Fibronectin distribution during somitogenesis in the chick embryo

Somite formation in vertebrates is a multi-stage process. From a relatively homogeneous rod of mesenchyme, the segmental plate, somites are formed in a repeating sequence. Cell-cell adhesion has been proposed as a causal factor in somitogenesis. This led to an analysis of fibronectin in the segmental plate with respect to the initiation of somitogenesis. The pattern of fibronectin distribution can be correlated with the initiation of somitogenesis in the anterior portion of the segmental plate. Fibronectin distribution was determined using a high resolution antibody localization technique. Differences in fibronectin distribution were verified with computer-assisted image analysis. The evidence presented supports the hypothesis that an increase in cell-cell adhesion is a significant factor in the initiation of somitogenesis.     

5-Aza-2'-deoxycytidine-induced cytotoxicity and limb reduction defects in the mouse

BACKGROUND: 5-Aza-2'-deoxycytidine (dAZA), causes hindlimb phocomelia in CD-1 mice. Studies in our laboratory have examined the hypothesis that compound- induced changes in gene expression may uniquely affect hindlimb pattern formation. The present study tests the hypothesis that dAZA causes limb dysplasia by inducing cytotoxicity among rapidly proliferating cells in the limb bud mesenchyme. METHODS: Pregnant CD-1 mice were given a teratogenic dose of dAZA (i.p.) at different times on GD 10 and fetuses evaluated for skeletal development in both sets of limbs by standard methods. Using general histology and BrdU immunohistochemistry, limb mesenchymal cell death and cell proliferation were then assessed in embryos at various times post dosing, shortly after initial limb bud outgrowth. The effect of dAZA on early limb chondrogenesis was also studied using Northern analysis of scleraxis and Alcian blue staining of whole mount limb buds. RESULTS: Compound related hindlimb defects were not restricted to a specific set of skeletal elements but consisted of a range of temporally related limb anomalies. Modest defects of the radius were observed as well. These results are consistent with a general insult to the limb mesenchyme. Mesenchymal cell death and reduced cell proliferation were also observed in both sets of limbs. The timing and location of these effects indicate a role for cytotoxicity in the etiology of dAZA induced limb defects. These effects also agree with the greater teratogenicity of dAZA in the hindlimb because they were more pronounced in that limb. The expression of scleraxis, a marker of early chondrogenesis, was reduced 12 hr after dAZA exposure, a time coincident with maximal cell death, as was the subsequent emergence of Alcian blue stained long bone anlagen. CONCLUSIONS: These findings support the hypothesis that cytotoxic changes in the limb bud mesenchyme during early limb outgrowth can induce the proximal limb truncations characteristic of phocomelia after dAZA administration.     

Position specific binding of a monoclonal antibody in chick limb buds

To analyze the molecular mechanism of the limb pattern formation, we have tried to make monoclonal antibodies against antigens from chick limb buds. We obtained one antibody named AV-1 which recognized a specific region of chick limb buds. AV-1 reacted with the distal portion of the anteroventral mesoderm of only developmentally early chick limb buds. Grafts of ZPA region tissue to an anterior site in an embryonic chick wing bud resulted in mirror-image dupliction of the AV-1 antigen region. These data show the possibility that this antigen plays some role in the limb pattern formation. This is the first evidence that a position specific substance really exists in developmentally early limb buds in which the pattern has been considered to be unspecified.     

Expression of DLX3 in chick embryos

Higher vertebrates appear to possess six genes encoding a homeodomain of the distal-less type. We report the cloning and expression pattern of the chicken DLX3 gene, a homeobox gene highly related to the DLX5 gene with regard to both the encoded protein structure and the expression pattern. DLX3 RNA was observed during the development of the olfactory and otic placodes, in the distal portion of the first and second visceral arch mesenchyme, in the growing limb buds, and in the tail tip. No expression occurs in the central nervous system.     

A novel device for long bone osteodistraction: description of device and case series

The purpose of this study was to present a novel intramedullary device (M-Bone; Phenix, Paris, France) that contains a mechanism for internal osteodistraction and bone transport in patients with segmental bone defects or limb length discrepancy after limb salvage operations. A total of five patients with primary bone tumors were enrolled in the study. After implantation, daily lengthening was performed in an outpatient setting either by the patient or with the help of a therapist, without the use of anesthesia. This unique device offers a totally new approach for the treatment of segmental bone defects or limb length discrepancy. It is designed to expand the remaining native bone by a magnetically activated drive system and induces new bone formation using osteodistraction and bone transport.     

Expressing hNF-LE397K results in abnormal gaiting in a transgenic model of CMT2E

Charcot-Marie-Tooth disease (CMT) is the most commonly inherited peripheral neuropathy. CMT disease signs include distal limb neuropathy, abnormal gaiting, exacerbation of neuropathy, sensory defects and deafness. We generated a novel line of CMT2E mice expressing an hNF-L(E397K) transgene, which displayed muscle atrophy of the lower limbs without denervation, proximal reduction in large caliber axons and decreased nerve conduction velocity. In this study, we showed that hNF-L(E397K) mice developed abnormal gait of the hind limbs. The identification of severe gaiting defects in combination with previously observed muscle atrophy, reduced axon caliber and decreased nerve conduction velocity suggests that hNF-L(E397K) mice recapitulate many of clinical signs associated with CMT2E. Therefore, hNF-L(E397K) mice provide a context for potential therapeutic intervention.     

Outgrowth by fin motor axons in wildtype and a finless mutant of the Japanese medaka fish

The outgrowth of motor axons to the developing pectoral fin of the Japanese medaka fish (Oryzias latipes) was investigated both in wildtype embryos and in the pectoral finless (pl) mutants in which adults are missing pectoral fins. Late in embryogenesis the pectoral fin is a simple limb which contains two antagonist muscles which are innervated by presumptive motor neurons from the first four spinal segments (S1-4). The pectoral fin develops from a fin bud located in S1 and S2 centered on the border between S1 and S2 and, as with other limbs, one of the earliest signs of differentiation is the apical ectodermal ridge (AER). By the time the AER is well formed the growth cones of the presumptive motor neurons have reached the base of the fin bud and formed a plexus by extending toward the fin bud upon emergence from the spinal cord. This is especially evident on the ventral surface of the metamerically arranged axial muscles. For example, growth cones from S2 extend in a diagonal direction (both anterior and lateral) towards the fin bud. One hypothesis which can account for the pattern of motor outgrowth is that growth cones are attracted to the base of the fin bud, perhaps via a long distance cue. This hypothesis was tested by examining outgrowth of segmental nerves in pl embryos in which the fin buds arrest early in development following the initial appearance of the AER. In pl, nerves from S1-4 converged to form a plexus at the base of the abnormal fin bud, but the pattern of outgrowth varied from wildtype in a way consistent with a diminished capacity of the fin bud to attract segmental nerves to it.     

Control of targeted flexor movements of the human arm at different levels of the possibility for their preprograming

Contribution of the processes of central preprograming of an equilibrium target position of the limb link was studied by testing two variants of motor task in humans. In the first variant, the tested person could obtain visual information about the target position before the movement initiation. In the second variant, such information initially was absent and was presented only in the course of the movement performance. It has been shown that in both variants the pattern of EMG activity of flexor muscles, which realize the movement (and, respectively, the pattern of motor commands, i.e., efferent activity of spinal motoneurons) demonstrated no fundamental differences. Therefore, it can be supposed that the attainment of a target level in both cases was preprogramed only to a limited extent; more probably, it was provided by successive current control of the limb link position. This control is based, first of all, on dynamic changes of the control signals. In general, data of the experiments are in agreement with the impulse—temporal hypothesis of control of targeted movements.     

The cross-groin flap for coverage of foot and ankle defects in children. Case reports.

The cross-groin flap is useful for covering soft tissue defects of the distal portion of the lower limb in a child.     

moz regulates Hox expression and pharyngeal segmental identity in zebrafish

In vertebrate embryos, streams of cranial neural crest (CNC) cells migrate to form segmental pharyngeal arches and differentiate into segment-specific parts of the facial skeleton. To identify genes involved in specifying segmental identity in the vertebrate head, we screened for mutations affecting cartilage patterning in the zebrafish larval pharynx. We present the positional cloning and initial phenotypic characterization of a homeotic locus discovered in this screen. We show that a zebrafish ortholog of the human oncogenic histone acetyltransferase MOZ (monocytic leukemia zinc finger) is required for specifying segmental identity in the second through fourth pharyngeal arches. In moz mutant zebrafish, the second pharyngeal arch is dramatically transformed into a mirror-image duplicated jaw. This phenotype resembles a similar but stronger transformation than that seen in hox2 morpholino oligo (hox2-MO) injected animals. In addition, mild anterior homeotic transformations are seen in the third and fourth pharyngeal arches of moz mutants. moz is required for maintenance of most hox1-4 expression domains and this requirement probably at least partially accounts for the moz mutant homeotic phenotypes. Homeosis and defective Hox gene expression in moz mutants is rescued by inhibiting histone deacetylase activity with Trichostatin A. Although we find early patterning of the moz mutant hindbrain to be normal, we find a late defect in facial motoneuron migration in moz mutants. Pharyngeal musculature is transformed late, but not early, in moz mutants. We detect relatively minor defects in arch epithelia of moz mutants. Vital labeling of arch development reveals no detectable changes in CNC generation in moz mutants, but later prechondrogenic condensations are mispositioned and misshapen. Mirror-image hox2-dependent gene expression changes in postmigratory CNC prefigure the homeotic phenotype in moz mutants. Early second arch ventral expression of goosecoid (gsc) in moz mutants and in animals injected with hox2-MOs shifts from lateral to medial, mirroring the first arch pattern. bapx1, which is normally expressed in first arch postmigratory CNC prefiguring the jaw joint, is ectopically expressed in second arch CNC of moz mutants and hox2-MO injected animals. Reduction of bapx1 function in wild types causes loss of the jaw joint. Reduction of bapx1 function in moz mutants causes loss of both first and second arch joints, providing functional genetic evidence that bapx1 contributes to the moz-deficient homeotic pattern. Together, our results reveal an essential embryonic role and a crucial histone acetyltransferase activity for Moz in regulating Hox expression and segmental identity, and provide two early targets, bapx1 and gsc, of moz and hox2 signaling in the second pharyngeal arch.     

Heat shock produces periodic somitic disturbances in the zebrafish embryo.

Environmental influences are known to produce segmental defects in a variety of organisms. In this paper we report upon segmental aberrations produced by brief heat shocks delivered to developing zebrafish embryos. The initial defects in the segmental pattern of somitic boundaries and motoneuron axon outgrowth were usually observed five somites caudal to the somite which was forming at the time of heat shock application. Segmental defects in zebrafish embryos exposed to a single heat shock treatment can occur in a periodic pattern similar to the multiple disturbances observed to occur in chick embryos. These data are discussed with regard to models involving cell cycle synchrony or 'clock and wavefront' schemes in the process of somitogenesis.     

Treatment of symptomatic diabetic neuropathy by surgical decompression of multiple peripheral nerves.

Symptomatic diabetic sensorimotor polyneuropathy is considered progressive and irreversible. The hypothesis that symptoms of diabetic neuropathy may be due to entrapment of peripheral nerves was investigated in a prospective study from 1982 to 1988 in which diabetics (38 type I, 22 type II) had surgical decompression of 154 peripheral nerves in 51 upper extremities and 31 lower extremities. Mean postoperative follow-up was 30 months (range 6 to 83 months). Considering the entire series, an excellent final result was noted for motor function in 44 percent and for sensory function in 67 percent of the decompressed nerves. Ten percent of the patients were not improved, and 2 percent were worse in sensorimotor function. Upper extremity nerve decompressions achieved better results than lower extremity nerve decompressions. Improvement in postoperative electrodiagnostic studies varied in relationship to the preoperative electrodiagnosis. Improvement was noted in 100 percent of those nerves with the preoperative diagnosis of "localized entrapment," 80 percent for "peripheral neuropathy with superimposed entrapment," and 50 percent for "peripheral neuropathy." Progressive neuropathy occurred in a nontreated limb of 50 percent of those patients whose surgically treated limb maintained improvement. The results of this study suggest that symptoms of sensorimotor diabetic neuropathy may be due partly to compression of multiple peripheral nerves. The results further suggest that surgical decompression of such nerves may result in symptomatic improvement.     

Development of the Pattern of Innervation of the Chick Limb

SYNOPSIS. There is a consistent repeatable pattern of innervationof the vertebrate limb by sensory and motor axons from the spinalcord. We have been looking at the factors which control thegeneration of this pattern in the developing chick limb. Thegross pattern of nerves in the limb is controlled by the limbtissue and arises as a result of the axon's response to itslocal environment. Each segmental root innervates a characteristic territory inthe limb. Each muscle is innervated by a characteristicallylocated motoneurone pool in the cord. Three models explaining the generation of these patterns arepresented: active selection, selective cell death, and passivedeployment. A review of recent experiments leads us to concludethat there is evidence for both passive deployment and local,short range, active selection.     

Loss of carpal elements in crocodilian limb evolution: morphogenetic model corroborated by palaeobiological data

The comparison of bone homology between the manus of an Early Cretaceous fossil crocodile and that of the extant species Alligator mississippiensis supports explicidy, for the first time, the hypothesis of carpal loss in crocodilian limb evolution. This hypothesis, based on a developmental model of the organization of the tetrapod limb, is in accordance with the fossil evidence, and may supersede traditional Haeckelian views based on recapitulatory paradigms. The homologous relationships of carpal elements reveal the existence of two carpal patterns- one plesiomorphic and one apomorphic-in the crocodilian lineage. Phylogenetic change is explained causally by alterations of the osteogenesis of the distal carpals 2 and 3, which remain unossified in extant crocodile adults. This implies that crocodilian limb evolution is constrained by a process of paedomorphosis. This modification of the architecture of the crocodilian hand is a terminal event of its evolutionary history, affecting only eusuchian crocodiles. The results of this study contest the traditional view that the skeletal pattern of the crocodilian limb has been conserved unchanged since the Triassic.     

Electromyographic assessment of rectus abdominis muscle function after deep inferior epigastric perforator flap surgery

SUMMARY: The authors evaluated rectus abdominis muscle function after deep inferior epigastric perforator (DIEP) flap elevation. Fifteen consecutive patients who were operated on for breast reconstruction with a free DIEP flap were included in the study. A turn-amplitude electromyographic analysis was used. For each patient, the muscle activity was recorded in the portion of the muscle that was split for the epigastric perforator vessel dissection, and also in the similar portion of the contralateral nondissected muscle. A first electromyographic examination was carried out soon after surgery (mean follow-up, 9 weeks), and a second electromyographic examination was carried out at a later date (mean follow-up, 15 months). The mean activity of the dissected muscles was 50 percent of the activity of the nondissected muscles at the first electromyographic examination and 70 percent at the second electromyographic examination. The authors suggest that the DIEP flap procedure induces a partial denervation of the rectus abdominis muscle in the area of dissection and that reinnervation occurs over time because the entire width of the muscle and sufficient segmental motor innervation are preserved.     

Relative variation in human proximal and distal limb segment lengths

The pattern of variation and covariation of proximal and distal limb segment lengths was examined within and between 20 geographically diverse skeletal samples of modern humans. Analyses of variance-covariance matrices (VCMs) of logarithmically transformed (ln) variates of humerus, radius, femur, and tibia length were performed to test the following hypotheses: first, within populations, the distal and proximal segments will have equal relative (i.e., size-independent) variability. However, between populations, the tibia is predicted to be more variable than the other segments. Tests of fit of computed VCMs to theoretical matrices by an iterative procedure (Anderson [1973] Ann. Stat. 1:135-141) reject the equal variance hypotheses, rather suggesting that the relative variances of the distal limb segments are greater than are those of the proximal. Males and females differ somewhat in that within females, the distal segments of both limbs have equal variance, while within males, the tibia has greater relative variance than the radius. The second hypothesis, regarding between-group variability, is somewhat supported in that between human populations, one cannot reject that the tibia has greater relative variance than the other limb segments. However, neither can one reject an alternative hypothesis that both distal limb segments (tibia and radius) are more variable than the proximal segments. Differential growth allometry is explored, and likely plays a major role in differences seen both within and between human populations.     

The phylogenetic relationships of the bilby, Macrotis lagotis (Peramelimorphia: Thylacomyidae), to the bandicoots- DNA sequence evidence

Partial sequencing of the 12S ribosomal RNA gene was used to test two competing hypotheses concerning the phylogenetic relationship of the bilby (Macrotis lagotis) to the Australian and New Guinean species of bandicoot. The first hypothesis proposes that the Australian and New Guinean bandicoots are in a monophyletic clade to the exclusion of the bilbies, whereas the second hypothesis proposes that the bilby is monophyletic with the Australian bandicoots to the exclusion of the New Guinean bandicoots. Phylogenies determined by both maximum-likelihood and neighbour-joining approaches supported the first hypothesis in which the bilby is excluded from the clade represented by the Australian and New Guinean bandicoots. Monophyly of the Australian and New Guinean bandicoots is consistent with the biogeographical scenario in which Australia and Papua New Guinea have undergone repeated connection and disconnection over the last 20 million years.     

A transgenic, mesodermal specific, Dkk1 mouse model recapitulates a spectrum of human congenital limb reduction defects

Congenital limb reduction defects occurring in isolation of other developmental abnormalities continue to be an important medical problem in which little progress has been made. Herein we generated transgenic mice expressing Dkk1 in an appendicular mesodermal pattern. Prx1-Dkk1 mice recapitulate a full spectrum of human congenital limb reduction defects, without other developmental issues, and have normal life-spans. Importantly, a close examination of the inheritance pattern suggests that there is a significant degree of incomplete penetrance as progeny of phenotypically positive or phenotypically negative, but genotypically positive Prx1-Dkk1 mice, consistently give rise to both phenotypically positive mice and phenotypically normal-appearing mice. Thus, this heterogeneous phenotype is reproducible with each generation regardless of the phenotype of the parents. We further go on to identify that mesenchymal stem cells from Prx1-Dkk1 mice have limited proliferative ability, but normal differentiation potential, which may explain the mechanism for the limb reduction defects observed. We believe Prx1-Dkk1 mice may prove useful in the future to study the mechanisms underlying the development of congenital limb reduction defects.