Confocal Microscopic Analysis of Merkel Innervation in the Cat Mystacial Vibrissa Follicles
Three-dimensional morphology of the Merkel innervation in very thick sections of the cat mystacial vibrissa follicles as well as in whole mounted tissues were demonstrated by using a confocal laser microscope with a powerful analyzing system for reconstru
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Summary Three-dimensional morphology of the Merkel innervation in very thick sections of the cat mystacial vibrissa follicles as well as in whole mounted tissues were demonstrated by using a confocal laser microscope with a powerful analyzing system for reconstruction. A sheet of densely arranged Merkel nerve endings usually having scattered gaps, was observed at the level of the ring sinus around a large sized follicle. The number of all Merkel nerve terminal disks was roughly estimated at about 3200 in the follicle. The gaps may suggest plasticity among the Merkel innervation.
Results and Discussion Vibrissa follicle-sinus complexes (FSCs) are exceptionally well-innervated, important tactile organs on the mystacial pads of most mammals (Rice et al. 1993). At least eight distinct endings including two kinds of Merkel innervation, at the mouth of the follicle and at the level of the ring sinus were clearly discriminated in the rat and cat FSCs by using confocal laser scanning microscopy (CLSM; Ebara et al. 2002). Recently, we developed a whole mount technique in order to observe the full innervation around the FSCs. In this study, we focused on the ring sinus Merkel innervation in the cat large FSCs and demonstrated the three-dimensional (3D) structures by using mainly the anti-protein gene product 9.5 antiserum as neuronal markers. Abundant CLSM stack images taken from the whole mounted FSCs by rotating around the shaft as well as from the 100200 f.lm-thick sections were made into 3D reconstructions by using a worldwidefirst highly sophisticated application, VG StudioMax 1.1 software (Volume Graphics, Germany). We could easily analyze and understand the detailed K. I. Baumann et al. (eds.), The Merkel Cell © Springer-Verlag Berlin Heidelberg 2003
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Fig. la-c. PGP immunopositive innervation in a 150-J.!m-thick section of cat FSC (#2 of row D). Merkel nerve endings situated in the outer root sheath were pseudo-colored in red. a View from the side of the vibrissa! shaft. b Back view of the middle part of a. c Left side view of b. A large caliber nerve fiber branches twice (open arrows), each branch penetrates the glassy membrane (GM) and terminates as the Merkel terminal disks (asterisks). Distorted and/or faintly labeled endings result in gaps in the total distribution of the endings (yellow areas ; cf. Fig. 2). Including these irregular endings, the number of all endings was about 592 in a. The dashed line indicates the position of the ring wulst (Rw). L Lanceolate endings, C club-like ring-wulst endings
morphology in spite of the highly complicated structure of innervation that is distributed in a very thin layer of the follicle (external root sheath) which is overlaid by a layer of dense lanceolate innervation in the mesenchymal sheath. In three dimensions, we could discriminate and segment all Merkel nerve endings that are situated in the external root sheath in both images taken from a thick sectioned FSC and a whole mounted FSC (Figs. I, 3). The distal ends of the Merkel terminal nerve fib
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