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[Cytochemical localization of an important adenyl-cyclase activity in Anuran amphibian embryo chordo-mesoderm by electron microscopy; its possible relations with the migration of primordial germ cells (author's transl)].

Chordo-mesoderm of Anura Amphibia embryos was investigated by electron microscopy and a cytochemical method was used to study the localization of adenyl-cyclase activity, in this part of embryo. It seem that, in Anura, at a given stage of development, there is some adenyl-cyclase activity, and, consequently, some liberation of AMPc. Pleurodeles waltlii (Urodela Amphibia) appears to be devoid of this adenyl-cyclase activity. AMPc could be the substance provided by chordo-mesoderm attracting germinal cells.

Adenylyl Cyclases↗

[A comparative electron microscopic study of the peripheral blood leukocytes of vertebrates].

The ultrastructure of leukocytes of the peripheral blood was studied in representatives of classes of fishes (Cyprinus carpio), amphibia (Rana redibunda, Bufo bufo), reptiles (Testudo horshfield), birds (Columba livia) and mammals (Rattus norvegicus). The granules are characterized by homogeneous or heterogeneous matrix. The granules of special leukocytes (turtle) and pseudoesinophils (pigeon) have larger dimensions than those in neutrophils of other animals. The matrix of the eosinophil granules of the carp and rat contain crystalloid structures while the granules of eosinophils of amphibia, turtles and pigeons are characterized by electron dense homogeneous matrix. The basophil granules are larger and in addition to homogeneous or fine granular structure (turtle, pigeon, rat) have lamellar structure (Cyprinus carpio, frog, toad). The ultrastructural organization of agranulocytes and plasma cells of the animals studied is also described.

Animals↗

[Studies on the occurrence of Listeria monocytogenes in fecal samples of domestic and companion animals].

Listeria (L.) monocytogenes was isolated from 33.3% fecal samples of 138 cattle (29 strains serovar 1/2b, 10 strains serovar 1/2a, 7 strains serovar 4ab), from 8% fecal samples of 100 hens (5 strains serovar 1/2b, 1 strain serovar 1/2a, 4ab and 4b each), from 8% fecal samples of 100 sheep (6 strains serovar 1/2a, 1 strain serovar 1/2b and 4ab each), from 5.9% fecal samples of 34 pigs (2 strains serovar 4ab), from 4.8% fecal samples of 400 horses (7 strains serovar 1/2a, 6 strains serovar 1/2b and 4ab each), from 1.3% fecal samples of 300 dogs (3 strains serovar 1/2b, 1 strain serovar 4ab), from 0.9% fecal samples of 350 pigeons (1 strains serovar 1/2a, 1/2b and 4ab each), from 0.4% fecal samples of 275 cats (1 strain serovar 4b) and from 5 respectively 2 fecal samples of 35 tortoises (3 strains serovar 4ab, 2 strains serovar 4b) respectively 5 amphibia (1 strain serovar 4ab and 4e each). Further 36.9% fecal samples of cattle, 15% fecal samples of hens, 6% fecal samples of horses, 5.9% fecal samples of pigs, 5% fecal samples of sheep, 4% fecal samples of dogs, 2.3% fecal samples of pigeons, 3 fecal samples of 35 tortoises and 1 fecal sample of 260 rabbits contained L. innocua. In further 2% fecal samples of hens, in further 1.5% fecal samples of horses and in further 0.6% fecal samples of pigeons was detected L. seeligeri. 3 fecal samples of horses and 1 fecal sample of dogs, lizards and amphibia each contained L. welshimeri, while in 1 fecal sample of tortoises and in 3 fecal samples of 33 lizards L. ivanovii was detected.

Amphibians↗

Distribution of choline acetyltransferase immunoreactivity in the brain of anuran (Rana perezi, Xenopus laevis) and urodele (Pleurodeles waltl) amphibians.

Because our knowledge of cholinergic systems in the brains of amphibians is limited, the present study aimed to provide detailed information on the distribution of cholinergic cell bodies and fibers as revealed by immunohistochemistry with antibodies directed against the enzyme choline acetyltransferase (ChAT). To determine general and derived features of the cholinergic systems within the class of Amphibia, both anuran (Rana perezi, Xenopus laevis) and urodele (Pleurodeles waltl) amphibians were studied. Distinct groups of ChAT-immunoreactive cell bodies were observed in the basal telencephalon, hypothalamus, habenula, isthmic nucleus, isthmic reticular formation, cranial nerve motor nuclei, and spinal cord. Prominent plexuses of cholinergic fibers were found in the olfactory bulb, pallium, basal telencephalon, ventral thalamus, tectum, and nucleus interpeduncularis. Comparison of these results with those obtained in other vertebrates, including a segmental approach to correlate cell populations, reveals that the cholinergic systems in amphibians share many features with amniotes. Thus, cholinergic pedunculopontine and laterodorsal tegmental nuclei could be identified in the amphibian brain. The finding of weakly immunoreactive cells in the striatum of Rana, which is in contrast with the condition found in Xenopus, Pleurodeles, and other anamniotes studied so far, has revived the notion that basal ganglia organization is more preserved during evolution than previously thought.

Animals↗

Organogenesis of the Harderian gland: a comparative survey.

Although research interest in the Harderian gland (HG) has increased during the last few years, only a small amount of information exists about its organogenesis. In mouse the HG appears in the posterior part of eye region, in the form of nonluminated tubules between the sixteenth and eighteenth days of gestation. At birth it is still not differentiated histologically. In birds the HG originates from the conjunctival epithelium at a late embryonic stage. In the English sparrow, Passer domesticus (incubation period of about 13 days), it appears between the seventh and the eighth days of incubation. In the chick embryo (incubation period of about 21 days) it originates between the eleventh and the twelfth days. Among reptiles the lizard Podarcis s. sicula has proved to be a useful model to clarify the embryological origin of the orbital glands since it possesses the anterior lacrimal gland contiguous to the HG in the medial corner of the orbit. The anlage of the orbital glands appears on about the twenty-second day of development (incubation period of about 43 days) in the form of a short tubule projecting from the conjunctival epithelium, at the time of development of the nictitating membrane. At this stage the mesenchymal cells surrounding the glandular blastema form a well-defined sac, later occupied by the orbital glands. From this stage until hatching the growth of the glandular blastema continues with the formation of acini which move posteriorly into the preformed mesenchymal sac. At the thirty-sixth day of development the more lateral acini differentiate into the HG. Only at the forty-first day do the more medial acini differentiate into the anterior lacrimal gland. At hatching the HG is fully differentiated. In anuran amphibia the primordium of the HG appears during the metamorphosis at the time of development of the nictitating membrane.

Amphibians↗

Structural diversity of the ordinary and specialized lateral line organs.

Lateral line organ, a superficial sensory system in amphibia and fish which provides the animal with information about its surrounding environment, is divided classically into two main different types, ordinary and specialized, whose functions are mechanoreceptive and electroreceptive, respectively. Although it has great diversity, the basic sensory unit, which is usually called "neuromast," is composed of sensory cells embedded in accessory cells. The functions of the latter are to support the sensory cells and to secrete the material that covers the organs, forming a cupular structure or filling a canal which enables the organ to communicate with the exterior. Sensory cells of mechanoreceptive neuromasts have a tuft of processes included in the cupular material; these are a kinocilium and a group of stereocilia with a typical staircase arrangement. The displacement of the stereocilia towards or away from the kinocilium produces different stimuli. The electroreceptive organs are more diverse. They include ampullary and tuberous organs. The latter can be subdivided into different types: knollenorgans, mormyromasts, gymnarchomasts, etc. All of these present a great diversity among species, but their morphology is less reported than that of the mechanoreceptive organs. This paper summarizes the structural features of the main different types of lateral line organs, as well as their taxonomic distribution and different patterns of distribution along the surface of the animal.

Amphibians↗

Ultrastructural studies of dorsal root axons regenerating through adult frog optic and sciatic nerves.

Optic nerves of adult fish and amphibia can successfully regenerate, in part because their glial cells, unlike those of mammals, provide an environment permissive to regrowth. We altered the environment of regenerating dorsal root axons in the frog, Rana pipiens, by grafting segments of optic nerve to test the permissiveness of CNS glial cells to other sensory neurons. We compared these preparations to grafts of segments of sciatic nerve. After allowing various times for survival, light and electron microscopy were used to evaluate the grafts. An agglomeration of astrocytes, tightly joined by desmosomes, initially formed in the center of the optic nerve grafts. Around this grew regenerating dorsal root axons, accompanied by Schwann cells. At early stages, some axons formed dilated terminal structures, which were not seen in peripheral nerve grafts. The appearance of blood vessels within the graft and the dispersion of cells allowed larger numbers of axons to grow through the graft. By eight weeks, 48% of dorsal root sensory axons had grown through optic nerve grafts, compared to 84% for sciatic nerve. These results suggest that astrocytes from optic nerve are not inhibitory to, and provide a suitable substrate for, regrowing sensory neurons.

Animals↗

Myelin protein zero and membrane adhesion.

Protein zero (P0) is a member of the immunoglobulin gene superfamily (IgCAM) that is expressed at high levels in myelinated vertebrates in central (fish and amphibia) and peripheral (all species) myelin. This glycoprotein is the major adhesive component of peripheral myelin, where it mediates self-adhesion of the Schwann cell plasma membrane. Although the expression of P0 is naturally limited to Schwann cells, the molecular mechanisms of P0-mediated adhesion can be considered general and "obligatory" because, when expressed in a variety of cell lines, P0 induces strong intercellular adhesion. Modeling studies, X-ray crystallographic analysis, and experimental site-directed mutagenesis have provided excellent working models for understanding how P0 mediates adhesion at the atomic level. These models remain to be experimentally tested. However, in humans, certain mutations in P0 produce dysmyelinating disease, possibly due to disruptions in the predicted P0 lattice.

Amino Acid Sequence↗

K+ Channel density increases selectively in the endfoot of retinal glial cells during development of Rana catesbiana.

The radial glial cells that span the retina, described by Müller in 1851, have a remarkable distribution of ion channels in adult amphibia that mediate extracellular K+ spatial buffering. 94% of the total membrane conductance of these cells resides in inward rectifier K+ channels in the endfoot processes apposed to the vitreous humour. We now report that this regional specialization is found in Müller cells isolated from adult (>120 day old) bullfrogs but to a far less extent in those from 10-20 day old tadpoles (stages 34-36). Using the cell attached configuration of the patch-clamp technique, we found, in agreement with previous studies in salamanders, that the endfoot of adult cells had 19.2+/-2.4 (mean +/- S.E., n = 81) channels/patch, whereas the soma had 1.81+/-0.28 (n = 21) channels/patch. In the tadpole, the respective values were 4.29+/-0.26 (n = 79) for the endfoot and 2.26+/-0.24 (n = 27) for the soma. The slope conductance of the inward rectifier K+ channel in 115 mM K+, 19.2+/-0.25 pS (n = 205), channel kinetics and the resting membrane potential (-69+/-2.7 mV, n = 224) were similar at both the endfoot and soma of both adults and embryos. We conclude that during development, the K+ conductance of the Müller cell endfoot, but not of the soma, increases due to a selective clustering of inwardly rectifying K+ channels in that specific region of the cell membrane. The properties of the channels change little during the transformation from tadpole to adult bullfrog.

Animals↗

Involvement of purines and phosphoinositides in spontaneous and progesterone-induced nuclear maturation of Bufo arenarum oocytes.

Although progesterone is the established maturation inducer in amphibia, it has been demonstrated that Bufo arenarum oocytes resume meiosis with no need of an exogenous hormonal stimulus if deprived of their enveloping follicle cells, a phenomenon called "spontaneous maturation." The present studies were designed to evaluate the participation of purines and phosphoinositides in the spontaneous and progesterone-induced maturation in Bufo arenarum full-grown oocytes. The presented data demonstrate that high intracellular levels of purines such as cAMP or guanosine can inhibit both spontaneous and progesterone-induced maturation in full-grown denuded Bufo arenarum oocytes. Moreover, the fact that the mycophenolic acid was able to induce maturation in denuded oocytes obtained during the nonreproductive period in a manner similar to that of the progesterone and also to increase the percentages of spontaneous maturation suggests that in Bufo arenarum, inosine monophosphate dehydrogenase inhibition is an important step in the resumption of meiosis. Inhibition of the phosphatidylinositol 4,5 bisphosphate hydrolysis by treatment of denuded oocytes with neomycin totally blocks spontaneous and progesterone-induced maturation, suggesting that the products of this hydrolysis (1,2 diacylglycerol and inositol 1,4,5 trisphosphate) may be involved in the maturation process of Bufo. In addition, our results indicate that the activation of protein kinase C is also involved in both types of maturation.

2,4-Dinitrophenol↗

Evolution of odorant receptors.

Odorant receptors (ORs) located in the nasal epithelium, at the ciliated surface of olfactory sensory neurons, represent the initial step of a transduction cascade that leads to odor detection. ORs form the largest and most diverse family of G-protein-coupled receptors (GPCRs). They are encoded by a multigene family that has been partially characterized in cyclostomes, teleosts, amphibia, birds and mammals, as well as in Drosophila melanogaster and the nematode Caenorhabditis elegans. As new sequence data emerge, it is increasingly clear that OR primary structure can vary dramatically across phyla. Some chemoreceptors are encoded by genes with little sequence similarity to the prototypical ORs originally isolated in mammals. A large number of sequences are now available allowing a detailed study of the evolutionary implications of OR diversity across species. This review discusses the evolutionary implications of the divergent primary structures of chemoreceptors with identical functions.

Animals↗

Effects of selective neurotoxins on eye growth in the young chick.

We have determined the extent of retinal cell damage and eye growth after treatment with different neurotoxins. Day-old chicks received an intraocular injection containing 2, 10, 50, 100 or 200 nmol of kainic acid (KA), an excitotoxic analogue of glutamate. After 21 days, with 2 nmol KA damage was confined to a small proportion of bipolar cells, whereas with 10-200 nmol KA there was dose-dependent damage to amacrine cells, ganglion cells and photoreceptors. There was an increase in eye weight and size of the vitreal chamber with 10 nmol KA or more. In a similar protocol using 5, 50 or 200 nmol quisqualic acid (QUIS), there was massive loss of amacrine cells and slight loss of horizontal cells, and a large increase in the anterior chamber depth. A single dose of 200 nmol N-methyl-D-aspartate lesioned amacrine cells, but did not alter eye size. Thus, excitotoxins which have different retinotoxic effects also have different effects on eye size. In another study, we examined the effects of KA and QUIS on occlusion-induced eye enlargement. Treatment with 200 nmol QUIS alone resulted in increased depth of the anterior chamber. QUIS combined with occlusion for 3 weeks did not interfere with occlusion-induced growth of the vitreal chamber. Therefore, the loss of a large proportion of amacrine cells did not interfere with occlusion-induced growth. KA results in eye growth in its own right, but restricts occlusion-induced growth. These effects are associated with damage to photoreceptors, amacrine and bipolar cells and a small proportion of ganglion cells. Finally, we examined the effects of an intraocular injection of tunicamycin, a purported photoreceptor-specific neurotoxin in amphibia. Tunicamycin (0.1 or 1 mumg/eye) resulted in a flattened anterior chamber but did not affect growth of the vitreal chamber. Tunicamycin inhibited occlusion-induced growth, and treated retinae displayed massive disruption and loss of all cell types apart from a single row of pigmented retinal epithelium. We conclude that (a) normal and occlusion-induced eye growth may have separate mechanisms of action, (b) the vitreous and anterior chambers have different growth mechanisms, and (c) photoreceptors may play a critical role in occlusion-induced growth of the eye.

Animals↗

Comparative histochemical study of alimentary tracts with special reference to the mucous neck cells of the stomach.

Combinations of recently developed paradoxical concanavalin-A staining (PCS) and other routine histochemical procedures have made it possible to classify mucosubstances more precisely. By taking advantage of these sequences, the present study was undertaken both to characterize the mucinous contents of alimentary tracts of several animal species, and to compare the variability of epithelial mucosubstances with special reference to the mucous neck cells of the stomach. The alimentary tracts obtained from 7 species of fish, 5 amphibians, 3 reptiles, 4 birds, and 6 mammals were used. Acidity of epithelial mucosubstances was highly variable even among the corresponding mucous cells. On the contrary, concanavalin-A reactivity seemed to be consistent with each cell and species. The mucous neck cells, which were observed in amphibians, reptiles, and mammals, consistently exhibited a characteristic mucosubstance with stable class-III reactivity by PCS. Inversely, stable class-III reactivity was found only in species possessing mucous neck cells, and was widely distributed in the esophageal glands of the frog, snake, and man; in esophageal mucous cells of the frog and skink; in pyloric glands of amphibia, reptiles, and mammals; and in Brunner's glands of mammals. These mucous cells seem to form an unique group with morphological and histochemical similarities. It is likely that the stable class-III reactivity by PCS is a hallmark of the mucous neck cells and related glands and that, in addition, concanavalin-A reactivity of mucosubstances is evolutionarily more fundamental characteristic.

Amphibians↗

Early development of the facial nerve in the chick embryo with special reference to the development of the chorda tympani.

The development of the facial nerve from Hamburger and Hamilton stage 17 to stage 28 is described in chick embryos by means of a new immunochemical nerve staining method that uses an antineurofilament protein (NFP) antibody. A postspiracular branch and an unknown transient posterior branch beneath the ostocyst were observed at stage 17. At stage 19, the primordia of the r. palatinus were observed. A prespiracular branch appeared at stage 21, and with the postspiracular nerve, it made a loop encircling the spiracle (spiracular loop). The first primordium of the ramus (r) hyoideus and transient rami (rr) dorsales appeared around stage 23. At stage 25, the chorda tympani was first observed to arise from the ventral end of the spiracular loop. At stage 26, a communicating branch, connexus cum nervo glossopharyngeo, was found along with the vena (v) capitis lateralis. The rr. dorsales seemed to represent the r. supratemporalis in lower animals. The communicating branches around the v. capitis lateralis seemed to correspond to the cutaneous nerve communications between the branchial nerves frequently encountered in Amphibia. It was found that the chorda tympani becomes a prespiracular nerve for the most part in the chick by the reduction of the postspiracular component of the spiracular loop. Thus, the nerve differs markedly from that in other animals, which is postspiracular. This difference explains the different passage of this nerve in the chick as compared with other amniotes.

Animals↗

Fine structure of the testis in the toad (Bufo arenarum hensel): a freeze-fracture study.

The present work reports novel findings in the toad testis using freeze-fracture techniques. Tight and gap junctions are observed in the nonfenestrated endothelial cells. Numerous gap junctions are present between interstitial cells. Sertoli-Sertoli junctional specializations in the toad are similar to those described in mammals although they appear to be less elaborate. The appearance of nuclear pores, in the seminiferous epithelium, ranges from a uniformly and apparently random distribution in Sertoli cells to large aggregations of closely spaced pores separated by free areas in the nuclear envelope of spermatocytes. Inasmuch as these features have also been described in mammals, the fact that they are present in amphibia might indicate that they represent a widespread pattern. Concomitantly with increasing chromatin condensation, nuclear pores diminish in round spermatids and seem to disappear in the more mature germ cells. The en face freeze-fracture views of annulate lamellae in the postnuclear cytoplasm and membranous features of the acrosome formation, flagellum, and undulating membrane of elongating spermatids are also described.

Animals↗

Redistribution of gastric K+-NPPase in vertebrate oxyntic cells in relation to hydrochloric acid secretion: a cytochemical study.

Gastric K+-NPPase represents a partial reaction of the (K+-H+)ATPase system, which is considered to be the proton pump in mammalian parietal cells. In the present paper, K+-NPPase activity was cytochemically studied by the method of Mayahara et al. (1980) in gastric glands of birds, amphibia, and mammals, either in the resting state induced by cimetidine or after stimulation of HCl secretion by histamine. The gastric K+-NPPase cytochemical reaction was localized only in oxyntic cells of the gastric mucosa in the three species tested. The subcellular distribution of the K+-NPPase reaction product drastically changes with the secretory state of HCl. In resting cells, the K+-NPPase staining is associated with the membranes of the endocellular tubular system while in HCl-secreting cells, it is associated with the plasma membrane of the elaborate secretory surface characteristic of this functional state. The above results demonstrate that the same enzymatic activity, which is associated with the gastric proton pump, is present in both membranous systems of the oxyntic cell secretory pole. This fact supports the proposal that the tubular system represents a membrane reserve that inserts the proton pump into the luminal plasma membrane in vertebrate oxyntic cells under the action of HCl secretagogues.

4-Nitrophenylphosphatase↗

Epicardial development in the axolotl, Ambystoma mexicanum.

Recent studies on avian and mammalian embryos have established that the epicardium is derived, not from the early heart tube, but from mesothelial tissue overlying the sinus venosus. We tested the validity of this concept for Amphibia by examining normal and cardiac lethal (c/c) mutant axolotl embryos (stages 35-43) by electron microscopy. In axolotl embryos, the myocardial surface of the heart remains exposed to the pericardial fluid through stage 39. At this stage the transverse septum releases into the pericardial cavity mesothelial cells that subsequently flatten over the adjacent ventricular myocardium. However, mesothelial cells observed on the developing epicardium always appear rounded and may extend a filopodium up to 75 microns. This apparent "substrate-dependent" difference in mesothelial cell shape may promote the extension of the epicardium over the rest of the myocardium. The initial site of epicardial formation persists in the adult as the ventricular pericardial stalk that connects the epicardium to the peritoneal lining of the transverse septum. Cardiac lethal (c/c) mutant embryos, despite the non-contractility of their myocardia, form their epicardia in the same way. This suggests that the c/c mutation does not impair those properties of the myocardium that render it a suitable substrate for epicardial spreading. The abnormal pattern of epicardial coverage of the edematous stage 41 c/c mutant heart could be the result of its abnormally large myocardial surface area, the abnormal proximity of the atrium to the transverse septum, and/or the absence of heart contractions which could aid the dispersion of mesothelial cells within the pericardial cavity. Despite species differences, epicardial development in the axolotl is similar to the general pattern described for higher vertebrate embryos.

Ambystoma↗

Morphogenesis of the frontal organ in Bufo bufo during development.

Morphogenesis of the frontal organ in Bufo bufo was examined under transmission electron microscope. Many remarkable similarities to the frontal organ of other Amphibia Anura are observed. It originates from a diverticulum in the dorsal region of the neural tube. It is egg-shaped, has an eccentric lumen, and is made up of three kinds of cells: 1) photoreceptors, which protrude into the lumen; 2) supportive cells; and 3) ganglion cells, which make synaptic contact with the photoreceptors. Peculiar to Bufo bufo is the melanin-like pigments around the light-sensitive part of the photoreceptors. These pigments may prevent light dispersion. The frontal organ in Bufo bufo starts degenerating during the early premetamorphic stages.

Animals↗