Control of Melanoblast Differentiation in Amphibia by α-Melanocyte Stimulating Hormone,a Serum Melanization Factor,and a Melanization Inhibiting Factor

A ventrally localized melanization inhibiting factor (MIF) has been suggested to play an important role in the establishment of the dorsal-ventral pigment pattern in Xenopus laevis [Fukuzawa and Ide: Dev. Biol., 129:25–36, 1988]. To examine the possibility that melanoblast expression might be controlled by local putative MIF and melanogenic factors, the effects of α-melanocyte stimulating hormone (α-MSH), a serum melanization factor (SMF) from X. laevis or Rana pipiens, and MIF on the “outgrowth” and “melanization” of Xenopus neural crest cells were studied. Outgrowth represents the number of neural crest cells emigrating from cultured neural tubes, and melanization concerns the percentage of differentiated melanophores among the emigrated cells. MSH or SMF stimulate both outgrowth and melanization. The melanogenic effect of Xenopus serum in this system is more than twice that of Rana serum. The actions of MSH and Xenopus serum on melanization seem to be different: 1) Stronger melanization is induced by Xenopus serum than by MSH, and the onset of melanization occurs earlier with Xenopus serum; 2) MSH stimulates melanization only in the presence of added tyrosine; and 3) MSH causes young melanophores to assume a prominent state of melanophore dispersion during culture, while Xenopus serum (10%) had only a slight dispersing effect and not until day 3. A fraction of Xenopus serum presumably containing molecules of a smaller molecular weight (MW <30 kDa) than that of a pigment promoting factor reported in calf serum [Jerdan et al.: J. Cell Biol., 100:1493–1498, 1985] produces the same remarkable melanogenic effects as does intact serum. While this fraction stimulates outgrowth, another fraction presumably containing larger molecules (MW > 100 kDa) does not. MIF contained in Xenopus ventral skin conditioned medium (VCM) inhibits both outgrowth and melanization dose dependently. When VCM is used in combination with MSH, the stimulating effects of MSH on both outgrowth and melanization are completely inhibited. In contrast, the stimulatory effects of Xenopus serum are not completely inhibited when combined with VCM, although melanization is reduced to approximately 40% that of controls. MIF activity was also found to be present in ventral, but not in dorsal, skin conditioned media of R. pipiens when tested in the Xenopus neural crest system. We suggest that ventrally localized MIF plays an important role in amphibian pigment pattern formation and that the interacting effects of MIF and melanogenic factors influence melanoblast differentiation, migration, and/or proliferation of neural crest cells to effect the expression of pigmentary patterns.     

Evaluations of permethrin-impregnated clothing and three topical repellent formulations of deet against tsetse flies in Zambia

Permethrin-impregnated clothing and three topical repellent formulations of deet (diethyltoluamide) were field tested against natural populations of tsetse flies, mostly Glossina morsitans centralis Machado, in central Zambia. Volunteers wore different combinations of clothing impregnated with permethrin 0.125 mg ai/cm2 and repellents while riding in a vehicle that was driven slowly (4-6 km/h), with the windows and rear door open, through fly-infested areas. The mean rate of tsetse bites was about twenty per 75 min for unprotected people. The treatment combination of permethrin-impregnated clothing (blue cotton coveralls) and either of two controlled-release deet formulations on exposed skin of face and arms provided 91% mean protection, but this was not significantly better (P greater than 0.05) than wearing deet repellent alone (76-87% protection). No significant differences of protection were observed between the three repellent treatments, although the two controlled-release formulations (intended to be more persistent) were applied at approximately half the dosage of the standard 75% deet. Wearing permethrin-impregnated coveralls alone provided relatively poor protection (34%) for the untreated and exposed skin of head and hands. However, olive drab mesh jackets treated with permethrin reduced the tsetse biting rate by 75%.     

Crustecdysone Mediated Changes in Crayfish

Molting in crayfish, essential for growth is preceded by a decreasein total organic content during premolt. Mineral, as calcium,is conserved in gastroliths after mobilization from the exoskeleton.Crustecdysone treatment results in gastrolith formation by intactcrayfish in intermolt, state C, but does not influence the rateof gastrolith formation in eyestalkless animals. Moreover, therate of gastrolith formation is not increased at higher hormonedoses; conversely, less dense, abnormal gastrohths result, whileapolysis is evident by 48 to 60 hours after treatment. It appearsthat crustecdysone promotes reabsorption of the organic matrixsince treatment with this hormone does not lead to a measureabledecrease in calcium content of shell based on dry weight. Premolt crayfish normally have a higher amino acid pool in alltissues than do those in intermolt, and eyestalk removal fromintermolt crayfish results in an increase in the free-aminoacid level of all tissues by 24 hours; crustecdysone treatmentresults in a significant increase in muscle amino acids by 12hours after a hormone dose of 5 µg/g body weight. In thecontext of integumentary growth and protein synthesis low dosesof crustecdysone (1 µg/g) increase the in vivo incorporationof amino acid into hypodermis, but not hepatopancreas, by 24hours, and the increase is significant by 36 hours; a higherdose (5 µg) does not change this rate of incorporation.Selective increase in hypodermis protein is not evident in acidphosphatase levels from 12 to 48 hours, but respiration of thistissue is significantly elevated within two hours after hormoneinjection.     

Alteration of the Tetrahymena Genome During Nuclear Differentiation*†

Synopsis.
The DNA of the macro- and the micronucleus of Tetrahymena thermophila has been compared by various biochemical methods. It became evident from their thermal denaturation temperatures and buoyant densities that the 2 DNAs were very similar in overall composition. Small differences were detected when the sequence complexities of these DNAs were compared by DNA renaturation studies. The studies suggested that ˜ 10% of the micronuclear genome was lost or underrepresented in the macronucleus. Comparison of individual gene levels revealed further differences. By using the technic of gene cloning a micronuclear sequence was isolated which hybridized only with micronuclear, but not with macronuclear DNA. These results indicated the occurrence of elimination or underreplication of this sequence in the macronucleus. Gene amplification was also shown to occur. In the micronucleus only a single copy of rDNA was found integrated into the chromosome. During macro-nuclear development, amplification was observed to occur, and the amount of rDNA to increase, until there were ˜ 200 copies per haploid genome in the mature macronucleus. all of them extrachromosomal and palindromic. The 3rd case of alteration involved a simple repeated sequence, (CCCCAA)n, present in the termini of rDNA and also in many other locations of the genome. Restriction endonuclease digestion studies revealed drastic differences in the organization of the repeats between macro-and micronucleus. These differences may be interpreted as the results of chromosome fragmentation which occurs at every cluster of the repeats during macronuclear development. The relationship between this event and gene amplification and elimination is discussed.     

Phylogeography and species discrimination in the Paracalliope fluviatilis species complex (Crustacea: Amphipoda): can morphologically similar heterospecifics identify compatible mates?

We examined the phylogeographic patterns of Paracalliope fluviatilis (Amphipoda) over its entire range and the prevalence of mate discrimination in laboratory mate choice tests using genetically distinct populations. We predicted that mate discrimination would increase as the level of genetic divergence increased between populations. Thirty different haplotypes were found with pairwise sequence divergences in the range 1–23.5% between locations. Individuals were selected from seven genetically distinct populations and males were presented with ‘local’ (same population) or ‘foreign’ (genetically divergent) females. Males were more likely to pair with local than foreign females but there was no evidence of a gradual increase in discrimination. Discrimination became most prominent (approximately 5 : 1 local : foreign) when genetic divergences exceeded approximately 20%; matings between divergent individuals also resulted in significantly fewer females producing eggs. We suggest that: (1) this abrupt shift in discrimination occurs because individuals from different, but similarly divergent, clades rely on different recognition cues (e.g. moulting pheromones with disparate chemical signatures) that trigger recognition and subsequent discrimination of incompatible mates; (2) geological history associated with sea level changes and a series of isolation events may be responsible for the patterns of discrimination that we observed; and (3) amphipods may be more genetically variable relative to other invertebrate taxa. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99 , 196–205.     

Induction of Unresponsiveness to Canine Renal Allografts by Total Body Irradiation and Bone Marrow Transplantation

RECENT progress in the definition of the DL-A antigens within a closely bred colony of beagles maintained at the Mary Imogene Basset Hospital (Cooperstown colony) has shown that these antigens are components of the principal system of histocompatibility (DL-A) in the dog^1–3. Dausset and associates⁴ have described the segregation of DL-A antigens in 679 offspring of 151 consecutive matings within the Coopers-town colony. Through methods of backcross analysis, 23 DL-A haplotypes (that is, specificities determined by the DL-A region of one single chromosome) have been identified and the DL-A genotypes of 1,302 offspring of 517 matings in this colony have been defined⁴. This combined genetic and serological information has provided an opportunity to reassess the reported survival rates of allogeneic bone marrow transplantation in serologically matched littermate dogs⁵. We describe here the long term survival of bone marrow transplants and renal allografts obtained from the same donors in nine littermate and three non-littermate recipients selected on the basis of genetic and serological criteria of DL-A compatibility in the Cooperstown colony. The results are contrasted with the survival of twenty-two renal allografts performed in untreated recipients selected on the basis of similar criteria.     

Immunofluorescent Localization of Myosin at the Anterior Pole of the Coccidian,Toxoplasma gondi1

Indirect immunofluorescence using anti-myosin rabbit sera showed myosin localized in a characteristic pattern at the anterior pole of Toxoplasma gondii. This polar fluorescent staining was abolished by pre-absorption of the anti-sera with myosin extracted from avian muscle. Both intracellular and extracellular T. gondii showed similar patterns when formaldehyde-fixed, but neither showed polar fluorescence when acetone was used as the sole fixative. Immunofluorescent staining of live T. gondii revealed no polar fluorescence, suggesting that myosin is not present on the outer parasite membrane. Anti-myosin serum did not prevent host cell invasion and plaque formation in the presence of human complement. Inhibition of contractile proteins with cytochalasin D inhibited T. gondii motility and infectivity in a plaque formation assay. The pattern of polar fluorescence described here resembles the IgM-associated polar staining frequently detected in human sera, but we believe it is a different phenomenon because human sera that showed such staining retained their activity after pre-absorption with avian myosin. The unusual localization of myosin at the anterior pole of T. gondii tachyzoites may play a role in the function of anterior organelles, which are thought to facilitate the invasion of host cells.     

Short-Term Effects of Fire and Other Fuel Reduction Treatments on Breeding Birds in a Southern Appalachian Upland Hardwood Forest

Abstract: We compared the effects of 3 fuel reduction techniques and a control on breeding birds during 2001-2005 using 50-m point counts. Four experimental units, each >14 ha, were contained within each of 3 replicate blocks at the Green River Game Land, Polk County, North Carolina, USA. Treatments were 1) prescribed burn, 2) mechanical understory reduction (chainsaw-felling of shrubs and small trees), 3) mechanical + burn, and 4) controls. We conducted mechanical treatments in winter 2001-2002 and prescribed burns in spring 2003. Tall shrub cover was substantially reduced in all treatments compared to controls. Tree mortality and canopy openness was highest in the mechanical + burn treatment after burning, likely due to higher fuel loading and hotter burns; tree mortality increased with time. Many bird species did not detectably decrease or increase in response to treatments. Species richness, total bird density, and some species, including indigo buntings (Passerina cyanea) and eastern bluebirds (Sialia sialis), increased in the mechanical + burn treatment after a 1-year to 2-year delay; eastern woodpewees (Contopus virens) increased immediately after treatment. Hooded warblers (Wilsonia citrina), black-and-white warblers (Mniotilta varia), and worm-eating warblers (Helmitheros vermivorus) declined temporarily in some or all treatments, likely in response to understory and (or) leaf litter depth reductions. Densities of most species affected by treatments varied with shrub cover, tree or snag density, or leaf litter depth. High snag availability, open conditions, and a higher density of flying insects in the mechanical + burn treatment likely contributed to increased bird density and species richness. In our study, fuel reduction treatments that left the canopy intact, such as low-intensity prescribed fire or mechanical understory removal, had few detectable effects on breeding birds compared to the mechanical + burn treatment. High-intensity burning with heavy tree-kill, as occurred in our mechanical + burn treatment, can be used as a management tool to increase densities of birds associated with open habitat while retaining many forest and generalist species, but may have short-term adverse effects on some species that are associated with the ground- or shrub-strata for nesting and foraging.