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L Alibardi

Publications and source records attributed to L Alibardi.

At least 37 records · Page 2Linked to original sources

Immunocytochemical and electrophoretic distribution of cytokeratins in the regenerating epidermis of the lizard Podarcis muralis.

Using immunocytochemistry at light- and electron-microscope levels, we studied the distribution of three monoclonal antibodies (AE1, AE2, AE3) specific for mammalian alpha-keratins in regenerating lizard epidermis. We also characterized the keratins expressed during this process by immunoblotting after electrophoretic separation. The AE1 antibody is localized in the basal and suprabasal layers of prescaling and scaling epidermis. During the first stages of scale neogenesis, the AE1 antibody also marks the differentiating oberhautchen and beta-layer, but it disappears from these layers as they mature. This antibody does not stain the prekeratinized and keratinized outermost layers in the hinge region. The AE2 antibody labels the superficial wound epidermis, prekeratinizing and keratinized beta- and alpha-layers, but not basal and suprabasal cells. The AE3 antibody labels all living and keratinized epidermal layers, although AE3 immunoreactivity decreases and disappears as the beta-layer matures. The ultrastructural study shows that the AE2 and AE3, but not the AE1, antibodies specifically label small electron-dense areas within the beta-layer, suggesting retention of alpha-keratins. In the stages of tail regeneration examined, immunoblotting with the three antibodies used for the immunolocalization gives a pattern similar to that of the normal epidermis, except distally, where the process of scale differentiation begins. In this region, in addition to the keratin forms discovered in the normal and in proximal regenerating epidermis, an intense low molecular weight band at 40-41 kDa, positive to all three antibodies, is clearly detectable. Furthermore, in the distal region AE1 and AE3 antibodies, but not the AE2, recognize a weak band at 77-78 kDa not present in the normal and proximal epidermis. The localization and the possible role of the different keratins in the regenerating epidermis is discussed.

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Organization and characterization of the keratin cytoskeleton in the previtellogenic ovarian follicle of the lizard Podarcis sicula raf.

The cytokeratin (CK) cytoskeleton, previously described by immunofluorescence in the ovarian follicle of Podarcis sicula, at the electron microscope results constituted by bundles of 10 nm thick intermediate filaments containing keratin. These bundles are better evident in the cytoplasm of the pyriform cell apex pointed toward the oocyte surface and mostly in the intercellular bridges connecting fully differentiated pyriform cells to the oocyte. During the differentiation of pyriform cells, the intermediate filament bundles first appear inside the intercellular bridge, when the small follicle cells progressively enlarge after their fusion with the oocyte and assume a morphology of "intermediate" cells. The present study also reports a comparative analysis by immunolabeling, SDS-PAGE, and immunoblotting with anticytokeratins CK8, CK18, and CK19 antibodies of both the ovarian follicle and the intestine of Podarcis sicula. These antibodies, specific to the keratins of monolayered intestinal cells, react also against those expressed in the oocytes of Xenopus laevis. This study shows the presence in the ovarian follicle of this reptile only of keratin forms of homologues to the CK8 and CK18 of mammals and the lack of CK19. The same analysis carried out utilizing AE1 and AE3 antibodies, which recognize most of the acidic and basic keratins in mammals, has shown additional forms of keratins specifically expressed in the ovarian follicle (50 kDa) and in both the examined tissues (49 and 60 kDa). The reported results indicate that in the ovarian follicle of this reptile, keratins have peculiar characteristics that can be explained by the unique structural function of the cytoskeleton in this system.

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Cytology, synaptology and immunocytochemistry of commissural neurons and their putative axonal terminals in the dorsal cochlear nucleus of the rat.

The first binaural integration within the auditory system responsible for sound localization depends upon commissural neurons that connect the two symmetrical cochlear nuclei. These cells in the deep polymorphic layer of the rat dorsal cochlear nucleus were identified with the electron microscope after injection of the retrograde tracer, Wheat Germ Agglutinin conjugated to Horseradish Peroxydase, into the contralateral cochlear nucleus. Commissural neurons are multipolar or bipolar with an oval to fusiform shape. Few commissural neurons, most inhibitory but also excitatory, connect most of the divisions of the rat cochlear nuclei. The most common type is a glycinergic, sometimes GABAergic, moderately large cell. Its ergastoplasm is organized into peripheral stacks of cisternae, and few axo-somatic synaptic boutons are present. Another type of commissural neuron is a medium-sized, spindle-shaped cell, glycine and GABA-negative, with sparse ergastoplasm and synaptic coverage. A giant, rare type of commissural neuron is glycine-positive and GABA-negative, with short peripheral stacks of ergastoplasmic cisternae. It is covered with synaptic boutons, many of which contain round synaptic vesicles. Another rare type of commissural neuron is a moderately large cell, oval to fusiform in shape, immunonegative for both glycine and GABA, and contacted by many axo-somatic boutons. It contains large dense mitochondria and numerous dense core vesicles of peptidergic type. Some labelled boutons, mostly inhibitory and probably derived from commissural neurons, contact pyramidal, cartwheel, giant and tuberculo-ventral neurons. The prevalent inhibition of electrical activity in a cochlear nucleus observed after stimulation of the contralateral cochlear nucleus may be due to commissural inhibitory terminals which contact excitatory neurons such as pyramidal and giant cells. Other inhibitory commissural terminals which contact inhibitory neurons such as cartwheel and tuberculo-ventral neurons, may explain the stimulation of electrical activity in the DCN after contralateral stimulation.

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Ultrastructural localization of alpha-keratins in the regenerating epidermis of the lizard Podarcis muralis during formation of the shedding layer.

In the epidermis of lizards, alpha- and beta-keratins are sequentially produced during a shedding cycle. Using pre- and post-embedding immunocytochemistry this study shows the ultrastructural distribution of 3 alpha-keratin antibodies (AE1, AE2, AE3) in the renewing epidermis and in the shedding complex of the regenerating tail of the lizard Podarcis muralis. The AE1 antibody that recognizes acidic low MW keratins is confined to tonofilament bundles in basal and suprabasal cells but is not present in keratinizing beta- and alpha-cells. The AE2 antibody that recognises higher MW keratins weakly stains pre-keratinized cells and intensely keratinized alpha-layers. A weak labeling is present in small electrondense areas within the beta-layer. The AE3 antibody, that recognizes low and high MW basic keratins, immunolabels tonofilament bundles in all epidermal layers but intensely the alpha-keratinizing and keratinized layers (mesos, alpha-, lacunar and clear). Keratohyalin-like granules, present in the clear cells of the shedding layer, are negative to these antibodies so that the cornified clear layer contains keratins mixed with non-keratin material. The AE3 antibody shows that the mature beta-layer and the spinulated folds of the oberhautchen are labeled only in small dense areas among the prevalent electron-pale beta-keratin material. Therefore, some alpha-keratin is still present in the beta-layer, and supports the idea that alpha-keratins (basic) function as scaffold for beta-keratin deposition.

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Identification of tuberculo-ventral neurons in the polymorphic layer of the rat dorsal cochlear nucleus.

The tuberculo-ventral tract represents a short nervous circuit within the auditory cochlear nuclei. Tuberculo-ventral neurons of the dorsal cochlear nucleus send isofrequency inhibitory inputs to bushy cells of the ventral cochlear nucleus. Injection of wheat germ agglutinin conjugated to horseradish peroxidase into the rat ventral cochlear nucleus, labelled tuberculo-ventral neurons retrogradely in the deep polymorphic layer of the ipsilateral dorsal cochlear nucleus. Five to 20% of the perimeter of these cells was covered by synaptic boutons, most of which contained flat and pleomorphic vesicles. These boutons contained glycine and sometimes GABA. Occasional small axo-somatic boutons contained round vesicles and were immunonegative for both glycine and GABA. This study shows that the synaptic profile of tuberculo-ventral neurons is different from that of other medium-size glycinergic neurons within the polymorphic layer or more superficial regions of the dorsal cochlear nucleus like cartwheel neurons. In fact the latter mostly receive boutons that contain pleomorphic vesicles.

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Light and electron microscopical localization of filagrin-like immunoreactivity in normal and regenerating epidermis of the lizard Podarcis muralis.

In the stratum granulosum of mammalian epidermis, keratohyalin contains the histidine-rich protein filagrin which determines the aggregation of keratin bundles in terminally differentiating keratinocytes. Among reptiles, the lizard epidermis possesses keratohyalin-like granules during a period preceding molting. Tritiated histidine is taken up at 3-22 h post-injection mainly in keratohyalin-like granules of the clear layer, and less intensely in oberhautchen cells, in pre-keratinizing cells of wound epidermis and in the alpha-layer. No labelling was observed in beta-cells and mesos cells. Single and double immunogold localization of filagrin and keratin, using mammalian antibodies, show that some filagrin-like immunoreactivity is localized in 0.15-0.40 microm diameter roundish keratohyalin-like granules in cells of wound epidermis, and lacunar and clear layers (soft alpha-layers). Pro-filagrin was not immuno-localized in these layers with the mammalian antibodies. The small granules merge with keratin bundles into mature keratinocytes where immunopositive keratin filaments predominate and the filagrin-like immunolabelling rapidly disappears. Little or no labelling is observed in the large keratohyaline-like granules of the clear layer. This may be due to lack of filagrin-like immunoreactivity but may also be due to epitope-masking or chemical degradation of filagrin-like molecules. No immunoreactivity is present in the beta-layer and mesos-layer but the immunolabelling reappears in the maturing alpha-layer and lacunar layer. This study suggests that histidin-rich protein with some filagrin-like immunoreactivity is initially present in those alpha-layers of lizard epidermis where keratohyalin-like granules are present, such as lacunar and clear cells, and that a filagrin-like molecule is degraded or altered in mature keratinocytes.

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Cytology of large neurons in the guinea pig dorsal cochlear nucleus contacting the inferior colliculus.

Large neurons in the dorsal cochlear nucleus of the guinea pig which project to the inferior colliculus were identified after injections of the neural tracer WGA-HRP. Retrograde labelled cells (pyramidal and giant neurons) in the dorsal cochlear nucleus were glycine and GABA immunonegative and showed a similar ultrastructure. Between 30 and 60% of their perimeter was covered by axo-somatic boutons, most of which (>50%) contained pleomorphic synaptic vesicles. Other boutons (about 40% of total) contained flat vesicles and few (5-6%) contained round vesicles, a characteristic of the excitatory cells innervating the inferior colliculus. Immunogold-cytochemistry, coupled to silver intensification, showed that more than 50% of axo-somatic pleomorphic boutons and over 90% of boutons containing flat and pleomorphic vesicles store glycine. Rare WGA-HRP labelled axo-somatic boutons containing flat-pleomorphic vesicles were seen on pyramidal and giant neurons. This suggests that a few inhibitory collicular terminals contact the excitatory large neurons in the dorsal cochlear nucleus.

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Putative commissural and collicular axo-somatic terminals on neurons of the rat ventral cochlear nucleus.

The type of synaptic terminals from the cochlear nucleus and inferior colliculus that terminate in the contralateral ventral cochlear nucleus are not known. These terminals were studied with the electron microscope and immunogold after injection of wheat germ agglutinin conjugated to horseradish peroxidase into the inferior colliculus or into the cochlear nucleus. The tracer anterogradely labelled boutons onto the main neurons of the contralateral ventral cochlear nucleus. Most of these cells (95%) were glycine immuno-negative and represent excitatory neurons. After injection of the tracer into the contralateral inferior colliculus few anterogradely labelled boutons were seen on spherical and multipolar cells of type II in the anteroventral cochlear nucleus. Rare labelled boutons were present on multipolar cells of type I and II, globular neurons and octopus cells in the posteroventral cochlear nucleus. After injection into the contralateral dorsal and ventral cochlear nucleus labelled boutons were seen more frequently than after injection into the inferior colliculus. These terminals contacted most of large neurons, especially multipolar cells of type II and less frequently of type I. Also globular and spherical cells were contacted by commissural terminals. Octopus cells received less frequently putative commissural terminals. Most boutons contained pleomorphic vesicles and stored GABA. A lower number of boutons with pleomorphic and flat vesicles contained glycine and sometimes GABA, both inhibitory neurotransmitters. Few boutons containing round vesicles were immuno-negative for both glycine and GABA, and were considered putative commissural excitatory terminals. The latter often contacted glycinergic neurons of type II so that also these terminals might elicit an inhibition with at least a disynaptic mechanism after contralateral stimulation.

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Epidermal differentiation in the developing scales of embryos of the Australian scincid lizard Lampropholis guichenoti.

Formation of the first epidermal layers in the embryonic scales of the lizard Lampropholis guichenoti was studied by optical and electron microscopy. Morphogenesis of embryonic scales is similar to the general process in lizards, with well-developed overlapping scales being differentiated before hatching. The narrow outer peridermis is torn and partially lost during scale morphogenesis. A second layer, probably homologous to the inner peridermis of other lizard species, but specialized to produce lipid-like material, develops beneath the outer peridermis. Two or three lipogenic layers of this type develop in the forming outer surface of scales near to the hinge region. These layers form a structure here termed "sebaceous-like secretory cells." These cells secrete lipid-like material into the interscale space so that the whole epidermis is eventually coated with it. This lipid-like material may help to reduce friction and to reduce accumulation of dirt between adjacent extremely overlapping scales. At the end of their differentiation, the modified inner periderm turns into extremely thin cornified cells. The layer beneath the inner peridermis is granulated due to the accumulation of keratohyalin-like granules, and forms a shedding complex with the oberhautchen, which develops beneath. Typically tilted spinulae of the oberhautchen are formed by the aggregation of tonofilaments into characteristically pointed cytoplasmic outgrowths. Initially, there is little accumulation of beta-keratin packets in these cells. During differentiation, the oberhautchen layer merges with cells of the beta-keratin layer produced underneath, so that a typical syncytial beta-keratin layer is eventually formed before hatching. Between one-fourth distal and the scale tip, the dermis under epidermal cells is scarce or absent so that the mature scale tip is made of a solid rod of beta-keratinized cells. At the time of hatching, differentiation of a mesos layer is well advanced, and the epidermal histology of scales corresponds to Stage 5 of an adult shedding cycle. The present study confirms that the embryonic sequence of epidermal stratification observed in other species is basically maintained in L. guichenoti.

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Epidermal differentiation during carapace and plastron formation in the embryonic turtle Emydura macquarii.

As part of a large comparative study on the development of reptilian skin, we provide the first ultrastructural description of differentiation of the epidermis of the carapace and plastron in the Chelonia, using the Australian pleurodiran turtle Emydura macquarii as a model. The epidermis is initially composed of an external flat peridermis and a basal layer of cuboidal cells. During differentiation, the peridermis darkens, flakes off and is partially lost before hatching. Four to 6 layers of flat cells containing lipids and mucus form from the basal layer beneath the external peridermis. Because such cells are found only during embryogenesis, we have referred to these layers as embryonic epidermis. They contain reticulate bodies made of a meshwork of coarse filaments similar to those described in the inner peridermis of lizard and bird embryos. In advanced embryos, cells of the embryonic epidermis condense into a thin dark stratum which is subsequently lost after hatching. The lowermost 2 layers of the embryonic epidermis keratinise, as for a typical lepidosaurian alpha-layer. A splitting zone is progressively formed beneath the alpha-layer to separate the embryonic epidermis from the underlying beta-layer. Patterns of cytodifferentiation of the beta-synthesising cells over the carapace and plastron essentially resemble those of the lepidosaurian epidermis. The beta-keratin matrix initially accumulates among ribosomes as round bodies not clearly surrounded by a membrane. These bodies appear not to be derived from the Golgi apparatus. Melanosomes and other dark granules of uncertain nature are present among early differentiating beta-cells. The round beta-keratin bodies merge with the dense bodies to produce the definitive variegated pattern of the mature beta-keratin layer. The histochemistry suggests that calcium combines with organic molecules within beta-keratinising cells to harden the tissue. In contrast to the beta-keratin cells of lizards and snakes, cells of the mature beta-keratin layer of E. macquarii maintain their cell boundaries in part or completely, a characteristics shared with the beta-keratin layer of Sphenodon and crocodilians.

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Characterization of tuberculo-ventral neurons in the dorsal cochlear nucleus of the guinea pig.

Tuberculo-ventral neurons in the deep polymorphic layer of the dorsal cochlear nucleus of the guinea pig are immunopositive to glycine, their main neurotransmitter, as revealed by post-embedding immunogold. The initial size of gold particles (10-20 nm) was increased (40-70 nm) by silver intensification for a better and rapid individuation of glycine positive cells at low magnification. The most specific ultrastructural characteristic of tuberculo-ventral neurons is their synaptic profile that shows most flat-pleomorphic axo-somatic boutons. This synaptic profile is different from that of other medium size neurons, such as cartwheel neurons in the superficial layer of the dorsal cochlear nucleus. The synaptic profile of tuberculo-ventral neurons together with other characteristics may explain their activity connected to the neural network responsible for echos suppression or/and sound localization.

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Fine structure, synaptology and immunocytochemistry of large neurons in the rat dorsal cochlear nucleus connected to the inferior colliculus.

Neurons in the rat dorsal cochlear nucleus that project to the inferior colliculus (pyramidal and giant) were retrograde labelled with wheat germ agglutinin conjugated to horseradish peroxydase. Both cell types showed a similar ultrastructural feature, particularly the rough endoplasmic reticulum was well developed and sometimes surrounded the nucleus. The synaptological profile was similar in pyramidal and giant cells. Axo-somatic terminals covered 40-70% of the perimeter of pyramidal cells and 35-60% of the perimeter of giant neurons. Giant neurons featured bipolar or multipolar shape and different orientation but they possessed a similar synaptic profile. Most axo-somatic terminals contained flat and pleomorphic synaptic vesicles, some pleomorphic vesicles. Few terminals contained round vesicles. These cells were consistently immuno-negative for both glycine and GABA and variably positive for glutamate. The immunoelectron microcopic study of thin sections showed that glycine immunoreactivity was constantly present in terminals enriched with flat vesicles, which often did not show GABA immunoreactivity. Few anterograde labelled boutons containing flat vesicles were in contact with the proximal dendrites and the cell bodies of pyramidal and giant neurons. The origin of these terminals is discussed. No other cells of the dorsal cochlear nucleus, in particular cartwheel and tuberculo-ventral neurons, were in contact with labelled boutons. The present results suggest that descending inhibitory collicular projections are essentially directed to the large excitatory neurons of the dorsal cochlear nucleus.

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Ultrastructural and immunocytochemical characterization of neurons in the rat ventral cochlear nucleus projecting to the inferior colliculus.

Neurons in the rat ventral cochlear nucleus which project to the inferior colliculus were identified after retrograde labelling of the neural tracer wheat germ agglutinin conjugated to horse radish peroxidase. After tracer injection into the Inferior Colliculus, electron microscopy and immunocytochemical localization of glycine, GABA and glutamate in retrograde labelled neurons were employed. In the acoustic root area and posterior ventral cochlear nucleus, most of the body surface of the neurons projecting to the inferior colliculus was 10-30% covered by axo-somatic boutons and appeared as multipolar cells of type I. These large to medium size cells with sparse stacks of ergastoplasmic cisternae organized in Nissl bodies, were heavily labelled and were the main projecting neurons to the inferior colliculus. Most of these cells were glycine and GABA negative but variably glutamate positive, suggesting that they are excitatory neurons. This result suggests the absence of an inhibitory innervation of the inferior colliculus from these cells. A few retrograde labelled, large to giant neurons showed an irregular surface, sparse short stacks of ergastoplasmic reticulum, numerous microtubules, cell bodies 60-80% covered by synaptic boutons, and they appeared as multipolar cells of type II. These cells were less strongly retrograde labelled than multipolar type I, and were occasionally glycine positive, presumably inhibitory. This suggests that at least a small caliber collateral axon stemming from these neurons can reach the inferior colliculus. Occasional glycine positive octopus cells not labelled with the tracer were also observed. The contribution of glycinergic axons to the innervation of the inferior colliculus appears therefore to by very limited. Occasional labelled cells were represented by apparently globular bushy neurons, but the weak labelling suggests that the tracer was taken up by collateral axons reaching the inferior colliculus and not by the main axon. Globular bushy cells were consistently negative for both glycine and GABA, and variably positive for glutamate. In the anteroventral cochlear nucleus, labelled multipolar type I and II showed similar immunocytological and ultrastructural characteristics to those in the posteroventral cochlear nucleus but their dimension was smaller. Cells identified as spherical bushy neurons were never retrograde labelled.

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Ultrastructural and immunocytochemical characterization of commissural neurons in the ventral cochlear nucleus of the rat.

Medium to large-giant multipolar neurons in the rat ventral cochlear nucleus were retrograde labelled after injection of the tracer Wheat Germ Agglutinin conjugated to Horse Radish Peroxidase into the contralateral cochlear nucleus. Light microscopy immunocytochemistry showed that 42.45% of these retrograde labelled neurons, generally strongly labelled with the tracer, were markedly glycine immunopositive, and that 57.55%, usually weakly retrograde labelled neurons, were immunonegative or weakly positive for glycine. These commissural neurons were generally GABA negative and variably immunopositive for glutamate. About 1/3rd of the commissural neurons had variably developed a rough endoplasmic reticulum whilst axo-somatic boutons covered 20-40% of the cell body. These cells were recognized as multipolar neurons of type I. Most of them were weakly glycine positive or even negative and a few appeared glycinergic. A little less than the remaining 2/3rds of the whole commissural population in the postero-ventral cochlear nucleus presented a surface which was 65-85% covered with synaptic boutons, among which some also appeared labelled. These cells were recognized as multipolar neurons of type II. Many microtubules and neurofilaments were present, free ribosomes being more numerous around Nissl bodies with short cisternae. A few low retrograde labelled type II were weakly or non glycinergic. A small number of large to giant neurons type II, strongly retrograde labelled, appeared to be glycine positive, consistently GABA negative and variably glutamate positive. A very small proportion of retrograde labelled neurons appeared having the characteristics of globular bushy neurons. Their weak labelling, however, suggests that they project by collaterals or thin axons to the contralateral cochlear nucleus. Spherical bushy cells in the rat anteroventral cochlear nucleus lack the nuclear capping of rough endoplasmic reticulum observed in the cat, and none was labelled after injection into the contralateral cochlear nucleus. Globular and spherical neurons were variably glutamate positive but glycine and GABA negative. In conclusion, the present study suggests that commissural neurons include a small number of strongly labelled large to giant glycinergic and presumably inhibitory type II and, less frequently type I. A large group of less heavily labelled commissural neurons of type I and II contain low levels or no glycine, which is probably used for metabolic purposes rather than as a neurotransmitter. This suggests that these neurons are presumably excitatory.

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Differentiation of the epidermis during scale formation in embryos of lizard.

The formation of the first epidermal generations and in particular of the shedding complex in the developing scales of the lizards Anolis lineatopus and Podarcis muralis was studied by electron microscopy. The initially linear and bilayered epidermis turned into symmetric papillae, which became asymmetric scales. The outer periderm was more electrondense than the following layer, provisionally named 'inner periderm' but probably derived from the basal layer. Coarse thick filaments progressively filled the cytoplasm of the inner periderm or formed reticulate bodies resembling avian peridermal granules. Peridermal cells cornified slightly and could be shed in ovo. From the basal layer various suprabasal epithelial layers were produced. The first layer contained keratohyalin-like granules and was identified as a clear layer. Beneath it a spinulated (Anolis) or serrated (Podarcis) oberhäutchen differentiated. The clear and 'oberhäutchen' layers constitute the first shedding complex. The first epidermal layers that were shed consisted of flaking periderm or periderm together with the clear layer. The differentiation of a mesos layer was under way before hatching, when the epidermal morphology resembled a stage 4-5 of the adult shedding cycle. Fibroblasts under the inner side of the scale made few contacts with the basement membrane and their cytoplasmic elongations were mostly oriented parallel to the dense lamina. Instead, fibroblasts under the basal layer of the outer scale surface (BLOS) made numerous contacts with the basement membrane, suggesting that more dermal-epidermal interactions take place on this side of the scale.

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Scale morphogenesis during embryonic development in the lizard Anolis lineatopus.

The development of scales was analysed in embryos of the Jamaican Iguanid lizard Anolis lineatopus after the injection of cell proliferation markers [3H]thymidine and 5-bromodeoxyuridine. Embryos were fixed at successive postinjection periods, from 2 h up to 13 d and sections of developing skin were studied by autoradiography and immunocytochemistry. The epidermis during the initial stages of morphogenesis (flat epidermis and symmetric scale stages) expands chiefly by the tangential proliferation of the basal layer. The superficial periderm took part in its own laminar expansion. The rates of proliferation in the epidermis and dermis were similar during the flat epidermal stage, but dermal proliferation decreased under the wave-like epidermis of symmetric scales. No specific localisation of proliferating cells was visible either in the epidermis and or the dermis at these 2 early stages. Scale asymmetry is brought about by cell multiplication and hypertrophy of the cells of the future outer surface of the scale that behaves as an epithelial placode capable of producing beta-keratinised cells. The growth of the outer surface determines the asymmetry of the scale. Conversely, the future inner side and hinge region showed a lower degree of cell proliferation and cells remained cuboidal or became flat. Also the proliferation of the dermis under the scaling epidermis was significantly diminished. During the asymmetric scale stage, labelled cells were more common in the hypertrophied basel epithelium from 2 h until 4 d postinjection, during which time a few cells moved into the upper keratinizing layers. At 6-13 d postinjection, labelled cells were commonly seen in the upper beta-keratinising layers.

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Observations on proliferating sheath cells in the regenerating nerves of lizard.

The proliferation of sheath cells (Schwann and endoneurium) of growing nerves has been studied by autoradiography after 3H-thymidine administration to lizards (Anolis carolinensis and Lampropholis delicata) with regenerating tails. Schwann cells of regenerating nerves derive from the multiplication of resident cells within the growing nerves, but labelled Schwann cells derived from the regenerative blastema also appear to ensheath the new axons. Endoneurium cells of growing nerves derive from the blastema around the nerves more than from dividing and migrating cells of the original endoneurium. After 6-20 days post-injection the number of labelled sheath cells in the older proximal regenerated nerves increased due to local division and migration. In the proximal spinal ganglia which innervate the regenerating tail some satellite cells, but not neurons, took up 3H-thymidine and multiplied.

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Autoradiographic ultrastructural observation on the meninges of the regenerating tail of lizards.

The formation of the meninges around the new spinal cord of the regenerating tail of the lizards Anolis and Lampropholis has been studied after injection of 3H-thymidine and 3H-proline. An optical and electron microscopic autoradiographical study revealed that many irregular 3H-thymidine labeled meningeal fibroblasts surround the apical regenerating spinal cord. Meningeal fibroblasts become flat at about 0.5-1 mm from the tail tip while they actively uptake 3H-proline involved in the synthesis of collagen and elastin. Elastic fibres are in intimate contact with the meningeal fibroblasts. One hour after injection of 3H-proline trace grains were seen most in the Golgi apparatus and secretory elastin-like vesicles. The importance of the elastic fibrils around the caudal spinal cord of lizard in relation to the body movements is discussed.

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