Presence Of Laminations Research Articles

Atypical structure of the nuclear membrane, distribution of nuclear pores and lamin B1 in spermatozoa of patients with complete and partial globozoospermia.

Globozoospermia is a form of male infertility characterized by spermatozoa with spherical heads lacking acrosomes. The aim of this study was to evaluate ultrastructural and molecular defects in different types of globozoospermia. Semen samples from 12 infertile patients (9 with complete globozoospermia and 3 with partial globozoospermia) and 10 normozoospermic men (control) were examined by transmission electron microscopy and immunocytochemistry with antibodies against lamin B1. The presence of lamin A and progerin was assessed by reverse transcription-PCR. Whole exome sequencing was performed in three patients. In semen samples with complete and partial globozoospermia, lamin B1 was observed at the periphery of sperm nuclei, whereas lamin A and progerin were absent. Nuclear envelope pores were found in spermatozoa from both patient groups, regardless of morphology and chromatin condensation, in contrast to the control group. Non-condensed chromatin was present in 51%-81% of cases of complete globozoospermia and in 36%-79% of cases of partial globozoospermia. Homozygous DPY19L2 and SPATA16 variants were identified in two patients with partial globozoospermia and one patient with complete globozoospermia. An atypical nuclear membrane with abnormal nuclear pore distribution and lamin B1 localization was observed in spermatozoa from patients with both complete and partial globozoospermia. The genetic defects in the DPY19L2 and SPATA16 genes detected in patients from both globozoospermic groups suggest a generalized disruption of nuclear structure in globozoospermia, highlighting the genetic and phenotypic similarities between complete and partial globozoospermia.

A novel anisotropy template for an improved interpretation of elastic anisotropy data

Tight unconventional rocks are characterized by the presence of laminations, preferentially oriented cracks, and an interconnected network of compliant minerals. Such anisotropic features can mechanically deform due to pressure depletion during production, leading to a human-induced change of elastic and fluid transport properties. Rock physics models allow us to better predict and assess stress- and direction-dependent elastic moduli of the rock, useful for horizontal stress estimates. However, elastic anisotropy can be challenging to measure and interpret. We have developed an anisotropy template that can be used to assess stress-dependent changes in elastic moduli and investigate rock textures. We present here the template construction using an effective medium model consisting of stiff and compliant layers and crack inclusions and evaluate the origin of stress-dependent stiffness changes in acoustic data from Berea, Bakken, Three Forks, and Mancos formations.

Deformation of the nucleus by TGF\u03b21 via the remodeling of nuclear envelope and histone isoforms

The cause of nuclear shape abnormalities which are often seen in pre-neoplastic and malignant tissues is not clear. In this study we report that deformation of the nucleus can be induced by TGFβ1 stimulation in several cell lines including Huh7. In our results, the upregulated histone H3.3 expression downstream of SMAD signaling contributed to TGFβ1-induced nuclear deformation, a process of which requires incorporation of the nuclear envelope (NE) proteins lamin B1 and SUN1. During this process, the NE constitutively ruptured and reformed. Contrast to lamin B1 which was relatively stationary around the nucleus, the upregulated lamin A was highly mobile, clustering at the nuclear periphery and reintegrating into the nucleoplasm. The chromatin regions that lost NE coverage formed a supra-nucleosomal structure characterized by elevated histone H3K27me3 and histone H1, the formation of which depended on the presence of lamin A. These results provide evidence that shape of the nucleus can be modulated through TGFβ1-induced compositional changes in the chromatin and nuclear lamina.

The effect of impact angle and bond strength on fragmentation in laminated materials

An experimental study was undertaken to explore the effects of weak laminations on the fragmentation of impacting rocks. In this work, the fragmentation behavior of manufactured disc specimens of laminated travertine marble is described according to variations in the lamination bond strength, the orientation of laminations at impact, and the impact energy. An experimental apparatus and procedure were developed so that the discs, 96mm in diameter and 25mm thick, could be impacted neatly on their circumferential edge, at a selected lamination impact angle, and subject to filming with a high speed camera. The results of the study generally indicated that weaker bonds cause fragmentation from lower drop heights, as expected. However, the results also indicate that the critical height for breakage is relatively lower and insensitive to the impact angle when the laminations are between 45 and 90 degrees relative to the impacting surface, but the critical height for breakage increases significantly (by a factor of between 2 and 4) as the orientation of the laminations becomes parallel to the impact surface. The presence of laminations and their orientation to the impact surface, directly influenced the failure mechanism of specimens upon impact. At low lamination impact angles, failure was by parting along laminations in direct tension (reflected compression). At intermediate impact angles (approximately 45 degrees), the fragmentation of the specimens was more explosive and fractures were a combination of steps across stronger sheets, and splits along weaker bonded laminations. For high impact angles, fragmentation occurred in indirect tension, along weak laminations.

Effect of Lamination on Shale Reservoir Properties: Case Study of the Montney Formation, Canada

The presence of lamination on sedimentary rocks is a distinct characteristic, particularly in shales. They are distinct due to the contrast between successive layers with regard to grain size, composition, color, and sedimentary structures, such as graded beds. Typically, the degree of lamination is controlled by the sedimentation rate and flow regime. Herein, we developed a mudstone classification scheme in terms of lamination because lamination-based shale facies are related to differing features in mineral composition, porosity, and Young’s modulus. This study also attempts to verify whether wireline log patterns are relevant to shale lithofacies. The relationship between the porosity and lamination of the Montney Formation can be used to estimate reservoir properties. Our results show that an increased silt lamina in mudstone leads to an increase in the quartz and calcite contents and a decrease in the clay content, which increases the porosity, permeability, and Young’s modulus. However, reservoir quality is not solely dependent on lamination because of the complex interaction between components. The degree of lamination affected the neutron, density, and sonic log responses. Furthermore, the presence of lamination tends to decrease the neutron percentage, with similar trends in density and sonic log box plots in the study area. When the percentage of clay or cement material decreases, the neutron and density log responses diminish. Meanwhile, when the rock texture variation increases with an increase in the degree of lamination, the sonic log response decreases.

Phosphogenesis during the Cenozoic transition from greenhouse to icehouse conditions: Upper Oligocene to lower Miocene siliceous, phosphate, and organic‐rich sediments near La Purísima, Baja California Sur, Mexico

AbstractUpper Oligocene and lower Miocene, siliceous, organic‐, and phosphate‐rich sediments are widespread in Baja California Sur (Mexico). A representative section at La Purísima was analysed for its sedimentology, stratigraphy, geochemistry, and mineralogy. A corresponding age model was obtained by dating zircons from ash layers (27.84 ± 0.33 to 21.21 ± 0.59 Ma). The sediments were deposited in an upwelling‐dominated, hemipelagic setting, for which the presence of lamination, the scarcity of benthic organisms (except for in gravity‐flow deposits), and redox‐sensitive trace‐element enrichments indicate oxygen‐depleted conditions. Anoxic conditions were particularly strong around 27 and 24–22 Ma. Gravity‐flow deposits are frequent and predominantly composed of phosphatic‐coated grains. They were generated by seismic and volcanic activity, as is indicated by the close association with volcanic ash layers. The phosphatic‐coated particles were formed in a more proximal, better‐oxygenated shelf environment. They precipitated also in situ within the hemipelagic sediments, where they were often concentrated by subsequent winnowing. In situ phosphogenesis also partly cemented the gravity‐flow deposits. At La Purísima, phosphogenesis occurred throughout the time interval investigated and was particularly important around 28–25.5 and 23.5–21.5 Ma. These two time intervals correspond to the late Oligocene glacial maximum and the Oligocene–Miocene and early Miocene glacial intervals Mi1 and Mi1a. This provides evidence for the increasing importance of glacial denudation during the Oligocene, which led to an enhanced phosphorus flux into the ocean. Cooler climates also promoted the efficient transfer of phosphorus to thermocline waters by increased upwelling. Subsidiary phases of phosphogenesis during the intervening warm periods are explained by the weathering of glacial legacy sediments. These observations suggest that during the transition from greenhouse to icehouse conditions in the Oligocene and Miocene, new and radical changes in the global phosphorus cycle affected and partly inverted feedback mechanisms between climate, geochemical cycles and life, and profoundly influenced the biosphere and its evolution.

Abstract 3672: The three-dimensional (3D) spatial lamin A/C organization in normal lymphocytes, and in Hodgkin and Reed-Sternberg cells

Abstract Classical Hodgkin Lymphoma (HL) is a B-Cell lymphoma comprising mononucleated Hodgkin cells (H) and diagnostic bi- to multi-nucleated Reed Sternberg cells (RS). Genomic instability in HL is characterized by telomere dysfunction caused by critically short telomeres and telomere uncapping. Super resolution microscopy showed significant increases in the amount of DNA-poor nuclear spaces in HL compared to normal primary lymphocytes. A direct 3D interaction between telomeres and shelterin protein TRF2 is progressively disrupted from H to RS cells by either massive upregulation or downregulation of TRF2. TRF2 interacts with nuclear matrix protein lamin A/C in the maintenance of the 3D genome organization, which is disturbed in H and RS cells. Therefore we hypothesized that the TRF2- telomere-lamin A/C interaction is also disturbed in H and RS cells and leads to organizational alterations of the genome in HL. To this end, we analyzed lamin A/C expression levels of lymphocytes, H and RS cells and found that lamin A/C is upregulated in cancer cells compared to both activated and resting lymphocytes. The regular homogeneous and round shaped pattern found in lymphocytes was replaced in H and RS cells by a more irregular one, characterized by the presence of lamin A/C invaginations and septa that subdivide the nuclei into multiple smaller compartments. Our data show no co-localization between telomeres and lamin A/C in the DNA-poor spaces. Lamin A/C bordered the DNA-poor spaces but was never found inside them or in association with inner telomere fragments. 3D TRF-2 staining showed upregulation of the protein compared to normal lymphocytes, in correlation with the observation made in EBV-negative patients with clinically aggressive disease. The analysis of pre-treatment Hodgkin's patients' samples will allow to determine whether the TRF2-telomere-lamin A/C binding is different in patients with recurrent versus non-recurrent disease. Citation Format: Fabio Contu, Aleksander Szczurek, Roberta Vanni, Hans Knecht, Sabine Mai. The three-dimensional (3D) spatial lamin A/C organization in normal lymphocytes, and in Hodgkin and Reed-Sternberg cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 3672.

Relationship of oil seep in Kudat Peninsula with surrounding rocks based on geochemical analysis

This study aims to investigate the relation of oil seepage at Sikuati area with the structural and petroleum system of Kudat Peninsula. The abundance of highly carbonaceous rocks with presence of lamination in the Sikuati Member outcrop at Kudat Peninsula may give an idea on the presence of oil seepage in this area. A detailed geochemical analysis of source rock sample and oil seepage from Sikuati area was carried out for their characterization and correlation. Hydrocarbon propectivity of Sikuati Member source rock is poor to good with Total Organic Carbon (TOC) value of 0.11% to 1.48%. and also categorized as immature to early mature oil window with Vitrinite Reflectance (VRo) value of 0.43% to 0.50 %Ro. Based on biomarker distribution, from Gas Chromatography (GC) and Gas Chromatography-Mass Spectrometry (GC-MS) analysis, source rock sample shows Pr/Ph, CPI and WI of 2.22 to 2.68, 2.17 to 2.19 and 2.46 to 2.74 respectively indicates the source rock is immature and coming from terrestrial environment. The source rock might be rich in carbonaceous material organic matter resulting from planktonic/bacterial activity which occurs at fluvial to fluvio-deltaic environment. Overall, the source rock from outcrop level of Kudat Peninsula is moderately prolific in term of prospectivity and maturity. However, as go far deeper beneath the surface, we can expect more activity of mature source rock that generate and expulse hydrocarbon from the subsurface then migrating through deep-seated fault beneath the Sikuati area.

Single plane illumination microscopy as a tool for studying nucleome dynamics.

Single plane illumination microscopy (SPIM) is a new optical method that has become extremely important in recent years. It is based on the formation of a "light slice" in the specimen in which fluorescently tagged molecules are observed. The spatial resolution is close to that of confocal optics, but without the disadvantages inherent to scanning or high laser irradiation doses. A recent development is light sheet fluctuation microscopy, which exploits the dynamic information contained in the fluorescence intensity fluctuations of each image pixel. Here we review the principles of this method and show some recent applications to the dynamics of transcription factors and chromatin. We show that the dimerization of Fos and Jun proteins is directly linked to their binding to DNA; that nuclear receptor activation changes their intranuclear dynamics; and that the viscoelastic behavior of interphase chromatin strongly depends on the presence of lamin A.

Complete archive of late Turonian to early Campanian sedimentary deposition in newly drilled cores from the Tarfaya Basin, SW Morocco

An expanded succession of organic-rich marlstones and limestones deposited in the Tarfaya Basin provides an outstanding opportunity to closely retrace climate evolution and sea-level changes during the Cretaceous greenhouse period. We present high-resolution X-ray fluorescence (XRF) scanning and bulk carbon- and oxygen-isotope records from two newly drilled sediment cores in the Tarfaya Atlantic coastal basin, which recovered a continuous Upper Turonian to Campanian succession of ∼290 m thickness. The XRF core scanning records reveal three long-term oscillations in the abundance of terrigenous elements (increase of Al, Ti, K, Si, and Fe normalized against Ca), which correspond to progressive transgressive phases followed by rapid regressions during the Coniacian and early Santonian. Sea-level highstands during this interval corresponding to the Coniacian−Santonian oceanic anoxic event 3 (OAE 3) are characterized by overall oxygen-depleted to anoxic conditions at the seafloor (indicated by the high organic carbon content, the presence of laminations, and low log[Mn/S], high log[V/Ca], and high log[Br/Ca]). The upper Santonian interval marks the transition from anoxic to oxic bottom-water conditions, prevalent through the early Campanian. The composite bulk carbonate δ 13 C curve exhibits strong similarities to the global stacked δ 13 C reference curve, characterized by negative excursions in the early Coniacian (Navigation and East Cliff events) and late Santonian (bracketed by the Haven Brow and Buckle events) and by positive excursions in the latest Turonian (Hitchwood event), middle Coniacian (Wight Fall event), and at the Santonian-Campanian boundary. During the early Campanian, enhanced accumulation of fine-grained carbonate and clay-rich hemipelagic sediments, increasing bulk carbonate δ 18 O, and low log(Br/Ca) and log(V/Ca) values indicate climate cooling, associated with a substantial improvement in bottom-water ventilation. Two long-term δ 13 C cycles of ∼2 m.y. duration, probably related to variations in Earth’s orbital eccentricity, are associated with the long-term cooling trend initiating the Campanian−Maastrichtian climate transition toward a cool greenhouse state.

Compression and flexural strength of bone cement mixed with blood.

To assess the compression and flexural strength of bone cement mixed with 0 ml, 1 ml, or 2 ml of blood. High viscosity polymethyl methacrylate (PMMA) loaded with or without gentamicin was used. Blood was collected from total knee arthroplasty patients. In the same operating room, one pack of cement each was mixed with 0 ml (control), 1 ml, or 2 ml of blood for 1 minute during the dough phase. The dough was extruded into cylindrical and rectangular moulds for 20 minutes of setting, and then cured in phosphate buffered saline at 37±1ºC for 7 days. The samples were visually inspected for fractures and areas of weakness, and then scanned using microcomputed tomography. 48 gentamicin-loaded and 59 non-gentamicin-loaded samples mixed with 0 ml (control), 1 ml, or 2 ml of blood were randomised for flexural and compression strength testing; each group had at least 6 samples. In samples loaded with or without gentamicin, the flexural and compressive strength was highest in controls, followed by samples mixed with 1 ml or 2 ml of blood. In samples mixed with 2 ml of blood, the flexural strength fell below the standard of 50 MPa. In samples mixed with 2 ml of blood and all gentamicin-loaded samples, the compressive strength fell below the standard of 70 MPa. Microcomputed tomography revealed areas of voids and pores indicating the presence of laminations and partitions within. The biomechanical strength of PMMA contaminated with blood may decrease. Precautions such as saline lavage, pack drying the bone, change of gloves, and prompt insertion of the implant should be taken to prevent blood from contaminating bone cement.

NUMERICAL PREDICTION OF THE EFFECTS OF LONGITUDINAL CRACK-LIKE LAMINATIONS ON THE TENSILE PROPERTIES OF WIRES

Carbon steel wires used for civil engineering applications may contain laminations. In the published literature, lamination was modelled as a separation between two faces with a finite distance. This technique is not suitable for modelling the line-type/crack-like laminations that may be present in the wires. In this paper, the effects of longitudinal line-type laminations on the tensile properties of carbon steel wires were investigated using Finite element (FE) analysis. Laminations were modelled as seams which truly simulate the line-type/crack-like laminations that have been reported to be instrumental to the failure of pre-stressing wires. FE analysis revealed that laminations do not significantly reduce the yield and ultimate loads of the wires. However, laminations cause a significant reduction in the displacement at fracture of the wires and the reduction is proportional to the length of the laminations. Consequently, the presence of laminations reduces the ductility of the wires, which reduces the ability of the wires to withstand overload the wires may experience in service without causing a catastrophic failure of structures where wires provide the required reinforcement.

Vivianite is a key sink for phosphorus in sediments of the Landsort Deep, an intermittently anoxic deep basin in the Baltic Sea

Phosphorus (P) is an essential nutrient for marine organisms. Its burial in hypoxic and anoxic marine basins is still incompletely understood. Recent studies suggest that P can be sequestered in sediments of such basins as reduced iron (Fe)-P but the exact phase and the underlying mechanisms that lead to its formation are unknown. In this study, we investigated sediments from the deepest basin in the Baltic Sea, the Landsort Deep (site M0063), that were retrieved during the Integrated Ocean Drilling Project (IODP) Baltic Sea Paleoenvironment Expedition 347. The record comprises the whole brackish/marine Littorina Sea stage including past intervals of extensive hypoxia in the Baltic Sea that occurred during the Holocene Thermal Maximum (HTMHI) and the Medieval Climate Anomaly (MCA1HI and MCA2HI). Various redox proxies (e.g. the presence of laminations and high Mo contents) suggest almost permanent bottom water hypoxia during the Littorina Sea stage in the Landsort Deep. The bottom waters were likely even seasonally anoxic or sulfidic during the MCA1HI and MCA2HI, and permanently sulfidic during the HTMHI. With the use of micro-analysis of sieved minerals (SEM-EDS, XRD and synchrotron-based XAS), we show that Mn- and Mg-rich vivianite crystals are present at various depths in the Littorina Sea sediments. We also have indications for vivianite in the MCA1HI, MCA2HI and HTMHI deposits. The formation of vivianite thus likely explains the high Fe-bound P fraction throughout the whole Littorina Sea stage. Shuttling of Fe and Mn from the shelves into the basin and high inputs of P in settling organic matter are likely key drivers for vivianite formation. Our study shows that vivianite can likely form in near-surface sediments under a broad range of bottom water redox conditions, varying from hypoxic and anoxic to sulfidic.

Characterization of anisotropy in the permeability of organic-rich shales

Abstract Accurate determination of permeability is essential and challenging in organic-rich shales due to the tight and heterogeneous nature of shale formations, the presence of lamination as well as induced or natural fractures and the sensitivity of fracture permeability to stress. The tight nature of shale formation makes the traditional steady-state method not to be practical. Therefore, unsteady state methods such as pulse decay or crushed-sample have been practiced for shales. However, the pulse decay method can be still time consuming and the crushed-sample method is size-dependent as our numerical simulations show. As an alternative method, the results of complex transient method are presented. Moreover, the presence of lamination and fractures causes directional dependency of permeability. To characterize this type of anisotropy, three-plug method is usually applied. However, the variation of permeability with location causes this method to be unreliable in heterogeneous formations. Similarly, the presence of artificial micro-fractures affects the reliability of the permeability measurement in shales. Using variety of methods to measure permeability, the effects of these anisotropic and heterogeneous features under stress are discussed.

Mutations in Lamin A/C Gene Causes Mechanosensing Defects in Human Myoblasts

The mechanisms underlying mechanosensitivity are critical for muscle development and functionality. The presence of lamin A/C is essential for intact nucleoskeleton and mechanical transmission between the nucleus and the extracellular matrix. Therefore, we tested the hypothesis that mutation in the LMNA gene (encoding for lamin A/C) cause mechanosensing defects in human myoblasts.Myoblasts with LMNA p.Lys32del mutation (Lmna) and normal human myoblasts (WT) were immortalized and cultured in a linear 3D fibrin matrix. At day 1, WT have spontaneously aligned along the gel axis whereas Lmna cells exhibited random orientation (p<0.001). Mutated myoblasts had larger and more abundant actin bundles, and vinculin adhesion sites were longer and thicker compared with WT myoblasts (each p<0.05). This was associated with higher protein and mRNA expression of vinculin in Lmna cells (p<0.01). In WT, uni-axial cyclic stretch of the gel (0.5 Hz, 10% for 4 hours) induced cytoskeletal reorganization characterized by thickening of actin stress fibers and elongation of vinculin adhesion sites. These changes were absent in Lmna and in blebbistatin- or Y-27632-treated WT. Similar results were obtained in primary myoblasts from another patient with similar mutation and in myoblasts with LMNA p.Arg249Trp mutation. In conclusion, these data provide first evidence that lamin A/C mutation causes defects in myoblast mechanosensing that may contribute to mechanical damage in striated muscles from L-CMD.

D.P.25 Mutation in lamin A/C gene causes mechanotransduction defects in human myoblasts

The mechanisms underlying myoblast response to mechanical forces are critical for muscle development and functionality. To initiate adequate mechanotransduction events, myoblasts must be able to sense strains in their extracellular cellular matrix (ECM) substrate, transmit forces to the cell interior and activate intracellular signaling. The presence of lamin A/C is critical for intact nucleoskeleton and mechanical transmission between the nucleus and the ECM. Whether mutation in the LMNA gene (encoding for lamin A/C) causes mechanotransduction defects in human myoblasts has never been directly tested. Myoblasts with LMNA p.Lys32del mutation (D32) were obtained from a 12year-old boy suffering from LMNA-related congenital muscular dystrophy (L-CMD). Normal sex- and age-matched human myoblasts (WT) were used as controls. Myoblasts were immortalized, cultured in a 3D fibrin matrix. 3D constructs were subjected to axial cyclic stretch (10% amplitude, 0.5Hz, 4h). Confocal microscopy imaging and expression of mechanosensitive genes (Iex-1 and Ccnd-1) were analyzed. Unstrained D32 myoblasts had larger and more abundant actin bundles and longer and thinner nuclei (p<0.01) compared with WT. Repetitive strains induced a thickening of actin stress fibers, and an alignment of cells along the longitudinal axis of the 3D matrix in WT absent in D32 cells, while a significant nuclear elongation was observed in D32 but not in WT. Addition of blebbistatin, a myosin inhibitor or Y-27632, a Rho kinase inhibitor, abolished the strain-induced WT cytoskeleton reorganization and myoblasts alignment without changing WT nuclear shape. Milder effects on actin bundles were observed in D32. Strain-induced changes in the expression of Iex-1 and Ccnd-1 were higher in WT than in D32 (p<0.001). We provide first evidence that lamin A/C mutation causes mechanotransduction and cytoskeleton organization defects in human myoblasts and may contribute to mechanical damage in striated muscles from L-CMD.

The role of Lamin A in cytoskeleton organization in colorectal cancer cells

Up-regulated expression of lamin A has been implicated in increased cell invasiveness and mortality in colorectal cancer. Here we use quantitative proteomics to investigate lamin A regulated changes in the cytoskeleton that might underpin increased cell motility. Using siRNA knockdown of lamin A in a model cell line (SW480/lamA) we confirm that the presence of lamin A promotes cell motility. Using an enhanced technique to prepare cytoskeleton fractions in combination with 2D DiGE we were able to accurately and reproducibly detect changes in the representation of protein species within the cytoskeleton as low as 20%. In total 64 protein spots displayed either increased or decreased representation within the cytoskeleton of SW480/lamA cells compared to controls. Of these the identities of 29 spots were determined by mass spectrometry. A majority were multiple forms of three classes of proteins, including components of the actin and IF cytoskeletons, protein chaperones and translation initiation and elongation factors. In particular our data reveal that the representation of tissue transglutaminase 2, which is known to modify elements of the cytoskeleton and is associated with cancer progression, was highly over-represented in the cytoskeleton fraction of SW480/lamA cells. Overall, our data are consistent with changed protein cross-linking and folding that favours the formation of dynamic actin filaments over stress fibres accounting for the altered cell motility properties in SW480/lamA cells.

Attenuated anabolic response to exercise in lamin A/C haploinsufficient mice

The ability of exercise to decrease fat mass and increase bone mass occurs through mechanical biasing of mesenchymal stem cells away from adipogenesis and toward osteoblastogenesis. The mechanism explaining this effect remains poorly understood. Lamin A/C knockdown inhibits osteoblastogenesis while favors adipogenesis in vitro. In this study, we hypothesized that the presence of lamin A/C is required for the anabolic response of bone during exercise. Three-month-old female lamin A/C haploinsufficient (Lmna(+/-)) mice were exposed to strenuous maximal exercise protocol (2 sessions/week, 40 min/session) for 6 weeks. Wild type (WT) (exercise and sedentary) and sedentary Lmna(+/-) mice were used as controls. To determine changes in bone microarchitecture and cell numbers, distal femur was analyzed by microCT and histomorphometry respectively. Finally, levels of expression of nuclear β-catenin and sclerostin, two proteins involved in the anabolic response to exercise, were determined by immunofluorescence. Histomorphometry analysis showed a significant increase in bone volume fraction (BV/TV) in exercised vs. sedentary WT mice. In contrast, exercised Lmna(+/-) mice showed a significant reduction in microarchitecture as compared with sedentary Lmna(+/-) controls including trabecular and cortical thinning. In addition, we found a significant increase in bone cells number in exercised vs. sedentary WT mice whereas exercised Lmna(+/-) mice showed a significant reduction in osteoblasts and osteocytes number as compared with sedentary Lmna(+/-) controls. Finally, levels of activated β-catenin in osteoblasts and osteocytes were significantly decreased while sclerostin expression was increased in exercised Lmna(+/-) mice as compared with exercised WT controls. In summary, our data indicate that the presence of lamin A/C is required for the anabolic effect of exercise on bone thus suggesting a new important role of lamin A/C in bone biology.

Decreased Bone Formation and Osteopenia in Lamin A/C-Deficient Mice

Age-related bone loss is associated with changes in bone cellularity with characteristically low levels of osteoblastogenesis. The mechanisms that explain these changes remain unclear. Although recent in vitro evidence has suggested a new role for proteins of the nuclear envelope in osteoblastogenesis, the role of these proteins in bone cells differentiation and bone metabolism in vivo remains unknown. In this study, we used the lamin A/C null (Lmna −/−) mice to identify the role of lamin A/C in bone turnover and bone structure in vivo. At three weeks of age, histological and micro computed tomography measurements of femurs in Lmna −/− mice revealed a significant decrease in bone mass and microarchitecture in Lmna −/− mice as compared with their wild type littermates. Furthermore, quantification of cell numbers after normalization with bone surface revealed a significant reduction in osteoblast and osteocyte numbers in Lmna −/− mice compared with their WT littermates. In addition, Lmna −/− mice have significantly lower osteoclast number, which show aberrant changes in their shape and size. Finally, mechanistic analysis demonstrated that absence of lamin A/C is associated with increase expression of MAN-1 a protein of the nuclear envelope closely regulated by lamin A/C, which also colocalizes with Runx2 thus affecting its capacity as osteogenic transcription factor. In summary, these data clearly indicate that the presence of lamin A/C is necessary for normal bone turnover in vivo and that absence of lamin A/C induces low bone turnover osteopenia resembling the cellular changes of age-related bone loss.

Emergence of Micronuclei and Their Effects on the Fate of Cells under Replication Stress

The presence of micronuclei in mammalian cells is related to several mutagenetic stresses. In order to understand how micronuclei emerge, behave in cells, and affect cell fate, we performed extensive time-lapse microscopy of HeLa H2B-GFP cells in the presence of hydroxyurea at low concentration. Micronuclei formed after mitosis from lagging chromatids or chromatin bridges between anaphase chromosomes and were stably maintained in the cells for up to one cell cycle. Nuclear buds also formed from chromatin bridges or during interphase. If the micronuclei-bearing cells entered mitosis, they either produced daughter cells without micronuclei or, more frequently, produced cells with additional micronuclei. Low concentrations of hydroxyurea efficiently induced multipolar mitosis, which generated lagging chromatids or chromatin bridges, and also generated multinuclear cells that were tightly linked to apoptosis. We found that the presence of micronuclei is related to apoptosis but not to multipolar mitosis. Furthermore, the structural heterogeneity among micronuclei, with respect to chromatin condensation or the presence of lamin B, derived from the mechanism of micronuclei formation. Our study reinforces the notion that micronucleation has important implications in the genomic plasticity of tumor cells.