Early onset of phenotype and cell patterning in the embryonic zebrafish retina

The regular arrangement of retinal cone cells in a mosaic pattern is a common feature of teleosts. In the zebrafish, Brachydanio rerio, the retinal cone mosaic comprises parallel rows consisting of a repeating motif of four cone types. In order to elucidate the temporal and spatial aspects of the genesis of the cone mosaic in the developing retina, we generated a monoclonal antibody that specifically binds to the double cone photoreceptor of the adult. We first saw staining in the developing retina with this antibody, FRet 43, at 48 hours postfertilization, the time at which the first photoreceptor cells undergo their final mitotic division. We then injected embryonic fish with the thymidine analog, 5-bromo-2'-deoxyuridine (BrdU), confirming with a double-labeling experiment that the onset of FRet 43 antigenicity occurs within three hours of the cellular division that generates the double cone photoreceptors. Then we stained tangential sections of the 54-hour embryonic retina with FRet 43, further showing that cells devoid of staining alternate with stained pairs of cells in a pattern that is consistent with the arrangement of photoreceptors in the adult cone mosaic. These results indicate that a marker of the double cone phenotype is expressed at approximately the same time as cellular birthday and that the mosaic patterning is present within 6 hours of this expression.     

Comparative avian phylogeography of Cameroon and equatorial Guinea mountains: implications for conservation

We illustrate the use of Faith's 'Phylogenetic Diversity' measure to compare the phylogeographic structure of two bird species with patterns of avian endemism across six mountains in Cameroon and Equatorial Guinea. The Mountain Greenbul and Cameroon Blue-headed Sunbird showed phylogeographic patterns that together defined three biogeographic regions: Bioko, Mt. Cameroon, and the northern mountains of Cameroon. In contrast, the distributions of endemic species were largely a function of geographical distance, with close mountains sharing more endemic species than distant mountains. Moreover, for both species, populations on Mt. Cameroon were distinctive with respect to the ecologically relevant character bill size. Our results, while preliminary, illustrate the utility of a comparative approach for identifying geographical regions that harbour evolutionarily distinct populations and caution against using only the distributional patterns of endemics to prioritize regions for conservation. Results show that patterns of endemism may not be concordant with patterns of phylogenetic diversity nor morphological variation in a character important in fitness. While incorporation of additional species from unrelated taxa will be necessary to draw definitive conclusions about evolutionarily distinct regions, our preliminary results suggest a conservation approach for the Afromontane region of the Gulf of Guinea that would: (i) emphasize protection of both Bioko and Mt. Cameroon, thereby maximizing preservation of within-species phylogenetic and morphologic diversity; (ii) emphasize protection within the northern mountains to further conserve intraspecific phylogenetic diversity and maximize protection of endemic species.     

Environmental heterogeneity and alternative mating tactics in the damselfly Protoneura amatoria

Conditional male mating strategies have been studied extensivelyin relation to male attributes, such as size and resource-holdingpotential, but few studies have considered the effects of environmentalheterogeneity on the use of alternative mating tactics. In somesystems, environmental heterogeneity may be the key to understandingthe evolution and maintenance of such polyphenisms. I examinedthe influence of the physical environment on the use of alternativetactics by the damselfly Protoneura amatoria. Male P. amatoriareversibly use 2 tactics to gain matings: 1) sit and wait inthe canopy for passing females or 2) hover over the water andattempt to grab females that are ovipositing in floating debris.Observations in 3 streams indicated that the use of the hoveringtactic was greater under high-light than low-light conditionsand at higher densities of ovipositing females. The densityof ovipositing females was correlated with both the light conditionsand the availability of oviposition substrate, indicating thatphysical factors exert indirect as well as direct influenceson tactic use. Experimental manipulations showed that both malesand females responded directly to light conditions and suggestedthat males responded directly to the density of ovipositingfemales. These results can be explained largely in terms ofthe cues and constraints inherent in different light environments.Thus, the conditional mating strategy of P. amatoria appearsto have evolved in response to, and been maintained by, fine-scalevariation in the physical environment. These findings are discussedin relation to flight dynamics and predation risk.     

Persistence of antipredator behavior in an island population of California quail

Island populations may provide unique insights into the evolution and persistence of antipredator behavior. If antipredator behavior is costly and islands have reduced predation risk, then we expect the reduction or loss of antipredator behavior on islands. However, if even a single predator remains, the multipredator hypothesis predicts that antipredator behaviors will be conserved. We compared the flight initiation distances (FID) of California quail (Callipepla californica) on Santa Catalina Island (a location with reduced predation pressure) with quail on the mainland. We found no differences in FID between mainland and island quail. However, despite employing consistent testing methods, the starting distance from which quail were approached was significantly reduced for quail studied on the island when compared with quail studied on the mainland. Our results are consistent with the multipredator hypothesis because, while the island population had substantially fewer predators, some predators remained and some antipredator behavior persisted.     

Inherent interfacial mechanical gradients in 3D hydrogels influence tumor cell behaviors

Cells sense and respond to the rigidity of their microenvironment by altering their morphology and migration behavior. To examine this response, hydrogels with a range of moduli or mechanical gradients have been developed. Here, we show that edge effects inherent in hydrogels supported on rigid substrates also influence cell behavior. A Matrigel hydrogel was supported on a rigid glass substrate, an interface which computational techniques revealed to yield relative stiffening close to the rigid substrate support. To explore the influence of these gradients in 3D, hydrogels of varying Matrigel content were synthesized and the morphology, spreading, actin organization, and migration of glioblastoma multiforme (GBM) tumor cells were examined at the lowest (<50 μm) and highest (>500 μm) gel positions. GBMs adopted bipolar morphologies, displayed actin stress fiber formation, and evidenced fast, mesenchymal migration close to the substrate, whereas away from the interface, they adopted more rounded or ellipsoid morphologies, displayed poor actin architecture, and evidenced slow migration with some amoeboid characteristics. Mechanical gradients produced via edge effects could be observed with other hydrogels and substrates and permit observation of responses to multiple mechanical environments in a single hydrogel. Thus, hydrogel-support edge effects could be used to explore mechanosensitivity in a single 3D hydrogel system and should be considered in 3D hydrogel cell culture systems.     

Pseudoloma neurophilia n. g., n. sp., a new microsporidium from the central nervous system of the zebrafish (Danio rerio)

An unusual xenoma-forming microsporidium was discovered in the central nervous system of moribund zebrafish from a laboratory colony in Eugene, Oregon. Infected fish were often emaciated and lethargic, and histological examination commonly revealed severe myelitis and myositis associated with the infection. Based on its structure, development, and small subunit ribosomal DNA sequence it is unique among fish microsporidia. Spores are uninucleate, ovoid to pyriform, with a prominent posterior vacuole. Spores average 5.4 x 2.7 microm with 13-16 coils of the polar filament. The microsporidium produces xenomas within the spinal cord and hindbrain of fish, and xenomas contained sporophorous vesicles with up to 16 spores. Sporoblasts and presporoblast stages (probably sporonts) are found occasionally in small aggregates dispersed randomly throughout xenomas. It clustered in the "Ichthyosporidium group" along with other fish microsporidian genera based on rDNA sequence analysis. The rDNA sequence of the zebrafish microsporidium was most similar to that of Ichthyosporidium, but showed only 12.1% similarity and therefore this microsporidium can be considered a distinct genus and species, which we have named Pseudoloma neurophilia n. g., n. sp.     

Comparative avian biodiversity of five mountains in northern Cameroon and Bioko

Larison, B., Smith, T.B., Fotso, R. & McNiven, D. 2000. Comparative avian biodiversity of five mountains in northem Cameroon and Bioko. Ostrich 71 (1 & 2): 269–276.

Endemism among birds is widespread in the montane forests of western Cameroon and the Gulf of Guinea. The region includes some of the rarest and most threatened species in Africa. We conducted avian surveys of four previously unsurveyed montane sites in northern Cameroon, including Mt. Ngang-Ha, Hoséré Vokré, Tchabal Gandaba, and Tchabal Mbabo, as well as the northern slope of Caldera de Luba on the island of Bioko, Equatorial Guinea. We report here on avian species richness and relative abundance, and evaluate the conservation potential of each site based on avifaunal richness. The montane forest on both Tchabal Mbabo and Caldera de Luba is extensive, while on the other mountains, the vegetation is not characteristic of montane forest, and consists primarily of small gallery forests embedded in savanna. Tchabal Mbabo and Caldera de Luba had the greatest species richness and abundance of montane birds, while Tchabal Gandaba had the greatest overall avian species richness and abundance. Few montane species were noted on Mt. Ngang-Ha and Hosere Vokre, and avian abundance was quite low on both mountains. Of the mountains surveyed, Tchabal Mbabo and Caldera de Luba exhibit the greatest potential for conservation based on extent of montane forest, and montane species richness and abundance.     

Detecting lacZ gene expression in living cells with new lipophilic, fluorogenic beta-galactosidase substrates.

Current methods for detecting lacZ expression in transformed cells are limited because they require such harsh conditions that viability of the cells after detection is drastically reduced. To overcome this problem, we developed a series of new substrates for detection of lacZ expression in living cells under standard culture or physiological conditions. After incubation with these fluorogenic substrates, cultured lacZ-positive mammalian cells appear morphologically normal, continue to divide, and retain the fluorescent product. Because the product is so well retained, fluorescence intensity can be quantitatively related to the level of gene expression. We have demonstrated this correlation using transformed yeast cells bearing various plasmids, each containing the lacZ gene and a unique promoter sequence with known capabilities for promoting gene expression in yeast.