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Biomedical subjects

D M Redenbach

Publications and source records attributed to D M Redenbach.

14 recordsLinked to original sources
PubMed2005

Granulocytosis and increased adhesion molecules after resistive loading of the diaphragm.

Upregulation of endothelial cell adhesion molecules, followed by an influx of granulocytes and macrophages, can contribute to exertion-induced skeletal muscle injury. The purpose of this study was to quantify circulating leukocyte subsets, diaphragm injury and infiltrating leukocyte subsets, and surface expression of vascular cell adhesion molecule (VCAM)-1 and intracellular adhesion molecule (ICAM)-1 in the diaphragm after inspiratory resistive loading (IRL). Eight New Zealand white rabbits underwent 1.5 h of IRL and seven control rabbits underwent a sham procedure. Blood samples, taken at baseline and 2, 6, 12, 24, 48 and 72 h after the onset of IRL or sham, showed that band cell counts had increased at 6 h post-IRL. Point counting of haematoxylin and eosin-stained cross-sections, sampled at 72 h post-IRL, showed greater injury in diaphragms from IRL rabbits compared with controls. Immunohistochemical processing showed increased expression of ICAM-1 and VCAM-1, and higher granulocyte and macrophage counts in IRL diaphragms than control diaphragms. Macrophages were the predominant inflammatory cells. Increased intracellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression, and infiltration of granulocytes and macrophages may contribute to inspiratory resistive loading-induced diaphragm injury.

Airway Resistance↗
PubMed1998

A comparative study of structures comprising the thoracic outlet in 250 human cadavers and 72 surgical cases of thoracic outlet syndrome.

OBJECTIVE: We have hypothesized that variations in fibrous, muscular and osseous structures with the potential to entrap the brachial plexus occur within the thoracic outlet of the normal population; and that these variations are different in pattern and frequency from those in patients presenting with thoracic outlet syndrome (TOS). METHODS: Structural anomalies with potential for entrapping elements of the brachial plexus were examined following dissections of the posterior triangle of the neck in 250 human cadavers (N = 500 thoracic outlet dissections) and catalogued jointly by an anatomist and a thoracic surgeon. The pattern and frequency of anomalies in the 250 cadavers was compared to that encountered in 72 surgical cases of removal of the first rib for relief of symptomatic TOS (N = 72 procedures, 55 patients). RESULTS: Relevant structural variations were encountered in 46% of cadavers, exhibiting no left right or gender preference overall. When compared with the surgical group in which 100% exhibited structurally relevant anomalies, significant differences in pattern of anomalous structures and gender distribution were revealed. Anomalies posterior to the brachial plexus, ranging from fibrous bands to cervical ribs in both groups, were prevalent in the surgical group. A 'scissors-like' pattern, with neural entrapment by anterior and posterior anomalies was frequently encountered in females. CONCLUSIONS: Based on these data and embryological considerations, we propose a revised and simplified classification of impingement mechanisms within the anatomic thoracic outlet. Comparing these data to radiological imaging and observations at surgery, we offer a new perspective for the investigation and management of patients with TOS.

Cadaver↗
PubMed1997

The nature of leukocyte shape changes in the pulmonary capillaries.

The size discrepancy between leukocytes [white blood cells (WBCs)] and pulmonary capillaries requires WBCs to deform. We investigated the persistence of this deformation on cells leaving the capillary bed and the role played by the cytoskeleton. Isolated rabbit lungs were perfused in situ via the pulmonary artery with effluent fractions collected from the left ventricle. Washout curves from cell counts in each fraction confirmed that WBCs are preferentially retained over erythrocytes. WBC deformation present on exit from the circulation was compared with that present after recovery in paired fractions, fixed either immediately or 60 min later. These cells were compared with cells recovered from the capillary in perfused fixative or fixed in peripheral blood. Our results show that leukocyte deformation persisted after the cells exited the pulmonary circulation. This deformation was associated with minimal submembranous F-actin staining, and microtubule distribution and cell polarization were unchanged. We conclude that cytoskeletal changes that occur during WBC deformation in the pulmonary capillaries are minimal and differ from those known to occur in actively migrating cells during chemotaxis.

Animals↗
PubMed1997

Effect of mechanical deformation on structure and function of polymorphonuclear leukocytes.

The present studies were designed to test the hypothesis that mechanical deformation of polymorphonuclear leukocytes (PMN) leads to functional changes that might influence their transit in the pulmonary capillaries. Human leukocytes were passed through 5- or 3-micron-pore polycarbonate filters under controlled conditions. Morphometric analysis showed that the majority of PMN were deformed and that this deformation persisted longer after filtration through 3-micron filters than through 5-micron filters (P < 0.05) but did not result in the cytoskeletal polarization characteristic of migrating cells. Flow cytometric studies of the filtered PMN showed that there was a transient increase in the cytosolic free Ca2+ concentration after both 3- and 5-micron filtration (P < 0.01) with an increase in F-actin content after 3-micron filtration (P < 0.05). Although L-selectin expression on PMN was not changed by either 5- or 3-micron filtration, CD18 and CD11b were increased by 3-micron filtration (P < 0.05). Priming of the PMN with N-formyl-methionyl-leucyl-phenylalanine (0.5 nM) before filtration resulted in an increase of CD11b by both 5 (P < 0.05)- and 3-micron (P < 0.01) filtration. Neither 5- nor 3-micron filtration induced hydrogen peroxide production. We conclude that mechanical deformation of PMN, similar to what occurs in the pulmonary microvessels, induces both structural and functional changes in the cells, which might influence their passage through the pulmonary capillary bed.

Actins↗
PubMed1995

Distribution of Sertoli cell microtubules, microtubule-dependent motors, and the Golgi apparatus before and after tight junction formation in developing rat testis.

Sertoli cells are polarized epithelial cells of the seminiferous epithelium which provide structural and physiological support for differentiating germ cells. They establish different basal and adluminal environments for the selective nurturing of pre- and post-meiotic germ cells within the seminiferous epithelium, segregated by the Sertoli-Sertoli cell tight junctional complex, the blood-testis barrier. Tight junction formation between epithelial cells in vitro is a critical polarizing event associated with changes in polarized targeting of membrane-specific proteins and reorganization of microtubules, centrioles, and the Golgi apparatus. To investigate whether tight junction formation is associated with organelle reorganization in Sertoli cells in vivo, we have characterized distribution patterns of Sertoli cell microtubules, the mechanoenzymes kinesin and cytoplasmic dynein, and the Golgi apparatus during tight junction formation in developing rat testis. Immunocytochemistry on samples taken at 5, 10, 15, 20, and 25 days of age was used to examine the distribution of these proteins during the extensive cellular reorganization that culminates in the formation of the blood-testis barrier at 19 days of age. Our data show that the distribution patterns reflect the extensive intercellular repositioning of tubule cells in developing seminiferous tubules, but that changes in intracellular organization are not temporally associated with formation of the blood-testis barrier.

Age Factors↗
PubMed1994

Microtubules with altered assembly kinetics have a decreased rate of kinesin-based transport.

Microtubules treated with the gamma-diketone 2,5-hexanedione (2,5-HD) have altered assembly behavior characterized by precocious nucleation and rapid elongation. By measuring the rate of microtubule transport, we have examined the potential functional significance of this 2,5-HD-induced microtubule modification. 2,5-HD-treated microtubules were transported at only 70% of the rate of control microtubules in a simple kinesin-based motility assay on glass coverslips using video and computer enhanced differential interference contrast microscopy. Since 2,5-HD is capable of forming both pyrrole adducts and crosslinks with tubulin, the contributions of pyrrole formation and crosslinking to slowed microtubule transport were determined. 3-Acetyl-2,5-hexanedione (AcHD), a pyrrole forming, non-crosslinking congener of 2,5-HD which does not alter microtubule assembly, did not produce slowed microtubule transport as occurs with 2,5-HD. However, glutaraldehyde, a pyrrole-independent crosslinking agent which alters microtubule assembly in the same way as 2,5-HD, slowed microtubule transport. These results indicate that a 2,5-HD-induced microtubule modification, possibly a crosslink-related conformational change, produces both an alteration in the kinetics of assembly and an alteration in the microtubule-motor interaction.

Animals↗
PubMed1994

Seminiferous tubule fluid secretion is a Sertoli cell microtubule-dependent process inhibited by 2,5-hexanedione exposure.

Mammalian Sertoli cells are responsible for the formation and secretion of seminiferous tubule fluid (STF) which provides the nutritional and hormonal microenvironment necessary for spermatogenesis. Exposure of rats to 2,5-hexanedione (2,5-HD) results in testicular injury characterized by a decrease of STF secretion which immediately precedes bulk germ cell necrosis. The earliest biochemical change in 2,5-HD-exposed rats is an alteration in testicular microtubule assembly kinetics. In this study, we investigate the relationship between microtubule-dependent processes and STF secretion in adult Sprague-Dawley CD rats by exposing seminiferous tubules to two types of toxicants: (1) those which alter microtubules (colchicine and 2,5-HD) and (2) an inhibitor of protein secretion (brefeldin A, [BFA]). Secretion of STF is quantitated by monitoring the rate of transport of a microinjected oil droplet in the lumen of isolated seminiferous tubules using time-lapse stereoscopic microscopy. The rate of oil droplet transport in seminiferous tubules isolated from testis pretreated in vivo for 2 hr with colchicine (40 micrograms/testis) was significantly decreased. Exposure of isolated seminiferous tubules to BFA (10 micrograms/ml) for 40 min also significantly decreased the transport of lumenal oil droplets. Exposure of rats to 1%, 2,5-HD in drinking water decreased transport of injected oil droplets in seminiferous tubules beginning at 3 weeks of exposure in the absence of significant alterations in testicular morphology. These data demonstrate that normal STF secretion requires an intact, microtubule-dependent intracellular membrane transport pathway and strengthen the association between 2,5-HD-induced disruption of Sertoli cell STF secretion and 2,5-HD-induced alterations in Sertoli cell microtubules.

Analysis of Variance↗
PubMed1994

Microtubules are oriented with their minus-ends directed apically before tight junction formation in rat Sertoli cells.

We have examined the polarity of microtubules in Sertoli cells of the seminiferous epithelium during testicular development to test the hypothesis that microtubules change their polarity during tight junction formation. Microtubules in a number of polarized epithelial cells, including Sertoli cells, are oriented with their minus-ends directed toward the apical surface of the cell. Indirect evidence from cultured epithelial cell models suggests that this orientation may be achieved during the relocation of centrioles, reorganization of microtubules, and repositioning of the Golgi that occurs during tight junction formation. Using the microtubule hook decoration technique, we have determined the polarity of Sertoli cell microtubules at 5 and 15 days postnatally, prior to the establishment of the tight junctional complex of the blood testis barrier at 19 days. Our results indicate that, although centrioles and the Golgi apparatus migrate from an infranuclear location at 5 days to a supranuclear location by 15 days, the minus-ends of microtubules are already directed toward the apical surface of the cell by 5 days of age. These data indicate that the establishment of the apically directed minus-end orientation of microtubules of mature Sertoli cells precedes rearrangement of the centrioles and Golgi and is not temporally related to the formation of the tight junctional complex of the blood testis barrier.

Animals↗
PubMed1992

Distribution of the microtubule-dependent motors cytoplasmic dynein and kinesin in rat testis.

To examine the possible role of microtubule-based transport in testicular function, we used immunofluorescent techniques to study the presence and localization of the microtubule mechanoenzymes cytoplasmic dynein (a slow-growing end-directed motor) and kinesin (a fast-growing end-directed motor) within rat testis. Cytoplasmic dynein immunofluorescence was observed in Sertoli cells during all stages of spermatogenesis, with a peak in apical cytoplasm during stages IX-XIV. Cytoplasmic dynein immunofluorescence was also localized within Sertoli cells to steps 9-14 (stages IX-XIV) germ cell-associated ectoplasmic specializations. In germ cells, cytoplasmic dynein immunofluorescence was observed in manchettes of steps 15-17 (stages I-IV) spermatids, and small, hollow circular structures were seen in the cytoplasm of step 17 and step 18 spermatids during stages V and VI. Kinesin immunofluorescence was observed in manchettes of steps 10-18 spermatids (stages X-VI). The stage-dependent apical Sertoli cell cytoplasmic dynein immunofluorescence, in conjunction with the previously reported orientation of Sertoli cell microtubules (slow-growing ends toward the lumen) and peak secretion of androgen-binding protein and transferrin, is consistent with the hypothesis that cytoplasmic dynein is involved in Sertoli cell protein transport and secretion. Further, the localization of cytoplasmic dynein and kinesin to manchettes is consistent with current hypotheses concerning manchette function.

Animals↗
PubMed1992

Binding between mammalian spermatid-ectoplasmic specialization complexes and microtubules.

Ectoplasmic specializations (ESs) are submembrane specializations that consist of Sertoli cell plasma membrane linked by an ordered array of actin filaments to a cisterna of endoplasmic recticulum (ESER). They are thought to function in the spermatid-Sertoli cell adhesion junction. Microtubules occur adjacent to the cytoplasmic face of the ESER and are oriented parallel to the long axis of the Sertoli cell, the direction of spermatid translocation during spermatogenesis. Our hypothesis that spermatid orientation and translocation in the seminiferous epithelium is microtubule dependent predicts that microtubules bind to ESs. To test for binding between microtubules and ESs, we have developed an in vitro assay in which spermatid-ES complexes were isolated from the seminiferous epithelium and incubated with bovine brain microtubules that were labeled with [3H]GTP and stabilized with taxol. Binding was determined by scintillation counts from gradient fractions enriched for spermatid-ES complexes and depleted of unbound microtubules by differential centrifugation. Our data indicate that microtubules bind to spermatid-ES complexes in a substrate concentration-dependent manner and can be released with 5 mM GTP or 10 mM MgATP. Binding is competitively reduced with excess unlabeled microtubules and is inhibited by 100 microM vanadate and 2 mM N-ethylmaleimide (NEM). The amount of binding is unchanged by 10 microM vanadate, 2 mM erythro-(2-hydroxy-3-nonyl)adenine (EHNA) or 1 mM 5'-adenylylimidodiphosphate (AMP-PNP). Immunofluorescence and autoradiographic data confirm that labeled microtubules bind to ES locations on spermatid-ES complexes. These data are consistent with the hypothesis that spermatid translocation is a microtubule-based transport event.

Actins↗
PubMed1991

Microtubule polarity in Sertoli cells: a model for microtubule-based spermatid transport.

Bundles of microtubules occur adjacent to ectoplasmic specializations (ESs) that line Sertoli cell crypts and support developing spermatids. These microtubules are oriented parallel to the direction of spermatid movement during spermatogenesis. We propose a model in which ESs function as vehicles, and microtubules as tracks, for microtubule-based transport of spermatids through the seminiferous epithelium. Microtubule polarity provides the basis for the direction of force generation by available mechanoenzymes. As part of a more general study designed to investigate the potential role of microtubule-based transport during spermatogenesis, we have studied the polarity of cytoplasmic microtubules of Sertoli cells. Rat testis blocks were incubated in a lysis/decoration buffer, with and without exogenous purified bovine brain tubulin. This treatment results in the decoration of endogenous microtubules with curved tubulin protofilament sheets (seen as hooks in cross section). The direction of curvature of the hooks indicates microtubule polarity; that is, clockwise hooks are seen when viewing microtubules from the plus to the minus end. We found that, in Sertoli cells, most of the hooks were orientated in the same direction. Significantly, when viewed from the base of the epithelium, hooks pointed in a clockwise direction. The clockwise direction of dynein arms on axonemes of sperm tails, in the same section, provided an internal check of the section orientation. Electron micrographs of fields of seminiferous epithelium were assembled into montages for quantitative analysis of microtubule polarity. Our data indicate that Sertoli cell cytoplasmic microtubules are of uniform polarity and are orientated with their minus ends toward the cell periphery. These observations have significant implications for our proposed model of microtubule-based transport of spermatids through the seminiferous epithelium.

Animals↗
PubMed1988

Effect of neonatal denervation on the distribution of fiber types in a mouse fast-twitch skeletal muscle.

This study was designed to assess the changes in fiber-type distribution of the extensor digitorum longus (EDL) muscle of the mouse during the first 21 days of age following neonatal sciatic neurectomy. Denervated and normal muscles were compared at 7, 14, and 21 days of age and the normal EDL was also studied at 1 day of age. Frozen sections of the EDL were treated histochemically to detect NADH-tetrazolium reductase and myosin ATPase reactions. Quantitative assessment included measurements of cross-sectional areas and fiber counting. Denervation resulted in muscle atrophy which was due primarily to a decrease in individual fiber area as opposed to fiber loss. Histochemical maturation of the EDL was severely affected by neonatal denervation during the first three postnatal weeks. By 21 days, two extrafusal fiber types which were both oxidative could be distinguished. One type was highly atrophied and resembled an immature fiber exhibiting myosin ATPase staining at both acid and alkaline preincubation conditions, whereas another type was less atrophied and showed myosin ATPase staining resembling fast-twitch (type IIA) fibers. These findings emphasize the importance of an intact nerve supply in determining the phenotypic expression of skeletal muscle, and point to the early postnatal period as a critical stage in fiber type differentiation.

Adenosine Triphosphatases↗
PubMed1988

Alterations in contractile properties of extensor digitorum longus muscle from C57BL/6J mice following denervation at 1 day of age.

Changes in contractile properties of developing fast-twitch skeletal muscle of the C57/BL6J mouse were studied following neonatal denervation. A sciatic neurectomy was performed at 1 day of age and then denervated and control muscles were examined at 7, 14, and 21 days postdenervation. In addition, normal muscles were studied at 1 day of age. The denervated muscles exhibited prolongation of time-to-peak twitch tension and half-relaxation time, a slowing of the maximum velocity of shortening, and a marked increase in resistance to fatigue compared with controls. Isometric tetanus tension was reduced compared to the control muscle both in absolute terms and when expressed relative to body weight at all ages studied. The absolute isometric twitch tension was reduced at 7 and 14 days, but was reduced only at 21 days when expressed as a fraction of the muscle weight. Post-tetanic twitch potentiation failed to appear in the denervated muscle. It would appear that neonatal denervation results in an uncoupling of the developmental pattern of skeletal muscle.

Animals↗