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PubMed1990

The Effects of Prenatal Intracranial Infusion of Tetrodotoxin on Naturally Occurring Retinal Ganglion Cell Death and Optic Nerve Ultrastructure.

In the developing vertebrate nervous system, cell death is known to play an important role in determining final neuron number. Retinal ganglion cells in the cat's visual system undergo a massive elimination by cell death during the prenatal period between E44 (age of embryo in days) and birth (= E65). We have examined whether neural activity contributes to ganglion cell death by infusing tetrodotoxin (TTX), a blocker of the voltage-sensitive sodium channel. TTX was infused intracranially via osmotic minipumps implanted in utero at E42. The effects of the TTX treatment on ganglion cell death and optic nerve ultrastructure were examined at either E49 or E57 by electron microscopy and quantitative analysis of optic axon number. The numbers of optic nerve axons counted in the optic nerves of animals after either 1 or 2 weeks of TTX treatment were not significantly different from the counts in normal animals at comparable ages: E49 TTX-3.2 x 105; E48 normal-3.3 x 105; E57 TTX-2.1 x 105; E59 normal-2.4 x 105. These results suggest that retinal ganglion cells cannot be rescued from death by blockade of neural activity central to the optic chiasma. However, the ultrastructure of optic nerves following 2 weeks of TTX infusion was quite abnormal. The usual packaging of axons into fascicles by glia was disrupted by the presence of many pale, organelle-poor processes that were about 10 times larger in their cross-sectional areas than axons in either normal or TTX-treated nerves. Examination of these processes in serial transverse or in longitudinal electron microscope (EM) sections of the nerve revealed that they were most likely glial in origin. The ultrastructural organization of the optic nerve following 1 week of TTX treatment was normal, indicating that this effect on glial ultrastructure is either cumulative or delayed in onset. These results suggest that while the conduction of action potentials to the terminals of retinogeniculate axons may not play a significant role in regulating ganglion cell number prenatally, it may affect the normal maturation of optic nerve glia.

Journal Article↗
PubMed1989

The spermatozoa of the polychaeta (Annelida): an ultrastructural review.

1. Polychaete sperm are divisible into ect-aquasperm, ent-aquasperm, and introsperm. 2. Ect-aquasperm are the commonest type of polychaete sperm and are considered plesiomorphic for the Polychaeta. Re-evolution of ect-aquasperm (as neo-aquasperm) is, nevertheless, tentatively hypothesized for some Sabellida. 3. In terms of ultrastructural studies of sperm in the investigated polychaete families, only ect-aquasperm have been demonstrated for 16 families; only ent-aquasperm for 3 families; ect- and ent-aquasperm for 3; ect- and intro-sperm for 2; ect-, ent- and intro-sperm for 1 family; and only introsperm for 11 families but investigations can only be regarded as preliminary. To date no family is known to have ent- and intro-sperm only. Sperm ultrastructure has yet to be examined in the orders Magelonida, Psammodrilida, Cossurida, Spintherida, Sternapsida, Flabelligerida and Fauvelopsida. 4. Much variation occurs in gross morphology, ultrastructure and configuration of the several components of ect-aquasperm: acrosome, nucleus, mitochondria, and centrioles and associated anchoring apparatus. A 9 + 2 axoneme is constant. 5. Group-specific sperm structure has been demonstrated for the Nereidae (chiefly ect-aquasperm), and for introsperm of the families Histriobdellidae, Questidae; Capitellidae, Spionidae and Protodrilidae. Species-specificity of all classes of spermatozoa is well established. 6. The very small size of ect-aquasperm is correlated with production of large numbers of sperm as an adaptation to broadcast spawning. Simplicity of structure may relate more to conservation of materials than to hydrodynamics. 7. Fertilization by ent-aquasperm requires fewer eggs than in external fertilization and is accompanied by a tendency to lecithotrophy. Elongation of the nucleus and development of asymmetry are seen in several of the few known examples of ent-aquasperm. Whether modifications are related to transfer or to other features, such as lecithotrophy, is uncertain. 8. Evident multiple origins of polychaete introsperm contraindicate their value in establishing relationship between families, in contrast with their utility in groups such as decapod crustacea. 9. At the intrafamilial level polychaete introsperm have taxonomic and phylogenetic value, as seen in the Spionidae, Capitellidae, and Histriobdellidae, and are distinctive of each of these and other families. 10. At higher taxonomic levels, the ultrastructure of the sperm of the oligochaetoid Questidae distinguishes this family from euclitellates, each class of which has its own distinctive subtype of the euclitellate introsperm. 11.(ABSTRACT TRUNCATED AT 400 WORDS)

Acrosome↗
PubMed1996

Prenatal development of the sheep pineal gland: an ultrastructural study.

The ultrastructure of the pineal gland of 32 sheep embryos was studied from day 54 of development through birth. Embryos were arranged in four age-groups, defined in terms of the most relevant histological features: group 1 (54 to 67 days of prenatal development), group 2 (71 to 92 days), group 3 (98 to 113 days), and group 4 (118 to 150 days). A primary cell type, designated the pinealoblast, was observed from 54 days until birth; ultrastructurally, this cell was found to contain all the organelles required for hormone synthesis. A second cell population, classified as interstitial cells by virtue of their location among pinealoblasts, appeared at 78 days gestation and persisted until birth. Interstitial cells were scarce and exhibited tropism for the perivascular space. From 118 days gestation until birth, a third cell type, termed the pigmented cell, was visible. Pigmented cells, whose ultrastructural characteristics differed from those of pinealoblasts, contained a large number of pigment granules of varying size and shape. The pineal gland of developing sheep embryos showed considerable innervation and abundant vascularization; this, together with certain ultrastructural characteristics, suggests that the gland has a secretory function in uterine life.

Animals↗
PubMed1980

The ultrastructure of bone marrow plasma cells obtained from patients with multiple myeloma during the clinical course of the disease.

The ultrastructure of plasma cells from 65 consecutive bone marrow specimens from 13 patients with multiple myeloma is described. Biopsies were taken from 12 of the patients prior to and after initiation of treatment. The study was undertaken with the aim of correlating ultrastructural characteristics of the myeloma cells with biochemical and clinical parameters, including the survival time after treatment of the patients with cytostatics. Intranuclear inclusions were only seen in the cells of four patients, all of whom had rather long survival times. Two of these are still alive and, furthermore, their plasma cells are characterized by a low mean number of mitochondria in each sectioned cell studied, i.e. 7 and 14, respectively. Nuclear bodies were found in about 7 per cent of the plasma cells in biopsies taken before treatment was initiated. During treatment this percentage increased significantly to about 16 (2P = 0.004). A nuclear/cytoplasmic asynchrony was a characteristic feature of the ultrastructure of the myeloma cells. During treatment a significant increase from 34 to 54 per cent (2P = 0.03) was observed in the number of plasma cells with slight asynchrony. No correlation could be established between any ultrastructural features and the values obtained with clinical tests considered of major prognostic significance at the time of diagnosis, e.g. the concentrations of serum creatinine, haemoglobin, serum albumin and serum calcium.

Adult↗
PubMed2004

Hematologic reference intervals for koi (Cyprinus carpio), including blood cell morphology, cytochemistry, and ultrastructure.

BACKGROUND: Hematologic data are used routinely in the health care of humans and domestic mammals. Similar data for fish are largely fragmentary or have not been collected. OBJECTIVES: The primary purpose of this study was to determine hematologic reference intervals for koi, an ornamental strain of the common carp (Cyprinus carpio). Secondarily, the morphology, cytochemical reactions, and ultrastructure of koi blood cells were characterized. METHODS: A CBC was performed manually on heparin-anticoagulated blood specimens using Natt and Herrick's diluent and a Neubauer-ruled hemacytometer. Leukocyte differential counts were done on Wright-Leishman- and Diff-Quik-stained blood smears. Cytochemical reactions of koi leukocytes were determined using commercial kits. Transmission electron microscopy was performed to characterize the ultrastructural features of koi blood cells. RESULTS: Hematologic reference intervals were established for healthy koi for PCV (30-34%), hemoglobin concentration (6.3-7.6 g/dL), RBC count (1.7-1.9 X 10(6)/ microL), WBC count (19.8-28.1 X 10(3)/ microL), RBC indices, and differential leukocyte counts. Lymphocytes were the predominant leukocyte (accounting for up to 80% of all leukocytes), whereas eosinophils were rare. Basophils were positive with PAS staining. Naphthol AS-D chloroacetate esterase activity was observed only in eosinophils. alpha-Naphthyl butyrate esterase and beta-glucuronidase activities were positive in monocytes. Some lymphocytes were reactive for alpha-naphthyl butyrate esterase and acid phosphatase activity. Ultrastructurally, leukocytes, erythrocytes, and thrombocytes were identified on the basis of cytoplasmic organelles and granule appearance. CONCLUSION: Hematologic reference intervals and knowledge of the cytochemical reactions and ultrastructural characteristics of koi leukocytes will help standardize hematologic studies in this species.

Animals↗
PubMedDate not supplied

The effects of amorolfine and oxiconazole on the ultrastructure of Trichophyton mentagrophytes. A comparison.

Amorolfin applied in concentrations of 0.1-100 micrograms ml-1 caused considerable damages to the ultrastructure of Trichophyton mentagrophytes: Electron-transparent areas appear in the cytoplasma. The cell wall increases in thickness. Extracytoplasmic membrane vesicles are formed and deposited in the cell wall. Starved fungal cells, with normal ultrastructure, can be found. Lysed, dead cells demonstrate the process of final, vigorous ultrastructural damage. The feature of the damage caused by amorolfin is comparable to that effected by azole antifungals. The characteristics of the ultrastructural changes are similar to those due to amorolfin in the yeast species Candida albicans.

Antifungal Agents↗
PubMed1984

Colloidal gold and biotin-avidin conjugates as ultrastructural markers for neural antigens.

The use of two new reagents, biotin-avidin conjugates and colloidal gold particles, for ultrastructural immunocytochemistry has been reviewed, and their potential for immunocytochemistry in the central nervous system has been discussed. Although these compounds have been used very little in the C.N.S. in the past, with their high sensitivity, versatility, and potential for simultaneous localization of two antigens with two electron dense markers, these reagents may prove very useful for answering a number of neural questions. Colloidal gold adsorbed to immunoglobulins can be used for light microscopic observation, and owing to its high electron density and ability to readily adsorb to immunoglobulins, protein A, and antigen-carrier complexes, it also serves as an excellent immunomarker at the ultrastructural level. Several procedures have been described for the use of pre- and post-embedding immunostaining with colloidal gold particles. Different size gold particles, adsorbed to different antibodies can be used for pre- and post-embedding staining of different antigens in the same tissue. While at the light microscopic level only the large particles are visible as a pink haze, a fluorescent step can be added to allow easy identification and characterization of neurones labelled with any size colloidal gold, even if the pink colour of the gold is not visible. These same sections can subsequently be studied ultrastructurally. Unlike enzymatic methods, gold particles have less tendency to obscure ultrastructure. Biotin can be easily conjugated to immunoglobulins and, due to its high affinity for avidin (10(15) M-1) which has four biotin binding sites, can be localized with avidin markers, or biotin-avidin-marker complexes. Some procedures such as the avidin-biotin-peroxidase complex are extremely sensitive. Biotin and avidin can be bound to a number of different markers including peroxidase, colloidal gold, and ferritin. Photomicrographs from our hypothalamic work have been used to illustrate the histological results of some of the procedures described.

Animals↗
PubMed1990

Ultrastructural alterations induced by two ergosterol biosynthesis inhibitors, ketoconazole and terbinafine, on epimastigotes and amastigotes of Trypanosoma (Schizotrypanum) cruzi.

We report the ultrastructural alterations induced during the proliferative stages of Trypanosoma (Schizotrypanum) cruzi, the causative agent of Chagas' disease, by two ergosterol biosynthesis inhibitors, ketoconazole and terbinafine, which had previously been shown to be potent growth inhibitors whose effects are potentiated when used in combination (J. A. Urbina, K. Lazardi, T. Aguirre, M. M. Piras, and R. Piras, Antimicrob. Agents Chemother. 32:1237-1242, 1988). Epimastigotes treated with a low concentration of ketoconazole (1 microM), which blocks ergosterol biosynthesis at the level of C-14 demethylation of lanosterol and induces cell lysis coincident with total ergosterol depletion, showed gross alterations of the kinetoplast-mitochondrion complex, which swelled and lost the organization of its inner membrane and the electron-dense bodies of its matrix. Thus, coincident with the beginning of cell lysis, the kinetoplast-mitochondrion complex occupied greater than 80% of the cell volume, while other subcellular structures such as the nucleus and subpellicular microtubules were not affected. Terbinafine, which blocks ergosterol synthesis in these cells at the level of squalene synthetase and thus leads to almost immediate arrest of growth at concentrations greater than 1 microM, produced proliferation of glycosomelike bodies, binucleated cells (arrest at cytokinesis), and eventually massive vacuolization. When the drugs were combined, the predominant effect was mitochondrial swelling, which was more drastic and took place earlier than that observed in cells treated with ketoconazole alone. In amastigotes proliferating in Vero cells, ketoconazole at the concentration required to eradicate the parasites (10 nM) produced mitochondrial swelling, the appearance of autophagic vacuoles containing partially degraded subcellular material, and finally a general breakdown of the subcellular structures. Terbinafine at 3 microM induced more limited ultrastructural damage to the amastigotes consistent with increased vacuolization of the cells and the appearance of occasional autophagic vacuoles. When the drugs were used in combination, just 1 nM was required for the total eradication of parasites, the ultrastructural effects were more extensive, and cell disintegration occurred earlier than when any of the drugs was used alone at a much higher concentration. No effect of the drugs on the ultrastructure of the host cells were observed at any of the concentrations tested.

Animals↗
PubMed1991

Occurrence and ultrastructural characterization of bacteria in association with and isolated from Azolla caroliniana.

The occurrence and ultrastructure of bacteria in leaf cavities of symbiotic Azolla caroliniana were examined by transmission electron microscopy. Bacteria were observed in all leaf cavities of Azolla cultures. Five ultrastructurally distinct types of bacteria were observed in each individual leaf cavity. Features used to characterize the bacteria included morphology, cell wall structure, and cytoplasmic organization. At least one gram-positive and as many as four gram-negative types of bacteria reside in leaf cavities of A. caroliniana. The morphological and ultrastructural characteristics of the gram-positive bacterium suggest that it is an Arthrobacter sp. The gram-negative bacteria could not be cultured; therefore, they have not been classified further. Bacterial cell shape and cell wall structure were similar in leaf cavities of different ages, but cell size and cytoplasmic composition varied. The relative contributions of each bacterial type to the total community within individual leaves was determined. Ultrastructural characteristics of bacterial isolates cultured from A. caroliniana in a free-living state were also examined.

Bacteria↗
PubMed1995

Ultrastructural diversity of the cellulase complexes of Clostridium papyrosolvens C7.

Transmission electron microscopy was used to investigate the ultrastructural features of diverse cellulase and cellulase-xylanase multiprotein complexes that are components of the cellulase-xylanase system of Clostridium papyrosolvens C7. The multiprotein complexes were separated by anion-exchange chromatography into seven biochemically distinguishable fractions (F1 to F7). Most individual F fractions contained, in relatively large numbers, an ultrastructurally recognizable type of particle that occurred only in smaller numbers, or not at all, in the other F fractions. It is suggested that these ultrastructurally distinct particles represent the biochemically distinct multiprotein complexes that constitute the cellulase-xylanase system of C. papyrosolvens C7. Some of the particles consisted of tightly packed globular components that appeared to be arranged in the shape of a ring with conical structures pointing out along its axis. Other particles had triangular, polyhedral, or star shapes. The major protein fraction (F4) almost exclusively contained particles consisting of loosely aggregated components, many of which appeared to be arranged along filamentous structures. The ultrastructural observations reported here support our previous conclusion that the cellulase-xylanase system of C. papyrosolvens C7 comprises at least seven different high-molecular-weight multiprotein complexes. Furthermore, results of this and earlier studies indicate that the interactions between C. papyrosolvens C7 and cellulose are different from those that have been described for Clostridium thermocellum.

Cellulase↗
PubMed2004

Increased DNA fragmentation and ultrastructural changes in fibromyalgic muscle fibres.

OBJECTIVE: To determine whether there is evidence of increased DNA fragmentation and ultrastructural changes in muscle tissue of patients with fibromyalgia (FM) compared with healthy controls. METHODS: Muscle tissues from 10 community residents with FM and 10 age and sex matched healthy controls were examined "blindly" for the presence of DNA fragmentation by two different methods: terminal deoxynucleotidyl transferase (TdT) staining (TUNEL) and the FragEL-Klenow DNA fragmentation detection kit. Ultrastructural analysis of tissue was performed by electron microscopy. RESULTS: DNA fragmentation was detected by both methods in 55.4 (SEM 2.5)% of the nuclei in muscle tissue of patients with FM compared with 16.1 (4.1)% (p<0.001) of the nuclei in healthy controls. Contrary to expectation, no typical features of apoptosis could be detected by electron microscopy. The myofibres and actin filaments were disorganised and lipofuscin bodies were seen; glycogen and lipid accumulation were also found. The number of mitochondria was significantly lower in patients with FM than in controls and seemed to be morphologically altered. CONCLUSION: The ultrastructural changes described suggest that patients with FM are characterised by abnormalities in muscle tissue that include increased DNA fragmentation and changes in the number and size of mitochondria. These cellular changes are not signs of apoptosis. Persistent focal contractions in muscle may contribute to ultrastructural tissue abnormalities as well as to the induction and/or chronicity of nociceptive transmission from muscle to the central nervous system.

Adult↗
PubMed1993

Immunological and ultrastructural studies in acute biphenotypic leukaemia.

AIMS: To compare the sensitivity of the ultrastructural method to detect myeloperoxidase (MPO) with light microscopy and immunocytochemistry using an anti-MPO antibody; to examine the expression of lymphoid antigens in relation to MPO activity in blast cells from cases of biphenotypic leukaemia. METHODS: Blast cells from 14 cases of biphenotypic acute leukaemia were analysed. Immunological markers were performed by single or double immunofluorescence staining on a flow cytometer. The presence of MPO was determined by light microscopy, electron microscopy on fixed and unfixed cells, and by immunoalkaline phosphatase with an anti-MPO antibody. The immunogold method was applied at the ultrastructural level to assess the expression of lymphoid and myeloid antigens at the same time as the MPO activity. RESULTS: Six of the 14 cases were initially classified as acute lymphoblastic leukaemia (ALL) and eight as acute myeloid leukaemia (AML). MPO activity was shown at the ultrastructural level in 4-99% blasts from all cases. Six of the 14 were MPO negative by light microscopy and three of these were negative with the antibody anti-MPO. Coexpression of lymphoid antigens (CD19, CD10, or CD2) and MPO was shown by the immunogold method in four out of 11 cases; in seven cases the blasts coexpressed myeloid antigens (CD13, CD33) and MPO. CONCLUSIONS: Electron microscopy is more sensitive for showing MPO than light microscopy and immunocytochemistry; the immunogold method combined with MPO used at the ultrastructural level can help to define the cell lineage involved in biphenotypic leukaemia by highlighting the myeloid component defined by MPO.

Acute Disease↗
PubMed1984

Cellular ultrastructure of Amphiuma distal nephron: effects of exposure to potassium.

The cellular ultrastructure of the renal distal nephron of the salamander, Amphiuma means, was examined by electron-microscopic and stereological techniques before and after exposure to potassium in the ambient environment. The distal nephron of Amphiuma is composed of three ultrastructurally distinct segments: early distal (or diluting segment), late distal, and collecting tubule. The early distal tubule structurally resembles the mammalian thick ascending limb of Henle's loop. Large renin-like granules are present in the smooth muscle cells of the afferent arteriole in the vicinity of the early distal tubule, suggesting the presence of a rudimentary juxtaglomerular apparatus. Late distal tubules are composed of one large cell type characterized by extensive basal membrane invaginations, often extending to the luminal membrane. Collecting tubules contain principal and intercalated cells that are ultrastructurally similar to cells of the mammalian cortical collecting tubule. Exposure to potassium had no effect on the ultrastructure of early distal cells but led to a sharp increase in the basolateral membrane surface density of principal cells in the collecting tubule (1.17 +/- 0.08-1.63 +/- 0.13 micron2/micron3). Potassium adaptation leads to a similar structural response in the mammalian collecting tubule. Since Amphiuma collecting tubules can be isolated and perfused in vitro and impaled with ion- and voltage-sensitive microelectrodes, the observed structural adaptation suggests that the collecting tubule may be a useful preparation to study the cellular mechanisms of potassium adaptation.

Animals↗
PubMed2003

Assessment of the ultrastructural and proliferative properties of human embryonic stem cell-derived cardiomyocytes.

Assessment of early ultrastructural development and cell-cycle regulation in human cardiac tissue is significantly hampered by the lack of a suitable in vitro model. Here we describe the possible utilization of human embryonic stem cell (ES) lines for investigation of these processes. With the use of the embryoid body (EB) differentiation system, human ES cell-derived cardiomyocytes at different developmental stages were isolated and their histomorphometric, ultrastructural, and proliferative properties were characterized. Histomorphometric analysis revealed an increase in cell length, area, and length-to-width ratio in late-stage EBs (>35 days) compared with early (10-21 days) and intermediate (21-35 days) stages. This was coupled with a progressive ultrastructural development from an irregular myofibrillar distribution to an organized sarcomeric pattern. Cardiomyocyte proliferation, assessed by double labeling with cardiac-specific antibodies and either [3H]thymidine incorporation or Ki-67 immunolabeling, demonstrated a gradual withdrawal from cell cycle. Hence, the percentage of positively stained nuclei in early-stage cardiomyocytes ([3H]thymidine: 60 +/- 10%, Ki-67: 54 +/- 23%) decreased to 36 +/- 7% and 9 +/- 16% in intermediate-stage EBs and to <1% in late-stage cardiomyocytes. In conclusion, a reproducible temporal pattern of early cardiomyocyte proliferation, cell-cycle withdrawal, and ultrastructural maturation was noted in this model. Establishment of this unique in vitro surrogate system may allow to examine the molecular mechanisms underlying these processes and to assess interventions aiming to modify these properties. Moreover, the detailed characterization of the ES cell-derived cardiomyocyte may be crucial for the development of future cell replacement strategies aiming to regenerate functional myocardium.

Cell Differentiation↗
PubMed1995

Choroid plexus electrolytes and ultrastructure following transient forebrain ischemia.

A temporal profile of lateral and fourth ventricle rat choroid plexus (LVCP and 4VCP, respectively) tissue injury and recovery was determined using alterations in K, Na, and H2O content and ultrastructure after 10 min of transient forebrain ischemia (TFI). At 0.5 h postischemia the LVCP displayed a maximum reduction in K content by 32% and a significant increase in Na content by 85% and H2O content by 22%. LVCP tissue K, Na, and H2O content returned to sham values by 24 h postischemia. Ultrastructural changes appeared more severe between 0.5 and 12 h postischemia, whereas by 24 h, normal ultrastructure was restored. Elevations in 4VCP tissue Na (P < 0.05) and H2O content, which were less than those in LVCP, gradually reached a maximum by 24 h compared with sham. No change in 4VCP tissue ultrastructure was observed. These results indicate that the LVCP tissue is more vulnerable than 4VCP in the bilateral carotid artery occlusion model but that it recovers in a timely manner after TFI. Furthermore, the ability of the LVCP tissue to rapidly recover suggests its functional importance in helping to restore brain homeostasis.

Animals↗
PubMed1987

Effects of flow rate on proximal tubule ultrastructure.

In this study we investigated the ultrastructure of the proximal tubule during alterations in fluid flow to determine whether previously observed changes in solute and water reabsorption may be related to alterations in cell ultrastructure. In each kidney, two proximal tubules (S2 segments) were perfused simultaneously at 5 and 45 nl/min. Subsequently, cell ultrastructure of control and experimental tubules was examined by electron microscopy in combination with morphometry. Changes in flow rate greatly modified the geometry of the tubule epithelium. Enhanced flow increased luminal diameter and decreased cell height. The number of microvilli per square micrometer of luminal epithelial surface area decreased with increased flow rate from a control value of 42.5 to 35.5 at high flow. However, the total number of microvilli per millimeter tubule length did not change. Thus the distance between microvilli was dependent on flow rate and was estimated to be 621, 741, and 904 A in low flow, control, and high flow tubules, respectively. We suggest that increased flow rate, perhaps by altering transepithelial hydrostatic pressure gradients, leads to an increase in the distance between the microvilli and to other alterations in cellular ultrastructure that may contribute to the augmentation of solute and water reabsorption.

Animals↗
PubMed1993

Vasopressin- and cAMP-induced changes in ultrastructure of isolated perfused inner medullary collecting ducts.

Studies were performed to correlate arginine vasopressin (AVP)-induced changes in epithelial ultrastructure with changes in osmotic water permeability in isolated perfused rat terminal inner medullary collecting ducts (tIMCD). The tubules were perfused in three time periods, i.e., a 40-min basal period, a 40-min period with 0.1 nM AVP in the bath, and a 60-min withdrawal period. In each phase, the osmotic water permeability (Pf) was measured, and the perfused tubules were fixed for electron microscopy. AVP caused a four- to eightfold increase in Pf and induced several ultrastructural changes as follows: increased cell height of IMCD cells, expansion of the intercellular spaces, formation of large vacuoles, and increased coated pit density in the apical plasma membrane [from 0.6 +/- 0.2 (n = 6) to 2.9 +/- 0.3 (n = 7) pits/100 microns membrane length]. During AVP withdrawal, Pf decreased toward the basal value in association with partial reversal of the ultrastructural changes including a decrease in coated pit density to 1.0 +/- 0.2 (n = 4). Stimulation with 8-bromoadenosine 3',5'-cyclic monophosphate (8-bromo-cAMP) (0.1 mM) produced similar changes in Pf. Coated pit density increased to 2.1 +/- 0.4 (n = 4) after cAMP stimulation and after cAMP withdrawal decreased to 1.2 +/- 0.2 (n = 6). In contrast to stimulation with AVP, cAMP stimulation did not result in dilated intercellular spaces or formation of large vacuoles. The only ultrastructural feature that directly correlated with the water permeability was the density of coated pits in the apical membrane. Organelles involved in the endocytic pathway were studied with cationized ferritin or albumin-gold in the luminal perfusate. At the end of 40 min basal perfusion or AVP stimulation, luminal tracer was found almost exclusively in large multivesicular bodies (MVB). Tubules perfused with tracer during AVP withdrawal demonstrated rapid tracer accumulation in small vesicles and small MVB within 3-5 min, a time point corresponding to the rapid phase of Pf decrease. Later (30-60 min) the label was mainly confined to large MVB. Occasionally during AVP stimulation or withdrawal, small coated vesicles and smooth vesicles with coated extensions were noted to contain tracer. The data demonstrate AVP-mediated coated pit formation and cellular changes and show very rapid internalization of apical membrane after AVP withdrawal.

Animals↗
PubMed2004

Exercise training increases oxidative capacity and attenuates exercise-induced ultrastructural damage in skeletal muscle of aged horses.

Exercise training improves functional capacity in aged individuals. Whether such training reduces the severity of exercise-induced muscle damage is unknown. The purpose of the present study was to determine the effect of 10 wk of treadmill exercise training on skeletal muscle oxidative capacity and exercise-induced ultrastructural damage in six aged female Quarter horses (>23 yr of age). The magnitude of ultrastructural muscle damage induced by an incremental exercise test before and after training was determined by electron microscopic examination of samples of triceps, semimembranosus, and masseter (control) muscles. Maximal aerobic capacity increased 22% after 10 wk of exercise training. The percentage of type IIa myosin heavy chain increased in semimembranosus muscle, whereas the percentage of type IIx myosin heavy chain decreased in triceps muscle. After training, triceps muscle showed significant increases in activities of both citrate synthase and 3-hydroxyacyl-CoA-dehydrogenase. Attenuation of exercise-induced ultrastructural muscle damage occurred in the semimembranosus muscle at both the same absolute and the same relative workloads after the 10-wk conditioning period. We conclude that aged horses adapt readily to intense aerobic exercise training with improvements in endurance, whole body aerobic capacity, and muscle oxidative capacity, and heightened resistance to exercise-induced ultrastructural muscle cell damage. However, adaptations may be muscle-group specific.

Adaptation, Physiological↗