Application of confocal laser scanning microscopy to the deep pineal gland and other neural tissues.
The study of the deep pineal gland of the Mongolian gerbil and other neuronal tissue from the rat by means of confocal laser scanning microscopy (CLSM) is described. Opical serial sectioning was performed on thick (100-200 microns) sections of the deep pineal gland of the Mongolian gerbil stained immunohistochemically using antisera to S-antigen and tyrosine hydroxylase (TH). Both dual-stained and single-stained material was examined using the fluorochromes fluorescein isothiocyanate (FITC) and Texas Red. High resolution images were obtained showing that pinealocytes have 1-3 processes that extend primarily to other pinealocytes or presumptive pinealocytes. Pinealocytes are located within the deep pineal gland as well as adjacent to the posterior aspect of the medial habenular nuclei. Pinealocyte processes were not seen extending into the habenular nuclei, but rather ended within the deep pineal gland a significant distance from their perikarya. The TH-immunopositive fibers were distributed throughout the deep pineal gland, often forming "baskets" of fibers around pinealocytes rather than being associated primarily with blood vessels. Other uses of the confocal microscope are demonstrated on rat neural tissue reacted with peroxidase/diaminobenzidine (DAB) immunohistochemistry and FITC fluorescence immunohistochemistry (paraventricular nucleus) as well as Golgi-stained neuronal tissue (cerebral cortex). The HRP/DAB and Golgi-stained images were visualized using the reflected image mode of the confocal system.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Anatomy, Cell Biology, and Neurosciences, School of Medicine, University of South Carolina, Columbia 29208.
M G
MG
Welsh
J M
JM
Ding
J
J
Buggy
L
L
Terracio
D015100:3,3'-Diaminobenzidine; D000818:Animals; D016650:Fluorescein-5-isothiocyanate; D005455:Fluorescent Antibody Technique; D005849:Gerbillinae; D006056:Golgi Apparatus / Q000737:chemistry / Q000648:ultrastructure; D006735:Horseradish Peroxidase; D007106:Immune Sera; D007150:Immunohistochemistry / Q000379:methods; D008297:Male; D008855:Microscopy, Electron, Scanning / Q000379:methods; D009412:Nerve Fibers / Q000201:enzymology / Q000648:ultrastructure*; D009474:Neurons / Q000201:enzymology / Q000648:ultrastructure*; D010870:Pineal Gland / Q000166:cytology / Q000201:enzymology / Q000648:ultrastructure*; D051381:Rats; D014446:Tyrosine 3-Monooxygenase / Q000032:analysis / Q000276:immunology
D007106:Immune Sera; D015100:3,3'-Diaminobenzidine; D006735:Horseradish Peroxidase; D014446:Tyrosine 3-Monooxygenase; D016650:Fluorescein-5-isothiocyanate