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Biomedical subjects

J M Hurle

Publications and source records attributed to J M Hurle.

At least 19 recordsLinked to original sources

Interdigital chondrogenesis and extra digit formation in the duck leg bud subjected to local ectoderm removal.

In the chick embryo the interdigital tissue in the stages previous to cell death exhibits in vitro a high chondrogenic potential, and forms extra digits when subjected in vivo to local ectodermal removal. In the present work we have analyzed the chondrogenic potential both in vivo and in vitro of the interdigital mesenchyme of the duck leg bud. As distinct from the chick, the interdigital mesenchyme of the duck leg bud exhibits a low degree of degeneration, resulting in the formation of webbed digits. Our results show that duck interdigital mesenchyme exhibits also a high chondrogenic potential in vitro until the stages in which cell death starts. Once cell death is finished chondrogenesis becomes negative and the interdigital mesenchyme forms a fibroblastic tissue. In vivo the interdigital mesenchyme of the duck leg bud subjected to ectoderm removal forms ectopic foci of chondrogenesis with a range of incidence similar to that in the chick. Unlike those of the chick the ectopic cartilages of the duck are rounded and smaller, and appear to be located at the distal margin of the interdigital mesenchyme. Formation of extra digits in the duck occurs with a lower incidence than in the chick. It is concluded that ectopic chondrogenesis and formation of extra digits is related to the intensity of interdigital cell death. The non-degenerating interdigital mesenchymal cells destined to form the interdigital webs of the duck appear to contribute very little to the formation of interdigital cartilages.

Animals

Structural arrangement of the extracellular matrix network during myocardial development in the chick embryo heart.

We analyzed the extracellular matrix and the connective tissue of the developing chick myocardium ("myocardial interstitium"). The importance of this myocardial element for heart function has been well documented both for the normal and pathologic adult hearts. However, little information is available on the organization of the embryonic myocardial interstitium and its modifications during development and increasing intracardiac pressure. In the present study we used light and scanning electron microscopic techniques, and lectin probes to study the interstitium of the ventricular myocardium of chick embryos from stage 29 (day 6 of development) until hatching. Our observations trace the progressive appearance and organization of the elements of the extracellular matrix, comprising the epimysium, perimysium and endomysium, which form a well-defined architectural network. Finally, we discuss the role of these elements of the extracellular matrix and their possible relation with the biomechanical properties of developing heart.

Animals

Histochemical and ultrastructural changes in the extracellular matrix of the developing chick semilunar heart valves.

The present study analyzes the composition and organization of the extracellular matrix (ECM) and its changes in the course of development of the chick embryo semilunar heart valves. In the present work we have employed chick embryos from stage 29 until hatching, using silver and picrosirius red staining, lectin probes and light and transmission electron microscopy. Our results show that during semilunar valve development a series of elements arise and are organized in the ECM which seem to be more closely related to the maintenance of the structural and biomechanical properties of the valvular leaflets than with morphogenetic processes per se.

Animals

Immunofluorescent localization of tenascin during the morphogenesis of the outflow tract of the chick embryo heart.

The cono-truncus constitutes a complex segment of the developing heart that gives rise to the outflow tract of the ventricles and root of the pulmonary and aortic arteries. Numerous studies have revealed that the extracellular matrix plays a relevant role in most morphogenetic processes modulating cell behaviour. By means of immunofluorescence, we studied the distribution and possible involvement of tenascin during morphogenesis of the conus and truncus in chick embryo hearts between days 4.5-10 of incubation. Tenascin is an extracellular matrix glycoprotein with a significant role in morphogenesis and cell and tissue differentiation. Our results reveal a specific distribution of tenascin in the areas of the cono-truncus undergoing significant structural changes during morphogenesis of this cardiac segment, appearing mainly in the mesenchymal layer subjacent to the myocardial layer, the cono-truncal ridges and the aorto-pulmonary septum. The distribution of tenascin was compared and contrasted with that of collagen type I, which constitutes a further component of the extracellular matrix common to most developing connective tissues.

Animals

Histogenesis of the semilunar valves: an immunohistochemical analysis of tenascin and type-I collagen distribution in developing chick heart valves.

The development of the semilunar valves takes place in association with septation of the outflow tract in the embryonic heart. Although numerous studies have focused on this process, the causal mechanisms of valvular development remain obscure. This paper reports an immunohistochemical analysis of tenascin and type-I collagen distribution in developing chick heart valves. Tenascin is a glycoprotein that is present on some embryonic extracellular matrices. It plays several significant roles in tissue differentiation, cell growth, and tissue interactions; it is also important for the formation of specific zones of connective tissue that fulfill mechanical functions. Our results show that tenascin is present during valvular morphogenesis and histogenesis, and that its distribution is associated with zones specialized in bearing mechanical loads.

Animals

Experimental analysis of the role of ECM in the patterning of the distal tendons of the developing limb bud.

We have shown previously that from stage 27 the distal growing region of the limb exhibits a tenascin-rich sheet of extracellular matrix termed the "mesenchyme lamina" (ML), which runs from the ectodermal basement membrane in a proximal direction until it contacts the distal tip of the muscle blocks. This study reports experimental evidence that the mesenchyme lamina is a pretendinous structure that controls the spatial organization of the flexor and extensor tendons of the distal part of the chick leg. Two sets of experiments were designed to alter the ML position and examine subsequent tendon pattern formation. In a first series of experiments limbs with digits lacking phalangeal elements were induced by AER removal at stages 26 and 27. This procedure induced an abnormal arrangement of the ML around the distal tip of each terminal phalange of the truncated digit, which was followed by the development of a precisely similar pattern of abnormal extensor and flexor tendons. In the second set of experiments, an extradigit was induced to form in the interdigital mesenchyme through surgical removal of the marginal ectoderm of the third interdigit of stage 29 leg buds. By day 4 post-operation, a chondrogenic extradigit had formed, together with a ML that ran from the cartilage to the normal ventral flexor and dorsal extensor tendons. By day 6 post-operation, the experimentally induced ML had transformed into a tendinous structure connecting with the adjacent normal tendon. Both experiments show that the position of the ML defines the position of subsequent tendon development, thus supporting its role as a pretendinous structure which might be responsible for the alignment of the pretendinous condensing cells.

Animals

Myocardial fiber architecture in the human heart. Anatomical demonstration of modifications in the normal pattern of ventricular fiber architecture in a malformed adult specimen.

Ventricular myocardial fiber architecture has been considered an important factor in heart dynamics. Most anatomical studies however have focussed on the analysis of normal hearts. The present study compares ventricular myocardial fiber architecture patterns in dissections of 5 normal hearts and a malformed human heart with membranous ventricular septal defect, overriding right aorta, pulmonic stenosis, with absent pulmonary valve and hypertrophied right ventricle. Qualitative and quantitative changes in ventricular myocardial fiber architecture were noted in the malformed heart.

Adolescent

Experimental analysis of the in vivo chondrogenic potential of the interdigital mesenchyme of the chick leg bud subjected to local ectodermal removal.

Current in vitro investigations suggest that ectoderm plays a major role in limb morphogenesis by producing a diffusible factor which inhibits the chondrogenesis of the underlying mesenchyme. In the present work we report evidence supporting such an ectodermal role in vivo. Surgical removal of the marginal ectoderm from the third interdigit of chick leg buds at stages 27 to 30 induces the formation of PNA-positive prechondrogenic mesenchymal condensations 15 hr after the operation. The incidence of prechondrogenic condensations achieved 47, 95.2, and 92.8 of the experimental embryos of stages 27, 28, and 29, respectively. This high rate of prechondrogenic aggregate formation contrasted with a lower incidence of ectopic cartilage formation detectable by Alcian blue staining 40 hr after the operation. The sequential analysis of the experimental interdigits by means of peanut lectin labeling suggests that a number of prechondrogenic condensations undergo disaggregation 20 and 30 hr after the operation failing to form fully differentiated cartilages. When ectoderm removal was accompanied by the elimination of a variable amount of interdigital mesenchyme the incidence of prechondrogenic aggregates showed little differences but the formation of fully differentiated cartilages was reduced at a rate proportional to the amount of interdigital mesenchyme removed. From this study it can be concluded that the ectoderm in vivo appears to inhibit the process of aggregation of the mesenchymal cells to form prechondrogenic condensations. Furthermore our results suggest that as observed in vitro (C. P. Cotrill, C. Archer, and L. Wolpert, 1987, Dev. Biol. 122, 503-515) the transformation of prechondrogenic aggregates into fully differentiated cartilage requires the involvement of a critical amount of mesenchymal cells.

Alcian Blue

The extracellular matrix architecture relating to myotendinous pattern formation in the distal part of the developing chick limb: an ultrastructural, histochemical and immunocytochemical analysis.

In the later developmental stages (Hamburger and Hamilton, 25-34) the distal part of the chick leg possesses a distinctive extracellular matrix (ECM) architecture which relates to myotendinous patterning. There are two components: firstly, a system of dorsoventrally oriented fibrils which link the two ectodermal surfaces through the undifferentiated distal mesenchyme and secondly, a 'mesenchyme lamina' originates at the basement membrane distally, but proximally runs through the mesoderm, subjacent and parallel to the basement membrane. The 'mesenchyme lamina' appears to be a precursor of developing tendons and is spatially related to the distal tips of the myogenic blocks. As developing tendons form on the inner surface of the lamina at its proximal end, it becomes less distinct and disappears. Further dorsoventral fibrils run from the 'mesenchyme lamina' into the developing condensations and chondrogenic elements of the phalanges. The architecture of the ECM was revealed by silver and lectin staining (peanut and Ricinus communis agglutinins, PNA and RCA I), by immunocytochemistry (for fibronectin, tenascin, collagen type I) and by ultrastructural analysis. Both components stain with silver, PNA following neuraminidase digestion, RCA I, tenascin and collagen type I. However, the dorsoventral fibrils are positive for fibronectin and negative for PNA, while conversely the mesenchyme lamina is positive for PNA but much less so for fibronectin. Tenascin has been shown to be a specialized mesenchyme component of tendons and myotendinous junctions (Chiquet and Fambrough, 1984). Such a basement membrane forming a 'mesenchyme lamina' appears to be unique in epithelial-mesenchymal developing systems and points to an ectodermal role in tendon pattern formation within the mesenchyme. We discuss the possible role of mechanical force in converting the dorsoventral tenascin-positive fibrils into the localized pattern of tendon insertions into the proximal parts of the phalanges. Distally the dorsoventral fibrils may shape the digital plate by pulling together the two ectodermal surfaces. A similar ECM architecture is found in corresponding stages in the developing wing.

Animals

Spatial and temporal changes in the pattern of glycosylation of the developing chick limb tissue components as revealed by fluorescent conjugated lectin probes.

The changing pattern of expression of glycoconjugates during the differentiation of the chick leg bud between stages 17 to 34 (days 3 to 8 of incubation) was studied using fluorochrome-labelled plant lectins. Limb buds were fixed in cold acetic-alcohol and wax-embedded. Agglutinins of peanut (PNA), soybean (SBA) and succinylated wheat germ (WGAs) revealed a specific binding pattern in the apical ectodermal ridge (AER) between Hamburger and Hamilton stages 19-32. These stages coincide with the period of elevation of the AER. This specific binding pattern was absent from the adjacent dorsal and ventral ectoderm. Prechondrogenic cells were positive for WGA and for PNA, and the PNA-binding capacity was intensified after neuraminidase treatment. Premyogenic cells at stage 23 can be identified as negative to PNA after neuraminidase, while the blood vessels became positive. PNA, SBA, WGA, WGAs and, in addition, Ricinus communis (RCA-I) lectins stained the basal membrane. Strands of extracellular matrix which connect with the basal membrane and cross the limb transversely between dorsal and ventral ectoderm were stained by RCA-I, SBA and PNA after neuraminidase.

Animals

Effects of taxol on endothelial cells of the developing semilunar heart valves in the chick embryo.

Recent ultrastructural studies have revealed that there are differences in endothelial cell shape and cytoskeletal architecture between the arterial and ventricular faces of developing semilunar valves. In the present work we have analyzed valvular endothelial cell response to taxol in the chick embryo. Following taxol administration, abnormal mitotic cells appear in the endothelium of both faces of the cusps. Interphase cells show few structural alterations. No differential response of the valvular endothelium to the drug was observed in the arterial or ventricular faces of the cusps.

Alkaloids

Cell death in developing systems.

The so called 'normal', 'physiological' or 'programmed' cell death constitutes now a major field of developmental and cell biology. In the present article we review recent information on the biological significance of cell death in development. Special attention is paid to the range of techniques available for studies on cell death, the role of cell death during normal and abnormal development and to the mechanisms controlling cell death, including epigenetic and genetic factors.

Animals

Ventricular myocardial architecture in marine fishes.

The fiber architecture of the ventricular myocardium has been studied in elasmobranch (Isurus oxyrhinchus, Galeorhinus galeus, Prionace glauca) and teleost (Xiphias gladius, Thunnus thynnus, Thunnus alalunga) fish species with hearts displaying mixed types of ventricular musculature (compact and trabecular). In all cases, the compact myocardium is organized in layers of fiber bundles with an orderly arrangement within the ventricular walls. The number of these layers appears to be dependent on the relative thickness of the compact myocardium. Differences in the pattern of myocardial fiber arrangement were observed among the different fish species. In elasmobranchs the compact myocardium at the level of the atrioventricular orifice is continuous with the trabeculated myocardium. Furthermore, in elasmobranchs the trabeculated myocardium displays a precise arrangement in arcuate trabeculae running from the auriculoventricular to the conoventricular orifices. In teleosts, the compact myocardium is independent of the trabeculated myocardium and a large number of fibers insert into the bulboventricular fibrous ring. The trabeculated myocardium in these species displays an anarchic arrangement except at the level of the bulboventricular orifice, where the fibers tend to be aligned longitudinally, also being inserted into the fibrous ring. Minor differences, consisting mainly of the presence of extra bundles of fibers, were also observed among different individuals of the same species. The possible relationship between myocardial fiber architecture and ventricular shape is discussed.

Animals

Effects of colchicine administration on the endothelial cells of the developing semilunar heart valves of the chick embryo.

Recent ultrastructural studies have revealed that differences exist in endothelial cell shape and cytoskeletal architecture between the arterial and ventricular faces of developing semilunar valves. In the present work we analyzed the morphologic response of the valvular endothelial cells of chick embryos to colchicine by light microscopy, scanning electron microscopy and transmission electron microscopy. The results show that colchicine administration during the stages of valve morphogenesis causes a very conspicuous disruption of the endothelial layer of the arterial face of the valves. The cells appear rounded and show massive surface blebbing. These alterations were not present in the endothelial cells on the ventricular face of the valves at the same stages. On the basis of these results we suggest that a difference in the degree of cell differentiation exists between the endothelial cells of the arterial and ventricular faces of the cusps and that this difference may have morphogenetic significance.

Animals

Formation of extra-digits induced by surgical removal of the apical ectodermal ridge of the chick embryo leg bud in the stages previous to the onset of interdigital cell death.

In the present work we have studied the mechanism of formation and the possible morphogenetic significance of the process of ectopic chondrogenesis induced by surgical removal of AER of the interdigital spaces of the chick leg bud at stage 28-30 (Hurle and Gañan 1986). Our results show that ridge removal causes condensation and rounding of the underlaying mesenchymal cells followed by chondrogensis. The long-term study of the fate of these ectopic cartilages shows that in a high percentage of the cases the cartilages undergo morphogenesis taking by day 10 of incubation the appearance of the two distal phalanges of an extra-digit. These extra-digits lack tendons and are joined by thin interdigital membranes to the neighboring digits.

Animals

Morphologic characteristics and structure of surface excrescences (Lambl's excrescences) in the normal aortic valve.

The incidence, morphologic characteristics and structure of surface valve excrescences (Lambl's excrescences) were studied by transmission and scanning electron microscopy of the aortic valve of 56 human subjects, age range birth to 91 years, without cardiac disease. Valve excrescences consisting of a core of connective tissue covered by the endocardium were observed in 90% of the subjects; the incidence was significantly lower in patients in the first decade of life. Two types of excrescences, lamellar and filiform, were found. Lamellar excrescences are located along the lower boundary of the lunulas and occurred more often in those younger than 30 years. Filiform excrescences appear most often in the nodulus Arantius and in the free-margin of the cusps. The excrescences of the nodulus are the most numerous. Free-margin excrescences are the least numerous and occur more frequently in persons older than 40 years. The connective tissue core of the filiform excrescences contains abundant collagen fibrils and elastic material arranged in apposed layers with different collagen fibril orientation. A circular zone devoid of identifiable connective tissue is present at the center of the filiform excrescences.

Adult

Cell shape and cytoskeletal organization of the endothelial cells of the semilunar heart valves in the developing chick.

The composition and arrangement of the cytoskeletal elements of the endothelium of the semilunar valves have been studied in the embryonic chick heart during the stages 30 to 38. Microtubules, vimentin intermediate filaments and actin microfilaments were constant components of the valvular endothelial cells in the studied stages. Scanning electron microscopy after Triton-X-100 extraction revealed significant differences in the tridimensional arrangement of the cytoskeleton in the course of valve development. In the ventricular face of the cusps the cytoskeletal elements displayed a progressive longitudinal alignment, while in the arterial face of the cusps the cytoskeleton maintained the appearance of a network. Transmission electron microscopy revealed that these differences were especially prominent for vimentin intermediate filaments, although a similar tendency was also observed for microtubules. Microfilaments were scarce in the endothelial cells of both faces of the cusps, and the stress fibers typical of the endothelial cells of the adult valves were not observed in the embryonic material. The significance of these results in valve morphogenesis and histogenesis and a possible linkage with differences in the local characteristics of the blood flow are discussed.

Animals