coding characteristics. In general SP acts as an excitatory neurotransmitter raising the spontaneous activity level of ganglion cell responses. The SP-IR ganglion cell is an OFF-center unit in the turtle retina and may be driven in the center of its receptive field by luminosity bipolar cells and in its surround by amacrine cells with color-opponent properties.

Animals↗
PubMed1993

Visual experience during postnatal development determines the size of optic nerve axons.

The size of any axon is generally related to an internal programme (probably common for all neurones) that determines the number and organization of its cytoskeletal components. Here we show that axons from visually deprived rats are smaller than controls, although the number of microtubules and neurofilaments remains unchanged. Moreover the distance between both microtubules and neurofilaments and the amounts of 200 kDa, 160 kDa and 68 kDa neurofilament proteins are also diminished in the deprived axons. We suggest that cytoskeletal organization and axonal calibre are not only determined by intrinsic (genomic) factors, but that environmental stimulation is important for normal growth of nerve cells.

Animals↗
PubMed1993

Dendrites of rod dominant ON-bipolar cells are coupled by gap junctions in carp retina.

Gap junctions are supposed to be the anatomical substrate for electrical coupling between neurons. In fish retina, bipolar cells are electrically coupled and their receptive field diameters are always larger than dendritic field size, however, gap junctions have not been described between dendrites of bipolar cells. In this paper, using immunostaining for protein kinase C, we show that every rod dominant ON-bipolar cell is connected with its neighboring dendrites by gap junctions forming a plexus in the outer plexiform layer. These dendritic processes provide the site of electrical coupling. We suggest that dendrites of bipolar cells could be involved in lateral pathways in retina.

Animals↗
PubMed1993

A compiled BASIC program for analysis of spatial point patterns: application to retinal studies.

The pattern of distribution of a population of cells is of considerable interest to biologists and neurobiologists. However, the labor involved in collecting and analyzing the data often requires a significant amount of time. This paper presents a compiled BASIC program written using the Microsoft QuickBasic compiler for Apple Macintosh to facilitate such studies. The program allows collection and analysis of data that can be introduced either with the aid of a digitizing tablet of directly imported as x,y coordinates from different sources as, for example, word processors or image analysis software. Subsequently the program provides a quick, easy and interactive way of access to statistical, mathematical and graphical techniques used in the analysis of spatial point patterns. These techniques include several measures of dispersion (quadrat count, nearest neighbor and a 2-dimensional point autocorrelogram analysis) and arrangement. Although the program has been tested on spatial organization of retinal cells, it can be used to study the distribution of other cells in the nervous system and for different projects, as for example the distribution of microtubules and neurofilaments inside the axons. This software is available from the authors.

Animals↗
PubMed1992

Axon types classified by morphometric and multivariate analysis in the rat optic nerve.

Calibers of the rat optic nerve axons distribute unimodally and it is difficult to distinguish groups among them. However, these fibers arose from 3 types of ganglion cells and showed 3 conduction velocities. Performing a cluster analysis over several ultrastructural parameters we found 3 main groups of fibers. These groups are present in a very similar proportion to the ganglion cells groups described in the rat retina.

Animals↗
PubMed1991

Postembedding immunocytochemistry for GABA and glycine reveals the synaptic relationships of the dopaminergic amacrine cell of the cat retina.

Postembedding electron microscope immunocytochemistry of glycine and GABA conjugated to colloidal gold has been applied to pre-embedded cat retina stained with the antibody against tyrosine hydroxylase (Toh+). Toh+ stained cells are the equivalent of A18 amacrine cells of Golgi descriptions (Kolb et al., '81). The dendrites of Toh+ cells synapse upon several different types of glycine-positive amacrine cell bodies. We suggest that these are the A8, A3/A4, and AII amacrine cell varieties by analogous immunocytochemical staining intensity, to glycine autoradiographic labeling intensity (Pourcho and Goebel, '85). The greatest number of synapses from Toh+ dendrites are directed at the least glycine-positive amacrine, which is the AII cell by all morphological criteria. A few glycine-positive profiles are also presynapatic to the Toh+ stained cell body itself. Toh+ profiles are also presynaptic to GABA-positive amacrine cell bodies. The commonest amacrine synapsed upon is very heavily labeled with GABA immunocytochemistry. We consider it to be the A17 amacrine cell, which is known to label strongly by [3H] muscimol autoradiography (Pourcho and Goebel, '83). The cell body of the Toh+ amacrine cell also receives many synapses, which appear to be GABA-positive, and Toh+ profiles running in stratum 1 of the inner plexiform layer (IPL) are both pre- and postsynaptic to GABA-positive amacrine cell profiles. In addition, the cell body and primary dendrites of the Toh+ cell receive input from a bipolar type and GABA- or glycine-negative profiles. GABA-positive profiles, belonging to the interplexiform cell (IPC), are synapsed upon by Toh+ profiles that run in the outer plexiform layer (OPL).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

A useful programme in BASIC for axonal morphometry with introduction of new cytoskeletal parameters.

Interest in the structure of axons and quantification of their components has been growing over the last years. However, the existing literature contains few reports of available computer programmes to facilitate such studies. This paper presents a fully comprehensive BASIC programme for the morphometric analysis of electron micrographs of cross-sectional nerve fibres. From drawings of fibre and axonal contours and dots of the microtubules and neurofilaments, the programme calculates the following parameters: area, diameter and form factor of the fibres and axons, number and density of microtubules and neurofilaments, proportion between microtubules and neurofilaments (R-proportion), myelin thickness and the diameter of the axon relative to its sheath (g-ratio). The programme also introduces three new parameters to analyse the degree of uniformity of microtubule and neurofilament distribution: distances between microtubules and between neurofilaments, equilateral index and cytoskeletal intermingling index. The programme is written in Microsoft BASIC Interpreter for Apple Macintosh (Microsoft Corporation) but can be used on other computers. Although the programme has been tested on adult rat optic nerve fibres, it can be used for different projects concerning axonal morphometry.

Animals↗
PubMed1990

Distribution of immunoreactivity to protein kinase C in the turtle retina.

Immunocytochemical staining procedures using the HRP-complexed antibody to protein kinase C (PKC) have been carried out on the turtle retina. Wholemounts and frozen sections of retina have been studied by light microscopy to evaluate PKC immunoreactivity after stimulation of the retina with light and neurotransmitters known to be active in the vertebrate retina. The most dramatically stained sites are cone synaptic pedicles and bipolar cells under all conditions. Ganglion cells stain weakly under certain conditions. Applying the antibody to a 'control' retina under dark adapted conditions results in uniform background staining of both hyperpolarizing and depolarizing bipolar pathways, while stimulating the retina with K+ under dim light conditions results in discretely stained bipolar cells and a prominent band of staining in stratum 4 of the inner plexiform layer. Stronger stimulation of bipolar cells with their terminals contributing to strata 3 and 4 and the continuous dominant band in stratum 4 can be elicited with incubation of the retina in neurotransmitter agonists, GABA and dopamine. Incubation with dopamine, in particular, brings out the putative dopaminergic amacrine cell. The only condition in which a strong band in stratum 2 can be demonstrated is under stimulation with a flashing bar of spot of light. Thus K+ and neurotransmitter stimulation elicit PKC staining in neurons contributing to the ON or depolarizing sublamina of the IPL, while intermittent flashing light stimulus is required to elicit PKC staining in the OFF or hyperpolarizing sublamina of the IPL.

Animals↗
PubMed1990

The synaptic organization of the dopaminergic amacrine cell in the cat retina.

The dopaminergic amacrine cells of the cat retina have been stained by immunocytochemistry using an antibody to tyrosine hydroxylase (Toh). The complete population of Toh+ cells has been studied by light microscopy of retinal wholemounts to evaluate morphological details of dendritic structure and branching patterns. Selected Toh+ amacrine cells have been studied by serial-section electron microscopy to analyse synaptic input and output relationships. The majority of Toh+ amacrine cells occur in the amacrine cell layer of the retina and have their dendrites ramifying and forming the characteristic rings in stratum 1 of the inner plexiform layer. A minority of Toh+ cells have cell bodies displaced to the ganglion cell layer but their dendrites also stratify in stratum 1. All Toh+ cells have some dendritic branches running in stratum 2 as well as in stratum 1, and frequently they have long 'axon-like' processes (500-1000 microns long) dipping down to run in stratum 5 before passing up to rejoin the major dendritic arbors in stratum 1. In addition Toh+ stained processes follow blood vessels in the inner plexiform layer and in the ganglion cell layer. A population of Toh+ cells found in the inferior retina appears to give rise to stained processes that pass to the outer plexiform layer and therein to run for as far as one millimeter. Electron microscopy reveals that Toh+ amacrine cells are postsynaptic to amacrine cells and a few bipolar cell terminals in stratum 1 of the inner plexiform layer and are primarily presynaptic to AII amacrine cell bodies and lobular appendages, and to another type of amacrine cell body and amacrine dendrites hypothesized to be the A17 amacrine cell. The Toh+ dendrites in stratum 2 are presynaptic to AII lobular appendages primarily. Stained 'axon-like' processes running in stratum 5 prove to be presynaptic to AII amacrine dendrites as they approach the rod bipolar axon terminals and they may also be presynaptic to the rod bipolar terminal itself. The Toh+ stained dendrites that have been followed in the outer plexiform layer run along the top of the B-type horizontal cell somata and may have small synapses upon them. The only clear synapses seen in the outer plexiform layer are from the Toh+ profiles upon vesicle filled amacrine-like profiles that are in turn presynaptic to bipolar cell dendrites in the outer plexiform layer. We presume the cells postsynaptic to the Toh+ dendrites in the outer plexiform layer are interplexiform cells.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1990

Development of morphological types and distribution patterns of amacrine cells immunoreactive to tyrosine hydroxylase in the cat retina.

Using an antibody against tyrosine hydroxylase on newborn to 30-day kitten retinas, we have been able to follow the development of the dopaminergic amacrine cells of the cat retina by light-microscopical investigations of retinal wholemounts. The Type 1 or large Toh+ amacrine cells described by others (Oyster et al., 1985; Törk & Stone, 1979) and named A18 from a Golgi study (Kolb et al., 1981), is at birth (P1) an immature neuron with a small cell body and two or three simple thick radiating dendrites stratifying in stratum 1 with many of the dendrites ending in enlarged growth cones. With increasing postnatal age, the cell body size increases from 12.5 microns diameter to reach 15.5 microns diameter at P30. The dendritic fields also increase in size and complexity. At P1, cells of the central area exhibit dendritic appendages which then develop progressively until at P13 (after eye opening) they are part of rudimentary rings and by P30 the dendritic plexus of Toh+ dendrites and rings in stratum 1, typical of the adult cells, are complete. Toh+ stained processes with growth cones that run deep in stratum 5 of the inner plexiform layer and processes passing to the outer plexiform layer first become apparent at P1 in cells of central inferior retina but not till after P13 are these processes clearly expressed. At P1, the total number of Toh+ Type 1 cells is approximately 4000 and this number remains unchanged to the adult retina. However, the retina increases in size over the P1-P30 stage and thus the mean density of Type 1 Toh+ cells decreases from 30/mm2 at P1 to 18/mm2 at P30. The maximum density of Type 1 Toh+ cells occurs in central retina 2-4 mm superior temporal to the area centralis, corresponding to the maximum rod photoreceptor concentration. A second type of small Toh+ amacrine cell can be visualized at P1. This Type 2 cell is characterized by a much smaller cell body than Type 1 cells (9 microns diameter), and with faintly stained dendrites located in stratum 3 of the inner plexiform layer. During later postnatal days, Type 2 cells gradually become unstainable and only few are still seen in far peripheral retina by P23. Type 2 Toh+ cells from a total population of 40,000 cells at P1 with their highest density occurring in peripheral retina. By P13, they cannot be seen in central retina and are reduced to a total population of cells staining for the antibody of 7400 cells in far peripheral retina.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1989

Morphology and distribution of neurons immunoreactive for substance P in the turtle retina.

Immunocytochemical staining procedures with the HRP-complexed antibody to substance P have been carried out on the turtle retina. Examination by light microscopy of wholemount retinas has allowed us to evaluate the morphology and distribution of the substance P immunoreactive cell types. Two amacrine cell types and two or more ganglion cell types are stained in our hands. Type A amacrines are tri-stratified wide-field amacrines. They have their major dendrites in S1 and S3 of the inner plexiform layer and they emit fine dendrites from the major dendrites that end in varicose boutons in S5 on and around cell bodies in the ganglion cell layer. Some of the dendrites in S1 radiate out in axon-like fashion for 1 mm across the retina. The type B amacrine cells are small to medium-field in dendritic extent. They have smaller cell bodies than type A and a single or, at most, two primary dendrites that pass directly to S3 before branching profusely into an intricate net-like dendritic field. The ganglion cells that are stained with substance P antibodies appear to be of several types but their exact morphologies are in doubt because only portions of their major dendrites are stained. Substance P immunoreactive axons are clearly seen to project from the cell bodies to the optic nerve head and axons are stained in the optic nerve itself. The substance P-stained ganglion cells occur in an irregular distribution that reaches a peak density in an elongated band parallel to and 1 mm below the visual streak. The type B amacrine cells reach a maximum density in the visual streak and are distributed in a highly regular mosaic decreasing in density in elliptical isodensity contours from the visual streak. In contrast the type A amacrine cells are rare or absent in the streak, being located in an irregular mosaic in peripheral retina.

Animals↗
PubMed1989

[Relevancy of the clinical subjects in medical training].

A judgment of the relevance of twelve clinical science courses for preparing the students to the practice of medicine and to the development of the scientific mind, was tested by the Pair Comparison and Equal-Appearing Interval methods. The test groups consisted of medical school faculty members, medical students and physicians. Three groups of clinical sciences, according its relevancy for preparing the students for a career as physicians, were identified Internal Medicine, Pediatrics, Surgery and Public Health, comprised the group of maximum relevancy; History of Medicine, Medicolegal and Radiological studies, formed a group of lowest relevancy. The remainder sciences (Ophthalmology, Obstetrics and Gynecology, Otorhinolaryngology, Dermatology and Psychiatry, formed a middle group. Few differences were found when we considered the relevancy to the development of the scientific mind.

Curriculum↗
PubMed1987

Postnatal development of microtubules and neurofilaments in the rat optic nerve: a quantitative study.

In this paper the postnatal changes in the cytoskeleton of the rat optic nerve fibers are described and quantified. These changes are also compared with other parameters such as myelination and axonal caliber with the aim of describing a general pattern of optic nerve maturation from a morphological point of view. The results showed that during the first postnatal week microtubules outnumbered neurofilaments but between days 8 and 20 the neurofilaments rapidly increased and on day 20 were about twice as numerous as microtubules. This proportion remained almost unaltered from the end of the third week to the 44th postnatal day. The comparison with other parameters suggested that the cytoskeleton, and in particular the proportion between its components, may be a more reliable index for measuring optic nerve maturation than other variables commonly used.

Animals↗