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71.
Structural analysis of human neutrophil migration: Centriole, microtubule, and microfilament orientation and function during chemotaxis 总被引:41,自引:21,他引:20 下载免费PDF全文
Orientation of nucleus, centriole, microtubules, and microfilaments within human neutrophils in a gradient of chemoattractant (5 percent Escherichia coli endotoxin-activated serum) was evaluated by electron microscopy. Purified neutropils (hypaque-Ficoll) were placed in the upper compartment of chemotactic chambers. Use of small pore (0.45 μm) micropore filters permitted pseudopod penetration, but impeded migration. Under conditions of chemotaxis with activated serum beneath the filter, the neutrophil population oriented at the filter surface with nuclei located away from the stimulus, centrioles and associated radial array of microtubules beneath the nuclei, and microfilament-rich pseudopods penetrating the filter pores. Reversal of the direction of the gradient of the stimulus (activated serum above cells) resulted in a reorientation of internal structure which preceded pseudopod formation toward the activated serum and migration off the filter. Coordinated orientation of the entire neutrophil population did not occur in buffer (random migration) or in a uniform concentration of activated serum (activated random migration). Conditions of activated random migration resulted in increased numbers of cells with locomotory morphology, i.e. cellular asymmetry with linear alignment of nucleus, centriole, microtubule array, and pseudopods. Thus, activated serum increased the number of neutrophils exhibiting locomotory morphology, and a gradient of activated serum induced the alignment of neutrophils such that this locomotory morphology was uniform in the observed neutrophil populayion. In related studies, cytochalasin B and colchicines were used to explore the role of microfilaments and microtubules in the neutrophil orientation and migration response to activated serum. Cytochalasin B (3.0 μg/ml) prevented migration and decreased the microfilaments seen, but allowed normal orientation of neutrophil structures. In an activated serum gradient, colchicines, but not lumicolchicine, decreased the orientation of nuclei and centrioles, and caused a decrease in centriole-associated microtubules in concentrations as low as 10(-8) to 10(-7) M. These colchicines effects were associated with the rounding of cells and impairment of pseudopod formation. The impaired pseudopod formation was characterized by an inability to form pseudopods in the absence of a solid substrate, a formation of narrow pseudopods within a substrate, and a defect in pseudopod orientation in an activated serum gradient. Functional studies of migration showed that colchicines, but not lumicolchicine, minimally decreased activated random migration and markedly inhibited directed migration, but had not effect on random migration. These studies show that, although functioning microfilaments are probably necessary for neutrophil migration, intact microtubules are essential for normal pseudopod formation and orientation, and maximal unidirectional migration during chemotaxis. 相似文献
72.
Worldwide patterns of mitochondrial DNA differentiation in the harbor seal (Phoca vitulina) 总被引:3,自引:0,他引:3
Stanley HF; Casey S; Carnahan JM; Goodman S; Harwood J; Wayne RK 《Molecular biology and evolution》1996,13(2):368-382
The harbor seal (Phoca vitulina) has one of the broadest geographic
distributions of any pinniped, stretching from the east Baltic, west across
the Atlantic and Pacific Oceans to southern Japan. Although individuals may
travel several hundred kilometers on annual feeding migrations, harbor
seals are generally believed to be philopatric, returning to the same areas
each year to breed. Consequently, seals from different areas are likely to
be genetically differentiated, with levels of genetic divergence increasing
with distance. Differentiation may also be caused by long-standing
topographic barriers such as the polar sea ice. We analyzed samples of 227
harbor seals from 24 localities and defined 34 genotypes based on 435 bp of
control region sequence. Phylogenetic analysis and analysis of molecular
variance showed that populations in the Atlantic and Pacific Oceans and
east and west coast populations of these oceans are significantly
differentiated. Within these four regions, populations that are
geographically farthest apart generally are the most differentiated and
often do not share genotypes or differ in genotype frequency. The average
corrected sequence divergence between populations in the Atlantic and
Pacific Oceans is 3.28% +/- 0.38% and those among populations within each
of these oceans are 0.75% +/- 0.69% and 1.19% +/- 0.65%, respectively. Our
results suggest that harbor seals are regionally philopatric, on the scale
of several hundred kilometers. However, genetic discontinuities may exist,
even between neighboring populations such as those on the Scottish and east
English coasts or the east and west Baltic. The mitochondrial data are
consistent with an ancient isolation of populations in both oceans, due to
the development of polar sea ice. In the Atlantic and Pacific, populations
appear to have been colonized from west to east with the European
populations showing the most recent common ancestry. We suggest the recent
ancestry of European seal populations may reflect recolonization from Ice
Age refugia after the last glaciation.
相似文献
73.
Hemodynamic responses to oxygen breathing in man 总被引:1,自引:0,他引:1
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Multidimensional annotation of the Escherichia coli K-12 genome 总被引:2,自引:0,他引:2
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To determine whether the increase in cardiac output during mild to moderate exercise is related to an increase in the tissue redox potential, we compared the responses of cardiac output, total body oxygen consumption, and arterial blood lactate-to-pyruvate ratio (a measure of NADH/NAD) to treadmill exercise between dogs treated with normal saline and those treated with a hydrogen acceptor, new methylene blue. Normal saline was infused into the left atrium in the first group of dogs at a rate of 0.38 ml/min throughout the treadmill exercise (2.5 mph and 5.0 mph on a 6% incline, each for 20 min). In the second group, methylene blue was administered as a loading dose (4 mg/kg) before exercise, followed by a continuous infusion (0.15 mg X kg-1 X min-1) throughout exercise. A similar infusion of methylene blue was given to a third group of dogs without exercise; it reduced the arterial lactate-to-pyruvate ratio from 6.70 +/- 0.35 to 4.12 +/- 0.27 but had no or little effects on cardiac output, heart rate, arterial pressure, and left ventricular dP/dt and (dP/dt)/P. Treadmill exercise doubled cardiac output and increased total body O2 consumption three- to fourfold in the first two groups but increased arterial blood lactate-to-pyruvate ratio only in group 1 (6.0 +/- 0.54 to 9.97 +/- 0.91). The relationship between cardiac output and total body O2 consumption was unaffected by the simultaneous administration of methylene blue during exercise. Groups 1 and 2 also did not differ in their heart rate, left ventricular dP/dt and (dP/dt)/P, and plasma catecholamine responses to exercise.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献
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