Fine Structural Alterations Research Articles

Neurocan Contributes to Perineuronal Net Development

Perineuronal nets (PNs) are matrix molecule assemblies surrounding neuronal somata, dendrites and axon initial segments in a lattice-like appearance. PN molecules are involved in many structural and physiological processes during development and in adulthood, suggesting a crucial role in normal brain function. Neurocan, as one of the main PN proteoglycans, is suggested to control important developmental processes of neuronal tissue. This statement relies on thorough and excellent experimental work mainly conducted in reduced systems, such as cell cultures. However, previous data collected in neurocan-deficient mice do not seem to support neurocan’s role in development since brain development in general and the formation of PNs especially in the hippocampus were reported to be undisturbed in neurocan-deficient mice. Here, we aim to re-address the role of neurocan in developmental processes by investigating the influence of neurocan on PN formation in the medial nucleus of the trapezoid body, a PN-enriched nucleus in the auditory brainstem, using neurocan-deficient mice. Immunohistochemical and biochemical analyses demonstrate that neurocan controls the regulation of PN development by influencing mRNA and protein quantity of various PN molecules. Resulting alterations in PN fine structure are critical for PN function as estimated by reduced amount of GAD65/67 and prolongation of synaptic transmission delay of calyx of Held synapses. Thus, neurocan contributes to proper PN formation and synapse physiology in the MNTB.

Double Bond Characterization of Free Fatty Acids Directly from Biological Tissues by Ultraviolet Photodissociation

Free fatty acids (FA) are a vital component of cells and are critical to cellular structure and function, so much so that alterations in FA are often associated with cell malfunction and disease. Analysis of FA from biological samples can be achieved by mass spectrometry (MS), but these analyses are often not capable of distinguishing the fine structural alterations within FA isomers and often limited to global profiling of lipids without spatial resolution. Here, we present the use of ultraviolet photodissociation (UVPD) for the characterization of double bond positional isomers of charge inverted dication·FA complexes and the subsequent implementation of this method for online desorption electrospray ionization (DESI) MS imaging of FA isomers from human tissue sections. This method allows relative quantification of FA isomers from heterogeneous biological tissue sections, yielding spatially resolved information about alterations in double bond isomers within these samples. Applying this method to the analysis of the monounsaturated FA 18:1 within breast cancer subtypes uncovered a correlation between double bond positional isomer abundance and the hormone receptor status of the tissue sample, an important factor in the prognosis and treatment of breast cancer patients. This result further validates similar studies that suggest FA synthase activity and FA isomer abundances are significantly altered within breast cancer tissue.

Neonatal tactile stimulation reverses alterations in fine structure of small, but not large myelinated fibers, from the optic nerve of iron-deficient rats: A size-based selectivity

Iron is the most common micronutrient deficiency in the world and it is most prevalent in young children, exposing their developing brain to inadequate iron levels. The damage related to neuroanatomical parameters is not reversed after iron treatment. However, evidence suggest that tactile stimulation (TS) may offer great therapeutic efficacy in cases of nutritional disorders postnatally, since the brain is remarkably responsive to its interaction with the environment. Recently, we shown that neonatal iron deficient rats achieved some remedial effect by exposing them to TS treatment early in life, reinforcing the fact that the TS approach is a positive enriching experience, therefore, here we ask whether exposure to TS treatment, could also be employed to prevent fine structural changes in the fibers from optic nerve of rats maintained on an iron-deficient diet during brain development. To elucidate the protective effect of tactile stimulation, our methods resulted in 10,859 analyzed fibers, divided into small and large fibers. We found that iron deficiency led to a decreased axon, fiber and myelin size of small fibers, however, TS completely reversed the iron-decifiency-induced alteration on those fiber measurements. Large fibers were disproportionately affected by iron deficiency and there was no remediating effect due to tactile stimulation treatment. The present study adds new information regarding different alterations between small and large fibers due to diet and TS, which suggest a size-based selectivity. These results emphasize the concept that compromised brain development can be mitigated at an early age by environmental factors, such as tactile stimulation.

Salinity stress response of the invasive dinoflagellate Prorocentrum minimum

Estuarine and coastal environments are vulnerable to alien species invasions. Many nonindigenous organisms demonstrate high ecological plasticity, which is particularly well expressed in unicellular eukaryotes. In this study, responses of the potentially toxic invasive dinoflagellate Prorocentrum minimum, initially acclimated to salinity 17 (control), to short-term stress by salinities 4, 8 and 35 were investigated experimentally by determination of mortality, alterations in cell cycle and chromosome fine structure, RNA synthesis and DNA replication. The percentage of dead cells within the P. minimum population after stress by critical salinity 8 was lower compared to the other tested conditions. Salinity stress caused only moderate impact on the cell cycle pattern; changes of chromosome fine structure at salinities 4 and 8 were evident though reversible. We disclosed the elevated RNA synthesis and DNA replication which can play a ‘compensational’ role by speeding up the metabolic (synthetic) activity of these protists at the critical salinity conditions. The increased RNA synthesis in phase G1 and additional DNA replication in phase S of the cell cycle likely enhance salinity tolerance and cause relatively low mortality of these micro-eukaryotes at salinity 8. We infer that the enhanced synthesis of DNA and RNA can be one of the advanced ecological strategies of P. minimum which along with the other cellular and physiological characteristics (small body size, planktonic mode of life, fast reproduction, high evolution rates, mixotrophy, ability to form cysts, etc.) helps these dinoflagellates to invade successfully and populate permanently coastal ecosystems, remaining highly competitive in the unstable brackishwater environments.

Reduced CA2–CA3 Hippocampal Subfield Volume Is Related to Depression and Normalized by l-DOPA in Newly Diagnosed Parkinson’s Disease

Hippocampal dysfunctions may play an important role in the non-motor aspects of Parkinson’s disease (PD), including depressive and cognitive symptoms. Fine structural alterations of the hippocampus and their relationship with symptoms and medication effects are unknown in newly diagnosed PD. We measured the volume of hippocampal subfields in 35 drug-naïve, newly diagnosed PD patients without cognitive impairment and 30 matched healthy control individuals. Assessments were performed when the patients did not receive medications and after a 24-week period of l-DOPA treatment. We obtained a T1-weighted 3D magnetization-prepared rapid acquisition gradient echo image at each assessment. FreeSurfer v6.0 was used for image analysis. Results revealed a selectively decreased CA2–CA3 volume in non-medicated PD patients, which was normalized after the 24-week treatment period. Higher depressive symptoms were associated with smaller CA2–CA3 volumes. These results indicate that the CA2–CA3 subfield is structurally affected in the earliest stage of PD in the absence of cognitive impairment. This structural anomaly, normalized by l-DOPA, is related to depressive non-motor symptoms.

Expression of an (Engineered) 4,6-α-Glucanotransferase in Potato Results in Changes in Starch Characteristics.

Starch structure strongly influences starch physicochemical properties, determining the end uses of starch in various applications. To produce starches with novel structure and exploit the mechanism of starch granule formation, an (engineered) 4, 6-α-glucanotransferase (GTFB) from Lactobacillus reuteri 121 was introduced into two potato genetic backgrounds: amylose-containing line Kardal and amylose-free mutant amf. The resulting starches showed severe changes in granule morphology regardless of genetic backgrounds. Modified starches from amf background exhibited a significant increase in granule size and starch phosphate content relative to the control, while starches from Kardal background displayed a higher digestibility, but did not show changes in granule size and phosphate content. Transcriptome analysis revealed the existence of a mechanism to restore the regular packing of double helices in starch granules, which possibly resulted in the removal of novel glucose chains potentially introduced by the (engineered) GTFB. This amendment mechanics would also explain the difficulties to detect alterations in starch fine structure in the transgenic lines.

The structural and functional changes indcuced by lithium on the renal cortex of growing albino rats: Ultrastructure and laboratory study

Introduction: Lithium is a therapeutic agent currently used for the treatment of affective disorders controlling a variety of neurotic and psychosomatic manic depressions. The main objective of the present work was to demonstrate the histopathological effects of the therapeutic doses of Lithium on the renal tubules and glomeruli in growing albino rats. Material and Methods: Thirty growing male Sprague - Dawley albino rats were used in this study. The rats were divided into a control group formed of 6 rats and an experimental group formed of 24 rats which received a daily therapeutic dose of 20 mg Lithium/kg body weight by the same route for 7 weeks. The renal cortex in all animals is examined by light and electron microscopes. Blood was collected from the sacrificed animals for serum creatinine, urea, sodium and potassium to access the effect of lithium administration in a therapeutic dose on renal function. Results: The present work revealed that the therapeutic doses of Lithium induced nephrotoxicity in the form of degeneration and necrosis in the renal tubules and glomeruli. Alteration in the cellular fine structure and degenerated cytoplasm and cytoplasmic organelles were found revealing cellular degeneration and necrosis. Glomerulosclerosis and congestion were the predominant effect on the renal glomeruli. Conclusion: Histological and ultrastructrual features of Lithium nephrotoxicity were detected in the current study with therapeutic doses of Lithium.

Effect O6-guanine alkylation on DNA flexibility studied by comparative molecular dynamics simulations.

Alkylation of guanine at the O6 atom is a highly mutagenic DNA lesion because it alters the coding specificity of the base causing G:C to A:T transversion mutations. Specific DNA repair enzymes, e.g. O(6)-alkylguanin-DNA-Transferases (AGT), recognize and repair such damage after looping out the damaged base to transfer it into the enzyme active site. The exact mechanism how the repair enzyme identifies a damaged site within a large surplus of undamaged DNA is not fully understood. The O(6)-alkylation of guanine may change the deformability of DNA which may facilitate the initial binding of a repair enzyme at the damaged site. In order to characterize the effect of O(6)-methyl-guanine (O(6)-MeG) containing base pairs on the DNA deformability extensive comparative molecular dynamics (MD) simulations on duplex DNA with central G:C, O(6)-MeG:C or O(6)-MeG:T base pairs were performed. The simulations indicate significant differences in the helical deformability due to the presence of O(6)-MeG compared to regular undamaged DNA. This includes enhanced base pair opening, shear and stagger motions and alterations in the backbone fine structure caused in part by transient rupture of the base pairing at the damaged site and transient insertion of water molecules. It is likely that the increased opening motions of O(6)-MeG:C or O(6)-MeG:T base pairs play a decisive role for the induced fit recognition or for the looping out of the damaged base by repair enzymes.

Muscle biopsy substantiates long‐term MRI alterations one year after a single dose of botulinum toxin injected into the lateral gastrocnemius muscle of healthy volunteers

Despite numerous clinical and experimental studies on botulinum toxin type A (BoNT/A), long-term alterations of muscle texture and fine structure following BoNT/A treatment have thus far not been studied in normal human skeletal muscle. After obtaining institutional review board approval, we performed a prospective, placebo-controlled, double-blinded follow-up study on two healthy adults using magnetic resonance imaging (MRI) and muscle biopsy to visualize long-term alterations after a single BoNT/A injection into the lateral head of the gastrocnemius muscle. MRI disclosed a high-signal-intensity pattern in short tau inversion recovery sequences, and a reduction of the cross-sectional area in the BoNT/A-injected, but not in the saline-injected contralateral control muscle (at 6 to 9 months in volunteer A: 73%, in B: 62%; at 12 months in A: 88%, and in B: 78%). Enzyme histochemistry, 12 months after injection, confirmed neurogenic atrophy of muscle fibers only in the BoNT/A-injected muscle. Electron microscopy revealed additional degenerative changes at the neuromuscular junction. The data confirm that MRI is a suitable tool to monitor the long-term effect of BoNT/A on skeletal muscle. Neurogenic muscle atrophy following a single BoNT/A injection should be taken into consideration when repeated BoNT/A injections into the same muscles are proposed.

Structural and functional changes of sulfated glycosaminoglycans in Xenopus laevis during embryogenesis

Xenopus laevis is an excellent animal for analyzing early vertebrate development. Various effects of glycosaminoglycans (GAGs) on growth factor-related cellular events during embryogenesis have been demonstrated in Xenopus. To elucidate the relationship between alterations in fine structure and changes in the specificity of growth factor binding during Xenopus development, heparan sulfate (HS) and chondroitin/dermatan sulfate (CS/DS) chains were isolated at four different embryonic stages and their structure and growth factor-binding capacities were compared. The total amounts of both HS and CS/DS chains decreased from the pre-midblastula transition to the gastrula stage, but increased exponentially during the following developmental stages. The length of HS chains was not significantly affected by development, whereas that of CS/DS chains increased with development. The disaccharide composition of GAGs in embryos also changed during development. The degree of sulfation of the HS chains gradually decreased with development. The predominant sulfation position in the CS/DS chains shifted from C4 to C6 of GalNAc during embryogenesis. Growth factor-binding experiments using a BIAcore system demonstrated that GAGs bound growth factors including fibroblast growth factors-1 and -2, midkine, and pleiotrophin, with comparable affinities. These affinities significantly varied during development, although the correlation between the structural alterations of GAGs and the change in the ability to bind growth factors remains to be clarified. The expression of saccharide sequences, which specifically interact with a growth factor, might be regulated during development.

Ultrastructural pathology of human hydrocephalic cerebral cortex

The ultrastructural pathology of cerebral cortex in human hydrocephalus is described. Cortical biopsies of 17 patients of ages ranging from 10 days to 21 years old examined in our laboratory are described. Nerve cells show moderate and severe swelling characterized by dilation of endoplasmic reticulum canaliculi and perinuclear cistern, edema and degenerative changes of Golgi apparatus, variable degrees of mitochondrial swelling, and fragmentation of plasma membrane. The neighboring neuropil exhibits notable enlargement of extracellular space and signs of synaptic plasticity and degeneration. The astrocytes display edematous changes and phagocytic activity. Glycogen rich- and glycogen-depleted astrocytes are observed. Oligodendroglial cells appear normal in certain cases and in others show moderate hydropic changes. Numerous myelin figures are observed in some unidentified nerve cell processes. The capillary wall shows evident signs of blood-brain barrier dysfunction featured by increased endothelial vesicular and vacuolar transport, closed and open interendothelial junctions, and in severely edematous areas thinning and breakdown of capillary basement membrane. The role of ischemia, oxidative stress, increased calcium concentration, activation of N-methyl-d-aspartate (NMDA) receptors, and disturbance of ion homeostasis are discussed in relationship with the fine structural alterations of hydrocephalic brain parenchyma.

Ultrastructural aspects of liver injury with special reference to small bile ducts in patients with ulcerative colitis.

Liver tissue specimens taken at colectomy from 29 patients with chronic ulcerative colitis were studied by electron microscope. The fine-structural alterations were correlated with light microscopy and with biochemical liver function tests. The purpose was to identify ultrastructural features which could explain the pathogenesis of sclerosing cholangitis. Severely injured bile-duct epithelial cells were seen in three out of the eight light-microscopically diagnosed cholangitis cases, in the two cases of non-specific reactive hepatitis, and in the two fatty livers. Four cholangitis cases had, in heavily thickened bile-duct basement membranes, translucent areas containing bile-like material. Bile-duct microvilli were often blunted, and reduced in number. Intracanalicular bile thrombi and bile inclusions in hepatocytes were seldom seen, mostly but not exclusively in cholangitis. The fine-structural alterations apparently represent various stages of liver injury. These findings do not appear to be specific, but their prominence seems to correlate with the progression of the disease, at least in the case of histological parameters, but also in serum enzyme activities indicative of cholestasis. The bile-like electron-dense material found in proliferating basement membranes, very possibly regurgitated into the injured bile-duct wall after epithelial injury, could enhance the development of periductal fibrosis, leading to progression of sclerosing cholangitis.

Alterations in Heparan Sulfate in the Vessel in Response to Vascular Injury in the Mouse

Heparan sulfate (HS) is ubiquitous throughout the human body. The backbone of HS is composed of many types of sugars. HS serves as a docking site for a vast array of protein ligands. Recent evidence suggests a unique diversity in HS structure that alters protein binding and protein function. This diversity in HS structure has been overlooked till now. The goal of this study was to determine whether femoral artery wire injury modified HS structure. Femoral artery wire injury was performed in 16-week-old male C57BL6 mice. Transcript levels of a panel of enzymes that regulate HS fine structure, including N-deacetylase-N-sulfotransferases (Ndst) 1 and 2, exostoses (Ext) 1 and 2, C5 epimerase, and 2-O and 6-O sulfotransferases, were quantified with real-time quantitative polymerase chain reaction at 7 and 14 days post injury. All enzymes showed significant alterations in messenger RNA expression in response to injury. Ndst1, the most prevalent isoform, exhibited a 20-fold increase in response to injury. Injury induced significant alterations in fine structure specially increases in N-sulfated disaccharides at 14 days post injury. Vascular injury invokes transcriptional regulation of the enzymes that regulate HS structure, as well as changes in the pattern of HS chains in the vessel wall 14 days post injury. These findings may be important as the foundation of altered growth factor and chemokine binding in the process of vascular remodeling.

Hepatocyte Syndecan 1 Mediates Triglyceride‐Rich Lipoprotein Clearance in the Liver

We previously provided genetic evidence that alterations in heparan sulfate fine structure result in an accumulation of plasma triglyceride‐rich lipoproteins (MacArthur et al, J. Clin. Invest. 117:153, 2007). In this study, we sought to determine which heparan sulfate proteoglycans play a role in triglyceride‐rich lipoprotein metabolism. RT‐PCR analysis of hepatocytes demonstrated the expression of the membrane proteoglycans syndecans‐1, ‐2, and ‐4, glypicans‐1 and ‐4, and the secreted proteoglycans agrin, perlecan, and collagen XVIII. Analysis of available mice deficient in the various proteoglycans showed that only syndecan 1 and collagen XVIII mutants accumulated plasma triglycerides. Furthermore the extent of accumulation was comparable to that observed in heparan sulfate deficient strains. Immunoelectron microscopy showed expression of syndecan‐1 on plasma membrane microvilli of hepatocyte basal membranes lining the space of Disse. Mice deficient in syndecan‐1 synthesized VLDL normally, but exhibited reduced uptake of injected VLDL. Retinyl ester excursion studies showed that clearance of intestinally derived lipoproteins also depended on syndecan‐1. In contrast, collagen XVIII deficient mice did not show a defect in hepatic clearance, but rather a change in metabolism of lipoproteins in the peripheral circulation. These findings show that syndecan 1 is an hepatic proteoglycan receptor mediating the clearance of both intestinally derived and hepatic lipoprotein particles.

Ultrastructure and X-ray microanalysis of Euglena gracilis (Euglenophyta) under chromium stress

I. Rocchetta, P.I. Leonardi, G.M. Amado Filho, M. del Carmen Ríos de Molina and V. Conforti. 2007. Ultrastructure and X-ray microanalysis of Euglena gracilis (Euglenophyta) under chromium stress. Phycologia 46: 300–306. DOI: 10.2216/06-49.1The effect of chromium on the biology of Euglena gracilis was studied. The ultrastructural modifications caused by this metal and its location within the cell were analyzed by TEM and EDXA, respectively. The effects of chromium on protein, pigment, and lipid contents were also studied in order to evaluate the metabolic responses to metal exposure. Two strains of Euglena gracilis, UTEX 753 (from the Culture Collection of Texas University) and MAT (isolated from the Matanza River), were used in this research. Both were grown in photoauxotrophic and photoheterotrophic conditions and exposed to different metal concentrations. In all treated cells, increases in total protein and lipid contents, changes in chlorophyll amount, and alterations in fine structure were observed, especially with the higher concentration tested. In photosynthetic treated cells, assays showed chloroplast thylakoid disorganization, the presence of cytoplasm lipid globules, and several vacuoles with electron-dense inclusions and remnants of membranes inside. Nuclei presented lobulations, and eventually total fragmentation in some cells treated with the highest chromium concentration was seen, suggesting that chromium cytotoxicity leads to cellular death. The EDXA spectrum showed well-defined Cr and S peaks in the vacuoles containing electron-dense inclusions and remnants of membranes from autotrophic MAT samples. These results indicate that the different defense mechanisms against chromium depend on strain type and culture conditions. The S peak detected in MAT would suggest that sulfur-rich proteins groups play an important role in the detoxification system inducing metal-complex accumulation into vacuoles.

A large german kindred with cold‐aggravated myotonia and a heterozygous A1481D mutation in the SCN4A gene

Muscle sodium-channel disorders cover a spectrum of rare myotonic diseases. In a German family with 17 affected individuals in four generations, we identified a heterozygous missense mutation in exon 24 A1481D (c.4442 C>A) of the voltage-gated sodium channel gene (SCN4A) alpha subunit. Phenotypes of 12 family members were characterized by a mild myotonia with cold sensitivity but without paramyotonia. The index patient presented with fluctuating cold- and exercise-induced stiffness of ocular, facial, and distal muscles. The myotonia became more severe at the age of 22 years. His father had had cold- and exercise-induced periodic weakness with fluctuating myotonia since age 10. Later he developed a more severe, purely exercise- and cold-aggravated myotonia of arms, hands, and facial muscles. The father's mother presented with cold-induced myotonia until age 65, when progressive weakness of proximal limb muscles developed. Her muscle biopsies revealed considerable myopathic changes with a variety of fine structural alterations. This study presents a family with cold-aggravated myotonia and progression of myopathic changes in the muscle biopsy with increasing age. In older patients, sodium channelopathies may mimic the phenotypic features of myotonic dystrophy type 2.

Morphofunctional changes of orofacial muscles in patients with unilateral or bilateral cleft lip, alveolus and palate

The facial musculature is part of a biocybernetic feedback system within the stomatognathic system, the continuity of which is disturbed by clefts of the lip, jaw, and palate (cheilognathopalatoschisis). This results in topographic, fine-structural and functional alterations of the facial musculature. In two heads with unilateral clefts and two heads with bilateral clefts, the facial musculature was dissected and the M. orbicularis oris macroscopically and topographically examined. We found changes in the attachment of the M. orbicularis oris. The modiolus, which is coresponsible for the proper topographic relation of the M. obicularis oris to the other facial muscles, was unchanged. Further, we examined the vascular system of the muscle, which likewise adapts to the altered situation. Lip/jaw/palatal clefts result in anatomically functional and fine-structural alterations of the M. orbicularis oris, while the rest of the facial musculature remains unchanged.

Activity-Independent Prespecification of Synaptic Partners in the Visual Map of Drosophila

Specifying synaptic partners and regulating synaptic numbers are at least partly activity-dependent processes during visual map formation in all systems investigated to date . In Drosophila, six photoreceptors that view the same point in visual space have to be sorted into synaptic modules called cartridges in order to form a visuotopically correct map . Synapse numbers per photoreceptor terminal and cartridge are both precisely regulated . However, it is unknown whether an activity-dependent mechanism or a genetically encoded developmental program regulates synapse numbers. We performed a large-scale quantitative ultrastructural analysis of photoreceptor synapses in mutants affecting the generation of electrical potentials (norpA, trp;trpl), neurotransmitter release (hdc, syt), vesicle endocytosis (synj), the trafficking of specific guidance molecules during photoreceptor targeting (sec15), a specific guidance receptor required for visual map formation (Dlar), and 57 other novel synaptic mutants affecting 43 genes. Remarkably, in all these mutants, individual photoreceptors form the correct number of synapses per presynaptic terminal independently of cartridge composition. Hence, our data show that each photoreceptor forms a precise and constant number of afferent synapses independently of neuronal activity and partner accuracy. Our data suggest cell-autonomous control of synapse numbers as part of a developmental program of activity-independent steps that lead to a "hard-wired" visual map in the fly brain.

Ultrastructure and Lipid Alterations Induced by Cadmium in Tomato (Lycopersicon esculentum) Chloroplast Membranes

The effects of cadmium (Cd) uptake on ultrastructure and lipid composition of chloroplasts were investigated in 28-day-old tomato plants (Lycopersicon esculentum var. Ibiza F1) grown for 10 days in the presence of various concentrations of CdCl2. Different growth parameters, lipid and fatty acid composition, lipid peroxidation, and lipoxygenase activity were measured in the leaves in order to assess the involvement of this metal in the generation of oxidative stress. We first observed that the accumulation of Cd increased with external metal concentration, and was considerably higher in roots than in leaves. Cadmium induced a significant inhibition of growth in both plant organs, as well as a reduction in the chlorophyll and carotenoid contents in the leaves. Ultrastructural investigations revealed that cadmium induced disorganization in leaf structure, essentially marked by a lowered mesophyll cell size, reduced intercellular spaces, as well as severe alterations in chloroplast fine structure, which exhibits disturbed shape and dilation of thylakoid membranes. High cadmium concentrations also affect the main lipid classes, leading to strong changes in their composition and fatty acid content. Thus, the exposure of tomato plants to cadmium caused a concentration-related decrease in the fatty acid content and a shift in the composition of fatty acids, resulting in a lower degree of fatty acid unsaturation in chloroplast membranes. The level of lipid peroxides and the activity of lipoxygenase were also significantly enhanced at high Cd concentrations. These biochemical and ultrastructural changes suggest that cadmium, through its effects on membrane structure and composition, induces premature senescence of leaves.

Morpho-dynamic changes of mitochondria during ageing of human endothelial cells

Mitochondrial morphology is regulated in many cultured eukaryotic cells by fusion and fission of mitochondria. A tightly controlled balance between fission and fusion events is required to ensure normal mitochondrial and cellular functions. During ageing, mitochondria are undergoing significant changes on the functional and morphological level. The effect of ageing on fusion and fission of mitochondria and consequences of altered fission and fusion activity are still unknown although theoretical models on ageing consider the significance of these processes. Human umbilical vein endothelial cells (HUVECs) have been established as a cell culture model to follow mitochondrial activity and dysfunction during the ageing process. Mitochondria of old and postmitotic HUVECs showed distinct alterations in overall morphology and fine structure, and furthermore, loss of mitochondrial membrane potential. In parallel, a decrease of intact mitochondrial DNA (mtDNA) was observed. Fission and fusion activity of mitochondria were quantified in living cells. Mitochondria of old HUVECs showed a significant and equal decrease of both fusion and fission activity indicating that these processes are sensitive to ageing and could contribute to the accumulation of damaged mitochondria during ageing.