Cytoarchitectonic subdivisions in the subtectal midbrain of the lizard Gallotia galloti

Contemporary study of molecular patterning in the vertebrate midbrain is handicapped by the lack of a complete topological map of the diverse neuronal complexes differentiated in this domain. The relatively less deformed reptilian midbrain was chosen for resolving this fundamental issue in a way that can be extrapolated to other tetrapods. The organization of midbrain centers was mapped topologically in terms of longitudinal columns and cellular strata on transverse, Nissl-stained sections in the lizard Gallotia galloti. Four columns extend along the whole length of the midbrain. In dorsoventral order: 1) the dorsal band contains the optic tectum, surrounded by three ventricularly prominent subdivisions, named griseum tectale, intermediate area and torus semicircularis, in rostrocaudal order; 2) a subjacent region is named here the lateral band, which forms the ventral margin of the alar plate and also shows three rostrocaudal divisions; 3) the basal band forms the basal plate or tegmentum proper; it appears subdivided into medial and lateral parts: the medial part contains the oculomotor and accessory efferent neurons and the medial basal part of the reticular formation, which includes the red nucleus rostrally; the lateral part contains the lateral basal reticular formation, and includes the substantia nigra caudally; 4) the median band contains the ventral tegmental area, representing the mesencephalic floor plate. The alar regions (dorsal and lateral) show an overall cellular stratification into periventricular, central and superficial strata, with characteristic cytoarchitecture for each part. The lateral band contains two well developed superficial nuclei, one of which is commonly misidentified as an isthmic formation. The basal longitudinal subdivisions are simpler and basically consist of periventricular and central strata.     

Aldosterone regulation of active sodium chloride transport in the lizard colon (Gallotia galloti)

Summary Bioelectrical parameters and unidirectional sodium and chloride fluxes were measured under voltageclamp conditions in groups of lizards submitted to single or chronic aldosterone treatment. Both acute (AT) and chronic (CT) treatment induced significant increases in the short-circuit current (I _sc), as well as in the mucosa-to-serosa (J _m-s ^Na ) and net sodium flux (J _net ^Na ). In AT tissues, aldosterone did not change net chloride flux (J _net ^Cl ) but did so in CT tissues. Amiloride reduced the aldosterone-increased I _sc in AT and CT tissues, inhibited J _net ^Na in AT tissues and abolished it in CT colons. J _net ^Cl was also reduced by the diuretic in the group of AT colons, whereas no changes were observed in the CT tissues. Addition of luminal DIDS reduced Na⁺ absorption and totally inhibited Cl^- absorption in the AT tissues, but did not change I _sc. However, in CT tissues neither Na⁺ nor Cl^- transport were affected by DIDS. A good relationship between I _sc and J _m-s ^Na was apparent after DIDS treatment in AT tissues. In this group, simultaneous addition of DIDS and amiloride totally abolished J _net ^Na and reduced I _sc to untreated control values. Addition of serosal ouabain abolished I _sc and Na⁺ absorption in AT and CT colons, but Cl^- absorption was only altered in AT tissues. These results support the hypothesis that aldosterone induces an electrogenic, amiloride-sensitive sodium absorption, and in a dose-dependent fashion suppresses electroneutral NaCl absorption in the lizard colon.Abbreviations AT acutely treated - CT chronically treated animals - DIDS 4-4-diisothiocyanatostibene-2-2-disulfonic acid - DMSO dimethylsulphoxide - G _t tissue conductance - I _sc short circuit current - PD transepithelial potential difference - SITS 4-acetamido-4-isothiocyanatostilbene-2-2-disulfonic acid - UC untreated controls Preliminary results of this paper were presented at the X ^th meeting of the European Intestinal Transport Group (EITG), Askov Hojskole, Denmark, 16–19 September 1990     

Relationship between cortical electrical and cardiac autonomic activities in the awake lizard, Gallotia galloti

ECG and EEG signals were simultaneously recorded in lizards, Gallotia galloti, both in control conditions and under autonomic nervous system (ANS) blockade, in order to evaluate possible relationships between the ANS control of heart rate and the integrated central nervous system activity in reptiles. The ANS blockers used were prazosin, propranolol, and atropine. Time-domain summary statistics were derived from the series of consecutive R-R intervals (RRI) of the ECG to measure beat-to-beat heart rate variability (HRV), and spectral analysis techniques were applied to the EEG activity to assess its frequency content. Both prazosin and atropine did not alter the power spectral density (PSD) of the EEG low frequency (LF: 0.5-7.5 Hz) and high frequency (HF: 7.6-30 Hz) bands, whereas propranolol decreased the PSD in these bands. These findings suggest that central beta-adrenergic receptor mechanisms could mediate the reptilian waking EEG activity without taking part any alpha(1)-adrenergic and/or cholinergic receptor systems. In 55% of the lizards in control conditions, and in approximately 43% of the lizards under prazosin and atropine, a negative correlation between the coefficient of variation of the series of RRI value (CV(RRI)) and the mean power frequency (MPF) of the EEG spectra was found, but not under propranolol. Consequently, the lizards' HRV-EEG-activity relationship appears to be independent of alpha(1)-adrenergic and cholinergic receptor systems and mediated by beta-adrenergic receptor mechanisms.     

Diencephalic neuronal populations projecting axons into the basal plate in a lizard (Gallotia galloti).

Lizard diencephalic populations sending axons into the basal plate were studied by the in vitro HRP technique in the lizard Gallotia. Retrograde labeled cells were concentrated in distinct neuronal groups within alar plates of prosomeres p1 and p3, whereas the alar plate of p2 was poorly labeled. Efferent fibers from alar p1 and p3 populations entered the basal plate of the diencephalon along topologically dorsoventral courses, bifurcating thereafter into longitudinal ascending (rostral) and descending (caudal) trajectories. Thus, diencephalic segments p1 and p3 have alar cell populations contributing to the longitudinal premotor connectivity of the neural axis , whereas the alar p2 segment projects via the fasciculus retroflexus, the efferent tract of the epithalamus. However, the axons from the habenular complex bifurcate within or adjacent to the floor plate and not within the basal plate.     

Geological history and within-island diversity: a debris avalanche and the Tenerife lizard Gallotia galloti

Several processes have been described that could explain geographical variation and speciation within small islands, including fragmentation of populations through volcanic eruptions. Massive landslides, or debris avalanches, could cause similar effects. Here we analyse the potential impact of the 0.8 million-year-ago (Ma) Güimar valley debris avalanche on the phylogeography of the lizard Gallotia galloti on the Canary Island of Tenerife. Distributions of mitochondrial DNA lineages (based on cytochrome b sequences) were analysed on a 60-km southeastern coast transect centred on this area. Three main clades were detected, which can be divided into northern (one clade) and southern (two clades) groups that introgress across the valley. Maximum-likelihood estimates of migration rates (scaled for mutation rate) revealed highly asymmetric patterns, indicating that long-term gene flow into this region from both the northern and the southern populations greatly exceeded that in the opposite directions, consistent with recolonization of the area. The ancestral Tenerife node on the G. galloti tree is estimated at 0.80 Ma, matching closely with the geological estimate for the debris avalanche. Morphological variation (body dimensions and scalation) was also analysed and indicated a stepped cline in female scalation across the valley, although the patterns for male scalation and male and female body dimensions were not as clear. Together these findings provide support for the hypothesis that the debris avalanche has shaped the phylogeography of G. galloti and may even have been a primary cause of the within-island cladogenesis through population fragmentation and isolation. Current estimates of timing of island unification mean that the original hypothesis that within-island diversity is explained by the secondary contact of populations from the two ancient precursor islands of Teno and Anaga is less plausible for this and some other Tenerife species. Large-scale landslides have occurred on many volcanic islands, and so may have been instrumental in shaping within-island diversities.     

S100 immunoreactive glial cells in the forebrain and midbrain of the lizard Gallotia galloti during ontogeny

We identified S100 immunoreactive cells in the brain of the lizard Gallotia galloti during ontogeny using immunohistochemical techniques for light and electron microscopy. In double labeling experiments with antibodies specific for S100A1 and S100B (anti-S100) and proliferative cell nuclear antigen (anti-PCNA), myelin basic protein (anti-MBP), phosphorylated neurofilaments (SMI-31), glial fibrillary acidic protein (anti-GFAP), or glutamine synthetase (anti-GS), we detected S100-like immunoreactivity in glial cells but never in neurons. Restricted areas of the ventricular zone were stained in the hypothalamus from E32 to postnatal stages, and in the telencephalon at E35, E36, and in adults. S100 immunoreactivity was observed predominantly in scattered PCNA-negative cells that increased in number from E35 to the adult stage in the myelinated tracts of the brain and had the appearance of oligodendrocytes. Quantitative analysis revealed that all of the S100-positive glial cells were GFAP-negative, whereas most of the S100-positive glial cells were GS-positive. Ultrastructurally, most of these S100-positive/GS-positive glial cells resembled oligodendrocytes of light and medium electron density. In adult lizards, a small subpopulation of astrocyte-like cells was also stained in the pretectum. We conclude that in the lizard S100 can be considered a marker of a subpopulation of oligodendrocytes rather than of astrocytes, as is the case in mammals. The S100-positive subpopulation of oligodendrocytes in the lizard could represent cells actively involved in the process of myelination during development and in the maintenance of myelin sheaths in the adult.     

Distribution of neurotrophin-3 during the ontogeny and regeneration of the lizard (Gallotia galloti) visual system

We have previously described the spontaneous regeneration of retinal ganglion cell axons after optic nerve (ON) transection in the adult Gallotia galloti. As neurotrophin-3 (NT-3) is involved in neuronal differentiation, survival and synaptic plasticity, we performed a comparative immunohistochemical study of NT-3 during the ontogeny and regeneration (after 0.5, 1, 3, 6, 9, and 12 months postlesion) of the lizard visual system to reveal its distribution and changes during these events. For characterization of NT-3(+) cells, we performed double labelings using the neuronal markers HuC-D, Pax6 and parvalbumin (Parv), the microglial marker tomato lectin or Lycopersicon esculentum agglutinin (LEA), and the astroglial markers vimentin (Vim) and glial fibrillary acidic protein (GFAP). Subpopulations of retinal and tectal neurons were NT-3(+) from early embryonic stages to adulthood. Nerve fibers within the retinal nerve fiber layer, both plexiform layers and the retinorecipient layers in the optic tectum (OT) were also stained. In addition, NT-3(+)/GFAP(+) and NT-3(+)/Vim(+) astrocytes were detected in the ON, chiasm and optic tract in postnatal and adult lizards. At 1 month postlesion, abundant NT-3(+)/GFAP(+) astrocytes and NT-3(-)/LEA(+) microglia/macrophages were stained in the lesioned ON, whereas NT-3 became downregulated in the experimental retina and OT. Interestingly, at 9 and 12 months postlesion, the staining in the experimental retina resembled that in control animals, whereas bundles of putative regrown fibers showed a disorganized staining pattern in the OT. Altogether, we demonstrate that NT-3 is widely distributed in the lizard visual system and its changes after ON transection might be permissive for the successful axonal regrowth.     

Time-related interdependence between low-frequency cortical electrical activity and respiratory activity in lizard, Gallotia galloti

Electroencephalograms of medial cortex and electromyograms of intercostal muscles (EMG-icm) were simultaneously recorded in the lizard, Gallotia galloti, during two daily time periods (at daytime, DTP: 1200-1600 h; by night, NTP: 0000-0400 h), to investigate whether a relationship exists between the respiratory and cortical electrical activity of reptiles, and, if so, how this relationship changes during the night rest period. Testing was carried out by studying interdependence between cortical electrical and respiratory activities, by means of linear and nonlinear signal analysis techniques. Both physiological activities were evaluated through simultaneous power signals, derived from the power of the low-frequency band of the electroencephalogram (pEEG-LF), and from the power of the EMG-icm (pEMG-icm), respectively. During both DTP and NTP, there was a significant coherence between both signals in the main frequency band of pEMG-icm. During both DTP and NTP, the nonlinear index N measured significant linear asymmetric interdependence between pEEG-LF and pEMG-icm. The N value obtained between pEEG-LF vs. pEMG-icm was greater than the one between pEMG-icm vs. pEEG-LF. This means that the system that generates the pEEG-LF is more complex than the one that generates the pEMG-icm, and suggests that the temporal variability of power in the low-frequency cortical electrical activity is driven by the power of the respiratory activity.     

Tenascin-R and axon growth-promoting molecules are up-regulated in the regenerating visual pathway of the lizard (Gallotia galloti)

It is currently unclear whether retinal ganglion cell (RGC) axon regeneration depends on down-regulation of axon growth-inhibitory proteins, and to what extent outgrowth-promoting substrates contribute to RGC axon regeneration in reptiles. We performed an immunohistochemical study of the regulation of the axon growth-inhibiting extracellular matrix molecules tenascin-R and chondroitin sulphate proteoglycan (CSPG), the axon outgrowth-promoting extracellular matrix proteins fibronectin and laminin, and the axonal tenascin-R receptor protein F3/contactin during RGC axon regeneration in the lizard, Gallotia galloti. Tenascin-R and CSPG were expressed in an extracellular matrix-, oligodendrocyte/myelin- and neuron-associated pattern and up-regulated in the regenerating optic pathway. The expression pattern of tenascin-R was not indicative of a role in channeling or restriction of re-growing RGC axons. Up-regulation of fibronectin, laminin, and F3/contactin occurred in spatiotemporal patterns corresponding to tenascin-R expression. Moreover, we analyzed the influence of substrates containing tenascin-R, fibronectin, and laminin on outgrowth of regenerating lizard RGC axons. In vitro regeneration of RGC axons was not inhibited by tenascin-R, and further improved on mixed substrates containing tenascin-R together with fibronectin or laminin. These results indicate that RGC axon regeneration in Gallotia galloti does not require down-regulation of tenascin-R or CSPG. Presence of tenascin-R is insufficient to prevent RGC axon growth, and concomitant up-regulation of axon growth-promoting molecules like fibronectin and laminin may override the effects of neurite growth inhibitors on RGC axon regeneration. Up-regulation of contactin in RGCs suggests that tenascin-R may have an instructive function during axon regeneration in the lizard optic pathway.     

Parasite local maladaptation in the Canarian lizard Gallotia galloti (Reptilia: Lacertidae) parasitized by haemogregarian blood parasite

Biologists commonly assume that parasites are locally adapted since they have shorter generation times and higher fecundity than their hosts, and therefore evolve faster in the arms race against the host's defences. As a result, parasites should be better able to infect hosts within their local population than hosts from other allopatric populations. However, recent mathematical modelling has demonstrated that when hosts have higher migration rates than parasites, hosts may diversify their genes faster than parasites and thus parasites may become locally maladapted. This new model was tested on the Canarian endemic lizard and its blood parasite (haemogregarine genus). In this host–parasite system, hosts migrate more than parasites since lizard offspring typically disperse from their natal site soon after hatching and without any contact with their parents who are potential carriers of the intermediate vector of the blood parasite (a mite). Results of cross-infection among three lizard populations showed that parasites were better at infecting individuals from allopatric populations than individuals from their sympatric population. This suggests that, in this host–parasite system, the parasites are locally maladapted to their host.     

Regeneration of retinal axons in the lizard Gallotia galloti is not linked to generation of new retinal ganglion cells

Using anterograde tracing with HRP and antibodies (ABs) against neurofilaments, we show that regrowth of retinal ganglion cell (RGC) axons in the lizard Gallotia galloti commences only 2 months after optic nerve transection (ONS) and continues over at least 9 months. This is unusually long when compared to RGC axon regeneration in fish or amphibians. Following ONS, lizard RGCs up-regulate the immediate early gene C-JUN for 9 months or longer, indicating their reactive state. In keeping with the in vivo data, axon outgrowth from lizard retinal explants is increased above control levels from 6 weeks, reaches its maximum as late as 3 months, and remains elevated for at least 1 year after ONS. By means of BrdU incorporation assays and antiproliferating cell nuclear antigen immunohistochemistry, we show that the late axon outgrowth is not derived from new RGCs that might have arisen in reaction to ONS: no labeled cells were detected in lizard retinas at 0.5, 1, 1.5, 3, 6, and 12 months after ONS. Conversely, numbers of RGCs undergoing apoptosis were too low to be detectable in TUNEL assays at any time after ONS. These results demonstrate that retinal axon regeneration in G. galloti is due to axon regrowth from the resident population of RGCs, which remain in a reactive state over an extended time interval. Neurogenesis does not appear to be involved in RGC axon regrowth in G. galloti.     

Nonlinearity and fractality in the variability of cardiac period in the lizard, Gallotia galloti: effects of autonomic blockade

Both nonlinear and fractal properties of beat-to-beat R-R interval variability signal (RRV) of freely moving lizards (Gallotia galloti) were studied in baseline and under autonomic nervous system blockade. Nonlinear techniques allowed us to study the complexity, chaotic behavior, nonlinearity, stationarity, and regularity over time of RRV. Scaling behavior of RRV was studied by means of fractal techniques. The autonomic nervous system blockers used were atropine, propranolol, prazosin, and yohimbine. The nature of RRV was linear in baseline and under beta-, alpha(1)- and alpha(2)-adrenoceptor blockades. Atropine changed the linear nature of RRV to nonlinear and increased its stationarity, regularity and fractality. Propranolol increased the complexity and chaotic behavior, and decreased the stationarity, regularity, and fractality of RRV. Both prazosin and yohimbine did not change any of the nonlinear and fractal properties of RRV. It is suggested that 1) the use of both nonlinear and fractal analysis is an appropriate approach for studying cardiac period variability in reptiles; 2) the cholinergic activity, which seems to make the alpha(1)-, alpha(2)- and beta-adrenergic activity interaction unnecessary, determines the linear behavior in basal RRV; 3) fractality, as well as both RRV regularity and stationarity over time, may result from the balance between cholinergic and beta-adrenergic activities opposing actions; 4) beta-adrenergic activity may buffer both the complexity and chaotic behavior of RRV, and 5) neither the alpha(1)- nor the alpha(2)-adrenergic activity seem to be involved in the mediation of either nonlinear or fractal components of RRV.     

Autonomic mediation in the interdependences between cardiocortical activity time variations and between cardiorespiratory activity time variations in the lizard, Gallotia galloti.

Multivariate nonlinear analysis methods were applied to variability time series extracted from electrocardiographic, electrocorticographic and respiratory activities of Gallotia galloti lizards, to study interdependences between cardio-cortical activity time variations, and between cardio-respiratory activity time variations. Autonomic nervous system involvement in the mediation of such interdependences was investigated through pharmacological blockade. Cardiac variability was evaluated from the R-R intervals of the electrocardiogram (RRIv). Cortical (CORTv) and respiratory (RESPv) activity time variations were evaluated from power-data signals derived from both electrocorticogram and respiratory signal segments obtained within each R-R interval, respectively. A nonlinear index N to measure interdependence between the signals, and a surrogate data test to measure the significance and nature of the interdependences were used. A nonlinear dependence of RRIv vs. CORTv and of RRIv vs. RESPv was found. Both dependences seem to be unconnected with the functioning of both alpha(1)-adrenoceptor and cholinoceptor systems, but appear to be mediated by beta-adrenoceptor mechanisms. A linear dependence of CORTv vs. RRIv and of RESPv vs. RRIv was also found. Both dependences seem to be unconnected with the operation of alpha(1)-adrenoceptor, beta-adrenoceptor and cholinoceptor systems. It is suggested that both the cardiocortical and cardiorespiratory synchronizations studied seem to be mediated by beta-adrenoceptor mechanisms.     

Proximate causes of altitudinal differences in body size in an agamid lizard

Body size is directly linked to key life history traits such as growth, fecundity, and survivorship. Identifying the causes of body size variation is a critical task in ecological and evolutionary research. Body size variation along altitudinal gradients has received considerable attention; however, the underlying mechanisms are poorly understood. Here, we compared the growth rate and age structure of toad‐headed lizards (Phrynocephalus vlangalii) from two populations found at different elevations in the Qinghai‐Tibetan Plateau. We used mark‐recapture and skeletochronological analysis to identify the potential proximate causes of altitudinal variation in body size. Lizards from the high‐elevation site had higher growth rates and attained slightly larger adult body sizes than lizards from the low‐elevation site. However, newborns produced by high‐elevation females were smaller than those by low‐elevation females. Von Bertalanffy growth estimates predicted high‐elevation individuals would reach sexual maturity at an earlier age and have a lower mean age than low‐elevation individuals. Relatively lower mean age for the high‐elevation population was confirmed using the skeletochronological analysis. These results support the prediction that a larger adult body size of high‐elevation P. vlangalii results from higher growth rates, associated with higher resource availability.     

Autonomic mediation in the interdependences between cardiocortical activity time variations and between cardiorespiratory activity time variations in the lizard, Gallotia galloti

Multivariate nonlinear analysis methods were applied to variability time series extracted from electrocardiographic, electrocorticographic and respiratory activities of Gallotia galloti lizards, to study interdependences between cardio-cortical activity time variations, and between cardio-respiratory activity time variations. Autonomic nervous system involvement in the mediation of such interdependences was investigated through pharmacological blockade. Cardiac variability was evaluated from the R-R intervals of the electrocardiogram (RRIv). Cortical (CORTv) and respiratory (RESPv) activity time variations were evaluated from power-data signals derived from both electrocorticogram and respiratory signal segments obtained within each R-R interval, respectively. A nonlinear index N to measure interdependence between the signals, and a surrogate data test to measure the significance and nature of the interdependences were used. A nonlinear dependence of RRIv vs. CORTv and of RRIv vs. RESPv was found. Both dependences seem to be unconnected with the functioning of both alpha(1)-adrenoceptor and cholinoceptor systems, but appear to be mediated by beta-adrenoceptor mechanisms. A linear dependence of CORTv vs. RRIv and of RESPv vs. RRIv was also found. Both dependences seem to be unconnected with the operation of alpha(1)-adrenoceptor, beta-adrenoceptor and cholinoceptor systems. It is suggested that both the cardiocortical and cardiorespiratory synchronizations studied seem to be mediated by beta-adrenoceptor mechanisms.     

Geographic variation in scalation of the lizard Gallotia stehlini within the island of Gran Canaria

The lacertid lizard ( Gallotia stehlini ), an endemic of Gran Canaria, shows no visually obvious geographic variation, yet all seven scalation characters that were examined exhibit significant geographic variation. The number of collar scales, scales along the ventral trunk and femoral pores are correlated with habitat type, while the number of femoral pores is correlated negatively with altitude. Mantel tests were used to compare simultaneously an observed pattern with three hypothesized patterns (habitat type, altitude and proximity). They indicate that, while several individual characters are significantly associated with the putative causal factors of habitat type and altitude, there is no association between an overall scalation distance matrix and habitat type, or altitude when the effect of proximity is removed. Consequently, one should consider the individual characters as well as the multivariate generalized distances. Some of the observed patterns of geographic variation in scalation are very similar to those of the small scincid lizard Chalcides sexlineatus on Gran Canaria and also parallel the altitudinal and latitudinal variation in the scalation of the Tenerife lacertid ( Galotia galloti ). The low level of congruence in patterns of geographic variation in individual characters (i.e. some vary with latitude, some with altitude and one varies with longitude) is consistent with the hypothesis that ecogenetically caused geographic variation may result in lower inter-character congruence than phylogenetically caused geographic variation.