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Membrane physiology of retinal glial (Müller) cells.

Electrophysiological techniques were used to determine the ion selectivity properties and the spatial distribution of the membrane conductance of amphibian Müller cells. Membrane potential changes recorded during ion substitution experiments in frog (Rana pipiens) retinal slices demonstrated that the Müller cell K+:Na+ membrane permeability ratio is approximately 490:1 and that cell Cl- permeability is extremely low. In frog retinal slices, Müller cell input resistance was 8.5 megohms when measured in the inner plexiform layer and 4.8 megohms when measured in the optic fiber layer. Intact, enzymatically dissociated salamander (Ambystoma tigrinum) cells had an input resistance of 7.9 megohms, whereas cells lacking their endfoot process (removed by surgical microdissection or by shearing force) had a resistance of 152 megohms. Pressure ejection of a 100 mM K+ solution near the proximal surface of the endfeet of dissociated salamander cells produced depolarizations 7 times greater than did ejections near the lateral face of the endfoot and 24 to 50 times greater than did ejections near other cell regions. Similar K+ ejection results were obtained from Müller cells in salamander and frog retinal slices. Taken together, these results demonstrate that in both the frog and the salamander, approximately 95% of the total membrane conductance of Müller cells is localized in the cell's endfoot process. In salamander, the specific membrane resistance of the endfoot membrane was estimated to be 32 ohm X cm2 whereas the specific resistance of the remainder of the cell was 7300 ohm X cm2. This remarkably nonuniform conductance distribution has important consequences for theories concerning K+ regulation in the retina and for mechanisms underlying electroretinogram generation.

Ambystoma↗

Identification of sugar residues in secretory glycoconjugates of olfactory mucosae using lectin histochemistry.

Lectin histochemistry at the light microscope level was used to determine the distribution of sugar residues in secretory cells of the olfactory mucosae of salamander, hamster, and mouse. Differences in sugar composition and distribution of glycoconjugates found in sustentacular cells and acinar cells of Bowman's glands of these three animals were characterized. Oligosaccharides in secretory products of sustentacular cells in salamander olfactory mucosa contained sialic acid, galactose (Gal), N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc), fucose, and mannose residues. Glycoconjugates of these cells lacked terminal galactosyl-beta-(1,3)N-acetylgalactose (Gal beta 1,3GalNAc) residues. The sequences Gal beta 1,3GalNAc, N-acetyllactosamine (Gal beta 1,4GlcNAc), and GalNAc were penultimate to sialic acid residues. Sustentacular cells of mouse and hamster did not appear to contain O-linked oligosaccharides but stained for mannose-containing N-linked oligosaccharides. Glycoconjugates of acinar and duct cells of Bowman's glands in the salamander, hamster, and mouse contained variable amounts of beta(1,4)GlcNAc residues, and terminal N-acetyllactosamine, Gal beta 1,3GalNAc, and GalNAc residues. In the salamander, glycoconjugates of acinar cells possessed terminal GlcNAc residues but were not sialylated, while those of hamster and mouse generally stained for sialic acid but did not possess terminal GlcNAc residues. Secretory products of a subpopulation of rodent acinar cells also contained penultimate Gal beta 1,3GalNAc residues. Staining for sialic acid, Gal, GalNAc, and GlcNAc in glycoconjugates of rodents was often limited to a sub-population of Bowman's glands. This was especially noticeable in the mouse.

Ambystoma↗

Fibroblast growth factors in regenerating limbs of Ambystoma: cloning and semi-quantitative RT-PCR expression studies.

Urodele amphibians (newts and salamanders) have the ability to regenerate amputated limbs throughout their life span. Because fibroblast growth factors (Fgfs) play important roles in developing limbs, we initiated studies to investigate these growth factors in regenerating limbs. Partial cDNAs of Fgf4, 8, and 10 were cloned from both the Mexican axolotl, Ambystoma mexicanum, and locally collected spotted salamander, Ambystoma maculatum, two salamanders well recognized for their regenerative capabilities. cDNAs from the two Ambystoma species were virtually identical, ranging from 97-100% nucleotide identity. Axolotl Fgf4, 8, and 10 showed nucleotide sequence identity with chick Fgf4, 8, and 10 of 79%, 83%, and 72%, respectively. RT-PCR showed that these growth factors are expressed in regenerating axolotl limbs as well as in developing salamander larvae at the three-digit forelimb stage. Fgf8 and 10 are upregulated during regeneration and thus may be involved in distal signaling similar to that of the developing chick limb. Fgf4, however, was undetectable by RT-PCR in the distal tips of regenerates, suggesting that it does not play the same role in limb regeneration that it does in limb development. We also investigated the role these Fgfs may have in the nerve-dependence of regeneration. They were expressed similarly in aneurogenic and innervated limbs, suggesting that they are not the neurotrophic factors responsible for nerve-dependence. Denervation prevented Fgf8 and 10 upregulation, suggesting Fgf pathways are downstream of nerve-dependence. These data highlight important similarities and differences in Fgf expression between limb development and limb regeneration. J. Exp. Zool. 290:529-540, 2001.

Ambystoma↗

Molecular evidence for the early history of living amphibians.

The evolutionary relationships of the three orders of living amphibians (lissamphibians) has been difficult to resolve, partly because of their specialized morphologies. Traditionally, frogs and salamanders are considered to be closest relatives, and all three orders are thought to have arisen in the Paleozoic (>250 myr). Here, we present evidence from the DNA sequences of four mitochondrial genes (2.7 kilobases) that challenges the conventional hypothesis and supports a salamander-caecilian relationship. This, in light of the fossil record and distribution of the families, suggests a more recent (Mesozoic) origin for salamanders and caecilians directly linked to the initial breakup of the supercontinent Pangaea. We propose that this single geologic event isolated salamanders and archaeobatrachian frogs on the northern continents (Laurasia) and the caecilians and neobatrachian frogs on the southern continents (Gondwana). Among the neobatrachian frog families, molecular evidence supports a South American clade and an African clade, inferred here to be the result of mid-Cretaceous vicariance.

Africa↗

Immunolocalization of the electrogenic Na+-HCO-3 cotransporter in mammalian and amphibian kidney.

Electrogenic cotransport of Na+ and HCO-3 is a crucial element of HCO-3 reabsorption in the renal proximal tubule (PT). An electrogenic Na+-HCO-3 cotransporter (NBC) has recently been cloned from salamander and rat kidney. In the present study, we generated polyclonal antibodies (pAbs) to NBC and used them to characterize NBC on the protein level by immunochemical methods. We generated pAbs in guinea pigs and rabbits by immunizing with a fusion protein containing the carboxy-terminal 108 amino acids (amino acids 928-1035) of rat kidney NBC (rkNBC). By indirect immunofluorescence microscopy, the pAbs strongly labeled HEK-293 cells transiently expressing NBC, but not in untransfected cells. By immunoblotting, the pAbs recognized a approximately 130-kDa band in Xenopus laevis oocytes expressing rkNBC, but not in control oocytes injected with water or cRNA for the Cl-/HCO-3 exchanger AE2. In immunoblotting experiments on renal microsomes, the pAbs specifically labeled a major band at approximately 130 kDa in both rat and rabbit, as well as a single approximately 160-kDa band in salamander kidney. By indirect immunofluorescence microscopy on 0.5-micrometer cryosections of rat and rabbit kidneys fixed in paraformaldehyde-lysine-periodate (PLP), the pAbs produced a strong and exclusively basolateral staining of the PT. In the salamander kidney, the pAbs labeled only weakly the basolateral membrane of the PT. In contrast, we observed strong basolateral labeling in the late distal tubule, but not in the early distal tubule. The specificity of the pAbs for both immunoblotting and immunohistochemistry was confirmed in antibody preabsorption experiments using either the fusion protein used for immunization or similarly prepared control fusion proteins. In summary, we have developed antibodies specific for NBC, determined the apparent molecular weights of rat, rabbit, and salamander kidney NBC proteins, and described the localization of NBC within the kidney of these mammalian and amphibian species.

Ambystoma↗

Juvenile amphibians do not avoid potentially lethal levels of urea on soil substrate.

We examined the effects of a forest fertilizer (urea) on newly metamorphosed terrestrial amphibians (Western toads, Bufo boreas; Cascades frogs, Rana cascadae; long-toed salamanders, Ambystoma macrodactylum; and roughskin newts, Taricha granulosa). We examined avoidance behavior of Western toads and Cascades frogs on both paper towel and soil substrates dosed with urea (control and 100 kg N/ha and an additional treatment of 50 kg N/ha for Western toads on soil substrate) and avoidance behavior of long-toed salamanders on soil substrate dosed with urea. We further examined the survival and feeding behavior of all four species exposed to urea on soil substrate (100 kg N/ha) for 5 d. Juvenile Western toads and Cascades frogs avoided paper towels dosed with urea but did not avoid urea-dosed soil substrate. However, Western toads and Cascades frogs both suffered significant mortality when exposed to urea on a soil substrate for 5 d. Furthermore, after adjusting for weight, we found that urea-exposed juvenile Western toads and Cascades frogs consumed significantly fewer prey items (crickets) compared with nonexposed control animals. Long-toed salamanders did not discriminate against soil substrate dosed with urea, and neither long-toed salamanders nor roughskin newts died or reduced prey consumption as a result of urea exposure. Juvenile amphibians may not be able to detect and avoid harmful levels of urea fertilizer on a natural substrate. Furthermore, anthropogenic stressors such as urea fertilizer can significantly reduce the survival and prey consumption of juvenile amphibians. These effects are important to consider in light of possible threats to the conservation status of many amphibian species.

Agriculture↗

Lamina-associated polypeptide 2 (LAP2) expression in fish and amphibians.

Somatic and germinal cells of 15 fish and 33 amphibian species were examined by SDS-PAGE followed by immunoblotting to determine the expression of LAP2 (lamina-associated polypeptide 2). LAP2 expression in frogs, salamanders and fish does not vary with the mode of reproduction. In fish and frog cells, a rim-like LAP2 positive region was detected around the nucleus by indirect immunofluorescence microscopy. The cell distribution and expression patterns of LAP2 in fish, frogs and salamanders are comparable with those found in Xenopus and zebrafish. The mammalian somatic cell pattern, which may also occur in gymnophione amphibians, includes LAP2alpha, beta and gamma as major isoforms, whereas LAP2alpha does not occur in cells of fish, frogs and salamanders. In fish, LAP2gamma is the major isoform of somatic cells, suggesting that LAP2gamma may be ancestral. However, in the rainbow trout, as in frogs and salamanders, LAP2beta was the major somatic isoform. Fish and frog sperm only express low molecular weight polypeptides. In contrast, fish and frog oocytes express an oocyte-specific LAP2 isoform of high molecular weight. In the toad Bufo marinus this isoform becomes upregulated in pre-vitellogenic oocytes of 150-200 microm in diameter. The absence of LAP2alpha and the differential expression of LAP2 isoforms in somatic and germ cells, as found in fish and frogs, may be ancestral vertebrate characters. In spite of differences in developmental time, the LAP2 isoforms of somatic cells are upregulated during gastrulation, suggesting that LAP2 may be implicated in the early development of fish and frog.

Animals↗

Postcranial myology of the California newt, Taricha torosa.

Salamanders are generally agreed to represent the primitive tetrapod body plan, as well as a postural analog for early tetrapods. Dissection and description of the muscles of the forelimb, trunk, and hindlimb of the California newt, Taricha torosa, were undertaken to provide baseline data on the locomotor structures in this species. Hypaxial trunk muscles are similar to those of other vertebrates. As in other salamanders, limb muscles show a simple parallel-fibered architecture and often span multiple joints. Several differences in limb musculature were also noted. The extensor iliotibialis muscle possesses a single head in T. torosa, rather than the two heads common in larger salamander species. The ischioflexorius muscle, while divided into proximal and distal sections, is not distinct from the puboischiotibialis in its proximal portion. The femorofibularis is enlarged in this species; it is suggested that the femorofibularis and ischioflexorius muscles may be functionally analogous systems. Forelimb and hindlimb musculature show similar morphological patterns, particularly in distal limb segments, which may provide insight into the primitive arrangement of tetrapod limb muscles.

Abdominal Muscles↗

Acceleration of Ambystoma tigrinum metamorphosis by corticotropin-releasing hormone.

Previous work of others and ours has shown that corticotropin-releasing hormone (CRH) is a positive stimulus for thyroid and interrenal hormone secretion in amphibian larvae and that activation of CRH neurons may mediate environmental effects on the timing of metamorphosis. These studies have investigated CRH actions in anurans (frogs and toads), whereas there is currently no information regarding the actions of CRH on metamorphosis of urodeles (salamanders and newts). We tested the hypothesis that CRH can accelerate metamorphosis of tiger salamander (Ambystoma tigrinum) larvae. We injected tiger salamander larvae with ovine CRH (oCRH; 1 microg/day; i.p.) and monitored effects on metamorphosis by measuring the rate of gill resorption. oCRH-injected larvae completed metamorphosis earlier than saline-injected larvae. There was no significant difference between uninjected and saline-injected larvae. Mean time to reach 50% reduction in initial gill length was 6.9 days for oCRH-injected animals, 11.9 days for saline-injected animals, and 14.1 days for uninjected controls. At the conclusion of the experiment (day 15), all oCRH-injected animals had completed metamorphosis, whereas by day 15, only 50% of saline-injected animals and 33% of uninjected animals had metamorphosed. Our results show that exogenous oCRH can accelerate metamorphosis in urodele larvae as it does in anurans. These findings suggest that the neuroendocrine mechanisms controlling metamorphosis are evolutionarily conserved across amphibian taxa.

Ambystoma↗

Cellular contribution to symmetrical forelimbs from triploid-marked "polarizing region" in the embryo of axolotl, Ambystoma mexicanum.

Grafts of posterior tissue placed anterior to the limb bud in the salamander embryo exert a polarizing influence. To explain this result, the idea that the anteroposterior axis of the developing forelimb is polarized by a diffusible morphogen has been proposed. An alternative hypothesis, and the working hypothesis of the present study, is that the polarization of the developing salamander forelimb is accomplished by short-range cellular interactions resulting in intercalation rather than by the more global influence of a diffusible morphogen. One prediction of this intercalation hypothesis is that cells will be contributed to the limb from the "polarizing tissue." To test this idea, grafts of triploid marked polarizing tissue were implanted anterior to the limb bud in 82 diploid axolotl embryos at stages 32-34 of development. A total of 27 (33%) of the limbs that resulted were symmetrical and ranged in complexity from one to seven digits. Histological analysis of a subgroup of the original symmetrical limbs revealed that mesodermally derived tissues in the anterior side of these limbs (the side which formed as a duplication in response to the influence of the graft) contained high percentages of trinucleolate cells (muscle, 12.1%; connective tissue tissue, 12.5%; and cartilage, 13.4%) when compared to similar tissues in the posterior side of the same symmetrical limbs (muscle, 1.8%; connective tissue , 0.7%; and cartilage, 0.6%). When symmetrical limbs were amputated, 73% regenerated symmetrical limbs. When these regenerated limbs were again amputated, 63% formed symmetrical secondary regenerates. Histological analysis of the first generation of regenerated limbs revealed that the pattern of distribution of trinucleolate cells in each regenerate was similar to the pattern seen in the original symmetrical limb. These results indicate that there is considerable cellular contribution to the anterior side of the symmetrical forelimb from the mesoderm of grafted "polarizing tissue." This result supports the idea that short-range cellular interaction are sufficient for formation of symmetrical forelimbs in salamander embryos.

Ambystoma↗

Effects of lung reduction on rheotactic performance in amphibian larvae.

In this study we test the hypothesis that lung reduction in amphibians is a larval adaptation to reduce buoyancy and facilitate rheotaxis in lotic environments. Larvae of the spotted salamander Ambystoma maculatum (family Ambystomatidae) were raised from eggs with (= control) and without (= experimental) access to the air-water interface. The experimental larvae developed significantly smaller and more solidified lungs than the controls. Although mean snout-vent length did not differ between the control and experimental groups, the latter had a significantly greater mean body mass, indicating greater specific gravity in experimental versus controls. To evaluate rheotactic behavior, larvae were subjected to a water current in an artificial stream. The distance and time required to establish a purchase on the stream bottom were recorded. Although the times did not differ, the distances displaced downstream were significantly less in experimental than in control larvae. We conclude that lung reduction and solidification reduce buoyancy and enhance rheotactic ability in salamander larvae. These findings suggest that lunglessness in plethodontid salamanders evolved as an adaptation to stream environments.

Adaptation, Biological↗

Distribution and morphology of motoneurons innervating different muscle fiber types in an amphibian muscle complex.

The distribution and morphology of motoneurons innervating specific types of muscle fibers in the levator scapulae superior (LSS) muscle complex of the bullfrog (Rana catesbeiana) and tiger salamander (Ambystoma tigrinum) were studied by retrograde labelling with cholera toxin-conjugated horseradish peroxidase (CT-HRP). The LSS muscle complex in both of these amphibians has a segregated pattern of muscle-fiber types (tonic; fast oxidative-glycolytic twitch [FOG]; fast glycolytic twitch [FG]) along an anteroposterior axis. The entire motor pool was labelled by injection of CT-HRP into the whole LSS muscle complex. The motoneurons innervating specific fiber types were labelled by injection of CT-HRP into certain muscle regions. The organization of the motoneuron pool of the LSS complex of both species was arranged in two columns--one ventrolateral and one medial. In bullfrogs, the ventrolateral column contains motoneurons innervating FG and tonic fiber types and the medial column contains motoneurons innervating FOG fiber types. In tiger salamanders, the ventrolateral column contains motoneurons innervating FG fiber types and the medial column contains motoneurons innervating FOG and tonic fiber types. The different motoneuron types also have different soma sizes and patterns of dendritic arborization. In both species, FG motoneurons are the largest, whereas FOG motoneurons are intermediate in size and tonic motoneurons are the smallest. In bullfrogs, the main dendrites of FG motoneurons extend into the dorsolateral and the ventrolateral gray matter of the spinal cord, whereas the dendrites of FOG motoneurons extend into the ventral and medial cord. In the tiger salamander, dendrites of FG motoneurons extend into the ventrolateral spinal cord and dendrites of the FOG motoneurons extend more generally into the ventral cord. Dendrites of tonic motoneurons in both amphibians were small and short, and difficult to observe. These results establish that motoneurons innervating different types of muscle fibers in the LSS muscle complex are segregated spatially and display consistent morphological differences.

Ambystoma↗

Pathogen transmission as a selective force against cannibalism.

Cannibalism is uncommon in most species despite being taxonomically widespread. This rarity is surprising, because cannibalism can confer important nutritional and competitive advantages to the cannibal. A general, but untested, explanation for why cannibalism is rare is that cannibals may be especially likely to acquire pathogens from conspecifics, owing to greater genetic similarity among conspecifics and selection for host specificity and resistance to host immune defences among pathogens. We tested this hypothesis by contrasting the fitness consequences of intra- versus interspecific predation of diseased and non-diseased prey. We fed cannibalistic tiger salamander (Ambystoma tigrinum) larvae diseased conspecifics, healthy conspecifics, diseased heterospecifics (a sympatric congener, small-mouthed salamanders, A. texanum), or healthy heterospecifics. Cannibals that ate diseased conspecifics were significantly less likely to survive to metamorphosis and grew significantly less than those that ate diseased heterospecifics, but none of the other groups differed. Tiger salamander larvae also preferentially preyed on heterospecifics when given a choice between healthy conspecifics and heterospecifics. These results suggest that pathogen transmission is an important cost of cannibalism and provide a general explanation for why cannibalism is infrequent in most species. Copyright 1998 The Association for the Study of Animal Behaviour Copyright 1998 The Association for the Study of Animal Behaviour.

Journal Article↗

Development of the amphibian oculomotor complex: evidences for migration of oculomotor motoneurons across the midline.

The development of the oculomotor nucleus in five species of salamanders and one anuran species was investigated with tracing techniques. The data presented support the hypothesis that oculomotor motoneurons innervating the superior rectus muscle migrate across the midline. In the salamander Pleurodeles waltl, only ipsilateral oculomotor motoneurons are labeled in early development. Later, these neurons extend dendrites toward the contralateral side into the ventral tegmental neuropil, after which there is displacement of their nuclei (neuronal somata) across the midline. Cell bodies can be observed directly at the midline. In adult Salamandra salamandra, motoneurons innervating the superior rectus muscle are seen occasionally at the midline and on the ipsilateral side, with dendrites toward the contralateral side. Motoneurons on the ipsilateral side do not display these features. In Pleurodeles, developmental brain processes are slowed down, and the sequence of development of the contralateral subnucleus, which can be clearly observed, supports the migration hypothesis. In Xenopus laevis and most other species of salamanders this process is accelerated.

Ambystoma mexicanum↗

Bulgy tadpoles: inducible defense morph.

Predator induced morphological defenses are marked morphological shifts induced directly by cues associated with a predator. Generally, remote cues, i.e., chemical substances emitted from predators or injured conspecifics, are considered to be ideal signals to induce morphological change in aquatic environments rather than close cues, i.e., close chemical or tactile cues, since chemical substances that can propagate over relatively long distances and persist for a long period may allow organisms to keep safe and to deliberately change their morph. In fact, most organisms adopting an inducible morphological defense utilize remote chemical cues to detect predation risk and to produce morphological defenses. In this paper, we report a unique and functionally well designed inducible morphological defense strategy where the induction process requires close cues from a predator. The tadpoles of Rana pirica exhibited a bulgy bodied morphology when threatened with predation by larval salamanders, Hynobius retardatus, in close proximity. Predation trials and a function experiment showed that the induced bulgy morph is an adaptive defense phenotype against the gape-limited predator larval H. retardatus. Furthermore, R. pirica tadpoles use two adaptive strategies in terms of cost saving, i.e., adjustment of the extent of bulginess according to predation risk and reversibility by actual shrink of bulgy body after removing the predation threat. In general, R. pirica hatch earlier than H. retardatus. In natural ponds, during the early developmental stage R. pirica tadpoles live in close proximity to young H. retardatus larvae. As they grow, the salamanders gradually become serious predators and the predator-prey interaction becomes intimate. After a while, predation, cannibalism and metamorphosis decrease the number of salamanders in the ponds, and the predator-prey interaction weakens. Such a phenology in the predator-prey interaction allows the evolution of a close-cue detection system and adaptive cost-saving strategies. Our results highlight that the characteristics of the inducible defense depend on the intensity and specificity of the predator-prey system.

Adaptation, Physiological↗

Studies of the voltage-dependent polyspermy block using cross-species fertilization of amphibians.

Fertilization of frog eggs by frog sperm is inhibited if the egg's membrane potential is positive (N.L. Cross and R.P. Elinson, 1980, Dev. Biol. 75, 187-198); however, fertilization of salamander eggs by salamander sperm does not depend on membrane potential (M. Charbonneau, M. Moreau, B. Picheral, J.P. Vilain, and P. Guerrier, 1983, Dev. Biol. 98, 304-318). Since salamander sperm can fertilize frog eggs, we have investigated whether this cross-fertilization is voltage dependent. If, during insemination with Notophthalmus sperm, Xenopus eggs were voltage clamped between +7 and +20 mV, fertilization proceeded in 7/10 (70%) of the clamped eggs, compared to 38/48 (79%) of the neighboring eggs. In control experiments in which voltage-clamped Xenopus eggs were inseminated with Xenopus sperm, fertilization proceeded in only 1/10 (10%) of the clamped eggs, compared to 59/60 (98%) of the neighbors. Similar results were obtained with cross-fertilization experiments between Notophthalmus sperm and Rana eggs. These experiments indicate that the voltage dependence of fertilization depends on the species of sperm.

Amphibians↗

Effects of environmental O2 on blood flow and diffusing capacity in amphibian skin.

The effects of local environmental PO2 on cutaneous blood flow (Q) and the membrane diffusing capacity of the skin (D) were investigated in the leopard frog, Rana pipiens, and the lungless salamander, Desmognathus quadramaculatus. Halothane anesthetized animals were equilibrated with Freon-22 (Fr) and acetylene (Ac) in a box. A gas mixture containing either 0, 20 or 40% O2, respectively, in N2 and initially free of Fr and Ac was drawn through a small sample chamber on the abdomen. The excretion of Fr and Ac into the chamber was analyzed with a mass spectrometer. These conditions allowed the determination of the cutaneous conductance to Fr (GFr) and Ac (GAc) at the 3 levels of local environmental O2. GFr and GAc of the isolated skin and the blood solubilities of the 2 gases were also determined. To estimate Q, DAc and DFr, the data were analyzed with a homogeneous single capillary model. In the frog, Q varied directly with sample chamber [O2], but chamber [O2] had no effect on D. In the salamander, O2 had no effect on either Q or D. The results indicate that regulation of cutaneous gas exchange in the frog by local environmental O2 only involves alterations in Q. Similar control of cutaneous gas exchange is absent in the lungless salamander.

Acetylene↗

Antagonistic maternal immune reactions (rejection and facilitation) to the embryo in the urodele amphibian Salamandra salamandra lin.

In vitro assays have been employed to demonstrate that pregnant salamanders mount an immune reaction against their embryos. Maternal spleen cells kill up to 85% of dissociated embryonic epidermal cells during a 48 h incubation period. The degree of killing depends upon the ratio of maternal to embryonic cells and on the number of embryos borne by the mother. The cytotoxicity shows considerable specificity for the embryos of a given mother although a weak degree of killing can occur with embryos from other mothers, presumably due to some form of cross-reactivity. The effect is inhibited by the addition of maternal serum to the cultures. The degree of protection is also a function of the number of embryos borne by the mother. Pre-incubation experiments indicate that the maternal serum has a protective action on the embryonic cells which is largely specific for the female's own embryos (and suggested to be antibody in nature) and an inhibitory action on the maternal spleen cells which occurs also with spleen cells of other females (and suggested to be either an immune complex or a nonimmunological substances). An increase in beta protein peaks is seen following electrophoresis of sera from pregnant (and also allografted) salamanders. These findings indicate that the pregnant salamander mounts a double immune reaction against her embryos, an aggressive (rejection) reaction and a protective (facilitation) reaction.

Absorption↗