The Anatomical record · 1996
Study of human cutaneous sensory corpuscles using double immunolabelling and confocal laser scanning microscopy.
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Vega J A, Llamosas M M, Huerta J J, García-Fernández J M
Аннотация
BACKGROUND: The main constituents of sensory corpuscles, i.e., the central axon, the periaxonic Schwann-related, cells, and the perineurial-related cells, can be identified light microscopically by simple immunohistochemistry using specific antibodies. This paper demonstrates the usefulness of double immunolabelling for light and confocal laser-scanning microscopy (CLSM) in the study of human cutaneous sensory corpuscles.
MATERIALS AND METHODS: Antibodies directed against neurofilament proteins (NFPs) and S-100 protein were used to label the central axon and the lamellar cells of Meissner corpuscles or the inner-core lamellae of digital cutaneous Pacinian corpuscles, respectively. Samples were obtained from subjects with normal sensitivity and from patients with paresthesia or absence of clinical sensitivity. Single and double immunolabelling was performed, and the sections were studied by light or CLSM microscopy.
RESULTS: Double immunolabelling was effective for simultaneous observation of the central axon (NFP-positive) and periaxonic Schwann-related (S-100 protein-positive) cells in sensory corpuscles from normal human digital skin. The images that were obtained with both methods were comparable, but the axonic profiles were sharper with diaminobenzidine (DAB) used as a chromogen rather than with Texas-red used as a fluorochrome. Nevertheless, the ability to manipulate the focal plane by using CLSM permits one to better analyze the intracorpuscular relationships of the axon. Double immunolabelling in sensory corpuscles from the skin of patients with nerve compression showed the presence of a central axon in the corpuscles, whereas it was absent in the sensory corpuscles of clinically denervated skin.
CONCLUSIONS: Double immunolabelling is a useful method with which to analyze simultaneously two of the corpuscular constituents, and it may be used in the study of denervated and reinnervated sensory corpuscles.
MATERIALS AND METHODS: Antibodies directed against neurofilament proteins (NFPs) and S-100 protein were used to label the central axon and the lamellar cells of Meissner corpuscles or the inner-core lamellae of digital cutaneous Pacinian corpuscles, respectively. Samples were obtained from subjects with normal sensitivity and from patients with paresthesia or absence of clinical sensitivity. Single and double immunolabelling was performed, and the sections were studied by light or CLSM microscopy.
RESULTS: Double immunolabelling was effective for simultaneous observation of the central axon (NFP-positive) and periaxonic Schwann-related (S-100 protein-positive) cells in sensory corpuscles from normal human digital skin. The images that were obtained with both methods were comparable, but the axonic profiles were sharper with diaminobenzidine (DAB) used as a chromogen rather than with Texas-red used as a fluorochrome. Nevertheless, the ability to manipulate the focal plane by using CLSM permits one to better analyze the intracorpuscular relationships of the axon. Double immunolabelling in sensory corpuscles from the skin of patients with nerve compression showed the presence of a central axon in the corpuscles, whereas it was absent in the sensory corpuscles of clinically denervated skin.
CONCLUSIONS: Double immunolabelling is a useful method with which to analyze simultaneously two of the corpuscular constituents, and it may be used in the study of denervated and reinnervated sensory corpuscles.