Geographic patterns of population genetic structure in Mytilocypris (Ostracoda: Cyprididae): interpreting breeding systems,gene flow and history in species with differing distributions

Samples from 83 populations of salt lake Ostracods belonging to the genus Mytilocypris were collected from 74 saline lakes and ponds in the semi-arid regions of Australia. These populations were examined for variation at six polymorphic enzyme loci to diagnose breeding systems and to measure population structure, to investigate relative levels of gene flow in species with differing distributions and hence different presumed dispersal capabilities. Despite the occurrence of some populations in disjunct, peripheral, and recent ephemeral habitats, all populations of each species were found to reproduce sexually. Gene flow does occur on a local basis and appears to be facilitated by occupation in the same drainage basin for some species. There was considerable gene pool fragmentation among peripheral populations of four of the five species. Only one species, M. mytiloides, was relatively homogeneous across its range. It may be that gene flow is non-existent into peripheral populations because of poor dispersal abilities, or it may not be frequent enough to overcome local selective pressures. Regardless of these possibilities, the observed gene pool fragmentation has implications for allopatric speciation.     

Complex patterns of mating revealed in a Eucalyptus regnans seed orchard using allozyme markers and the neighbourhood model

The neighbourhood model apportions offspring of individual mother plants to self-fertilization, outcrossing to males within a circumscribed area around the mother plant (the neighbourhood), and outcrossing to males outside the neighbourhood. Formerly the model was applied only to haploid pollen gametes in the offspring of conifers, but is extended so that it can be used with genotypic data from diploid offspring of both angiosperms and gymnosperms. In addition, it is shown that the mating parameters can be estimated without independent estimates of allele frequencies in the pollen pools outside the neighbourhood; thus the model might be applied effectively to natural populations exposed to unknown external pollen sources. Parameters of the neighbourhood mating model were estimated for a 10-year-old seed orchard population of the insect-pollinated tree, Eucalyptus regnans, in southeast Australia, which contained a mixture of two geographical provenances (Victoria and Tasmania). The mating patterns revealed were complex. Crosses between trees of the same provenance occurred three times more often than crosses between trees of different provenances. Levels of self-fertilization and patterns of mating within neighbourhoods were influenced by provenance origin, crop fecundity and orchard position (central vs. edge) of mother trees. Gene dispersal, however, was extensive, with approximately 50% of effective pollen gametes coming from males more than 40 m away from mother trees (average distance between neighbouring trees was 7.4 m). Thus, insect pollinators are efficient promoters of cross-fertilization in this orchard, with the result that the effective number of males mating with each female is large.     

Microsatellite diversity and genetic structure of fragmented populations of the rare,fire-dependent shrub Grevillea macleayana

Recent habitat loss and fragmentation superimposed upon ancient patterns of population subdivision are likely to have produced low levels of neutral genetic diversity and marked genetic structure in many plant species. The genetic effects of habitat fragmentation may be most pronounced in species that form small populations, are fully self-compatible and have limited seed dispersal. However, long-lived seed banks, mobile pollinators and long adult lifespans may prevent or delay the accumulation of genetic effects. We studied a rare Australian shrub species, Grevillea macleayana (Proteaceae), that occurs in many small populations, is self-compatible and has restricted seed dispersal. However, it has a relatively long adult lifespan (c. 30 years), a long-lived seed bank that germinates after fire and is pollinated by birds that are numerous and highly mobile. These latter characteristics raise the possibility that populations in the past may have been effectively large and genetically homogeneous. Using six microsatellites, we found that G. macleayana may have relatively low within-population diversity (3.2-4.2 alleles/locus; Hexp = 0.420-0.530), significant population differentiation and moderate genetic structure (FST = 0.218) showing isolation by distance, consistent with historically low gene flow. The frequency distribution of allele sizes suggest that this geographical differentiation is being driven by mutation. We found a lack mutation-drift equilibrium in some populations that is indicative of population bottlenecks. Combined with evidence for large spatiotemporal variation of selfing rates, this suggests that fluctuating population sizes characterize the demography in this species, promoting genetic drift. We argue that natural patterns of pollen and seed dispersal, coupled with the patchy, fire-shaped distribution, may have restricted long-distance gene flow in the past.     

Nestmate relatedness and population genetic structure of the Australian social crab spider Diaea ergandros (Araneae: Thomisidae)

We characterized the population genetic structure of the Australian social spider Diaea ergandros using polymorphic allozyme markers. Our main objectives were to understand the social organization of D. ergandros and discern patterns of gene flow across distantly separated geographical areas. Spiders were sampled from nests located within 100 m wide locales, which were distributed within larger 50 km wide regions. Our results indicated that nestmates could have been produced by a single mother and father in 88.9% of D. ergandros nests. The remainder of nests contained spiders that were probably produced by polyandrous females or were immigrants from foreign nests. Nestmate relatedness was relatively high (r = 0.44) and did not differ significantly between the sexes or among juvenile, subadult and adult life stages. We also discovered that D. ergandros populations were highly structured, with significant differentiation detected among locales (FLR = 0.23) and regions (FRT = 0.081). Spiders within locales were also substantially inbred (FIL = 0.15). Overall, our data show that significant population subdivision exists in D. ergandros populations, and we suggest that the poor dispersal ability of Diaea spiders can account for the observed genetic structure.     

Neuropeptide Organization of the Nociceptive and Antinociceptive Brain Systems in the Cat

Using radioimmune techniques, we studied in detail the concentrations of -endorphin (-En), met- and leu-enkephalins (mE and lE, respectively), and substance P (SP) in a number of structures of the brainstem and forebrain of the cat. According to the proposed concept, these structures comprise the noci- and antinociceptive brain systems (NS and ANS). The above indices were measured in intact animals and in animals after nociceptive electrocutaneous stimulation (NECS) of the limb, stimulation of the ventrolateral zone of the midbrain central gray (vl SGC, a nociceptive midbrain structure), stimulation of the dorsolateral part of the above region and dorsal raphe nucleus (dl SGC and Rd, antinociceptive midbrain structures), and after combined stimulations (NECS preceded by conditioning stimulation of one of the above midbrain zones). We found that in the norm maximum SP concentrations were observed in the NS structures, while those of -En and mE were the highest in the hypothalamic nuclei belonging to the ANS and in its midbrain centers (dl SGC and Rd). Nociceptive ECS, stimulations of the studied midbrain zones, and combinations of these stimulations could result in specific and, in some cases, very significant (by an order of magnitude and more) shifts in the concentrations of the mentioned neuropeptides in the studied set of the central structures. After NECS and its combination with vl SGC stimulation, SP concentrations in the NS structures considerably increased, while -En and mE concentrations in the ANS components dropped. Stimulations of the dl SGC and Rd were accompanied by increases in the mE and -En levels and simultaneous drops in the SP concentrations in the ANS components; reciprocate shifts were observed within the NS. Changes in the lE level, which were related to the influences used, were less specific and mostly appeared as increases in this index in the structures of both the NS and ANS. Combinations of NECS with conditioning stimulations of the vl SGC, dl SGC, or Rd demonstrated that the latter exert significant modulatory effects on the NECS-induced shifts in the concentrations of the studied neuropeptides. Considering the obtained data, a hypothetical scheme of neuropeptide organization of the cerebral NS and ANS has been proposed. In the examined brain structures, there are neuronal populations belonging to the two main neurochemical systems. One of them is SP-ergic, while another consists of mE- and -En-ergic neurons; these systems are in antagonistic relations. Changes in the levels of mE and -En always induce the attended opposite shifts in the SP levels, and vice versa. The lE-ergic neuronal populations, which co-exist with the above neurochemical systems, are relatively nonspecifically activated by either (noci- and antinociceptive) drives, but, according to the pattern of its responses, the lE-ergic system is closer to the SP-ergic one. It is supposed that pain signals, when coming to the vl SGC, activate SP- and lE-ergic neuronal populations; later on, the posterior and lateral hypothalamic nuclei and preoptic region are involved in the transmission of the above signals. When released by the corresponding neuronal populations in the vl SGC, lE activates the key ANS structures (dl SGC and Rd), and the latter, in turn, activate other components of this system, which form its ascending compartment (ventromedial, dorsomedial, and paraventricular hypothalamic nuclei, septum, basolateral amygdala, hippocampal fields 3 and 4, and cingular cortex). In the ANS,-En and mE function as transmitters.     

Timing and ecdysteroid regulation of the molt in last instar greenhouse whiteflies (Trialeurodes vaporariorum)

A system of markers has been devised to track the development of 3rd and 4th instar/pharate adult greenhouse whiteflies. Instars were identified based on measurements of body width and body length. Depending upon the host plant, the product of the two measurements was exceptionally useful in distinguishing between instars. Body depth was used to divide the 3rd instar into eight stages and body depth and color and appearance of the developing adult eye were used to divide the 4th instar/pharate adult into nine stages. Under conditions of L:D 16:8 and a temperature of 26±2°C, the body depth of 3rd instars reared on greenbean increased from 0.025 (stage 1) to 0.2 mm (stage 8) and the instar duration was approximately 3 days. The body depth of 4th instars increased from approximately 0.1±0.02 (Stage 1) to 0.3±0.03 mm (Stage 5) and then remained constant or decreased slightly during adult development. Ecdysteroid titers peaked at approximately 120 fg/μg protein during Stages 3 through 6 of the 4th instar. Based on an external examination of developing 4th instars and the fluctuations in ecdysteroid titer, it appears that adult development is initiated in Stage 4 or 5 4th instars. Results from histological studies support this view. In Stage 4 nymphs, a subtle change was observed in the corneagenous cells of the eye. However, most Stage 4 4th instars possessed wing development characteristic of earlier, immature stages. In all Stage 5 insects, wing development had been initiated and the corneagenous cells had become quite distinct. In Stage 6 whiteflies, the wing buds were deeply folded and by Stage 7, spines were observed on the new cuticle, indicating that the adult cuticle was well-formed by this stage. Our study is the first to investigate the timing and regulation of the molt, to monitor ecdysteroid titers in precisely staged 4th instar whiteflies and to examine the internal anatomical changes associated with metamorphosis in these tiny homopteran insects.     

Effects of Bucephalus sp. (Trematoda: Bucephalidae) on Perna perna mussels from a culture station in Ratones Grande Island,Brazil

This study reports the prevalence of Bucephalus sp. in Perna perna populations from a culture station of southern Brazil and its effect on the mussel reproductive tissue and immune system. The prevalence of Bucephalus sp. in P. perna (n = 1871) was considered low (3.1%) and did not seasonally vary. Histological sections of the mantle of infected mussels revealed a marked (80%) reduction of the reproductive tissue that was severe even in mussels exhibiting a moderate infection degree. The total (THC) and differential (DHC) hemocyte counts were lower in infected mussels (3.9 x 10(6) hem/ml; granular hemocytes = 33%) as compared with non-infected animals (5.5 x 10(6) hem/ml; granular hemocytes = 40%). The plasma protein concentration did not vary upon infection. Hemocyte infiltration was significantly higher only in mussels with a very heavy infection degree. The parasite sporocysts were never seen encapsulated by the host hemocytes. Our results indicate that Bucephalus sp. promotes a severe castration in its host and apparently evades the mussel immune system.     

The degenerative effect of a single intranigral injection of LPS on the dopaminergic system is prevented by dexamethasone,and not mimicked by rh-TNF-alpha,IL-1beta and IFN-gamma

It is becoming widely accepted that the inflammatory response is involved in neurodegenerative disease. In this context, we have developed an animal model of dopaminergic system degeneration by the intranigral injection of lipopolysaccharide (LPS), a potent inductor of inflammation. To address the importance of the inflammatory response in the LPS-induced degeneration of nigral dopaminergic neurones, we carried out two different kinds of studies: (i) the possible protective effect of an anti-inflammatory compound, and (ii) the effect of the intranigral injection of inflammatory cytokines (TNF-alpha, IL-1beta and IFN-gamma) on dopaminergic neurones viability. Present results show that dexamethasone, a potent anti-inflammatory drug that interferes with many of the features characterizing pro-inflammatory glial activation, prevented the loss of catecholamine content, Tyrosine hydroxylase (TH) activity and TH immunostaining induced by LPS-injection and also the bulk activation of microglia/macrophages. Surprisingly, injection of the pro-inflammatory cytokines failed to reproduce the LPS effect. Taken together, our results suggest that inflammatory response is implicated in LPS-induced neurodegeneration. This damage may be due, at least in part, to a cascade of events independent of that described for TNF-alpha/IL-1 beta/IFN-gamma.     

Direct measurement of renal sympathetic nervous activity in high-fat diet-related hypertensive rats

The elevation of renal sympathetic nervous activity (SNA) is a possible cause of blood pressure (BP) elevation. Although a high-fat diet (FAT) often induces BP elevation in animals, the effect of FAT on renal SNA in animals is not consistent between studies. Thus, we compared the basal levels of efferent renal SNA and BP in FAT- or high-carbohydrate diet (CHO)-fed rats. Twenty-four male Sprague-Dawley rats were fed FAT (P/F/C=20/45/35% cal) or CHO (20/5/75) from 5 weeks of age. After 20-21 weeks of feeding, a 24-h urine sample was collected to measure sodium excretion. The next day, blood (0.2 ml) was withdrawn from a femoral artery, and basal efferent renal nerve discharges and mean arterial pressure (MAP) were recorded under anesthesia. Immediately after the experiment, abdominal (epididymal, perirenal and mesenteric) adipose tissues were dissected. Total abdominal fat weight was significantly greater in the FAT group than in the CHO group. The plasma level of leptin was significantly higher in the FAT group, but blood glucose and plasma insulin levels did not differ between the two groups. MAP and renal SNA were significantly higher in the FAT group. In addition, the ratio of urinary sodium excretion to dietary sodium intake was significantly lower in the FAT group than in the CHO group. The data suggest that the increased renal SNA may contribute to BP elevation in FAT-fed rats. The present study firstly demonstrated that renal SNA was elevated with FAT-related BP elevation.