MUSCULATURE AND NEUROTRANSMITTERS IN THE DIGESTIVE SYSTEM OF TREMATODES

In this paper we analyzed the results of our own and published data concerning the presence of muscle elements in various parts of the digestive system in adult and larval forms of trematodes. The data on the localization of the circular and longitudinal muscle fibers in the pharynx, esophagus, and intestine of various representatives of trematodes are presented. The results of immunocytochemical studies indicate the presence the serotonergic and peptidergic (FMRFamidergic) nerve elements in the parts of the digestive system of trematodes. The available literature date is supplemented by the studies conducted on Prodistomum alaskense, a representative of the family Lepocreadiidae, an intestinal parasite of deep-sea fish (Zaprora silenus and Aptocyclus ventricosus). The localization of the serotoninergic and FMRFamidergic nervous structures was identified using immunocytochemical methods and the confocal scanning laser microscopy. For musculature staining the TRITC (tetramethylrhodamine isothiocyanate) – conjugated phalloidin was used. The preparations were examined using a fluorescence microscope and a confocal scanning laser microscope. The analysis of the data obtained and the information available in the literature suggests that the muscular system of the digestive tract is well developed in trematodes of various taxonomic groups. The musculature of the digestive system of trematodes is innervated by serotonergic and peptidergic (FMRFamidergic) nerve elements, which are involved in the regulation of the contractile activity of various parts of the digestive system of trematodes.

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3-D image analysis of serial focal sections from confocal scanning laser microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100152732 ◽

1989 ◽

Vol 47 ◽

pp. 152-153 ◽

Cited By ~ 1

Author(s):

J. Christian Russ ◽

John C. Russ

Keyword(s):

Fluorescent Dyes ◽

Monochromatic Light ◽

Confocal Imaging ◽

Scanning Laser ◽

Confocal Scanning Laser Microscopy ◽

Image Point ◽

Confocal Scanning Laser Microscope ◽

Confocal Scanning ◽

Confocal Scanning Laser ◽

Focus Information

The Confocal Scanning Laser Microscope (CSLM) is used in two quite different imaging modes: reflection and transmission. Most instruments maintain their confocal optics only when operated in reflection mode, in which light reflected or emitted from points in the sample or on its surface, lying in the focal plane of the microscope, is detected to form an image. Point elevations can be measured by scanning through a range of focal distances. Recording in memory the maximum brightness at each pixel forms an image containing the “in-focus” information for the entire irregular surface or object (Figure 1). These are powerful capabilities and account for much of the current use of confocal microscopes for metrology of rough surfaces, and for use with fluorescent dyes.True transmission confocal imaging can be achieved by passing the light from the source through the specimen to a mirror, reflecting it back through the specimen again (maintaining the focus in the same plane), and thence to the detector. This is shown schematically in Figure 2. This is only possible with monochromatic light, because of sample-induced chromatic aberrations.

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Three-dimensional images of DAB-labeled neurons by confocal scanning laser microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100152690 ◽

1989 ◽

Vol 47 ◽

pp. 144-145

Author(s):

JS Deitch ◽

KL Smith ◽

C Lee ◽

JW Swann ◽

JN Turner

Keyword(s):

Three Dimensional ◽

Neuronal Morphology ◽

Scanning Laser ◽

Confocal Scanning Laser Microscopy ◽

Confocal Scanning Laser Microscope ◽

Basal Dendrites ◽

Confocal Scanning ◽

Three Dimensional Images ◽

Confocal Scanning Laser ◽

Areas Of Interest

The ability to correlate neuronal morphology and physiology has been greatly advanced by intracellular labeling through the recording pipette. However, visualizing the filled neuron required physically sectioning and reconstructing areas of interest, often resulting in figures that are two-dimensional. We have visualized the three-dimensional morphology of filled neurons reacted with nickel-intensified diaminobenzidine (DAB/Ni) using the confocal scanning laser microscope (CSLM).Neurons in slices of rat hippocampus were filled with biocytin, fixed in 4% paraformaldehyde, incubated in avidin-HRP (1:200) in 0.5% Triton X-100, and reacted with DAB in 0.04% nickel ammonium sulfate. Optical sections and three-dimensional images were recorded by using a Bio Rad MRC-500 CSLM with an argon ion laser.Biocytin filled all aspects of the neuron, including fine axons and spines. Fig. 1 is a conventional micrograph of a single neuron labeled with DAB/Ni. Figs. 2a and b are stereo pairs of the apical and basal dendrites of the neuron in Fig. 1.

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Fading of the diaminobenzidine reaction product in the confocal scanning laser microscope

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100152707 ◽

1989 ◽

Vol 47 ◽

pp. 146-147

Author(s):

JS Deitch ◽

KL Smith ◽

JW Swann ◽

JN Turner

Keyword(s):

Pyramidal Neurons ◽

Light And Electron Microscopy ◽

Scanning Laser ◽

Laser Exposure ◽

Confocal Scanning Laser Microscope ◽

Fluorescent Markers ◽

Before And After ◽

Staining Protocol ◽

Confocal Scanning ◽

Confocal Scanning Laser

Neurons labeled with horseradish peroxidase and reacted with diaminobenzidine (DAB) can be imaged using a confocal scanning laser microscope (CSLM) in the reflection mode. In contrast to fluorescent markers, the DAB reaction product is thought to be stable and can be observed by both light and electron microscopy. We have investigated the sensitivity of the DAB reaction product to laser irradiation, and present the spectrophotometric properties of DAB before and after exposure in the CSLM.Pyramidal neurons in slices of rat hippocampus were injected with biocytin (a biotin-lysine complex), fixed overnight in 4% paraformaldehyde, and vibratome sectioned at 75 μm. Biocytin was detected with avidin-HRP (1:200) in 0.5% Triton X-100, incubated in DAB (0.5 mg/ml) with or without 0.04% nickel ammonium sulfate (Ni), dehydrated, and imaged in a Bio Rad MRC-500 CSLM with an argon ion laser (488 and 514 nm). Spectrophotometric measurements of the soma were made on a Zeiss microspectrophotometer, as a function of laser exposure (100-1000 scans) and staining protocol.

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