Loss Of Regenerative Ability Research Articles

Cellular nucleic acid binding protein facilitates cardiac repair after myocardial infarction by activating β-catenin signaling

The regenerative capacity of the adult mammalian heart is limited, while the neonatal heart is an organ with regenerative and proliferative ability. Activating adult cardiomyocytes (CMs) to re-enter the cell cycle is an effective therapeutic method for ischemic heart disease such as myocardial infarction (MI) and heart failure. Here, we aimed to reveal the role and potential mechanisms of cellular nucleic acid binding protein (CNBP) in cardiac regeneration and repair after heart injury. CNBP is highly expressed within 7 days post-birth while decreases significantly with the loss of regenerative ability. In vitro, overexpression of CNBP promoted CM proliferation and survival, whereas knockdown of CNBP inhibited these processes. In vivo, knockdown of CNBP in CMs robustly hindered myocardial regeneration after apical resection in neonatal mice. In adult MI mice, CM-specific CNBP overexpression in the infarct border zone ameliorated myocardial injury in acute stage and facilitated CM proliferation and functional recovery in the long term. Quantitative proteomic analysis with TMT labeling showed that CNBP overexpression promoted the DNA replication, cell cycle progression, and cell division. Mechanically, CNBP overexpression increased the expression of β-catenin and its downstream target genes CCND1 and c-myc; Furthermore, Luciferase reporter and Chromatin immunoprecipitation (ChIP) assays showed that CNBP could directly bind to the β-catenin promoter and promote its transcription. CNBP also upregulated the expression of G1/S-related cell cycle genes CCNE1, CDK2, and CDK4. Collectively, our study reveals the positive role of CNBP in promoting cardiac repair after injury, providing a new therapeutic option for the treatment of MI.

Hepatocyte integrity depends on c-Jun-controlled proliferation in Schistosoma mansoni infected mice

Schistosomiasis is a parasitic disease affecting more than 250 million people worldwide. The transcription factor c-Jun, which is induced in S. mansoni infection-associated liver disease, can promote hepatocyte survival but can also trigger hepatocellular carcinogenesis. We aimed to analyze the hepatic role of c-Jun following S. mansoni infection. We adopted a hepatocyte-specific c-Jun knockout mouse model (Alb-Cre/c-Jun loxP) and analyzed liver tissue and serum samples by quantitative real-time PCR array, western blotting, immunohistochemistry, hydroxyproline quantification, and functional analyses. Hepatocyte-specific c-Jun knockout (c-JunΔli) was confirmed by immunohistochemistry and western blotting. Infection with S. mansoni induced elevated aminotransferase-serum levels in c-JunΔli mice. Of note, hepatic Cyclin D1 expression was induced in infected c-Junf/f control mice but to a lower extent in c-JunΔli mice. S. mansoni soluble egg antigen-induced proliferation in a human hepatoma cell line was diminished by inhibition of c-Jun signaling. Markers for apoptosis, oxidative stress, ER stress, inflammation, autophagy, DNA-damage, and fibrosis were not altered in S. mansoni infected c-JunΔli mice compared to infected c-Junf/f controls. Enhanced liver damage in c-JunΔli mice suggested a protective role of c-Jun. A reduced Cyclin D1 expression and reduced hepatic regeneration could be the reason. In addition, it seems likely that the trends in pathological changes in c-JunΔli mice cumulatively led to a loss of the protective potential being responsible for the increased hepatocyte damage and loss of regenerative ability.

Cryptic carnivores: Intercontinental sampling reveals extensive novel diversity in a genus of freshwater annelids

Freshwater annelids are globally widespread in aquatic ecosystems, but their diversity is severely underestimated. Obvious morphological features to define taxa are sparse, and molecular phylogenetic analyses regularly discover cryptic diversity within taxa. Despite considerable phylogenetic work on certain clades, many groups of freshwater annelids remain poorly understood. Included among these are water nymph worms of the genus Chaetogaster (Clitellata: Tubificida: Naididae: Naidinae). These worms have diverged from the detritivorous diet of most oligochaetes to become more predatory and exist as omnivores, generalist predators, parasites, or symbionts on other invertebrates. Despite their unusual trophic ecology, the true diversity of Chaetogaster and the phylogenetic relationships within the genus are uncertain. Only three species are commonly referenced in the literature (Chaetogaster diaphanus, Chaetogaster limnaei, and Chaetogaster diastrophus), but additional species have been described and prior molecular data suggests that there is cryptic diversity within named species. To clarify the phylogenetic diversity of Chaetogaster, we generated the first molecular phylogeny of the genus using mitochondrial and nuclear sequence data from 128 worms collected primarily across North America and Europe. Our phylogenetic analyses suggest that the three commonly referenced species are a complex of 24 mostly cryptic species. In our dataset, Chaetogaster “diaphanus” is represented by two species, C. “limnaei” is represented by three species, and C. “diastrophus” is represented by 19 species. North American and European sequences are largely interspersed across the phylogeny, with four pairs of clades involving distinct North American and European sister groupings. Overall, our study demonstrates that the species diversity of Chaetogaster has been underestimated and that carnivory has evolved at least twice in the genus. Chaetogaster is being used as a model for symbiotic evolution and the loss of regenerative ability, and our study indicates that researchers must be careful to identify which species of Chaetogaster they are working with in future studies.

Fu Fang Zhen Zhu Tiao Zhi Capsules Protect against Myocardial Ischemia by Inhibiting Cardiomyocyte Pyroptosis

Fu Fang Zhen Zhu Tiao Zhi (FTZ) is a traditional Chinese herbal prescription widely used to treat dyslipidemia, metabolic diseases, and diabetic coronary disorders. Cardiomyocyte death and loss of regenerative ability cause cardiac dysfunction and heart failure. FTZ can effectively treat diabetic cardiomyopathy and macrovascular diseases; however, the mechanism behind the phenomenon is still unclear. Here, we determined the mechanism of action of FTZ in treating myocardial infarction. Male C57BL/6 mice were treated with 2.4 or 1.2 g/kg FTZ, or administered saline by oral gavage daily for four weeks, and a 24-hour ligation was administered to the artery. Echocardiography was used to evaluate cardiac function. Hematoxylin and eosin and Evans blue/triphenyltetrazolium chloride staining were carried out by staining the cardiac tissue, used to evaluate cardiac function and infarct size. Using western blotting and reverse transcriptase-polymerase chain reaction, we determined the relative levels of NOD-like receptor protein (NLRP) 3, ASC, cleaved caspase-l (C-Caspase-1), GSDMD, and GSDMD-N. TUNEL, immunohistochemical, and immunofluorescence staining were used to determine cell death and NLRP3 expression. An enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of interleukin (IL)-1β and IL-18. FTZ reduced ischemia-induced cardiomyocyte cell death in vivo and H2O2-induced cell death in vitro by maintaining cardiac architecture and restoring cardiac function. FTZ decreased the NLRP3 expression and inhibited pyroptosis-correlated genes, including NLRP3, ASC, GSDMD, C-Caspase-1, and GSDMD-N. NLRP3 overexpression impaired the efficacy of FTZ by inducing pyroptosis. FTZ could preserve cardiac function resulting from ischemic insult by inhibiting pyroptosis, which was partially reversed by NLRP3 overexpression, indicating that NLRP3 could be a potential target of FTZ in treating myocardial infarction.

4D imaging analysis of the aging mouse neural stem cell niche reveals a dramatic loss of progenitor cell dynamism regulated by the RHO-ROCK pathway.

SummaryIn the adult ventricular-subventricular zone (V-SVZ), neural stem cells (NSCs) give rise to transit-amplifying progenitor (TAP) cells. These progenitors reside in different subniche locations, implying that cell movement must accompany lineage progression, but the dynamic behaviors of adult NSCs and TAPs remain largely unexplored. Here, we performed live time-lapse imaging with computer-based image analysis of young and aged 3D V-SVZ wholemounts from transgenic mice with fluorescently distinguished NSCs and TAP cells. Young V-SVZ progenitors are highly dynamic, with regular process outgrowth and retraction and cell migration. However, these activities dramatically declined with age. An examination of single-cell RNA sequencing (RNA-seq) data revealed age-associated changes in the Rho-Rock pathway that are important for cell motility. Applying a small molecule to inhibit ROCK transformed young into old V-SVZ progenitor cell dynamic behaviors. Hence RHO-ROCK signaling is critical for normal adult NSC and TAP movement and interactions, which are compromised with age, concomitant with the loss of regenerative ability.

An in vivo method to study post‐transcriptional regulation in germ stem cells

Stem cells in regenerative tissues must balance factors promoting self‐renewal with those driving differentiation. Faulty regulation of this balance can lead to tumor growth or loss of regenerative ability. The tractable germline of the nematode C. elegans has allowed the identification of a wealth of key stem cell regulatory factors and remarkably, virtually all are RNA binding proteins (RBP). Despite this insight, a mechanistic understanding of how these RBPs regulate their target mRNAs is lacking. Here, we develop an in vivo method to investigate their mechanisms. Our assay probes how a given RBP affects expression of a reporter mRNA when recruited to its 3′ untranslated region (3′UTR). To perform the assay, an RBP of interest is genetically engineered to contain a small λN peptide. The λN peptide binds a specific RNA hairpin, called boxB, with high affinity. Thus, a reporter mRNA engineered to contain boxB hairpins in its 3′UTR will bind the λN tagged RBP and either increase or decrease expression, depending on the RBP regulatory function. By imaging reporter expression in germline stem cells, we can gain insight into how the RBP regulates target transcripts in its normal biological context. The simple and rapid readout of the assay allows it to be used to probe how an RBP accomplishes its RNA regulation or screen for other factors necessary for RBP function. We have applied this assay to several of the key stem cell RBP regulators to test their functions in vivo. Our results demonstrate the tractability and ease of this method to study RBPs in the C. elegans germline as well as providing mechanistic insights into how the RBPs regulate stem cell fate decisions.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Kinesin KIF4A transports integrin β1 in developing axons of cortical neurons.

CNS axons have poor regenerative ability compared to PNS axons, and mature axons regenerate less well than immature embryonic axons. The loss of regenerative ability with maturity is accompanied by the setting up of a selective transport filter in axons, restricting the types of molecule that are present. We confirm that integrins (represented by subunits β1 and α5) are present in early cortical axons in vitro but are excluded from mature axons. Ribosomal protein and L1 show selective axonal transport through association with kinesin kif4A; we have therefore examined the hypothesis that integrin transport might also be in association with kif4A. Kif4A is present in all processes of immature cortical neurons cultured at E18, then downregulated by 14days in vitro, coinciding with the exclusion of integrin from axons. Kif4a co-localises with β1 integrin in vesicles in neurons and non-neuronal cells, and the two molecules co-immunoprecipitate. Knockdown of KIF4A expression with shRNA reduced the level of integrin β1 in axons of developing neurons and reduced neurite elongation on laminin, an integrin-dependent substrate. Overexpression of kif4A triggered apoptosis in neuronal and non-neuronal cells. In mature neurons expression of kif4A-GFP at a modest level did not kill the cells, and the kif4A was detectable in their axons. However this was not accompanied by an increase in integrin β1 axonal transport, suggesting that kif4A is not the only integrin transporter, and that integrin exclusion from axons is controlled by factors other than the kif4A level.

Evolutionary Loss of Animal Regeneration: Pattern and Process

The ability to regenerate lost or damaged body parts is widespread among animals and provides obvious potential benefits. It is therefore perplexing that this ability has become greatly restricted or completely lost in many lineages. Despite growing interest in the cellular and molecular basis of regeneration, our understanding of how and why regenerative abilities are lost remains rudimentary. In an effort to develop a framework for studying losses of regeneration, here I outline an approach for rigorously identifying such losses, review broad patterns of regenerative ability across animals, describe some of the clearest examples of regeneration loss, discuss some possible scenarios by which regeneration may be lost, and review recent work in annelids that is providing new insights into loss of regenerative ability.

Hind-Limb Regeneration in the Dwarf African Clawed Frog, Hymenochirus boettgeri (Anura: Pipidae)

The ability to regenerate lost or amputated limbs varies greatly among amphibians. To gain a better understanding of phylogenetic and ontogenetic factors influencing regenerative ability, regeneration in tadpoles of Hymenochirus boettgeri is described for the first time, then compared to data from closely related Xenopus laevis. Hind limbs of tadpoles of H. boettgeri were amputated at two different levels along the proximo-distal axis of the limb (at the knee and ankle joints), at stages ranging from 52 to 59. Tadpoles were allowed to recover for 15 or 20 days, at which times the amount of regeneration was examined. Observations were made through external morphology and whole-mount histological staining. Hymenochirus, like X. laevis and indeed like most anurans, gradually loses the ability to regenerate hind-limb structures as it progresses through metamorphosis, and this loss occurs in a proximo-distal direction; the ability to regenerate is retained longer, distally. Hymenochirus and Xenopus show similar patterns of limb and digit regeneration. In both pipid species, loss of regenerative ability appears to be correlated with the onset of ossification in the hind limb, although timing of this event differs between the two species.

Hind-Limb Regeneration in the Dwarf African Clawed Frog, Hymenochirus boettgeri (Anura: Pipidae)

The ability to regenerate lost or amputated limbs varies greatly among amphibians. To gain a better understanding of phylogenetic and ontogenetic factors influencing regenerative ability, regeneration in tadpoles of Hymenochirus boettgeri is described for the first time, then compared to data from closely related Xenopus laevis. Hind limbs of tadpoles of H. boettgeri were amputated at two different levels along the proximo-distal axis of the limb (at the knee and ankle joints), at stages ranging from 52 to 59. Tadpoles were allowed to recover for 15 or 20 days, at which times the amount of regeneration was examined. Ob- servations were made through external morphology and whole-mount histological staining. Hymenochirus, like X. laevis and indeed like most anurans, gradually loses the ability to regenerate hind-limb structures as it progresses through metamorphosis, and this loss occurs in a proximo-distal direction; the ability to regenerate is retained longer, distally. Hymenochirus and Xenopus show similar patterns of limb and digit regeneration. In both pipid species, loss of regenerative ability appears to be correlated with the onset of ossification in the hind limb, although timing of this event differs between the two species.

Intrinsic and extrinsic control of growth in developing organs.

The growth rate and final size of developing organs is controlled by organ-intrinsic mechanisms as well as by hormones and growth factors that originate outside the target organ. Recent work on Drosophila imagined discs and other regenerating systems has led to the conclusion that the intrinsic growth-control mechanism that controls regenerative growth depends on position-specific interactions between cells and their neighbors, and that these interactions also control pattern formation. According to this interpretation, local growth by cell proliferation is stimulated when cells with disparate positional information are confronted as a result of grafting or wound healing. This local growth leads to intercalation of cells with intervening positional values until the positional information discontinuity is eliminated. When all discontinuities have been eliminated from a positional field, growth stops. In this article we consider the possibility that organ growth during normal development may be controlled by an intercalation mechanism similar to that proposed for regenerative growth. Studies of imaginal disc growth are consistent with this suggestion, and in addition they show that the cell interactions thought to control growth are independent of cell lineage. Developing organs of vertebrates also show intrinsic growth-control mechanisms, as demonstrated by the execution of normal growth programs by immature organs that are transplanted to fully grown hosts or to hosts with genetically different growth parameters. Furthermore, these organ-intrinsic mechanisms also appear to be based on position-specific cell interactions, as suggested by the growth stimulation seen after partial extirpation or rearrangement by grafting. In organs of most adult vertebrates, the organ-intrinsic growth-control system seems to be suppressed as shown by the loss of regenerative ability, although it is clearly retained in the limbs, tails and other organs of salamanders. The clearest example of an extrinsic growth regulator is growth hormone, which plays a dominant role along with insulin-like growth factors, thyroid hormone and sex hormones in supporting the growth of bones and other organs in postnatal mammals. These hormones do not appear to regulate prenatal growth, but other hormones and insulin-like growth factors may be important prenatally. The importance of other growth factors in regulating organ growth in vivo remains to be established. It is argued that both intrinsic and extrinsic factors control organ growth, and that there may be important interactions between the two types of control during development.