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PubMed1992

Aging alters ornithine decarboxylase and decreases polyamines in regenerating rat liver but putrescine replacement has no effect.

Aging decreases rat liver regeneration. We (1) compared the expression of ornithine decarboxylase (ODC), a critical enzyme for liver regeneration, and polyamine levels in regenerating liver of 6-week-old and 1-year-old rats and (2) evaluated the effect of exogenous putrescine supplementation on liver regeneration in 1-year-old rats. ODC messenger ribonucleic acid (mRNA) transcript sizes were the same in rats of both ages. ODC mRNA content and enzyme activity were higher in the younger rats; however, magnitudes of increase after partial hepatectomy were greater in the older rats. From peak levels, the rate of decline of the mRNA was slower in the older rats, but enzyme activity declined at the same rate in both ages. ODC apoenzyme content was significantly less in normal liver tissue from 1-year-old rats, but there was little change after partial hepatectomy in rats of either age. No change in ODC transcriptional activity was found. Hepatic putrescine levels were lower in 48 hours regenerating liver tissue from 1-year-old rats. To determine whether supplemental putrescine would increase liver regeneration in 1-year-old rats, putrescine (600 mumol/kg IP every 4 hours) was administered beginning 4 days before or at the time of partial hepatectomy. This raised polyamine levels and decreased ODC activity significantly, but there was no change in regenerating liver weight, total DNA and RNA content, and tritiated thymidine incorporation at 48 hours. These results indicate that ODC expression is different and polyamine levels are lower in 1-year-old rats than in 6-week-old rats. However, putrescine supplementation that is sufficient to decrease ODC activity has no apparent effect on regeneration.

Aging↗
PubMed1990

Immediate-early gene expression differs between regenerating liver, insulin-stimulated H-35 cells, and mitogen-stimulated Balb/c 3T3 cells. Liver-specific induction patterns of gene 33, phosphoenolpyruvate carboxykinase, and the jun, fos, and egr families.

Immediate-early genes, whose expression increases independent of de novo protein synthesis during the transition from quiescence to proliferation, are postulated to play important regulatory roles in the growth response. The complement of immediate-early genes expressed must depend on the milieu of preexisting transcription factors in the quiescent cell as well as the type of mitogenic stimulation and, thus, may differ between cell types. We have begun characterizing the immediate-early response in regenerating liver and insulin-stimulated Reuber H-35 hepatoma cells in comparison with previously published results from mitogen-stimulated Balb/c 3T3 fibroblasts. The proliferating H-35 and regenerating liver cells maintain their similarity to quiescent liver as demonstrated by their continued production of the liver-specific albumin, CCAAT/enhancer binding protein, and phosphoenolpyruvate carboxykinase messenger RNAs (mRNA). Surprisingly, the phosphoenolpyruvate carboxykinase gene, which undergoes down-regulation in insulin-treated H-35 cells, was cloned by differential screening of a subtraction-enriched regenerating liver cDNA library and is an immediate-early gene in regenerating liver. H-35 cells treated with either insulin or phorbol 12-myristate 13-acetate express elevated levels of the jun genes, and phorbol 12-myristate 13-acetate pretreatment fails to abolish the insulin response, indicating that it does not depend on protein kinase C. jun family gene expression in regenerating liver differs from that in mitogen-treated fibroblasts in that the time course of expression of c-jun and junB is prolonged, and junD mRNA levels distinctly increase. Additionally, although c-fos and egr-1 mRNAs are expressed at elevated levels in stimulated liver cells, fos-B, fra-1, and egr-2 are not, which suggests that factors in addition to the serum response factor participate in the regulation of immediate-early gene induction. Interestingly, gene 33, which was cloned from a regenerating liver cDNA library by differential screening and lacks a recognizable serum response element, functions as an immediate-early gene in regenerating liver and in mitogen-treated H-35 and Balb/c 3T3 cells. These results suggest that gene 33 participates in the transition from quiescence to proliferation in many mitogen-treated cells in addition to its previously reported involvement in hormone responses. Overall, the results presented here suggest that the immediate-early response varies considerably between regenerating liver and mitogen-stimulated fibroblasts and could involve multiple, preexisting, tissue-specific, transcription-activating proteins.

Animals↗
PubMed1990

Cell interactions and regeneration control.

This paper is a review of the main findings of our laboratory on the control of regeneration by cell interactions. These include results related to the role of both cell contact and local soluble factors in regeneration of the legs of insects and newts and of the parapodia and segments of nereis. The pattern of these structures is considered to be defined by positional information distributed as longitudinal and transverse positional value sequences carried by epidermal (insect) or mesenchymal (newt) cells. By associating tissues to create transverse and longitudinal discontinuities in these sequences, single or multiple regenerating structures were obtained. These structures are formed by the intercalation of cells characterized by intermediate positional values which fill the gap between the tissues in contact. Positional information may also be changed during regeneration by the nerve cord in nereis and retinoids in the newts. We describe additional cases where morphogenesis occurs without any overt discontinuity in positional information, such as from a locally injured or non-injured insect trochanter, or after deflection of nerves in nereis and newt. Regeneration following an amputation may be considered as a special case of intercalary regeneration, the first stage being the juxtaposition of normally non-contiguous cells resulting in a longitudinal or/and a transverse gap. We also report studies on local factors produced by nerves and the blastema during newt limb regeneration. The nerve factor is necessary for the division of blastemal cells. After denervation, mesenchyme differentiates in an abnormal way. The mitogenic signal from the nerves is mediated by the PKC pathway. Its production is enhanced by regeneration of cut nerve fibers. The blastema also produces growth factors. We show that the epidermal cap and mesenchyme contain acidic FGF-like factor, and that the proliferating mesenchyme stimulates nerve fibers to regrow into the blastema.

Amphibians↗
PubMed1988

Regenerating afferent fibers stimulate the recovery of mauthner cell dendritic branching in the axolotl.

In the medulla of the axolotl (Ambystoma mexicanum), Mauthner cells (M-cells) occur as a pair of large identifiable neurons at the level of entry of the vestibular nerve (nVIII). Each receives synapses from the ipsilateral nVIII; the terminals can be identified as club endings and are restricted to a specific set of M-cell dendritic branches. We have examined these branches for morphologic changes following long-term deafferentation in the presence and absence of nerve regeneration. Deafferentation was brought about in a group of young larvae by unilaterally severing nVIII. The nerve was allowed to regenerate in half of the larvae. In those remaining, the nVIII ganglion was damaged to preclude or limit nerve regeneration. The contralateral side served as control. After 3 months survival, the larvae were killed and the medullae prepared for microscopy. To estimate the extent of nerve regeneration, axons in the experimental nVIII tract were counted and compared with the number in the control. The mean number of axons in the nVIII tract ipsilateral to intact ganglia indicated that 69% of the fibers had regenerated. In contrast, only 31% regenerated in larvae with damaged ganglia. Electron microscopic analysis of selected sections revealed that the mean number of nVIII terminals per section through M-cells ipsilateral to destroyed ganglia was significantly less than the mean number in analogous sections through either control cells or cells ipsilateral to intact ganglia. Control and experimental M-cells were reconstructed from serial sections. Deprived M-cells had significantly reduced dendritic branching patterns in the region that normally receives nVIII input. On the other hand, the extent of branching on cells receiving regenerated afferents from intact ganglia was like that of their contralateral controls. The distribution of dendritic branches on many reinnervated M-cells, however, was broader than that on control cells. Electron microscopic examination of the displaced dendritic branches (those extending into adjacent tracts) revealed that they received vestibular synapses. Thus, in some animals, regenerated vestibular fibers were not restricted to the nVIII tract. Deafferentation of the M-cells results in a reduction of dendritic branches in the region deprived of vestibular contacts.(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗
PubMed1986

Regeneration from half lower arms in the axolotl.

A technique involving grafting of pieces of skin from the head onto the limb in order to isolate halves of the limb is described. This technique was used to isolate posterior, anterior, dorsal and ventral halves of the lower arm. All halves produced regenerates but no part of the limb was able to produce a high proportion of regenerates with a complete pattern of skeletal structures. Posterior half stumps regenerated limbs with a mean digit number of 2.7 and had a normal dorsoventral muscle pattern. Anterior half stumps produced a high proportion of single-digit regenerates and had a mean digit number of 1.3. Dorsal and ventral half stumps regenerated limbs with a mean digit number of 2.8 and 2.3 respectively. Hypomorphic regenerates from dorsal and ventral half stumps often had only dorsal or ventral muscle. These results are in contrast to those from the upper arm (Wigmore & Holder, 1985) where a complete skeletal and muscular pattern regenerated from posterior and dorsal halves and hypomorphic regenerates were obtained from anterior and ventral half limbs.

Ambystoma↗
PubMed1986

Pathways of regenerated retinotectal axons in goldfish. I. Optic nerve, tract and tectal fascicle layer.

This study investigates the order of regenerating retinal axons in the goldfish. The spatiotemporal pattern of axon regrowth was assessed by applying horseradish peroxidase (HRP) to regenerating axons in the optic tract at various times after optic nerve section and by analysing the distribution of retrogradely labelled ganglion cells in retina. At all regeneration stages labelled ganglion cells were widely distributed over the retina. There was no hint that axons from central (older) ganglion cells might regrow earlier, and peripheral (younger) ganglion cells later, as occurs in normal development. The absence of an age-related ordering in the regenerated optic nerve was demonstrated by labelling a few axon bundles intraorbitally with HRP (Easter, Rusoff & Kish, 1981) caudal to the previous cut. The retrogradely labelled cells in retina were randomly distributed in regenerates and not clustered in annuli as in normals. Tracing regenerating axons which were stained anterogradely from intraretinal HRP applications or retrogradely from single labelled tectal fascicles illustrated the fact that the regenerating axons coursed in abnormal routes in the optic nerve and tract. On the surface of the tectum regenerated fibres re-established a fascicle fan. The retinal origin of tectal fascicles was assessed by labelling individual peripheral, intermediate and rostral fascicles with HRP. The retrogradely labelled ganglion cells in the retina were often more widely distributed than in normals, but were mostly found in peripheral, intermediate and central retina, respectively. The order of fibre departure from each tectal fascicle was revealed by placing HRP either on the fascicle's proximal or on its distal half. With proximal labelling sites labelled ganglion cells were found in the temporal and nasal retina, and with distal labelling sites labelled ganglion cells were confined to nasal retina only. Further, the axonal trajectories of anterogradely labelled dorsotemporal retinal ganglion cells were compared to those of dorsonasal retinal ganglion cells in tectal whole mounts. Dorsotemporal axons were confined to the rostral tectal half, whereas dorsonasal axons followed fascicular routes into the fascicle's distal end and reached into caudal tectum. This suggests that the fibres exited along their fascicle's course in a temporonasal sequence. Thus in the tectum, fibres in fascicles restore a gross spatial and age-related order and tend to follow their normal temporonasal sequence of exit.

Animals↗
PubMed1985

Decreased transfer of oligosaccharide from oligosaccharide-lipid to protein acceptors in regenerating rat liver.

The transfer of [14C]glucose from UDP-[14C]glucose to lipid intermediates and glycoproteins was decreased in regenerating rat liver microsomes 24 h after partial hepatectomy. In regenerating liver microsomes, the concentration of free dolichyl phosphate (Dol-P) was significantly decreased. However, it was only about 10% of total Dol-P, which was not significantly changed. On the addition of exogenous Dol-P, the transfer of [14C]glucose to glycoproteins was still decreased, while the decrease of the transfer to lipid intermediates was no longer observed. These results suggest that the glycoprotein synthesis is not regulated by the amount of Dol-P in regenerating liver microsomes. Oligosaccharide obtained from [14C]glucosyl-oligosaccharide-lipid was not distinguishable between regenerating liver and control by paper chromatography. The oligosaccharide transfer to protein in microsomes was compared by using [14C]glucosyl-oligosaccharide-lipid as oligosaccharide donor. The transfer of oligosaccharide to endogenous proteins decreased to 77% of control in regenerating liver and the transfer to exogenously added denatured alpha-lactalbumin decreased to 59% of control. Therefore, it is unlikely that the acceptor capacity of endogenous protein is decreased in regenerating liver. Neither the change in oligosaccharide-lipid under the condition for oligosaccharide transfer assay nor the stability of oligosaccharide transferase was different between regenerating liver and control. These results strongly suggest that the decrease in the activity of the oligosaccharide transferase in microsomes causes the decrease of glycoprotein synthesis in regenerating liver, which was shown in our previous studies.

Animals↗
PubMed1985

Microporous, complaint, biodegradable vascular grafts for the regeneration of the arterial wall in rat abdominal aorta.

Microporous, complaint, biodegradable vascular grafts prepared from mixtures of polyurethane (PU) and poly-L-lactic acid (PLLA) can function as temporary scaffolds for the regeneration of the arterial wall in small-caliber arteries. This study was undertaken to determine the most suitable composition for PU/PLLA vascular grafts to ensure an optimal regeneration. Four types of PU/PLLA vascular grafts differing in percent weight of the PU/PLLA mixture, molecular weight of PLLA, and pore size were implanted into the abdominal aorta of rats (n = 32). Six weeks after implantation two implants of each graft type were evaluated by means of scanning electron microscopy and six implants were evaluated by means of light microscopy. In two types of the PU/PLLA vascular grafts, both of which were prepared from a 95%/5% weight PU/PLLA mixture with PLLA of molecular weight 500,000 but which had a different pore size, there was (I) absence of aneurysm formation and maintenance of arterial implant pulsations, (II) regeneration of a complete antithrombogenic neointima, (III) regeneration of a neomedia of comparable thickness to the media of normal rat abdominal aorta with the regeneration of elastic laminae almost throughout its thickness, and (IV) regeneration of a sufficiently supporting neoadventitia. These results demonstrate that a 95%/5% weight PU/PLLA mixture with PLLA of molecular weight 500,000 is the most suitable composition for PU/PLLA vascular grafts to ensure an optimal regeneration of a neoarterial wall that is of sufficient strength, compliance, and thromboresistance to function as a small-caliber arterial substitute. Pore size of these PU/PLLA grafts does not affect regeneration.

Animals↗
PubMed1983

Tip regeneration and positional information in the slug of Dictyostelium discoideum.

We show in this paper that in the case of the slug of the cellular slime mould Dictyostelium discoideum the time which it takes for a new tip to regenerate at a given level can be used as a measure of positional information at that level. Our basic experiment consists of amputating slugs at various distances from the existing tip and thereby inducing the regeneration of a fresh tip; the time needed for regeneration is estimated by two independent methods. An identical operation, when performed in the anterior portion of a previously cut slug, tells us how this position-dependent regeneration time adjusts to a sudden change in the size of the slug. The reasons which lead us to conclude that tip regeneration times are in one-to-one correspondence with positional information are as follows. (i) Depending on their positions, the cells in a slug take different times to regenerate a new tip following amputation; (ii) regeneration times are scaled in relation to the total length of the slug and increase monotonically with the length cut off; and (iii) as judged by the regeneration time, cells can assess and remember their positions relative to the length of the slug. Our results highlight the importance of two rate processes. One might think of the slower process as being related to the setting up of a system of positional information in the slug, and the faster process as being a reflection of the kinetics of the positional value changing locally till it reaches the level appropriate to a tip.

Dictyostelium↗
PubMed1995

Impaired peripheral nerve regeneration in a mutant strain of mice (Enr) with a Schwann cell defect.

Schwann cell-axon interactions in the development, maintenance, and regeneration of the normal peripheral nervous system are complex. A previously described transgene-induced insertional mutation (BPFD#36), now referred to as Enervated (Enr), results in disrupted Schwann cell-axon interactions. In this report, after a crush or transection injury to Enr peripheral nerves, we demonstrate impaired nerve regeneration. There are fewer myelinated fibers per mm2 and thinner myelin sheaths surrounding regenerating axons in the nerves of homozygous mutant mice compared to wild type mice at 28 d after crush injury to the sciatic nerve. Abnormal Schwann cell-axon interactions remain in Enr/Enr animals as evidenced by the relatively frequent ultrastructural finding of unmyelinated large diameter axons in the regenerating nerves. Additionally, nerve graft experiments indicate that the impairment in regeneration is due to a Schwann cell defect. Morphologic and morphometric findings in conjunction with molecular analysis of regenerating nerves suggest that the Enr defect causes a disruption in the ability of "early" Schwann cells to differentiate to a more mature phenotype. In mutant homozygous and wild type nerves at 7 d after crush injury there are similar levels of mRNA for the low-affinity nerve growth factor receptor, but in the mutant homozygous regenerating nerves there is 11-fold less mRNA for glial fibrillary acidic protein, a more mature phenotypic marker of Schwann cells. This Schwann cell differentiation defect likely accounts for both the peripheral neuropathy and impaired nerve regeneration observed in Enr mice.

Animals↗
PubMedDate not supplied

[Identification of new genes and analysis of their expression during lens and retinal regeneration in adult newts].

During lens regeneration in Pleurodeles waltl, the dorsal iris zone is the cell source of the lens regeneration, while the ventral iris zone can serve as the cells' source of lens regeneration only under certain experimental conditions. The method of subtractive hybridization was used for the identification of genes responsible for the different proliferative potential of these zones. Differential screening of the enriched cDNA libraries, which were obtained as a result of subtractive hybridization of the cDNA samples of the ventral and dorsal iris zones 14 days after lens removal, revealed four clones specific to the dorsal iris and six clones specific to the ventral iris. Two of these, LeR-1 and VeR-1, were structurally characterized. Comparison of their primary structure with data from the Gene Bank showed no essential homology with the known sequences. Time-related changes in LeR-1 and VeR-1 expression were shown during lens regeneration. LeR-1 and VeR-1 expression was activated at the early stages of lens regeneration. The peaks of LeR-1 and VeR-1 expression were observed on the 14th day of lens regeneration in the dorsal and ventral iris zones, respectively. Furthermore, LeR-1 is activated during retina regeneration. The results of Southern hybridization suggest the presence of sequences complementary to LeR-1 in the genomes of Pleurodeles waltl and Rana temporaria. We propose that the activation of LeR-1 expression is related to the triggering of lens regeneration, while the activation of VeR-1 expression accompanies the inhibition of proliferative activity in the ventral iris zone.

Animals↗
PubMed1995

Changes of liver-resident NK cells during liver regeneration in rats.

To determine the role of NK cells in regulation of tissue growth, the phenotype and function of liver-resident NK cells were studied after 70% partial hepatectomy in rats. The process of liver regeneration was generally completed by day 14. In contrast, the number of liver-resident NK cells (NKR-P1bright) was restored as early as day 3 after partial hepatectomy. However, spontaneous functions of liver-resident NK cells, including killing of YAC-1 and P815 targets, Ab-dependent cellular cytotoxicity, and redirected killing via NKR-P1, were continuously suppressed throughout the entire period of liver regeneration (from 3 h to 14 days). Augmentation of NK cytotoxicity against P815 targets and induction of NK cell adherence to plastic following 24 h of IL-2 stimulation showed a similar pattern of suppression. However, IL-2-induced augmentation of YAC-1 killing, proliferation and generation of adherent NK cells, and LAK activity in 5- to 7-day cultures were found to be suppressed only during the first 24 h and increased between days 2 and 7 after hepatectomy. Sorted NK cells (> or = NKR-P1bright) from liver-resident mononuclear leukocytes 24 h after partial hepatectomy showed the same pattern of suppression as unsorted mononuclear leukocytes. In contrast to liver-resident NK cells, no significant changes were detected in peripheral blood or spleen NK cells of rats following partial hepatectomy. Of particular interest, in normal liver, hepatocytes were resistant to NK lysis, while resident NK cells were cytotoxic for various NK-sensitive targets. In contrast, during the early period of liver regeneration, when hepatocytes were sensitive to lysis by liver-resident NK cells of normal rats, NK cells obtained from regenerating liver tissues were unable to mediate cytotoxicity. At the final phase of liver regeneration (days 7-14 after hepatectomy), both resistance of hepatocytes to killing by NK cells and cytotoxicity of liver-resident lymphocytes against hepatocytes from regenerating liver were simultaneously restored. In vivo depletion of NK cells by injection of rats with anti-NKR-P1 mAb resulted in a significant augmentation of liver regeneration subsequent to partial hepatectomy. Our data suggest that liver-resident NK cells may be involved in regulation of the extent of liver regeneration.

Animals↗
PubMed1994

[Electron microscopic study on the degeneration and regeneration of the rat liver nerve fibers].

With advances in liver transplantation, attention has been directed to the regeneration of hepatic nerves. In this study, examination is made of ultrastructural degeneration and regeneration of hepatic nerves following severance at a point 8 mm proximal to the portal fissure in the rat liver. Immediately after the operation, distal nerve axons swelled and showed the disappearance of microtubules and neurofilaments. Newly formed axons, small in diameter and containing many microtubules and neurofilaments, could been seen at 2 days after the operation among those that had degenerated. The regenerated axons were surrounded completely by Schwann cells. The Schwann cell processes became shorter at days and regenerated axons were situated in the space between Schwann cell and the basement membrane. Degenerated axons could no longer be seen 14 days after the operation, and regenerating axons increasing in diameter filled the nerve fibers. At 21-30 days, Schwann cell processes gradually extended to enclose regenerating axons, nerve fiber regeneration was complete 42-56 days after the operation. The manner of nerve regeneration was consistent with the results of a previous immunohistochemical study using anti GAP-43 (growth associated protein-43) antibody.

Animals↗
PubMed1993

Phosphoprotein phosphatase activity in regenerating forelimbs of adult newts, Notophthalmus viridescens.

Traditional signal transduction pathways appear to be operating during amphibian forelimb regeneration, as in other developing systems. A consistent picture appears to be developing that suggests that growth factors promoting proliferation during regeneration act through cAMP-independent mechanisms while factors that modulate differentiation and morphogenesis include signaling pathways in which cAMP participates. Although our understanding of these pathways is far from complete, it is becoming increasingly evident that further characterization of signal transduction events will facilitate (a) identifying major signaling substances, (b) determining the cellular events in regeneration that they modulate, and (c) defining the mechanisms through which they act. The present study demonstrates that PPtases, and in particular PTPases, can be studied in regenerating newt limbs. More importantly, this investigation demonstrates that there are progressive and significant increases in PPtase activity during regeneration. Moreover, the observed patterns of PPtase activity during regeneration conform to an emerging picture that ligands acting through receptors possessing intrinsic tyrosine kinase activity promote proliferation within the regeneration blastema. However, the substrates used in this study precluded acquiring insights about modulating effects of serine/threonine kinases (e.g., PKA and PKC). Nevertheless, these data suggest that activation of PPtases contribute to the orchestration of the diverse cellular activities required to regenerate a vertebrate appendage.

Animals↗
PubMed1993

Skeletal muscle regeneration and plasticity of grafts.

The sequence of molecular and cellular events of muscle ontogeny leads to the proliferation, fusion, and differentiation of myoblasts to muscle cells. This sequence is closely paralleled in the grafting-ischemia model in which adult myoblast-satellite cells function as the muscle precursor cells. The study of skeletal muscle regeneration is a fertile and promising area of research in myogenesis. The early regenerative development and maturation of muscle is similar regardless of the perturbation that induced the degeneration-regeneration sequelae. In light of this, we maintain that the skeletal muscle graft model is useful to rigorously evaluate many regulatory aspects of skeletal muscle development and maturation in an adult animal host. One advantage of the graft model is that manipulation of the adult host, such as with exercise or hormone treatment, allows insight into their regulatory roles in muscle development and maturation. These approaches are often not possible for developing skeletal muscle in utero or in ovo. After skeletal muscle grafting, many structural and functional characteristics change with time until they reach a stable value. Successful regeneration requires revascularization, cellular infiltration, phagocytosis of necrotic muscle fibers, proliferation and fusion of muscle precursor cells, reinnervation, and recruitment and loading. The time taken to reach stable values varies among different structural and functional variables, and many reach stable values that are less than those of control skeletal muscle. There are differences in the degree of regenerative success because of the size of muscle mass grafted. In small and large grafts, regeneration is enhanced by facilitation of the reinnervation. Regeneration is evident without vascular repair in grafts of up to approximately 6 g, although in all but the 100 to 150-mg grafts in rats, a significant necrotic core is present. Regeneration is typically unsuccessful when muscle masses greater than 6 g are grafted without vascular repair. Large muscles can be grafted with vascular repair, and in this case, the cellular response is quite different, as the majority of fibers survive rather than degenerate and regenerate. Changing the components of physical activity during skeletal muscle regeneration can alter several attributes of the graft phenotype. The consensus of several experiments supports the interpretation that proper recruitment and force development by grafts are essential variables in the regulation of the development and maturation of muscle grafts. Morphological and physiological attributes of grafts adapt to changes in the habitual level of physical activity in a qualitatively similar fashion to control muscle.(ABSTRACT TRUNCATED AT 400 WORDS)

Adaptation, Physiological↗
PubMed1995

Modulation of retinoblastoma and retinoblastoma-related proteins in regenerating rat liver and primary hepatocytes.

Protein expression of the retinoblastoma (Rb) tumor suppressor gene product was examined by immunoblot analysis of nuclei isolated from regenerating rat liver after 70% partial hepatectomy (PH). Levels were almost undetectable in quiescent 0-h livers but increased 15- to 60-fold 3 to 24 h post-PH, 105-fold at 30 h, and 20- to 50-fold at 60 to 72 h post-PH. Expression returned to near baseline levels at 18, 42, and 48 h post-PH. A similar pattern of Rb protein expression in the regenerating liver was observed by indirect immunofluorescence microscopy, with peak nuclear expression at 30 h post-PH. Rb-related proteins with apparent molecular masses of 300, 156, and 74 kDa were detected in regenerating liver using mAbs to the Rb protein. Their expression increased 6- to 8-fold during regeneration, and only p156 returned to baseline levels at 60 h post-PH. Rb and its related proteins were detected in cultured primary hepatocytes, and although total protein levels did not change appreciably, there was a dramatic shift from cytosol into nuclei through 96 h. The half-life of the Rb protein was determined to be 1.9 h in regenerating liver and 2.2 h in cultured primary hepatocytes. Rb protein abundance in synchronized HuH-7 human hepatoma cells was cell cycle dependent and exhibited peak nuclear expression during S phase. Rb protein was detected primarily in its hyperphosphorylated state during liver regeneration and through the cell cycle of the HuH-7 cells. In vivo administration of transforming growth factor beta 1, an inhibitor of DNA synthesis in regenerating liver, resulted in reduced expression of Rb as well as its protein partners, cell cycle-dependent kinase 4 and cyclin E. The results suggest that in the regenerating rat liver and in synchronized HuH-7 cells, expression of Rb protein is modulated in a cell cycle-dependent fashion, remains primarily in a hyperphosphorylated state, and exhibits a relatively short half-life. The inhibition of Rb protein expression by transforming growth factor beta 1 may be linked to its simultaneous suppression of cell cycle-dependent kinase 4 and cyclin E protein levels.

Animals↗
PubMedDate not supplied

Fibrillations in regenerating muscle in dystrophic myopathies.

In the presented study we attempted to demonstrate a correlation between muscle regeneration and fibrillations in electromyography in dystrophic myopathies. Especially in Emery-Dreifuss muscular dystrophy there is much abnormal spontaneous activity, and NCAM (neural cell adhesion molecule)and cytoskeletal protein vimentin expressing myocytes are predominantly seen. Therefore, definitely regenerating fibers are identified apart from only a few remnants of previous necrosis. Moreover, in the other biopsies of dystrophic myopathies there are also scattered and clustered NCAM and vimentin expressing regenerating myofibers. Here, regressive fiber changes, like necrosis, are more prominent. Furthermore, most regenerating fibers show pseudo-cholinesterase activity indicating innervation. Interestingly, motor end-plate changes in regeneration and in disuse atrophy are very similar. They predominantly consist of terminal sprouting and pseudo-cholinesterase spread. However, in disuse atrophy there is no abnormal spontaneous spread in electromyography. Therefore, in regenerating muscle not innervation, but regeneration itself is likely to be the cause of fibrillations. In conclusion, a correlation is evident between regenerating muscle and fibrillations in electromyography.

Adolescent↗
PubMed1996

Effect of amniotic fluid in corneal sensitivity and nerve regeneration after excimer laser ablation.

To examine the effect of topical application amniotic fluids on the recovery of corneal sensitivity and nerve regeneration after excimer laser photokeratectomy, excimer laser was applied to 18 rabbits (VISX 20/20, 5 Hz, 7 microns depth: nine rabbits; 100 microns depth: nine rabbits). Human amniotic fluid (AF) was topically applied to the right eyes (AF group), and a balanced salt solution (BSS) was applied to the left eyes (BSS group). Corneal sensitivity was measured by using a Cochet-Bonnet aesthesiometer after weeks 1, 2, 4, 5, 8, and 12. Nerve-regeneration status was evaluated after weeks 2, 4, 5, 8, and 12 by gold chloride staining. Corneal sensitivity was initially subnormal and recovered close to the normal level at week 8. The sensitivity was higher in the AF group than in the BSS control group, except at week 2. Subepithelial nerve regeneration at the laser site was detected both at week 4 in the AF group and at week 5 in the BSS group. There was no significant correlation between the nerve-regeneration state and sensitivity changes. Long striated nerve regeneration from the deep stromal nerve was remarkable at the third month, especially in the BSS group. The BSS group showed more marked scarring of the superficial stroma, compared with the AF group. The nerve regeneration at the scar site was discontinued and delayed. Subepithelial leash nerves in the AF group were more abundant than those in the BSS group. The recovery of sensitivity and nerve regeneration were faster in the AF cornea than in the BSS cornea. These results suggest that the factors in AF helped the recovery of corneal sensitivity, nerve regeneration, and reduced scar formation.

Administration, Topical↗