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Experimental studies on hemopoiesis in the pronephros of Rana pipiens.

Embryogenesis of hemopoietic cell populations in the pronephros of Rana pipiens was examined during embryonic and early larval development. Differential cell counts of Wright-Giemsa-stained cell suspensions demonstrated that granulopoiesis is the predominant hemopoietic activity in the pronephros, erythropoiesis accounts for a minor component of the hemopoietic activity (less than 10%), and lymphopoiesis within the organ is negligible. Microdensitometric analysis of Feulgen-DNA stained granulocyte populations in pronephroses from larvae that had received chromosomally labeled pronephric analgen transplants between 84 and 96 h of development demonstrated that hemopoiesis in this organ is dependent on colonization by an extrinsic hemopoietic stem cell. A similar analysis of pronephric hemopoiesis in larvae which had received chromosomally labeled, presumptive ventral blood island transplants between 62 and 67 h of development, indicates that granulopoietic cells are not derived from the embryonic blood islands. It is proposed that the pronephros may be the initial site of granulocyte differentiation during early embryogenesis. Although the embryonic origin of the hemopoietic stem cell is unknown, indirect evidence from this study indicates a dorsal stem cell compartment.

Animals

Modulatory effect of metal ions on the immune response of fish: in vivo and in vitro influence of MnCl2 on NK activity of carp pronephros cells.

The in vivo and in vitro influence of MnCl2 on carp pronephros cells was investigated. Increased cytotoxicity against both YAC-1 and P 815 target cells was observed following an intraperitoneal injection of 40, 80, or 120 micrograms MnCl2/g body wt administrated 24 hr prior to the in vitro 51Cr release assay. Similarly, in vitro treatment of carp pronephros cells, at a final concentration of 60 micrograms/culture, resulted in an increase of NK cell activity in both YAC-1 and P 815 target cell lines. However, a significant decrease in this activity was shown with lower doses of MnCl2 (40 and 20 micrograms/culture).

Animals

Pronephros and mesonephros--Cohnheim revisited.

Erroneous quotations in the literature and Cohnheim's statement, in his Lectures on General Pathology, that the Wolffian body or mesonephros is the first anlage of the urogenital system prompted this description of the growth of our knowledge of the early development of the kidney. Some of the pertinent older literature is reviewed, and the concept of the holonephros, as opposed to the trinephric view of kidney development, is discussed. Emphasis is placed on the decreasing functional significance of the pronephros with evolutionary development, to the extent that the role of the pronephros in the human is questioned. Cohnheim's seemingly erroneous reference to the development of the kidney is shown to be merely a reflection of the views current at his time.

History, 19th Century

[Differentiation of the epithelium of the pronephros and the primary kidney in frogs].

The kidneys of tadpoles of different developmental stages were examined in preparations processed histologically and histochemically. It was found that differentiation of the provisory excretory organ tubules in frogs was "shortened" or "accelerated" after P. P. Ivanov's terminology, and developed differently as compared with differentiation of tubules of the definitive organ of excretion -- the primary kidney. When differentiating the epithelium of the proximal portion of the primary kidney nephron passes the stage of the high prismatic false-stratified epithelium. The pronephros tubules do not pass this stage and the epithelium becomes a strict monolayer from the very beginning. No mitoses are observed in the pronephros tubule epithelium even at the earliest differentiation stages. Later on, the beginning of tubule functioning, and with the reduction, and later disappearance of yolk granules in the epithelium solitary mitoses make their appearance. The mitotic activity of the primary kidney tubule epithelium is very high (70%) at the early stage of differentiation. Then its mitotic activity decreases (30%), and after the beginning of the tubule functioning mitoses in its epithelium become solitary.

Animals

The anionic charge barrier in the renal corpuscle of the pronephros in the lamprey, Petromyzon marinus L.

The charge barrier within the renal corpuscle of the pronephric kidney of the lamprey, Petromyzon marinus, was investigated at two life cycle intervals using cationized ferritin and polyethyleneimine. In the larval renal corpuscle the endothelium of the glomerular capillaries and the laminae rarae externa and interna of the glomerular basement membrane show regularly-spaced deposits of the tracers. The lamina densa remains unstained. Concomitant with a loss of major processes of the visceral epithelial podocytes and development of an extensive mesangial matrix in late adult life are alterations in the distribution of the anionic sites. The lamina rara interna is no longer a distinct entity and the mesangium contains irregularly-distributed anionic sites surrounding electron-dense deposits. The results indicate that the distribution of the anionic sites during adult life most likely affects the ability of the renal corpuscle to act as an efficient filtration device. This charge distribution is consistent with that seen during some renal pathologies of higher vertebrates.

Aging

Leukocytes of rainbow trout (Oncorhynchus mykiss) pronephros: cell types producing superoxide anion.

Fish pronephric and blood leukocytes yield a chemiluminescent response (CL) when stimulated appropriately. This response reflects the production of highly reactive oxygen derivatives which contribute to oxygen-dependent killing of targets such as pathogens and parasites. From suspensions of pronephric cells of the Rainbow trout, Oncorhynchus mykiss, we have obtained populations enriched for CL-positive cells. Four bands of cells were obtained using continuous gradients generated with 60% Percoll. The leukocytes of band II showed a very strong PMA-induced CL response, the magnitude of which was several times higher than that observed with equivalent numbers of cells form unseparated pronephric cell suspensions. Cells present in other bands were not significantly chemiluminescent. Flow cytometric analysis showed that band II contained large granular cells and small granular cells. Cytochemical analysis showed that this subpopulation was greatly enriched with neutrophils. Many band II cells adhere to glass whereas few band III cells do so. The glass-adherent cells loose their CL potential after a few days in vitro, whereas the nonadherent cells retain their CL responsiveness for at least a week in vitro.

Animals

Primed allogeneic reactions to leukocytes and epithelial cells in the bicolor damselfish.

The studies presented here describe allogeneic reactions in the bicolor damselfish, Pomacentrus partitus, a tropical marine teleost. Damselfish were immunized twice at 2-week intervals either by placement of reciprocal allografts or with primary cultures of epithelial cells. Two weeks after the second immunization, recipient splenocytes were tested for alloreactivity toward donor and third-party pronephros cells. For those animals immunized with cultured cells, reactivity toward donor and third-party epithelial cells was also examined. Unprimed animals responded to allogeneic pronephros between days 8 and 10 (28 degrees C). Fish immunized with epithelial cells showed accelerated mixed leukocyte reactions, with optimal responses occurring between days 4 and 6. Moreover, responses to the immunogen preceded responses to donor pronephros tissue by 2 days, occurring on day 2 or day 4. In addition, primed responses were of significantly greater magnitude than primary reactions. Reactivity toward third-party pronephros tissue paralleled responses of naive animals, indicating the specificity of response. No reactivity toward epithelial cells was observed in the absence of immunization, suggesting that accessory cell functions of damselfish pronephros and epithelial cells differ.

Animals

[Correlation of the differentiation of the epithelium of provisory and definitive kidneys in vertebrates].

Opisthonephros-the primary kidney of the frog-has features similar in its development to both primary and secondary kidneys of the rabbit, and at the same time they greatly differ from the frog's pronephros in the course of its development. It is in the development of the frog's pronephros where the regularity of shortened and accelerated differentiation of tissue of provisory organs as compared with definitive ones formulated by P.P. Ivanov has distinct manifestation. It is in agreement with the fact that the pronephros of lower vertebrates is considerably more primitive in its anatomical and physiological features than the primary and secondary kidneys of higher vertebrates which are in general similar with each other though functioning in different developmental periods.

Anatomy, Comparative

Xlcaax-1 is localized to the basolateral membrane of kidney tubule and other polarized epithelia during Xenopus development.

Xlcaax-1 is a novel, maternally expressed, 110-kDa, CAAX box containing protein that undergoes isoprenylation and palmitoylation through which it associates with the plasma membrane. We report here the cellular and subcellular localization of the xlcaax-1 protein during development of Xenopus laevis. Whole-mount immunocytochemistry and immunoperoxidase staining of tissue sections show that during development the xlcaax-1 protein accumulation is coincident with the differentiation of the epidermis, pronephros, and mesonephros. In the pronephros and mesonephros the xlcaax-1 protein is localized to the basolateral membrane of differentiated tubule epithelial cells. Thus, the xlcaax-1 protein serves as a marker for tubule formation and polarization during Xenopus kidney development. Xlcaax-1 may also be used as a marker for the functional differentiation of the epidermis and the epidermally derived portions of the lens and some cranial nerves. Western blot analysis shows that in the adult the xlcaax-1 protein is most abundant in kidney. Immunogold EM analysis shows that the xlcaax-1 protein is highly enriched in the basal infoldings of the basolateral membrane of the epithelial cells in adult kidney distal tubules. In addition, immunoperoxidase staining of tissue sections detected low levels of xlcaax-1 protein in the epithelial cells of skin, urinary bladder, gall bladder, and parietal glands of the stomach. The localization pattern of xlcaax-1 suggests that the protein may function in association with an ion transport channel or pump.

Animals

The cellular proliferative response, humoral antibody response, and cross reactivity studies of Tetrahymena pyriformis with Ichthyophthirius multifiliis in juvenile carp (Cyprinus carpio L.).

The immune response of 10-12-week-old juvenile carp to T. pyriformis [CCAP 1630/w, 1939 (w)] was investigated. The humoral antibody response following one and two intraperitoneal injections of whole live T. pyriformis (protein concentration of 25 micrograms/g body weight), separated by an interval of 4 weeks, was monitored over 12-16 weeks. Peak antibody titres were detected 6 weeks following antigen administration. Antibody titres were elevated following the second injection, and relatively high levels were still maintained by week 16. Proliferative responses measured by autoradiography following intraperitioneal injection of methyl 3H thymidine (1-2 microCi/g body weight) were recorded in the pronephros, opisthonephros, and spleen at weekly intervals for 4 weeks following each injection of T. pyriformis. Immunised fish had higher counts than nonimmunised controls, with the greatest counts occurring at 2 and/or 3 weeks following one injection of antigen and 2 weeks following two injections. Activity was recorded prior to peak antibody production with the pronephros and opisthonephros having greater proliferative responses than the spleen. Cross antigenicity studies to examine protection of carp against I. multifiliis following administration of T. pyriformis, showed little evidence in support of protection using this strain of T. pyriformis. Further, antisera raised in either carp, rabbits, or rats failed to show any cross antigenicity between these two species of protozoa.

Animals

[Metameric components of the occipital region of the polypterids (Pisces)].

Investigations into the development of the pronephros in Polypterus senegalus senegalus Cuvier have shown several details on the composition of the occipital region of this fish. It has been shown that the first six metotic metameres contribute to the structuring of the occipital region. These six metameres also are responsible for the existence of the pronephros. The epimeres of metamere I atrophies in the beginning of larval life. The muscular structures derived from the four following metameres become englobed in the occipital region. In the myoseptum between the previous metameres V and VI the post-occipital semi-vertebra, a pecularity of Polypterus, arises. The first vertebra of the column arises between structures that are derived from the previous metameres VI and VII. Such statements have been possible by a close examination of the disposition of the pectoral fin nerves in several larval stages. Because these nerves have been labelled with different names in several studies, a comparison of these names has been necessary.

Animals

[Differentiation of hematopoietic cells in explants of embryonal organs of Rana temporaria L].

The potencies of isolated embryonic hemopoietic organs (pronephros and liver) of Rana temporaria L. to the formation of the foci of hemopoiesis were studied. The pronephros and liver rudiments were explanted at the early developmental stages (late neurula and early tail bud) and cultivated in vivo in the diffusion chambers. The blast hemopoietic elements and differentiated blood cells are found in the explants within 7 to 10 days of cultivation. A suggestion is put forward that the differentiation of hemopoietic cells in the embryonic hemopoietic organs proceeds from the local cells-precursors.

Animals

N-cadherin transcripts in Xenopus laevis from early tailbud to tadpole.

Cadherins are Ca(++)-dependent cell adhesion molecules which play a key role in morphogenesis and histogenesis. Two mRNAs clones (8 and 9) corresponding to two N-cadherin pseudo-allelic genes are present in Xenopus laevis. We report here that these transcripts share a highly homologous coding region but diverge in the non-coding region. We have determined the pattern of N-cadherin expression at the mRNA level by in situ hybridization with a riboprobe complementary to the EC5 domain of Xenopus N-cadherin clone 8. This part of the sequence is the least conserved in the cadherin gene family, minimizing the risk of cross-hybridization to other cadherins. N-cadherin transcripts are not detectable in the first stages of development. Expression first appears in the neural plate and reaches its maximum level in the CNS at tailbud stage. From early tadpole, it diminishes, so that a very weak signal is detected in the premetamorphic frog brain. N-cadherin expression is not uniform within the CNS, with some areas such as the roof of the rhombencephalon and the olfactory bulbs expressing higher levels of the transcripts. N-cadherin is present in several mesodermal derivatives such as the notochord, the pronephros, and the heart. It is, however, virtually absent from the myotomes and appears in skeletal muscles at later stages of differentiation. All placodes express high levels of N-cadherin. The non-neural ectoderm and the endoderm are always negative. In the brain and the heart, high levels of hybridization are observed with probes corresponding to both copies of the N-cadherin pseudo-allelic genes in their 5' non-coding region, indicating that both alleles are transcribed.

Animals

Experimental studies on a lethal gene (1) in the Mexican axolotl, Ambystoma mexicanum.

1. Gene ł is a recessive lethal factor found in the white strain of axolotls. Animals heterozygous for the gene are phenotypically normal. When mated with each other they give offspring 25% of which exhibit the lethal effects of the gene. 2. The ł/ł homozygotes develop normally to an advanced embryonic stage (Harrison stage 40) before the effects of the gene are first manifested. They then come to display a characteristic combination of abnormalities, including a disproportionately small head, small and poorly developed eyes, abnormal poorly developed gills, undifferentiated limb buds, and reduced overall growth rate. They may feed briefly, but soon stop and invariably die within a few weeks of the time of hatching. 3. The action of gene ł has been analyzed by parabiosing mutant and normal embryos, and by grafting various organ primordia reciprocally between mutant and normal embryos. Parabiosis to normal embryos fails to correct the abnormalities of the mutants, although their survival may be somewhat prolonged. Grafts of mutant organ primordia (eye, limb, gill, pronephros, gonad, head) also invariably fail to show improved development or to survive on normal hosts; normal organ primordia develop normally on mutant hosts so long as the mutant survives. These experiments indicate that gene ł is a recessive autonomous cell lethal affecting all of the organ systems during late embryonic and early larval development.

Ambystoma

The marginal zone of the 32-cell amphibian embryo contains all the information required for chordamesoderm development.

The formation of the amphibian organizer is evidenced by the ability of cells of the dorsal marginal zone (DMZ) to self-differentiate to form notochord and to induce the formation of other axial structures from neighboring regions of the embryo. We have attempted to determine when these abilities are acquired in the urodele, Ambystoma mexicanum (axolotl), and in the anuran, Xenopus laevis, by removing the mesodermalizing influence of the vegetal hemisphere at different stages of development and culturing the animal hemisphere isolate. This was possible, even at the 32 and 64-cell stage, through the use of embryos with rare cleavage patterns. Cultured isolates were analyzed for morphological differentiation of mesodermal and neural structures, and for biochemical differentiation of the tissue-specific enzyme, acetylcholinesterase (AChE). Large amounts of mesodermal and neural structures, and normal expression of AChE were found in isolates made as early as the 32-cell stage in both species. Only a small increase in the percentage of isolates developing mesoderm was detected when isolations were made at later cleavage or blastula stages. The amount of mesoderm formed did not depend on the stage of isolation. Mesoderm differentiation was usually limited to the notocord and muscle. The isolates rarely formed pronephros, mesothelium, or mesenchyme, derivatives of ventral mesoderm, during normal development. The results indicate that the marginal zone of the cleavage-stage embryo contains all of the information needed for the formation of the organizer. The formation of dorsal mesoderm does not require subsequent interaction with the cells of the vegetal hemisphere, although the presence of those cells is likely to play a role in normal pattern formation.

Acetylcholinesterase

The distribution of mesenchyme proteoglycan (PG-M) during wing bud outgrowth.

This study utilizes immunofluorescence to describe the distribution of several extracellular matrix molecules in the chick embryo during the process of limb outgrowth and the formation of precartilage condensations. A large chondroitin sulfate proteoglycan (PG-M) is detected at the wing level at Hamburger and Hamilton stage 14 in and under the dorsal ectoderm, and is associated with the basement membranes around the neural tube, notochord and pronephros, but not with other basement membranes. The galactose-specific lectin, peanut agglutinin (PNA), has a similar distribution except that it also binds to the dorsal side of the neural tube. PG-M is not detected in the limb mesenchyme until after stage 17, when it is present in the distal region, as is PNA-binding material. With further development of the wing bud, PG-M is present in the subectodermal mesenchyme, the mesenchyme at the distal tip and in the prechondrogenic core. After stage 22 PNA-binding material becomes localized in the prechondrogenic core, the basement membranes under the apical ectodermal ridge, and the ventral sulcus. The distribution of these components (PG-M and PNA binding material) overlaps, but differs from that of type I collagen and fibronectin and basement membrane components, such as laminin, basement membrane heparan sulfate proteoglycan, and type IV collagen. Tenascin, on the other hand, is not detected in the limb bud until stage 25, after the appearance of cartilage matrix components such as type II collagen and cartilage proteoglycan (PG-H). These results are considered in relation to the formation of precartilage aggregates, and indicate that PNA binds to components in precartilage aggregates other than PG-M or tenascin.

Aggrecans

Mapping of neural crest pathways in Xenopus laevis using inter- and intra-specific cell markers.

This study examines the pathways of migration followed by neural crest cells in Xenopus embryos using two recently described cell marking techniques. The first is an interspecific chimera created by grafting Xenopus borealis cells into Xenopus laevis hosts. The cells of these closely related species can be distinguished by their nuclear dimorphism. The second type of marker is created by microinjection of lysinated dextrans into fertilized eggs which can then be used for intraspecific grafting. These recently developed fluorescent dyes are fixable and identifiable in both living and fixed embryos. After grafting labeled donor neural tubes into unlabeled host embryos, the distribution of neural crest cells at various stages after grafting was used to define the pathways of neural crest migration. To control for possible grafting artifacts, fluorescent lysinated dextran was injected into a single blastomere which gives rise to a large number of neural crest cells, thereby labeling the neural crest without grafting. By all three techniques, Xenopus neural crest cells were observed along two predominant pathways in the trunk. The majority of neural crest cells were observed along a "ventral" route, between the neural tube and somite, the notochord and somite, and along the dorsal mesentery. A second group of neural crest cells was observed "dorsally" where they populated the dorsal fin. A third minor "lateral" pathway was observed primarily in borealis/laevis chimerae and in blastomere-injected embryos; some neural crest cells were observed underneath the ectoderm lateral to the neural tube. Along the rostrocaudal axis, neural crest cells were not continuously distributed but were primarily located across from the caudal two-thirds of the somite. Fewer than 3% of the neural crest cells were observed across from the rostral third of each somite. When grafted to ventral locations, neural crest cells were not able to migrate dorsally but migrated laterally along the dorsal mesentery. Labeled neural crest cells gave rise to cells of the spinal, sympathetic, and enteric ganglia as well as to adrenal chromaffin cells, Schwann cells, pigment cells, mesenchymal cells of the dorsal fin, and some cells in the integuments and in the region of the pronephros. These results show that the neural crest migratory pathways in Xenopus differ from those in the avian embryo. In avians NC cells migrate as a closely associated sheet of cells while in Xenopus they migrate as individual cells. Both species exhibit a metamerism in the neural crest cell distribution pattern along the rostrocaudal axis.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Effects of organophosphorus insecticides: effects of trichlorfon and dichlorvos on the immune response of carp (Cyprinus carpio). III. In vitro effects on lymphocyte proliferation and phagocytosis and in vivo effects on humoral response.

Cells isolated from the pronephros of carp were incubated in vitro with the organophosphorus insecticide trichlorfon or dichlorvos, each of which is used in aquaculture to eliminate fish ectoparasites. Dose-dependent suppressive effects were observed in assays for lymphocyte proliferation and myeloid cell respiratory burst activities. Dichlorvos given by bath in vivo did not affect antibody production against Yersinia ruckeri even if the spleen and kidney were obviously contaminated according to acetylcholinesterase activity analysis. Some blood parameters of nonspecific immunity (ceruloplasmin, lysozyme, hemagglutinins) were slightly affected.

Animals