Despite numerous literature reports on the morphometry of the myelinated fibers of phrenic nerves in rats, a systematic study of the longitudinal and lateral symmetry of the unmyelinated fibers morphometry is not available. In this study, we have undertaken ultrastructural and morphometric studies of the phrenic nerve in adult rats, assessing two different levels (proximal and distal) from both right and left sides. Phrenic nerves of adult male Wistar rats were prepared for epoxy resin embedding and transmission electron microscopy. Morphometric analysis was performed with the aid of computer software, which took into consideration the unmyelinated fiber number, density, area, and diameter, as well as ratio between myelinated and unmyelinated fibers, and the percentage of the fascicular area occupied by the myelinated and unmyelinated fibers. Comparison of data from proximal and distal segments on the same side and from the same levels between sides was performed. Differences were considered significant when P < 0.05. The most important finding is that morphometric parameters of the phrenic nerve unmyelinated fibers in adult rats are both longitudinally and laterally symmetric. This study adds important morphometric information about the unmyelinated fibers of the phrenic nerves in adult rats for proximal and distal levels on both sides of the animal.
We have demonstrated that phrenic nerves' large myelinated fibers in streptozotocin (STZ)-induced diabetic rats show axonal atrophy, which is reversed by insulin treatment. However, studies on structural abnormalities of the small myelinated and the unmyelinated fibers in the STZ-model of neuropathy are limited. Also, structural changes in the endoneural vasculature are not clearly described in this model and require detailed study. We have undertaken morphometric studies of the phrenic nerve in insulin-treated and untreated STZ-diabetic rats and non-diabetic control animals over a 12-week period. The presence of neuropathy was assessed by means of transmission electron microscopy, and morphometry of the unmyelinated fibers was performed. The most striking finding was the morphological evidence of small myelinated fiber neuropathy due to the STZ injection, which was not protected or reversed by conventional insulin treatment. This neuropathy was clearly associated with severe damage of the endoneural vessels present on both STZ groups, besides the insulin treatment. The STZ-diabetes model is widely used to investigate experimental diabetic neuropathies, but few studies have performed a detailed assessment of either unmyelinated fibers or capillary morphology in this animal model. The present study adds useful information for further investigations on the ultrastructural basis of nerve function in diabetes.
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