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PubMed1990

Effects of modified chromophores on the spectral sensitivity of salamander, squirrel and macaque cones.

1. Chemically modified retinal chromophores were used to investigate the mechanisms that produce the characteristic spectral absorptions of cone pigments. Spectral sensitivities of single cones from the salamander, squirrel and macaque retina were determined by electrical recording. The chromophore was then replaced by bleaching the pigment and regenerating it with a retinal analogue. 2. Exposing a bleached cone to 9-cis-retinal for a brief period (less than 20 min) caused its flash sensitivity to recover to about 0.2 of the pre-bleach value. Similar exposure to a locked 6-s-cis, 9-cis analogue gave a recovery to about 0.03 of the pre-bleach value. 3. Unlike the flash sensitivity, the saturating photocurrent amplitude often recovered completely after bleaching and regenerating the pigment. 4. When the 3-dehydroretinal chromophore in the salamander long-wavelength-sensitive (red) cone was replaced with 11-cis-retinal, shortening the conjugated chain in the chromophore, the spectral sensitivity underwent a blue shift of 67 nm. 5. Pigments containing the planar-locked 6-s-cis.9-cis-retinal analogue absorbed at substantially longer wavelength than those containing unmodified 9-cis-retinal. The opsin shift, a measure of the protein's ability to modify the chromophore's absorption was larger for the locked analogue than for 9-cis-retinal. This suggests that the native chromophore assumes a twisted 6-s-cis conformation in these pigments. 6. The spectral sensitivities of red and green macaque cones containing 9-cis-retinal or planar-locked 6-s-cis.9-cis-retinal retained the 30 nm separation characteristic of the native pigments. This suggests that the different absorptions of of the 6-7 carbon bond in the retinal chromophore.

Animals↗
PubMed1993

GABAA and glutamate receptor involvement in dendrodendritic synaptic interactions from salamander olfactory bulb.

1. Whole-cell patch clamp and optical recording techniques were applied to the same in vitro salamander olfactory bulb preparations to study the postsynaptic responses of single mitral/tufted cells in the context of the surrounding neural activity in which they are embedded. Mitral/tufted cells were identified by intracellular filling with biocytin. 2. Single mitral/tufted cells were under a tonic GABAA receptor-mediated inhibitory influence as revealed by the recording of bicuculline methiodide (BMI)/picrotoxin-sensitive inhibitory postsynaptic currents (IPSCs) in symmetrical chloride conditions at a holding potential of -70 mV. Depolarizing voltage steps (100 ms) applied to single cells or electrical stimulation of the olfactory nerve or medial olfactory tract evoked a prolonged increase in the frequency of GABAergic IPSCs. 3. The frequency of spontaneous and driven IPSCs was reduced with application of the glutamate receptor antagonists 6-cyano-2,3-dihydroxy-7-nitro-quionoxaline (CNQX) or 2-amino-5-phosphonopentanoic acid (AP5) whereas olfactory nerve- or medial olfactory tract-driven IPSC frequency was enhanced with removal of bathing Mg2+, indicating that GABAergic interneurones were driven by mitral/tufted cells at both non-NMDA and NMDA receptors. 4. Olfactory nerve or medial olfactory tract stimulation evoked widely distributed changes in fluorescence in preparations stained with the voltage-sensitive dye RH414. The optical response predominantly consisted of a decrease in fluorescence, indicative of depolarization. The presence of the dye did not obviously affect mitral/tufted cell postsynaptic responses. 5. BMI enhanced the amplitude and duration of optical signals related to depolarization within the bulb and in regions central to the bulb. In the presence of BMI, depolarizing activity appeared to spread hundreds of micrometres into regions of the bulb not activated in control conditions showing explicitly that GABAA receptors in the bulb participate in lateral inhibition. 6. CNQX and AP5 attenuated the optical signals within the bulb supporting the contention that in these conditions, optical signals arise mainly from granule cell dendritic activity. Furthermore, AP5 or removal of bath Mg2+ reduced or enlarged the spatial distribution of activity respectively, suggesting that in some cases the NMDA receptor may be involved in generating or stabilizing spatial patterns of activity. 7. It is concluded that in the salamander olfactory bulb, both GABAA- and glutamate receptor-mediated synaptic transmission shape the different temporal and spatial patterns of neural activity associated with olfactory coding.

2-Amino-5-phosphonovalerate↗
PubMed1994

GABAergic and glutamatergic synaptic input to identified granule cells in salamander olfactory bulb.

1. Whole-cell patch clamp recording techniques were applied to granule cells in an in vitro salamander olfactory bulb preparation to study their morphology, membrane properties and pharmacology of postsynaptic responses to electrical stimulation of either the olfactory nerve (ON) or medial olfactory tract (MOT). Optical recordings of the same preparations stained with the voltage-sensitive dye RH414 were also made. 2. Anatomical reconstructions of biocytin-filled granule cells showed that they extend widespread spine-bearing dendrites and an axon-like process that branched within the external plexiform layer. 3. ON or MOT stimulation evoked a long-lasting depolarization, usually generating only a single action potential, in granule cells studied under standard recording conditions. Bath application of bicuculline methiodide (BMI, a GABAA receptor antagonist, 20 or 25 microM) enhanced the spontaneous and electrically evoked excitatory drive to granule cells. 4. The electrically evoked synaptic responses consisted of both excitatory and inhibitory synaptic inputs. Using symmetrical Cl- conditions inside and outside the cell to enhance Cl- currents, spontaneous and electrically driven BMI-sensitive inhibitory postsynaptic currents (IPSCs) were revealed, indicating that granule cells receive GABAergic synaptic input. 5. Bath application of GABA (250 microM to 1 mM) shunted and hyperpolarized granule cells as observed directly from whole-cell recordings and indirectly from cell-attached patch single channel recordings. 6. Bath application of the glutamate receptor antagonists 6-cyano-2,3-dihydroxy-7-nitroquinoxaline (CNQX, 10 microM) and/or DL-2-amino-5-phosphonopentanoic acid (DL-AP5, 100 microM) showed that granule cell dendrodendritic EPSPs are shaped by both non-NMDA and NMDA receptors. 7. The time course and pharmacological sensitivity of both single granule cell responses and ensemble responses recorded optically in the deeper layers of the bulb correlated well. 8. It is concluded that salamander granule cells integrate several types of synaptic input, may have both dendritic and axonal output, and play a major role in generating voltage-sensitive dye signals in the olfactory bulb.

Afferent Pathways↗
PubMed2005

Physiological properties of rod photoreceptor electrical coupling in the tiger salamander retina.

Using dual whole-cell voltage and current clamp recording techniques, we investigated the gap junctional conductance and the coupling coefficient between neighbouring rods in live salamander retinal slices. The application of sinusoidal stimuli over a wide range of temporal frequencies allowed us to characterize the band-pass filtering properties of the rod network. We found that the electrical coupling of all neighbouring rods exhibited reciprocal and symmetrical conductivities. On average, the junctional conductance between paired rods was 500 pS and the coupling coefficient (the ratio of voltage responses of the follower cell to those of the driver cell), or K-value, was 0.07. Our experimental results also demonstrated that the rod network behaved like a band-pass filter with a peak frequency of about 2-5 Hz. However, the gap junctions between adjacent rods exhibited linearity and voltage independency within the physiological range of rods. These gap junctions did not contribute to the filtering mechanisms of the rod network. Combined with the computational modelling, our data suggest that the filtering of higher frequency rod signals by the network is largely mediated by the passive resistive and capacitive (RC) properties of rod plasma membranes. Furthermore, we found several attributes of rod electrical coupling resembling the physiological properties of gene-encoded Cx35/36 gap junctions examined in other in vitro studies. This indicates that the previously found Cx35/36 expression in the salamander rod network may be functionally involved in rod-rod electrical coupling.

Action Potentials↗
PubMed1999

Proteinaceous pheromone affecting female receptivity in a terrestrial salamander.

A 22-kilodalton protein was isolated from the submandibular (mental) gland of the male terrestrial salamander, Plethodon jordani (family: Plethodontidae). This proteinaceous pheromone, termed plethodontid receptivity factor (PRF), was experimentally delivered to the female during courtship and shown to increase female receptivity. In most plethodontid salamanders, ovulation occurs weeks or months after insemination, so the pheromone-induced change in receptivity is the only known function of PRF. The messenger RNAs corresponding to isoforms of PRF were transcribed into complementary DNA, cloned, sequenced, and shown to have homology with cytokines of the interleukin-6 family. Pheromone activity would represent a previously unrecognized function for cytokines.

Amino Acid Sequence↗
PubMed1985

Morphological novelty in the limb skeleton accompanies miniaturization in salamanders.

Salamanders of the genus Thorius (Plethodontidae) are among the smallest tetrapods. Hypotheses of limb skeletal evolution in these vertebrates were evaluated on the basis of estimates of natural variation, comparisons of skeletal homology, and analysis of molecular phylogeny. Nine carpal arrangements occur in Thorius, more than in all twelve related genera of typically larger salamanders; six of these arrangements are unique. They represent a trend toward a decrease in the number of separate cartilages that is independent of locomotor and ecological specialization. Miniaturization may be an important source of morphological novelty, distinct from local adaptation, in vertebrates.

Animals↗
PubMed1994

Primary culture of salamander intestinal epithelial cells from nests: whole cell Na+ currents induced by valine.

Methods are described for isolating the cell nests, subepithelial clusters of germinative cells, from salamander intestinal mucosa and for growing the nests in culture into polarized monolayers of intestinal epithelial cells. Cells were viable in culture for up to 3 wk. The capacity of the monolayer cells to engage in membrane transport was evaluated using the patch-clamp technique in the whole cell mode. L-Valine (25 mM) induced an inward current in small intestinal cells of 25.8 +/- 5.7 pA and depolarized the cell membrane 14.5 +/- 1.6 mV. L-Alanine and L-phenylalanine were similarly effective, whereas D-valine was ineffective. The Km of the transporter for valine was 90 mM. Replacement of bath Na with tris(hydroxymethyl)aminomethane eliminated the inward current induced by valine. The basal (solute-independent) inward current was also reduced by Na+ replacement. Glucose did not induce a Na+ current. In contrast to the effect of valine on small intestinal cells, large intestinal cells were unresponsive to valine. It is concluded that the cultured small intestinal cells possess Na-amino acid but not Na-sugar cotransport. This profile of behavior is characteristic of undifferentiated small intestinal cells. Primary cultures of salamander small intestinal cells should be useful for studying enterocyte function and the developmental biology of the small intestinal mucosa.

Animals↗
PubMed1986

Locomotion without lungs: energetics and performance of a lungless salamander.

Lungless salamanders (4.1 g) were exercised on a treadmill enclosed in a Plexiglas respirometer at a range of speeds (0.05-0.24 km/h). O2 consumption (VO2) was determined continuously by open-flow respirometry. At the onset of exercise VO2 increased to a "steady state" in approximately 2-5 min. VO2 (ml O2 X g-1 X h-1) increased linearly with speed (S), VO2 = 2.3 (S) + 0.1 (r2 = 0.74). The minimum cost of transport (2.3 ml O2 X g-1 X km-1) was three-fourths of that predicted for a vertebrate of the same mass. Maximum O2 consumption (VO2max) was attained at 0.16 km/h (maximum aerobic speed), where this VO2 was six to nine times above standard rates. The net rate of whole-body lactate production (WBL) was insignificant at slow speeds (less than 46% VO2max) but did increase at submaximal work loads (85% VO2max). The highest WBL (0.07 mg X g-1 X min-1) was measured at a speed (0.20 km/h) that exceeded the maximum aerobic speed. Salamanders sustained exercise at slow speeds for over 2 h. The greatest decline in endurance (i.e., from 120 to 10 min) occurred at speeds below the maximum aerobic speed, where being lungless may have little consequence. At fast rates of locomotion a limited VO2max was associated with a low maximum aerobic speed and a modest capacity for sustained activity.

Animals↗
PubMed1981

Isolated perfused salamander proximal tubule: methods, electrophysiology, and transport.

Techniques are presented for the isolation and perfusion of renal proximal tubules from the neotenic salamander Ambystoma tigrinum. Methods are described for a determination of normal values for fluid transport and electrophysiological parameters. Stable cellular microelectrode recordings are reported that constitute the first intracellular measurements in an isolated perfused tubule preparation. With identical solutions in lumen and bath, fluid reabsorption averaged 0.28 nl.min-1.mm-1, transepithelial potential difference averaged -4.5 mV, transepithelial resistance was 52.1 omega.cm2, and the transepithelial chloride-to-sodium transference number ratio was 3.4. The basolateral cell membrane potential difference averaged -59.6 mV, and the ratio of apical-to-basolateral cell membrane resistance was between 3.9 and 5. Viability of the isolated perfused salamander proximal tubule preparation is demonstrated by a detailed comparison of the present data with results of in vivo micropuncture experiments on both Necturus and intact Ambystoma kidneys. In addition to being an advantageous preparation for long-term intracellular recordings, the Ambystoma kidney is unique in that proximal tubules can be studied both in isolation and by conventional micropuncture.

Ambystoma↗
PubMed1986

Electrophysiology of salamander proximal tubule. II. Interspace NaCl concentrations and solute-coupled water transport.

The role of the paracellular interspace in solute-coupled water transport was investigated in isolated perfused salamander (Ambystoma) proximal tubules using a null-point technique to estimate interspace NaCl concentrations. Constant composition of luminal fluid was maintained by rapid (200 nl/min) perfusion of tubules 600,micron or less in length. Inhibition of active transport by a decrease in bath temperature from 22 to 0 degrees C in 400 ms produced rapid depolarizations of both the transepithelial (Vte) and basolateral (Vbl) potential, followed by slower changes in potential that occurred at low temperature. During this period, the time course of Vbl was independent of small changes in bath NaCl concentration, whereas the time course of Vte at low temperature varied from a slow depolarization to a slow repolarization depending on whether the concentration of NaCl in the bath equaled or exceeded that in the perfusate. Absence of a slow change in Vte at low temperature indicated a match between the NaCl concentration of the interspace and the test concentration of NaCl in the bath. Using this technique with 12 tubules, the normal interspace NaCl concentration appeared to be approximately 4% above the NaCl concentration of either the lumen or bath, demonstrating that the interspace of the salamander proximal tubule can function as a local hyperosmotic compartment to facilitate fluid transport between solutions of identical composition.

Ambystoma↗
PubMed2005

Functional organization of ganglion cells in the salamander retina.

Recently, we reported a novel technique for recording all of the ganglion cells in a retinal patch and showed that their receptive fields cover visual space roughly 60 times over in the tiger salamander. Here, we carry this analysis further and divide the population of ganglion cells into functional classes using quantitative clustering algorithms that combine several response characteristics. Using only the receptive field to classify ganglion cells revealed six cell types, in agreement with anatomical studies. Adding other response measures served to blur the distinctions between these cell types rather than resolve further classes. Only the biphasic off type had receptive fields that tiled the retina. Even when we attempted to split these classes more finely, ganglion cells with almost identical functional properties were found to have strongly overlapping spatial receptive fields. A territorial spatial organization, where ganglion cell receptive fields tend to avoid those of other cells of the same type, was only found for the biphasic off cell. We further studied the functional segregation of the ganglion cell population by computing the amount of visual information shared between pairs of cells under natural movie stimulation. This analysis revealed an extensive mixing of visual information among cells of different functional type. Together, our results indicate that the salamander retina uses a population code in which every point in visual space is represented by multiple neurons with subtly different visual sensitivities.

Action Potentials↗
PubMed1993

Relative contribution of rod and cone inputs to bipolar cells and ganglion cells in the tiger salamander retina.

1. The relative contribution of rod and cone inputs to bipolar and ganglion cells were studied by comparing the response-irradiance relations, spectral sensitivities, and response waveforms of these neurons recorded from the isolated, flat-mounted tiger salamander retina under dark-adapted conditions. 2. Bipolar cells could be differentiated both on the basis of the polarity of the light response and on their relative rod/cone input. Thus some depolarizing bipolar cells appeared more strongly influenced by rod input (DBCR), whereas others were more influenced by cone input (DBCC). Similarly, hyperpolarizing bipolar cells could be divided into those that received rod-dominant input (HBCR) or cone-dominant input (HBCC). 3. The light onset response of sustained-ON ganglion cells reflected both rod-dominant input from DBCRs and cone-dominant input from DBCCs. 4. OFF ganglion cells displayed both a rod-dominant sustained light offset response and a cone-dominant transient light offset response, suggesting input from both HBCRs and HBCCs. 5. In ON-OFF ganglion cells, the light onset response was strongly rod dominated and was presumably mediated by DBCRs, whereas the light offset response displayed both rod and cone influence, suggesting input from HBCRs and HBCCs. The contribution of cones to the light onset response of ON-OFF ganglion cells was only observed in the presence of a rod-adapting background light. 6. A suppression of the light offset responses of OFF and ON-OFF ganglion cells was observed, which was dependent both on the wavelength and irradiance of the light stimulus. 7. These results indicate that the photoreceptor inputs to bipolar cells in the tiger salamander retina are segregated such that they form separate rod-dominant and cone-dominant pathways. Thus the response properties of the different types of ganglion cells are influenced not only by the excitatory and inhibitory inputs they receive from the bipolar and amacrine cells but also whether these inputs are provided through rod-dominant or cone-dominant pathways. The functional implications of these findings are discussed.

Ambystoma↗
PubMed1993

Identification of glutamate receptor subtypes mediating inputs to bipolar cells and ganglion cells in the tiger salamander retina.

1. The effects of glutamate receptor agonists and antagonists on bipolar cells and ganglion cells were studied with the use of intracellular and extracellular recording in the superfused, isolated, flat-mounted tiger salamander retina. The goal of the experiments was to correlate glutamate receptor subtypes with their localization at specific synaptic sites in the tiger salamander retina. The drugs tested were the kainate/alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), the N-methyl-D-aspartate (NMDA) receptor antagonist 3-(C+/-)-2-carboxy-piperazin-4-yl)-propyl-1-phosphonic acid (CPP) and L-2-amino-4-phosphonobutyrate (L-AP4). 2. The light responses of hyperpolarizing bipolar cells were suppressed by 20 microM CNQX, whereas L-AP4 had no effect on their light responses. In contrast, 20 microM CNQX had no effect on depolarizing bipolar cells, whereas L-AP4 abolished the light responses of these cells. 3. The light offset responses of OFF and ON-OFF ganglion cells were completely blocked by concentrations of CNQX as low as 5 microM. The light onset responses of ON-OFF ganglion cells were blocked when the concentration of CNQX was raised to 20 microM. In addition, 30 microM CPP partially blocked the light onset responses of ON-OFF ganglion cells but had a lesser effect on the light offset responses. 4. Twenty micromolars of CNQX blocked a transient component, and 20 microM CPP blocked a sustained component of the light response of sustained-ON ganglion cells.(ABSTRACT TRUNCATED AT 250 WORDS)

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1996

Response sensitivity and voltage gain of the rod- and cone-horizontal cell synapses in dark- and light-adapted tiger salamander retina.

1. Rods, cones, and horizontal cells (HCs) were recorded in superfused, flat-mounted isolated retinas of the larval tiger salamander, Ambystoma tigrinum, under dark- and light-adapted conditions. 2. Under dark-adapted conditions, HC responses to dim 500-nm light stimuli were mediated only by rods. In the linear voltage range (near the dark potentials), the average response to a light step of 0.5 s (500 nm, 0.438 photons per micron2 per s) was 0.41 +/- 0.06 (SD) mV for rods and 1.86 +/- 0.52 mV for HCs. The step sensitivity of rods was approximately 0.94 mV per photon micron2 s, or 0.032 mV per activated rhodopsin molecule (Rh*) rod second, and the step sensitivity of HCs was approximately 4.25 +/- 1.19 mV per photon micron2 s or 0.14 +/- 0.04 mV per Rh* rod second. The chord voltage gain of the rod-HC synapse had an average value of 4.54 and a range from 2.68 to 7.32. 3. By the use of the spectral subtraction method, we found that the average cone-mediated HC response to a 750-nm light step that elicited an average cone response of 0.73 +/- 0.20 mV was 1.15 +/- 0.31 mV. The step sensitivity of cones under dark-adapted conditions was 0.0012 mV per photon micron2 s, and that of the cone-mediated-HC response was 0.0019 mV per photon micron2 s. The chord voltage gains of the cone-HC synapses under dark-adapted conditions had an average value of 1.58 and a range from 0.82 to 2.05. 4. Under light-adapted conditions (with a 500-nm/-2.40 background light, which desensitized rod responses but did not substantially reduce the cone responses), the cones had an average response to a light step of 0.5 s (500 nm/-3.3) of 0.78 +/- 0.09 mV, and this response did not vary with time. The HC response to the same light step had an average value of 3.95 +/- 3.41 mV 3 min after the background light onset, and it increased with time until reaching a steady-state value of 5.95 +/- 3.63 mV approximately 15 min after the background light onset. The average chord voltage gain of the cone-HC synapse under such light-adapted conditions was 5.06 at 3 min after background light onset and 7.63 at 15 min after background light onset. These values are approximately 3-5 times higher than the chord voltage gain of the cone-HC synapse under dark-adapted conditions. 5. The background-induced increase of the chord voltage gain of the cone-HC synapse suggests that similarly to the rod-HC synapse, the voltage gain of the cone-HC synapse in the tiger salamander retina can also be modulated by light. Additionally, our results suggest that certain time-dependent process(es) in the synaptic cleft or postsynaptic membrane may be responsible for such modulation. 6. In addition to determining the average values of response sensitivity and chord voltage gains of the rod- and cone-HC synapses, we studied the variation of these parameters among different HCs. HCs with higher rod-HC synaptic gain had lower cone-HC synaptic gain, and HCs with lower rod-HC synaptic gain exhibited higher cone-HC synaptic gain under both dark- and light-adapted conditions. This suggests that the rod-HC and cone-HC synaptic gains in HCs are complementary to each other, and voltage responses of all HCs under dark- or light-adapted conditions are of comparable amplitudes.

Adaptation, Physiological↗
PubMed1997

Motor patterns and kinematics during backward walking in the pacific giant salamander: evidence for novel motor output.

Kinematic and motor patterns during forward and backward walking in the salamander Dicamptodon tenebrosus were compared to determine whether the differences seen in mammals also apply to a lower vertebrate with sprawling posture and to measure the flexibility of motor output by tetrapod central pattern generators. During treadmill locomotion, electromyograms (EMGs) were recorded from hindlimb muscles of Dicamptodon while simultaneous high-speed video records documented movement of the body, thigh, and crus and allowed EMGs to be synchronized to limb movements. In forward locomotion, the trunk was lifted above the treadmill surface. The pelvic girdle and trunk underwent smooth side-to-side oscillations throughout the stride. At the beginning of the stance phase, the femur was protracted and the knee joint extended. The knee joint initially flexed in early stance and then extended as the foot pushed off in late stance, reaching maximum extension just before foot lift-off. The femur retracted steadily throughout the stance. In the swing phase, the femur rapidly protracted, and the leg was brought forward in an "overhand crawl" motion. In backward walking, the body frequently remained in contact with the treadmill surface. The pelvic girdle, trunk, and femur remained relatively still during stance phase, and most motion occurred at the knee joint. The knee joint extended throughout most of stance, as the body moved back, away from the stationary foot. The knee flexed during swing. Four of five angles showed significantly smaller ranges in backward than in forward walking. EMGs of forward walking showed that ventral muscles were coactive, beginning activity just before foot touchdown and ceasing during the middle of stance phase. Dorsal muscles were active primarily during swing. Backward locomotion showed a different pattern; all muscles except one showed primary activity during the swing phase. This pattern of muscle synergy in backward walking never was seen in forward locomotion. Also, several muscles demonstrated lower burst rectified integrated areas (RIA) or durations during backward locomotion. Multivariate statistical analysis of EMG onset and RIA completely separated forward and backward walking along the first principal component, based on higher RIAs, longer durations of muscle activity, and greater synergy between ventral muscles during early stance in forward walking. Backward walking in Dicamptodon uses a novel motor pattern not seen during forward walking in salamanders or during any other locomotor activity in previously studied tetrapods. The central neuronal mechanisms mediating locomotion in this primitive tetrapod are thus capable of considerable plasticity.

Analysis of Variance↗
PubMed2006

Identification of intergenomic recombinations in unisexual salamanders of the genus Ambystoma by genomic in situ hybridization (GISH).

Unisexual salamanders in the genus Ambystoma (Amphibia, Caudata) are endemic to eastern North America and are mostly all-female polyploids. Two to four of the bisexual species, A. laterale, A. jeffersonianum, A. texanum and A. tigrinum, contribute to the nuclear genome of unisexuals and more than 20 combinations that range from diploid to pentaploid have been identified in this complex. Because the karyotypes of the four bisexual species are similar, homologous and homoeologous chromosomes in the unisexuals can not be distinguished by conventional or banded karyotypes. We chose two widespread unisexual genomic combinations (A.laterale-2 jeffersonianum [or LJJ] and A. 2 laterale-jeffersonianum [or LLJ]) and employed genomic in situ hybridization (GISH) to identify the genomes in these unisexuals. Under optimum conditions, GISH reliably distinguishes the respective chromosomes attributed to both A.laterale and A. jeffersonianum. Of four populations examined, two were found to have independently evolved homoeologous recombinants that persist in both LJJ and LLJ individuals. Our results refute the previous hypothesis of clonal integrity and independent evolution of the genome combinations in these unisexuals. Our data provide evidence for intergenomic interactions between maternal chromosomes during meiosis in unisexuals and help to explain previously observed non-homologous bivalents and/or quadrivalents among lampbrush chromosomes that were possibly initiated by partial homosequential pairing among the homo(eo)logues. To explore the utility of GISH in other members of the complex, probes developed from A. laterale were also applied to unisexuals that contained A. tigrinum and A. texanum genomes. GISH is an effective tool that can be used to identify and to quantify genomic constituents and to investigate intergenomic interactions in unisexual salamanders. GISH also has potential application to examine possible genomic evolution in other unisexuals.

Ambystoma↗
PubMed1989

Evolution of motor patterns: aquatic feeding in salamanders and ray-finned fishes.

Patterns of muscle activity (motor patterns) have generally been found to be strongly conserved during the evolution of aquatic feeding behavior within closely related groups of fishes and salamanders. We conducted a test of the generality of motor pattern conservation with a much broader phylogenetic scope than has been done previously. Activity patterns of three cranial muscles were quantified from electromyographic (EMG) recordings made during suction feeding in a salamander (Ambystoma mexicanum) and 4 widely divergent species of ray-finned fishes (Amia calva, Notopterus chitala, Micropterus salmoides and Lepomis macrochirus). General features of the motor pattern were the same in all species, but multivariate and univariate analyses of variance revealed highly significant differences among the 5 species in the average muscle activity pattern, indicating that the motor pattern has not been precisely conserved among these 5 taxa. Five of eight EMG variables that describe the intensity and timing of muscle activity differed among species. Only the intensity of activity of the adductor mandibulae appears to be a strongly conserved feature of the suction feeding motor pattern in anamniotes. A discriminant function analysis of the 8 EMG variables successfully classified about two thirds of the feeding incidents as belonging to the correct species. In contrast to the results of previous studies of closely related taxa, we found that numerous quantitative differences exist among species, indicating that functionally significant details of suction feeding motor patterns have changed during evolution, whereas several general features of the pattern have been conserved.

Ambystoma↗
PubMed2004

From biomedicine to natural history research: EST resources for ambystomatid salamanders.

BACKGROUND: Establishing genomic resources for closely related species will provide comparative insights that are crucial for understanding diversity and variability at multiple levels of biological organization. We developed ESTs for Mexican axolotl (Ambystoma mexicanum) and Eastern tiger salamander (A. tigrinum tigrinum), species with deep and diverse research histories. RESULTS: Approximately 40,000 quality cDNA sequences were isolated for these species from various tissues, including regenerating limb and tail. These sequences and an existing set of 16,030 cDNA sequences for A. mexicanum were processed to yield 35,413 and 20,599 high quality ESTs for A. mexicanum and A. t. tigrinum, respectively. Because the A. t. tigrinum ESTs were obtained primarily from a normalized library, an approximately equal number of contigs were obtained for each species, with 21,091 unique contigs identified overall. The 10,592 contigs that showed significant similarity to sequences from the human RefSeq database reflected a diverse array of molecular functions and biological processes, with many corresponding to genes expressed during spinal cord injury in rat and fin regeneration in zebrafish. To demonstrate the utility of these EST resources, we searched databases to identify probes for regeneration research, characterized intra- and interspecific nucleotide polymorphism, saturated a human - Ambystoma synteny group with marker loci, and extended PCR primer sets designed for A. mexicanum / A. t. tigrinum orthologues to a related tiger salamander species. CONCLUSIONS: Our study highlights the value of developing resources in traditional model systems where the likelihood of information transfer to multiple, closely related taxa is high, thus simultaneously enabling both laboratory and natural history research.

Ambystoma↗