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N Buffinger

Publications and source records attributed to N Buffinger.

4 recordsLinked to original sources

Myogenic specification of somites is mediated by diffusible factors.

Specification of myogenesis in early chicken somites and segmental plate was studied using transfilter explant cultures to determine if myogenic specification by axial structures is mediated by cell-cell contact. Formation of muscle fibers that express myosin heavy chain was assessed in somites transfilter from neural tube and notochord. Either the neural tube or the notochord from early chick embryos (ED 2) induces myogenesis in unspecified somites (Hamburger and Hamilton (HH) stages 11-14) when these tissues are separated by a 0.2- or a 0.05-micron pore filter. The ventral neural tube is found to be a strong inducer of myogenesis, while the dorsal neural tube is found to have low inducing activity. The reduced myogenic inducing activity of the dorsal neural tube is associated with an activity that inhibits myogenic differentiation in specified somites, somites containing cells already committed to myogenesis. Only 6-8 hr of transfilter exposure to neural tube is required to initiate somite myogenesis that is sustained in the absence of neural tube. Other tissues such as specified somites, heart, or ectoderm from the same aged embryo do not induce myogenesis in unspecified somites. Removal of the prospective floor plate of the caudal neural plate at HH stage 8 or 9 does not eliminate the inducing activity of the neurectoderm on unspecified somites (HH stage 11-14). Recombination of somites with neural tube or spinal cord from progressively older embryos (ED 4-20) showed myogenesis inducing activity at all ages, though the activity waned as development proceeded. Paraxial mesoderm need not segment into somites to respond to the inducing activity of the neural tube. We conclude that induction of myogenesis in somites does not require cell-cell contact between either the neural tube or the notochord and therefore induction is mediated by diffusible factor(s); that the inducing activity is localized principally to the notochord and ventral half of the neural tube, but not necessarily to the floor plate; that the myogenic response is specific to neural tube and notochord; and that the dorsal neural tube weakly induces myogenesis and contains an activity that inhibits myogenic cells from differentiating.

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Myogenic specification in somites: induction by axial structures.

Specification of the myogenic phenotype in somites was examined in the early chick embryo using organotypic explant cultures stained with monoclonal antibodies to myosin heavy chain. It was found that myogenic specification (formation of muscle fibers in explants of somites or segmental plates cultured alone) does not occur until Hamburger and Hamilton stage 11 (12-14 somites). At this stage, only the somites in the rostral half of the embryo are myogenically specified. By Hamburger and Hamilton stage 12 (15-17 somites), the three most caudal somites were not specified to be myogenic while most or all of the more rostral somites are specified to myogenesis. Somites from older embryos (stage 13-15, 18-26 somites) showed the same pattern of myogenic specification--all but the three most caudal somites were specified. We investigated the effects of the axial structures, the notochord and neural tube, on myogenic specification. Both the notochord and neural tube were able to induce myogenesis in unspecified somites. In contrast, the neural tube, but not the notochord, was able to induce myogenesis in explants of segmental plate, a structure which is not myogenic when cultured alone. When explants of specified somites were stained with antibodies to slow or fast MyHC, it was found that myofiber diversity (fast and fast slow fibers) was established very early in development (as early as Hamburger and Hamilton stage 11). We also found fiber diversity in explants of unspecified somites (the three most caudal somites from stage 11 to 15) when they were recombined with notochord or neural tube. We conclude that myogenic specification in the embryo results in diverse fiber types and is an inductive process which is mediated by factors produced by the neural tube and notochord.

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Fibroblast growth factor in the extracellular matrix of dystrophic (mdx) mouse muscle.

Polyclonal antibody F547 reacts with a bovine basic fibroblast growth factor (bFGF) and a human recombinant bFGF, but not with bovine acidic fibroblast growth factor. This antibody localized bFGF in the extracellular matrix of mouse skeletal muscle, primarily in the fiber endomysium, which includes the heparin-containing basal lamina. In mdx mouse muscle, which displays persistent regeneration, FGF levels in the extracellular matrix are higher than those in controls. Overabundance of matrix FGF in mdx muscles may be related to an increase in both satellite cell and regenerative activity in the dystrophic muscle and may help explain the benign phenotype of mdx animals compared with the genetically identical human Duchenne muscular dystrophy.

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Fibroblast growth factor is stored in fiber extracellular matrix and plays a role in regulating muscle hypertrophy.

A polyclonal antibody specific to bovine basic fibroblast growth factor (bFGF) localizes bFGF in the extracellular matrix of rat skeletal muscle fibers, where it is bound by the heparin-rich basal lamina. FGF accumulation appears to be augmented in hypertrophied compared to control fibers. In addition, FGF-like growth activity can be isolated from hypertrophied muscle. This growth activity binds to heparin and can be purified by heparin-affinity chromatography. When assayed in a chick myogenic cell culture system, the purified rat growth activity stimulates myobenic cell replication and stimulates myosin accumulation and synthesis. The function, in vivo, of basal lamina-localized FGF in rat muscle is not known, but it is most probably related to increased satellite cell replication seen during normal fiber maturation, during work-induced hypertrophy, and during regeneration. During hypertrophy, we observe the appearance of new fibers in addition to growth of preexisting fibers. It is hypothesized that muscle hypertrophy involves activation of satellite cells which may then fuse with existing fibers and, in addition, also form new fibers.

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