1 Number of text pages (47) and figures (8).A short running head; Role of OPN in tendon tissue remodeling. Keywords AbstractIt has been shown that musculoskeletal tissues undergo dynamic tissue remodeling by a process that is quite sensitive to the mechanical environment. However, the detailed molecular mechanism underlying this process remains unclear. We demonstrate here that after denervation-induced mechanical stress deprivation, tendons undergo dynamic tissue remodeling as evidenced by a significant reduction of the collagen fibril diameter.Importantly, the transient up-regulation of osteopontin (OPN) expression was characteristic during the early phase of tendon tissue remodeling. Following this dynamic change of OPN expression, matrix metalloproteinase (MMP)-13 expression was induced, which presumably accounts for the morphological changes of tendon by degrading tendon collagen fibrils. The modulation of MMP-13 expression by OPN was specific, since the expression of MMP-2, which is also known to be involved in tissue remodeling, did not alter in the tendons under the absence nor presence of OPN. We also demonstrate that the modulation of MMP-13 expression by OPN is due to the signaling through cell surface receptors for OPN. Thus, we conclude that OPN plays a crucial role in conveying the effect of denervation-induced mechanical stress deprivation to the tendon fibroblasts to degrade the extracellular matrices by regulating MMP-13 expression in tendon fibroblasts.3
The effects of in situ freezing and the combination of in situ freezing and stress-shielding on the microstructure and ultrastructure of the patellar tendon were studied with use of 20 mature rabbits. The patellar tendon was frozen in situ with liquid nitrogen to kill fibroblasts and then was completely released from stress by chronically pulling a stainless-steel wire installed between the patella and the tibial tubercle. Microstructurally, the freezing treatment induced separation of collagen fiber bundles and fibroblast necrosis at 3 weeks, although the separation disappeared at 6 weeks. Ultrastructurally, small collagen fibrils with a diameter of less than 90 nm were predominant; at 6 weeks, the area occupied by collagen fibrils had decreased. In the frozen-shielded tendon, numerous large spaces were observed in the matrix at 3 weeks. This treatment increased the number of fibrils with a diameter greater than 360 nm and decreased the number of collagen fibrils per unit of area and the area occupied by collagen fibrils at 3 weeks. This study demonstrated that in situ freezing and the combination of in situ freezing and stress-shielding leads to a smaller volume of collagen fibrils per unit of cross section of the patellar tendon by mechanisms that remain to be defined.
The contribution of sympathetic efferent outflow through the central nervous system (CNS) during prolonged hemorrhagic hypotension of 50 mmHg was examined in anesthetized dogs. Mean blood pressure, heart rate, and renal nerve activity (RNA) were recorded simultaneously after arterial bleeding for 2 h. In animals with an intact neuraxis, hemorrhage to approximately 50 mmHg increased RNA to 180 +/- 12% of control level at 5 min after bleeding, but returned close to control level after 10 min. A secondary increase in RNA followed, with its maximum (280 +/- 12%) at 30 min after bleeding. Then RNA gradually decreased so that at end of experiment (120 min after bleeding) RNA was 10 +/- 9% of control. The initial increase in RNA was abolished by denervation of afferents from carotid sinus and cardiopulmonary regions, but secondary RNA response during hemorrhagic hypotension was similar to that in the intact group and occurred when the head region of animals was exposed to a hypotension of 50 mmHg, and perfusion to peripheral regions of the body was maintained near normal range. However, when perfusion to the head was maintained at a steady level and peripheral regions were exposed to hypotension of 50 mmHg, RNA response did not change significantly. These results provide evidence that prolonged hemorrhagic hypotension, which induces severe brain ischemia, causes biphasic sympathetic outflow via the CNS. The increase in sympathetic activity was followed by decrease with time. This decrease may contribute to the pathogenesis of vasomotor paralysis occurring at the irreversible stage of hemorrhagic shock.
The distribution of substance P (SP) immunoreactivity and the colocalization of SP with other bioactive substances in chromaffin cells and nerve fibers were investigated in the rat adrenal gland at the light microscopic level. In the capsule and cortex, SP immunoreactivity was seen in some nerve fibers around blood vessels and in thick nerve bundles passing through the cortex directly into the medulla. In the medulla, the SP immunoreactivity was observed in a small number of chromaffin cells; these SP-immunoreactive chromaffin cells were either phenylethanolamine N-methyltransferase (PNMT) immunoreactive or immunonegative, indicating that they were either adrenaline cells or noradrenaline (NA) cells. SP-immunoreactive varicose nerve fibers were also found in the medulla and were in contact with a cluster of the NA cells showing catecholamine fluorescence, which suggests that SP from medullary nerve fibers may regulate the secretory activity of the NA cells. Because no SP-immunoreactive ganglion cell was present in the rat adrenal gland, the intra-adrenal nerve fibers were considered to be extrinsic in origin. The double-immunostaining method further revealed that the SP-immunoreactive chromaffin cells also exhibit immunoreactivities for calcitonin gene-related peptide (CGRP), and neuropeptide tyrosine (NPY), suggesting that these peptides can also be released from the chromaffin cells by certain stimuli. The intra-adrenal nerve fibers in the medulla were composed of SP-single immunoreactive, and SP/CGRP-, SP/choline acetyltransferase (ChAT)-, SP/nitric oxide synthase (NOS)-, SP/pituitary adenylate cyclase activating polypeptide (PACAP)-, ChAT/NOS-, and ChAT/PACAP-immunoreactive nerve fibers, which may affect the secretory activity of the NA cells. In the adrenal capsule, the nerve fibers were present around blood vessels and showed immunoreactivities for SP/ CGRP, SP/NPY, SP/NOS, and SP/vasoactive intestinal polypeptide, suggesting that the origin of nerve fibers in the capsule may differ from those in the medulla.
The mode of participation of three vascular beds of the kidney, intestine and skeletal muscles during blood pressure oscillation elicited by what is called a "side pressure exertion experiment" was investigated in anesthetized rabbits.
From postnatl-day-0 to postnatal-day-2, a few acetylcholinesterase (AChE)-active and choline acetytransferase (ChAT)-immunoreactive nerve fibers and relatively numerous vesicular acetylcholine transporter (VAChT)-immunoreactive puncta were observed in the rat adrenal medulla. Despite relatively numerous clear vesicles in the nerve fibers, the synthesis and hydrolysis of acetylcholine may not be fully activated until postnatal-day-2. The number of AChE-active and ChAT-immunoreactive nerve fibers dramatically increased and that of VAChT-immunoreactive puncta gradually increased from postnatal-day-3 to postnatal-week-1. The synthesis and hydrolysis of acetylcholine may be dramatically activated in the nerve fibers of the medulla until postnatal-week-1. From postnatalweek-2 to postnatal-week-3, the number of AChE-active and the ChATimmunoreactive nerve fibers gradually increased and reached the adult levels. The VAChT-immunoreactive puncta per unit area was maximum number at postnatal-week-2. The synthesis and hydrolysis of acetylcholine in the nerve fibers of the medulla may be completed between postnatal-week-2 to postnatal-week-3. The diameter of the VAChT-immunoreactive puncta gradually increased from postnatal-day-0 with aging. However, the number of the VAChT-immunoreactive puncta gradually decreased from postnatal-week-2 onwards. In electron-microscopy, the VAChT-immunoreactive deposits were seen in clusters of clear vesicles, and the diameter of the nerve fibers and the number of clear vesicles at postnatal-week-8 increased compared with those at postnatal-week-2. The AChE-active, ChAT-immunoreactive, and VAChT-immunoreactive nerve THE ANATOMICAL RECORD 292:371-380 (2009) fibers observed around noradrenaline (NA) cells were denser than those around adrenaline (A) cells in the medulla at postnatal-week-8. These suggest that the preferential innervation of NA and A cells may cause the differential secretion NA and A.
Immunocytochemical studies of the rat carotid body during postnatal development revealed neuropeptide Y (NPY), tyrosine hydroxylase (TH), and dopamine beta-hydroxylase (DBH) immunoreactivities. In adult rats (at postnatal week 10), NPY and DBH immunoreactivities were shown in a few small chief cells (cell number/section shown as mean +/- SD: NPY 3.4+/-2.6, DBH 3.2+/-2.3), in large ganglion cells, and in numerous varicose nerve fibers of the carotid body. TH immunoreactivity was found in almost all chief cells, in a few ganglion cells, and in numerous varicose nerve fibers in the carotid body. By using the double-immunostaining technique, most NPY-immunoreactive chief cells, ganglion cells, and nerve fibers exhibited DBH immunoreactivity. The NPY- and DBH-immunoreactive chief cells in the rat carotid body were numerous from birth (NPY 93.8+/-14.9, DBH 89.7+/-12.3) to postnatal week 1 (NPY 65+/-14.5, DBH 61.6+/-11.3), but decreased quickly from postnatal week 2 (NPY 6.1+/-3.5, DBH 3.6+/-2.8) onwards. A few NPY- and DBH-immunoreactive ganglion cells were found in the periphery or in the center of the rat carotid body during postnatal development. TH immunoreactivity was observed in almost all chief cells and in a few ganglion cells in all developmental stages. NPY- and DBH-immunoreactive nerve fibers were very scarce in the carotid body from birth to postnatal week 1, began to increase gradually after postnatal week 2, and reached the adult level by postnatal week 5. The present study suggests that the expression of NPY and noradrenaline in chief cells and in the nerve fibers of the rat carotid body may be regulated during postnatal development.
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