Biomedical subjects

J C Mira

Publications and source records attributed to J C Mira.

At least 19 recordsLinked to original sources

PubMed2005

Variation in nerve autograft length increases fibre misdirection and decreases pruning effectiveness: an experimental study in the rat median nerve.

OBJECTIVES: In the clinical set, autologus nerve grafts are the current option for reconstruction of nerve tissue losses. The length of the nerve graft has been suggested to affect outcomes. Experiments were performed in the rat in order to test this assumption and to detect a possible mechanism to explain differences in recovery. METHODS: The rat median nerve was repaired by ulnar nerve grafts of different lengths. Rats were evaluated for 12 months by behavioural assessment and histological studies, including ATPase myofibrillary histochemistry and retrograde neuronal labelling. RESULTS: It was demonstrated that graft length interferes in behavioural functional recovery that here correlates to muscle weight recovery. Short nerve grafts recovered faster and better. Reinnervation was not specific either at the trunk level or in the muscle itself. The normal mosaic pattern of Type I muscle fibres was never restored and their number remained largely augmented. An increment in the number of motor fibres was observed after the nerve grafting in a predominantly sensory branch in all groups. This increment was more pronounced in the long graft group. In the postoperative period, about a 20% reduction in the number of misdirected motor fibres occurred in the short nerve graft group only. CONCLUSION: Variation in the length of nerve grafts interferes in behavioural recovery and increases motor fibres misdirection. Early recovery onset was related to a better outcome, which occurs in the short graft group.

Adenosine Triphosphatases↗

PubMed2001

Expression and neural control of myogenic regulatory factor genes during regeneration of mouse soleus.

Given the importance of the myogenic regulatory factors (MRFs) for myoblast differentiation during development, the aims of this work were to clarify the spatial and temporal expression pattern of the four MRF mRNAs during soleus regeneration in mouse after cardiotoxin injury, using in situ hybridization, and to investigate the influence of innervation on the expression of each MRF during a complete degeneration/regeneration process. For this, we performed cardiotoxin injury-induced regeneration experiments on denervated soleus muscle. Myf-5, MyoD, and MRF4 mRNAs were detected in satellite cell-derived myoblasts in the first stages of muscle regeneration analyzed (2--3 days P-I). The Myf-5 transcript level dramatically decreased in young multinucleated myotubes, whereas MyoD and MRF4 transcripts were expressed persistently throughout the regeneration process. Myogenin mRNA was transiently expressed in forming myotubes. These results are discussed with regard to the potential relationships between MyoD and MRF4 in the satellite cell differentiation pathway. Muscle denervation precociously (at 8 days P-I) upregulated both the Myf-5 and the MRF4 mRNA levels, whereas the increase of both MyoD and myogenin mRNA levels was observed later, in the late stages of regeneration (30 days P-I). This significant accumulation of each differentially upregulated MRF during soleus regeneration after denervation suggests that each myogenic factor might have a distinct role in the regulatory control of muscle gene expression. This role is discussed in relation to the expression of the nerve-regulated genes, such as the nAChR subunit gene family. (J Histochem Cytochem 49:887-899, 2001)

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PubMed2000

Neural and hormonal control of expression of myogenic regulatory factor genes during regeneration of Xenopus fast muscles: myogenin and MRF4 mRNA accumulation are neurally regulated oppositely.

With the aim to investigate the influence of both innervation and thyroid hormone, on the expression of the MRFs during muscle regeneration, we performed cardiotoxin injury-induced regeneration experiments on fast muscles of adult Xenopus laevis subjected to different experimental conditions, including denervation and T3 treatment, and analyzed the accumulation of the four myogenic regulatory factors (MRFs) using RT-PCR and in situ hybridization. We show here that manipulation of hormone levels or innervation resulted in differential alterations of MRF expression. Denervation and T3 treatment transiently down-regulated Myf-5 mRNA levels at the beginning of the regeneration process. Myf-5 was the only myogenic factor subject to thyroid hormone influence. Muscle denervation persistently reduces the levels of MRF4 transcripts as early as the first stages of regeneration, whereas the levels of myogenin mRNA were increased in the late stages of regeneration. This suggests that MRF4 expression may be induced by innervation and hence may be involved in mediating transcriptional responses to innervation and that myogenin expression may compensate for the down-regulation of MRF4 gene. This switch in MRF gene expression following denervation could have important consequences for the ability of Xenopus regenerating muscles to recover function after denervation.

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PubMed1999

Long-term denervation modulates differentially the accumulation of myogenin and MRF4 mRNA in adult Xenopus muscle.

In adult Xenopus laevis, we analyzed, using reverse transcriptase-polymerase chain reaction (RT-PCR) and in situ hybridization, the influence of long-term muscle denervation on the accumulation of MRF4 and myogenin transcripts. The brachial muscle was denervated by cutting the brachial nerve and was examined after 4 months. MRF4 mRNA levels decreased about two-fold in denervated muscle as compared with contralateral muscle. Myogenin mRNA levels, by contrast, were induced about five-fold by denervation. This report shows that muscle denervation persistently reduces the levels of MRF4 transcripts suggesting that MRF4 expression may be induced by innervation and hence may be involved in mediating transcriptional responses to innervation. The up-regulation of myogenin by denervation suggests that myogenin expression may compensate for the down-regulation of MRF4 gene.

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PubMed1999

Experimental median nerve repair by fresh or frozen nerve autografts and xenografts.

The authors described the reconstruction of a terminal branch of the brachial plexus (the median nerve) by different kinds of peripheral nerve grafts, in rats. Fresh or frozen autografts from Sprague-Dawley rats and fresh or frozen xenografts from Beagle dogs were used. Three, six, nine and twelve months after grafting, rats underwent histological assessment (muscle, nerve and spinal cord) and simple functional assessment by the grasping test. The immune reaction was prevented by the freezing and thawing method that had rendered xenografts acellular. This process allowed a satisfactory reinnervation of the flexor carpi radialis muscle (FCR) and a function recovery about 75% of control value. Nevertheless, the force recovery in rats that received frozen grafts was slower than those received fresh autografts. Probably, the destruction of cellular elements by freezing produced a deficient environment for nerve regeneration. However, this gap was partially compensated at twelve months after surgery by the maturation and the secondary adaptation of regenerated nerve fibers. Theses results showed that the force recovery is directly correlated to the capability of the nerve fibers to reproduce, histologically, a next to normal nerve pattern.

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PubMed1998

Proximal sequences of the aldolase A fast muscle-specific promoter direct nerve- and activity-dependent expression in transgenic mice.

Muscle activity is known to modulate the muscle fiber phenotype. Changes in muscle activity (normal or experimentally induced) lead to modifications of the expression status of several muscle-specific genes. However, the transcription regulatory elements involved in the adaptative response are mainly unknown. The aldolase A muscle-specific promoter, pM, is expressed in adult fast twitch muscle with a preferential expression in fast glycolytic-2B fibers. Its activity is induced during postnatal muscle maturation, suggesting a role of nerve and/or muscle activity. Indeed, denervation of gastrocnemius in newborn mice prevented the activation of the promoter in this muscle, despite the nerve-independent formation of 2B fibers. Although the nerve was necessary for pM onset during development, denervating the gastrocnemius in adults had only mild effects on pM activity. By contrast, a transgene including the pM proximal regulatory sequences that are sufficient to reproduce the 2B fiber-specific expression of the endogenous promoter was shown to be highly sensitive to both neonatal and adult denervation. Transgenes containing muscle-specific pM proximal promoter elements were used to delineate the regulatory elements involved in this response to innervation and changes in the contractile activity pattern. Nerve- and activity-dependent elements could be localized in the 130-base pair-long proximal promoter region of the human aldolase A gene.

Animals↗

PubMed1998

Localization of Myf-5, MRF4 and alpha cardiac actin mRNAs in regenerating Xenopus skeletal muscle.

We have analysed the spatial and temporal expression patterns of Myf-5, MRF4 and alpha cardiac actin mRNAs during muscle regeneration following cardiotoxin injury in adult Xenopus laevis using in situ hybridization. Myf-5 transcripts began to be detected in the activated satellite cells as early as the beginning of the regeneration process, then dramatically decreased in young plurinucleated myotubes. MRF4 mRNA was detected later, just before the young myotube stage, and was strongly expressed during the different stages of the maturation of myotubes. Like Myf-5, alpha cardiac actin mRNA began to accumulate early in activated satellite cells. These results, which contribute to an overview of the expression of the genes coding for myogenic bHLH proteins during muscle regeneration, are discussed in relation to the expression of these factors during development.

Actins↗

PubMed1998

Selective restoration of sensation by peripheral nerve grafts directly implanted into the contralateral C7 dorsal root ganglion: an experimental study in rat brachial plexus.

OBJECTIVE: In brachial plexus injuries, when the dorsal root ganglion (DRG) is avulsed from the spinal cord there is no possibility of direct repair. Therefore, in the present report the median nerve was connected directly to the contralateral C7 DRG, to restore forepaw sensation. METHODS: The ulnar nerve was sectioned, and a 15-mm segment of nerve was removed. The median nerve was sectioned, transposed dorsally, and repaired using a sural nerve graft directly implanted into the C7 contralateral DRG. Rats were then assessed 6 and 12 months later by behavioral, histological, and plasma extravasion techniques. RESULTS: All animals recovered sensation in the forepaw. Retrogradely labeled sensory neurons were demonstrated in the C7 DRG, and the whole forepaw was labeled after plasma extravasion induction by median nerve antidromic stimulation. Several myelinated and positively neurofilament-stained fibers were demonstrated in the grafted median nerve. CONCLUSION: The surgical strategy proposed might be a useful alternative to selective sensory repair in the emerging field of brachial plexus reconstruction by direct spinal cord surgery.

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PubMed1997

Selective motor hyperreinnervation using motor rootlet transfer: an experimental study in rat brachial plexus.

Misdirection of sensory fibers into motor pathways is, in part, responsible for the poor results obtained after peripheral nerve repair. After avulsion of the C-5 root in rats, the authors connected a C-4 ventral rootlet to the musculocutaneous nerve by means of a sural nerve graft. In this way, they were able to increase the number of regenerating motor fibers and avoid growth of sensory fibers into the nerve grafts. Functional recovery was evaluated electrophysiologically and histologically. The origin of the axons that reinnervated the nerve graft was analyzed by means of morphological studies including retrograde labeling procedures. Motor neurons survived and regenerated after the rootlet transfer and there was no functional impairment. Many neurons were retrograde labeled in the ventral horn and widespread biceps muscle reinnervation was demonstrated with recovery of nearly normal electrophysiological properties. Motor hyperreinnervation of the musculocutaneous nerve was observed. This high degree of reinnervation in a long (40-mm) graft was attributed to the good chance that a muscle fiber can be reinnervated by a motor fiber when the number of regenerating motor neurons is increased and when competitive sensory fibers are excluded from reinnervation.

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PubMed1997

[Post-traumatic reconnection of the cervical spinal cord with skeletal striated muscles. Study in adult rats and marmosets].

In an attempt at repairing the injured spinal cord of adult mammals (rat, dog and marmoset) and its damaged muscular connections, we are currently using: 1) peripheral nerve autografts (PNG), containing Schwann cells, to trigger and direct axonal regrowth from host and/or transplanted motoneurons towards denervated muscular targets; 2) foetal spinal cord transplants to replace lost neurons. In adult rats and marmosets, a PNG bridge was used to joint the injured cervical spinal cord to a denervated skeletal muscle (longissimus atlantis [rat] or biceps brachii [rat and marmoset]). The spinal lesion was obtained by the implantation procedure of the PNG. After a post-operative delay ranging from 2 to 22 months, the animals were checked electrophysiologically for functional muscular reconnection and processed for a morphological study including retrograde axonal tracing (HRP, Fast Blue, True Blue), histochemistry (AChE, ATPase), immunocytochemistry (ChAT) and EM. It was thus demonstrated that host motoneurons of the cervical enlargement could extend axons all the way through the PNG bridge as: a) in anaesthetized animals, contraction of the reconnected muscle could be obtained by electrical stimulation of the grafted nerve; b) the retrograde axonal tracing studies indicated that a great number of host cervical neurons extended axons into the PNG bridge up to the muscle; c) many of them were assumed to be motoneurons (double labelling with True Blue and an antibody against ChAT); and even alpha-motoneurons (type C axosomatic synapses in HRP labelled neurons seen in EM in the rat); d) numerous ectopic endplates were seen around the intramuscular tip of the PNG. In larger (cavitation) spinal lesions (rat), foetal motoneurons contained in E14 spinal cord transplants could similarly grow axons through PNG bridges up to the reconnected muscle. Taking all these data into account, it can be concluded that neural transplants are interesting tools for evaluating both the plasticity and the repair capacities of the mammalian spinal cord and of its muscular connections.

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PubMed1995

Relationship between muscle myosin isoforms and contractile features in rabbit fast-twitch denervated muscle.

The effects of 8-day-old rabbit fast-twitch gastrocnemius denervation on the type of myosin isoforms and on contractile features (maximum velocity Vmax and contraction time (CT) of the muscle were followed between 15 and 60 days postnatal. The myosin isoforms and the Vmax and CT values of the denervated gastrocnemius displayed large changes during this period. These changes, which led at 2 months postnatal to a muscle displaying the properties of a slow-twitch muscle did not occur in synchrony: complete conversion to slow-type myosin isoforms occurred only at 60 days postnatal, whereas complete conversion to slow-twitch Vmax and CT values occurred as soon as 35 days postnatal. The results address a new question concerning the relationship between muscle myosin and contractile features.

Aging↗

PubMed1995

The rat brachial plexus and its terminal branches: an experimental model for the study of peripheral nerve regeneration.

Despite the introduction of microsurgical techniques into clinical practice, the results of surgical procedures involving the brachial plexus and peripheral nerves are still far from spectacular. We therefore studied the rat brachial plexus and its terminal branches in 203 rats. Detailed anatomic and morphologic analyses of the biceps brachii and musculocutaneous nerve, finger flexors, flexor carpi radialis, and the median nerve were performed. Various sources of conventional and vascularized nerve grafts were explored. After musculocutaneous nerve section or median nerve section, there were no articular contractures or automutilations, which constitutes an advantage for these experimental models over the sciatic nerve model. The brachial plexus and its terminal branches provide a good experimental model which can be used to assess the development and normal control of muscle function, examine the mechanisms underlying functional recovery, and test the effects of treatments to enhance recovery.

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PubMed1995

The grasping test: a simple behavioral method for objective quantitative assessment of peripheral nerve regeneration in the rat.

Behavioral recovery is the major goal of neuroscience research on the peripheral nervous system. However, behavioral methods of assessing peripheral nerve regeneration are still lacking. In the present work we used the median nerve and finger flexors as the experimental model. After resection or crush injury of the median nerve, rats were submitted daily to the grasping test. For this, they were gently lifted by the tail and allowed to grasp a grid connected to an ordinary electronic balance. While grasping the animal continued to be lifted by the tail with increasing firmness until it lost its grip. At this precise moment the value shown by the balance was recorded. In the crushed median nerve, recovery of function was clearly demonstrated by the grasping test. The test also indicated the exact day on which recovery began and its improvement with time. This very simple objective behavioral method provides a sensitive quantitative technique for assessing recovery. The experimental model using the median nerve seems to be better than the sciatic nerve model, as no articular contractures or autotomies were noted in the present experimental animals.

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PubMed1995

Response to denervation of rabbit soleus and gastrocnemius muscles. Time-course study of postnatal changes in myosin isoforms, fiber types, and contractile properties.

In contrast to general belief, the response of rabbit muscles to denervation is maturation to slow-like type muscles [7]. We report now an investigation by biochemical, morphological, and mechanical studies of the time course effects of muscle denervation on the slow-type soleus and fast-type gastrocnemius to help elucidate the mechanism of maturation of rabbit denervated muscles to slow-like muscles. In both muscles, denervation induced selective progressive atrophy of most fast fibers and hypertrophy of many slow fibers which displayed wide Z-lines; this was accompanied by the appearance of hybrid LC1F- and LC1E-associated slow myosins. The percentage of slow myosins increased with age similarly in the contralateral and denervated soleus. On the other hand, the percentage of slow myosins remained low in the contralateral gastrocnemius, whereas it increased to 95% in the denervated gastrocnemius; in the denervated gastrocnemius, the percentage of slow myosins reached 50% at about 35 days postnatal. At this age, the maximal shortening velocity of the denervated gastrocnemius and its twitch contraction time were already those of a slow-type muscle. This suggests that in addition to myosin, other proteins contributed to the mechanical properties of the denervated gastrocnemius. Transformation of rabbit denervated muscles to slow-like type muscles, which are associated with a lower energy requirement and higher muscle endurance than fast-type muscles, may constitute an adequate model for human neuromuscular pathology.

Aging↗

PubMed1994

The effect of denervation on myosin isoform synthesis in rabbit slow-type and fast-type muscles during terminal differentiation. Denervation induces differentiation into slow-type muscles.

The soleus and gastrocnemius medialis of eight-day-old rabbits were denervated and the effects were examined after fifty-two days by biochemical, cytochemical and mechanical methods. The contralateral soleus exhibited the properties of slow-type muscle, namely a predominance of slow-type myosin isoforms and slow-type oxidative fibers, slow twitch and low maximal velocity for shortening. The contralateral gastrocnemius exhibited the properties of fast-type muscle, namely a predominance of fast-type myosin isoforms and fast-type non-oxidative fibers, fast twitch and high maximal velocity of shortening. Denervation of muscles caused the differentiation of the two muscles towards slow-type muscles. Both denervated soleus and gastrocnemius muscles exhibited a predominance of slow-type myosins (either the normal type, made up of slow heavy and light chains, or the hybrid type, made up of slow heavy and regulatory light chains and fast essential light chains), a predominance of slow-type fibers, and slow mechanical properties. Thus, innervation in rabbit appears to be a determining factor for differentiation into fast-type muscle, but it is not necessary for differentiation into slow-type muscle. This conclusion contradicts the findings of previous studies in rat and thus raises new questions concerning the role of nerves in controlling the expression of myosin isoforms and the differentiation of muscle fibers.

Animals↗

PubMed1994

Median nerve neurotization by peripheral nerve grafts directly implanted into the spinal cord: anatomical, behavioural and electrophysiological evidences of sensorimotor recovery.

Over the years, peripheral nerve grafts, a favorable environment to support axonal elongation, have given rise to increasing interest as a possible solution for promoting spinal cord repair. In the experiments described here, following an avulsion injury of the rat brachial plexus, the median nerve was repaired by a peripheral nerve graft (PN) inserted directly into the dorsal side of the spinal cord. Eight months later the animals were submitted to behavioral tests, electrophysiological and histological studies. Regrowth of axons from both motoneurons and ganglionic neurons was demonstrated following a single superficial dorsal implantation of a PN. Sensorimotor peripheral reinnervation allowed most of the studied animals to recover enough flexor activity for grasping. Reinnervation was achieved even without prior root avulsion suggesting that the presence of a PN is sufficient to induce sprouting in the spinal cord from axotomized and non-axotomized neurons.

Animals↗

PubMed1994

Electrophysiological evaluation of the effects of naftidrofuryl on skeletal muscle reinnervation in the rat.

The effects of naftidrofuryl on the reinnervation of the rat gastrocnemius muscle after its denervation by localized freezing of the sciatic nerve were tested with electrophysiological techniques. Daily intraperitoneal injections of 30 mg/kg of naftidrofuryl do not increase the rate of axonal regeneration since early signs of reinnervation appeared as in controls around the 10th day after surgery. However, axonal sprouting is markedly increased since the percentage of muscle fibers with polyneuronal innervations was almost twice as high as in controls at the 15 and 21 day postoperative stages. The promoting effects of naftidrofuryl on polyneuronal innervation which gives rise to a redundant innervation during the first period of reinnervation constitutes an improvement of motor function which might be efficient for treatment of nerve injury and neuropathies.

Animals↗

PubMed1994

Vascular freezing--a new method for immediate and permanent vasospasm relief: an experimental study in the rat.

Although much work has been done on the etiology and prevention of vasospasm, a spasmolytic agent capable of firmly protecting against or reversing vasospasm has not yet been found. In this paper, we describe a new physical method of treating vasospasm. It consists of directly freezing the arterial walls. Experiments were performed on the epigastric artery of 40 female Sprague-Dawley rats. A vasospasm was created by mechanical vessel manipulation. The vasospasm was treated by freezing the vessel walls with a dermatologic hand-held liquid nitrogen spray. Vascular spasm and patency controls were made immediately after thawing and 30 minutes and 3, 10, and 120 days after freezing. Epigastric island flaps were harvested based on the frozen artery at 3, 10, and 120 days after the freezing procedure. Histologic studies also were effectuated 3, 10, and 120 days after the freezing procedure. Relief from vasospasm was instantaneous and permanent. After freezing, almost all the cells in the media and endothelium died and the adrenergic fibers degenerated. This was followed by a phagocytic debridement, complete regeneration of the endothelium, and a limited regeneration of the muscle fibers in the media. The adrenergic innervation was recovered, and no alterations in the adrenergic fibers distal to the lesion were noted at any time. None of the arteries examined underwent thrombosis, and all the flaps harvested survived very well, indicating that the vascular bed distal to the site of the freezing was normal. This vascular freezing technique may well have clinical applications for microsurgical transfers and for the prophylactic treatment of vasospasm disease. However, its indications still require clarification.

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