Defective retinal depolarizing bipolar cells in regulators of G protein signaling (RGS) 7 and 11 double null mice

Two members of the R7 subfamily of regulators of G protein signaling, RGS7 and RGS11, are present at dendritic tips of retinal depolarizing bipolar cells (DBCs). Their involvement in the mGluR6/Gα(o)/TRPM1 pathway that mediates DBC light responses has been implicated. However, previous genetic studies employed an RGS7 mutant mouse that is hypomorphic, and hence the exact role of RGS7 in DBCs remains unclear. We have made a true RGS7-null mouse line with exons 6-8 deleted. The RGS7(-/-) mouse is viable and fertile but smaller in body size. Electroretinogram (ERG) b-wave implicit time in young RGS7(-/-) mice is prolonged at eye opening, but the phenotype disappears at 2 months of age. Expression levels of RGS6 and RGS11 are unchanged in RGS7(-/-) retina, but the Gβ5S level is significantly reduced. By characterizing a complete RGS7 and RGS11 double knock-out (711dKO) mouse line, we found that Gβ5S expression in the retinal outer plexiform layer is eliminated, as is the ERG b-wave. Ultrastructural defects akin to those of Gβ5(-/-) mice are evident in 711dKO mice. In retinas of mice lacking RGS6, RGS7, and RGS11, Gβ5S is undetectable, whereas levels of the photoreceptor-specific Gβ5L remain unchanged. Whereas RGS6 alone sustains a significant amount of Gβ5S expression in retina, the DBC-related defects in Gβ5(-/-) mice are caused solely by a combined loss of RGS7 and RGS11. Our data support the notion that the role of Gβ5 in the retina, and likely in the entire nervous system, is mediated exclusively by R7 RGS proteins.     

Facts From Feces: Nitrogen Still Measures Up as a Nutritional Index for Mammalian Herbivores

Abstract: Fecal nitrogen (FN) has been applied widely as an index of dietary quality in studies of nutritional ecology of free-ranging and captive vertebrate herbivores, particularly ruminants. Three related articles in the Journal of Wildlife Management (JWM; Leslie and Starkey 1985, 1987; Hobbs 1987) have been cited (n = 150) in 87 publications and 39 peer-reviewed journals. The critique by Hobbs (1987) and the reply by Leslie and Starkey (1987) on limitations and appropriate applications of FN have been used to justify use of FN or negate its value as a nutritional proxy. We undertook a retrospective analysis of FN applications since 1985, largely because we sensed that methodological cautions noted in the 3 JWM publications were not being followed, leading to faulty conclusions and management applications, and that application protocols needed updating. From January 1986 through July 2007, the 107 species-by-continent applications of FN, citing the 3 JWM publications singly or in any combination, were diverse; FN was used in various ways on 5 continents and for 50 wild and domestic species. Cumulative rates of departure from recommended FN applications increased in recent years, largely in studies that compare different species while failing to fully acknowledge that differences likely reflected digestive capabilities rather than differences in some aspect of dietary intake. Post-1985 research on plant secondary compounds (e.g., tannins) has refined limitations to the application of FN, permitting more straightforward protocols than were possible in 1985. Although use does not necessarily reflect value, the number of published applications during the past 22 years indicates that peer reviewers from a variety of scientific disciplines view FN as a suitable proxy for nutritional status, which can be used to contrast study units when carefully defined by the study design. Any index can have shortcomings, and there are still circumstances when application of FN is problematic. Precise prediction of intake with FN under field conditions is still hampered by inherent variability, but contrasts of comparable study units and species can be appropriate. Published protocols for FN, as amended herein, should be adhered to, and conclusions are strengthened by the use of multiple nutritional indices.     

The Avian Shoulder: An Experimental Approach

SYNOPSIS. This essay is in two parts. The first describes functionalstudies of the shoulder in modern vertebrates that led to theformulation of the hypotheses that motor patterns of homologousmuscles have been maintained during the evolution of the tetrapodshoulder, and that a primitive organization of the neural controlcomponents has persisted in derived groups. The second part of this essay focuses upon a longstanding questionin vertebrate evolution: what neuromuscular and musculoskeletalchanges in the tetrapod shoulder accompanied the evolution offlight in birds? The lack of empirical data on shoulder functionin extant birds limited our insight into this question, andprompted our initiation of experimental studies. Preliminarykinematics of the furcula and humerus of European starlings(Sturnus vulgaris) flying in a wind tunnel, as revealed by highspeed cineradiography, are presented. The two halves of thefurcula, which contact the coracoids dorsally, are bent laterallyduring downstroke and medially during upstroke by as much as60% of the intrafurcular resting distance. High speed film andelectromyographic studies of freeflying pigeons (Columba livia)reveal that the supracoracoideus muscle is strongly activatedduring wing elevation and, as predicted from studies of Varanusand Didelphis, an additional activation burst occurs at mid-downstrokein 48% of the recordings.     

The effects of injection of amphetamine on female insemination in the black blow fly, Phormia regina (Diptera: Calliphoridae)

Abstract. An earlier study on the blowfly Phormia regina (Meigen) demonstrated that the injection of amphetamine (12 μg) depletes biogenic amine levels in the CNS. In the present study, P. regina females were injected with amphetamine (12 μg), each female was placed with three males and insemination success was determined. Amphetamine inhibited female insemination by 43.3% at 2–90 min post-injection and by 70% at 10–60 min post-injection. At 180–270 min post-injection, there was no significant inhibition of female insemination. This study indicates a possible role in insects for the biogenic amines in female insemination.     

Azure a-Schiff, Alcian Blue, HIO4-Schiff, Naphthol Yellow S—A Sequential Staining Method for Paraffin Sections

Paraffin sections from tissue fixed 4-12 hr in 10% formalin containing 0.5% cetyl pyridinium chloride, and washed 2 hr, were stained as follows: (1) Hydrolyze in 5 N HCl at room temperature for 8.5-9 min, or use standard Feulgen hydrolysis at 60 C. (2) Stain in azure A-Schiff, 0.5% in bisulfite bleach (1 N HCl, 5; 10% Na₂S₂O₅, 5; and distilled water 90—parts by volume) for 10 min. (3) Place in bisulfite bleach 2 changes, 2 min each; wash in water, 1-2 min. (4) Stain in Alcian blue (0.1% in 0.01 2V HCl, pH 2.0) for 10 min. (5) Place in 0.01 N HCl for 2-3 min; wash in water for 1-2 min. (6) Oxidize in 0.5% HIO₄ for 5 min; wash in water, 1-2 min. (7) Stain in Schiff's leucofuchsiu, 10 min. (8) Treat with bisulfite bleach as in step 3; wash in running water, 10 min. (9) Stain in naphthol yellow S (0.01% in 1% acetic acid) for 1-2 min. (10) Place in 1% acetic acid for 2 min, dehydrate in tertiary butanol, clear and cover. Result: DNA is deep blue; acidic mucins are light blue; neutral polysaccharides, red to magenta; and proteins, yellow. Proper timing of the hydrolysis for the Feulgen reaction is the most critical step. Overhydrolysis results in green nuclei (staining by naphthol yellow S) whereas purplish nuclei are the results of insufficient hydrolysis.     

Occupancy of Mountain Plover and Burrowing Owl in Colorado

Abstract: Concern over the decline of grassland birds has spurred efforts to increase understanding of grassland bird-habitat relationships. Previous studies have suggested that black-tailed prairie dogs (Cynomys ludovicianus) provide important habitat for shortgrass prairie avifauna, such as mountain plover (Charadrius montanus) and western burrowing owl (Athene cunicularia hypugaea), although such studies are lacking in Colorado (USA). We used methods to estimate occupancy (ψ) of mountain plover and burrowing owl on prairie dog colonies and other shortgrass prairie habitats in eastern Colorado. Mountain plover occupancy was higher on prairie dog colonies (ψ = 0.50, 95% CI = 0.36–0.64) than on grassland (ψ = 0.07, 95% CI = 0.03–0.15) and dryland agriculture (ψ = 0.13, 95% CI = 0.07–0.23). Burrowing owl occupancy was higher on active prairie dog colonies (ψ = 0.80, 95% CI = 0.66–0.89) compared with inactive colonies (ψ = 0.23, 95% CI = 0.07–0.53), which in turn was much higher than on grassland (ψ = 0.01, 95% CI = 0.00–0.07) and dryland agriculture (ψ = 0.00, 95% CI ψ 0.00–0.00). Mountain plover occupancy also was positively correlated with increasing amounts of prairie dog colony in the landscape. Burrowing owl occupancy was negatively correlated with increasing amounts of prairie dog colony in the surrounding landscape. Our results suggest that actions to conserve mountain plovers and burrowing owls should incorporate land management to benefit prairie dogs. Because managing for specific colony attributes is difficult, alternative management that promotes heterogeneity may ensure that suitable habitat is available for the guild of grassland inhabitants.     

Historical and Ecological Factors in the Biogeography of Aposematic Neotropical Butterflies

SYNOPSIS. Quantitative analysis of low-level (mostly subspecific)geographic differentiation of aposematic color patterns in threegroups of forest butterflies (Heliconiini, Ithomiinae, and Troidini),in 1,550 half-degree quadrants throughout the Neotropics, indicatesfour principal species-endemicity regions, 44 principal subspecies-endemicitycenters, and 300 quadrants of subspecies hybridization. Theendemism centers are different in outline but nearly identicalin position for each of the three ecologically different groups.Basic data on present and past physical environments (climate,soils, geomorphology, and vegetation structures), when integratedinto maps, show 1) regions of favorable ecological conditionsfor forest organisms in the present, 2) regions of environmentalconformities and rapid transitions in the present, and 3) regionsof high probability for humid forest persistence during themajor landscape changes at the end of the last glacial period,13,000–20,000 yr ago. Species diversity (a local phenomenon)is readily related to favorable modern ecological conditionsand environmental microheterogeneity, promoted by unpredictablemild disturbance. Biogeographical patterns of evolution at thelevel of regional subspecies correlate very well with presumedpaleoecological conditions. The biogeographical data also supportan hypothesis of differentiation resulting from areas of rapidenvironmental change; most of the hybridization quadrants fallwithin transition zones. There is lower correlation betweencenters of endemism and zones of en vironmental conformity,but some individual cases of differentiation in the butterfliesmay be adequately explained by modern ecological factors. Speciespresently undergoing differentiation in isolated habitat islands(Amazonian savannas and high-altitude Andean forests and paramos)may offer the best opportunities for observation of evolutionaryprocesses in the present.     

Biomechanical and Morphometric Differences in Triticum aestivum Seedlings Differing in Rht Gene-Dosage

Biomechanical and morphometric comparisons among coleoptilesfrom wheat seedlings differing in Rht gene-dosage (Rht = 0,2, 4 doses) are presented in an effort to evaluate the influenceof Rht on the mechanics of soil penetration by this organ. Rhtis known to reduce seedling establishment compared to the wildtype. Data from 3–7-day-old seedlings indicate that Rhtreduces tissue elastic modulus E, increases the second momentof area I, and decreases the slenderness ratio (l/r) of coleoptiles.Rht-relatedchanges in E and I are such that the flexural stiffness of coleoptilesfrom Rht plants does not differ significantly from the wildtype-hence the growing coleoptiles of all three genotypes haveequivalent biomechanical capacity to penetrate the soil. Rhtreduction of coleoptile slenderness ratios confers a capacityto safely sustain higher axial compressive loads compared tocoleoptiles with equivalent flexural stiffness but higher ratios.However, wild type seedlings produce longer coleoptiles andlonger subcrown internodes than Rht seedlings. Longer coleoptilesdeliver the crown node closer to the top of the soil beforethe crown node extends beyond the lateral confinement of thecoleoptile. This reduces the potential for buckling of the subcrowninternode and leaves due to the compressive loading of soil.Rht affects a variety of mechanical features whose influenceis dependent upon the stage of seedling growth and the degreeof soil compaction. However, at equivalent depths of burialwhich exceed the maximum length of coleoptiles and moderatesoil compaction, Rht is biomechanically disadvantageous to seedlingestablishment. Wheat, germination, biomechanics, Rht-gene