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Biomedical subjects

T J Sims

Publications and source records attributed to T J Sims.

102 records · Page 6Linked to original sources

The generation of neurons involved in an early reflex pathway of embryonic mouse spinal cord.

The generation of lateral motor neurons (LMNs), interneurons and dorsal root ganglion (DRG) neurons of the cervical mouse spinal cord has been investigated by [3H]thymidine autoradiographic techniques. This investigation has two main objectives: (a) to determine on which embryonic days these three neuronal populations are born, and (b) to investigate the possibility that the neurons comprising early reflex circuits might be formed by a retrograde temporal sequencing of generation. LMNs are the first neurons generated in the cervical spinal cord. They arise between E8.8 and E11.5, and approximately 90% of these cells are born within a 36-hour period between E9 and E10.5. The earliest time of origin for interneurons is on E9.5, and those cells which are generated between E9.5 and E10.5 cluster in two distinct regions of the adult spinal cord. One of these regions is the lateral portions of laminae IV through VI; this appears to be the location of many ipsilateral association neurons. DRG neurons begin to arise on E9.5 and their generation is completed by E14. There is a trend within the DRG population for large neurons to be born before small neurons. Those cells with diameters of 40 micron or greater reach their generation peak on E10.5, while those smaller than 40 micron arise in the greatest numbers on E12. The findings of other investigations have provided evidence for a retrograde sequence of synaptic closure in the formation of the early disynaptic forelimb reflex pathway. The temporal difference in synapse formation in the terminal fields of DRG and association neurons is discussed in terms of our observation that both of these populations appear to have similar generation times. We suggest that factors responsible for the delayed synaptic closure of DRG afferents include the greater distances and the degree of collateralization which these afferents must undergo in order to establish their terminal fields. Finally, we discuss the possibility that the temporal sequence of neuronal generation and factors involved with the growth of neurites combine to produce a retrograde sequence of synaptic closure in the early disynaptic forelimb reflex pathway of mouse spinal cord.

Animals↗

The development of monamine-containing neurons in the brain and spinal cord of the salamander, Ambystoma mexicanum.

The distribution of monoamine-containing neurons in the CNS of the developing and adult axolotl, Ambystoma mexicanum, has been investigated using the histochemical fluorescence technique of Falck and Hillarp combined with microspectrofluorimetry. The earliest catecholamine-containing neurons to be detected are located in the ventral ependymal zone of the spinal cord at the time of hatching (Stage 41). Between stages 43 and 46, catecholamine fluorescence can be detected in neurons in the following regions: nucleus preopticus, the hypothalamic-infundibular region, and the brain stem reticular formation. 5-HT-containing neurons are only observed in the midbrain raphe region and are first detected at stage 44. In contrast to these early monoamine fluorescing groups, catecholamine-containing neurons are not routinely detectable in the nucleus interpeduncularis until six months of age. All monoamine-containing neuronal groups detected in developing axolotls are also present in both sexes of the adult. However, the fluorescence intensity is less in monoamine-containing neurons observed in adults than in early developing subjects. All catecholamine-containing neuronal groups, with the exception of those located in the midbrain region (nucleus interpeduncularis, reticular zone) have fluorescent processes that contact the cerebrospinal fluid (CSF). The presence of CSF-contacting processes in the hypothalamic and spinal cord regions suggest that the CSF may act as a medium through which bioactive substances are transported from one brain region to another. Intense catecholamine fluorescence is observed in cells of the notochord prior to the detection of the monoamine-containing neurons in the CNS. A possible involvement of catecholamines in the inductive effects of the notochord during development is discussed.

Age Factors↗

Collagen of Dupuytren's disease.

1. In contrast to collagen from the aponeurosis of normal adult subjects, the nodules, contractures and apparently unaffected aponeurosis from patients with Dupuytren's disease contained substantial amounts of type III collagen. 2. The presence of type III collagen supports the previous proposal that the initial response to injury is the synthesis of an increased proportion of this form of collagen. The increased amounts in the apparently unaffected aponeurosis indicate the disease is not strictly focal but more systemic than is usually considered.

Collagen↗

Presence of type III collagen in guinea-pig dermal scar.

Guinea-pig dermal scar was shown to contain type III collagen, and, from densitometric analysis of gel electrophoretograms, it was shown to have a higher concentration than the surrounding dermis. This finding is consistent with the 'embryonic' nature of newly formed dermal wound tissue, reflected in increased hydroxylation of collagen lysine and the presence of dihydroxylysinonorleucine (after reduction) as the major cross-link.

Animals↗

Chemistry of the collagen cross-links. Nature of the cross-links in the polymorphic forms of dermal collagen during development.

Both the type I and type III collagens present in embryonic dermis are stabilized by the intermolecular cross-link, hydroxylysino-5-oxonorleucine, derived from hydroxylysine-aldehyde, although the type I collagen possesses a significant proportion of dehydrohydroxylysinonorleucine. However, concurrent with the change in the proportion of the two types of collagen during postnatal development there is a change-over with both type I and III collagens to the labile cross-link, dehydrohydroxylysinonorleucine, derived from lysine aldehyde. The results indicate that the change in the nature of the cross-link with development is determined primarily by the change in the extent of hydroxylation of the lysine residues in the terminal non-helical regions rather than being due to the change in the type of collagen.

Amino Acids↗

Characterization of the collagen of human hypertrophic and normal scars.

The collagen produced in response to an injury of human skin is initially stabilized by a cross-link derived from hydroxyallysine, and characteristic of embryonic skin. In normal healing there is a change over with time to the cross-link derived from allysine, which is typical of young skin collagen. In contrast, hypertrophic scars fail to follow the time-related changes of normal skin, but retain the characteristics of embryonic collagen, indicating a continued rapid turnover of the collagen. This is further supported by the high proportion of the embryonic Type III collagen present in hypertrophic scars.

Adolescent↗

Patterns of Schwann cell myelination of axons within the spinal cord.

Patterns of Schwann cell myelination of long-projecting axons in the spinal cord were studied. The goal was to determine if such axons arising from neurons whose somata and processes are normally confined to the central nervous system can interact effectively with Schwann cells, the myelinating cells of the peripheral nervous system. In one paradigm Schwann cells develop in the dorsal funiculi of the lumbar spinal cord subsequent to radiation-induced alterations in development of the glial populations. Light and electron microscopic evaluations were made in the region of the corticospinal tracts (CSTs), which in the rat occupy the base of the dorsal funiculi. At 90 days following irradiation, larger axons of these tracts (> 1.5 microns in diameter) were myelinated by Schwann cells, and smaller axons were ensheathed by them. In the second paradigm cultured Schwann cells were injected into the medial portions of the ventral funiculi at 13 days post-irradiation when the glial population was markedly reduced. Earlier investigations from this laboratory demonstrated that Schwann cells do not develop in the irradiated ventral funiculi, as they do dorsally. When placed in proximity to long-projecting axons in the medial portion of the ventral funiculi, the Schwann cells either formed compact myelin sheaths or ensheathed axons, depending upon their diameter. Fasciculation and presence of collagen were characteristic of this paradigm but were absent from the Schwann cell-occupied regions of the CSTs. This probably relates to the presence of fibroblasts in the injected cultures.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Polymer scaffolds fabricated with pore-size gradients as a model for studying the zonal organization within tissue-engineered cartilage constructs.

The zonal organization of cells and extracellular matrix (ECM) constituents within articular cartilage is important for its biomechanical function in diarthroidal joints. Tissue-engineering strategies adopting porous three-dimensional (3D) scaffolds offer significant promise for the repair of articular cartilage defects, yet few approaches have accounted for the zonal structural organization as in native articular cartilage. In this study, the ability of anisotropic pore architectures to influence the zonal organization of chondrocytes and ECM components was investigated. Using a novel 3D fiber deposition (3DF) technique, we designed and produced 100% interconnecting scaffolds containing either homogeneously spaced pores (fiber spacing, 1 mm; pore size, about 680 microm in diameter) or pore-size gradients (fiber spacing, 0.5-2.0 mm; pore size range, about 200-1650 microm in diameter), but with similar overall porosity (about 80%) and volume fraction available for cell attachment and ECM formation. In vitro cell seeding showed that pore-size gradients promoted anisotropic cell distribution like that in the superficial, middle, and lower zones of immature bovine articular cartilage, irrespective of dynamic or static seeding methods. There was a direct correlation between zonal scaffold volume fraction and both DNA and glycosaminoglycan (GAG) content. Prolonged tissue culture in vitro showed similar inhomogeneous distributions of zonal GAG and collagen type II accumulation but not of GAG:DNA content, and levels were an order of magnitude less than in native cartilage. In this model system, we illustrated how scaffold design and novel processing techniques can be used to develop anisotropic pore architectures for instructing zonal cell and tissue distribution in tissue-engineered cartilage constructs.

Animals↗

Bi-zonal cartilaginous tissues engineered in a rotary cell culture system.

In this study, we aimed at validating a rotary cell culture system (RCCS) bioreactor with medium recirculation and external oxygenation, for cartilage tissue engineering. Primary bovine and human culture-expanded chondrocytes were seeded into non-woven meshes of esterified hyaluronan (HYAFF-11), and the resulting constructs were cultured statically or in the RCCS, in the presence of insulin and TGFbeta3, for up to 4 weeks. Culture in the RCCS did not induce significant differences in the contents of glycosaminoglycans (GAG) and collagen deposited, but markedly affected their distribution. In contrast to statically grown tissues, engineered cartilage cultured in the RCCS had a bi-zonal structure, consisting of an outgrowing fibrous capsule deficient in GAG and rich in collagen, and an inner region more positively stained for GAG. Structurally, trends were similar using primary bovine or expanded human chondrocytes, although the human cells deposited inferior amounts of matrix. The use of the presented RCCS, in conjunction with the described medium composition, has the potential to generate bi-zonal tissues with features qualitatively resembling the native meniscus.

Adult↗