Aspidoscelis laredoensis and A. gularis hybridization.

13 pages : illustrations (some color) ; 26 cm.Karyotypes and allozyme data for 32 genetic loci overwhelmingly support the conclusion that Aspidoscelis laredoensis is a diploid all-female species that had a hybrid origin between A. gularis x A. sexlineatus. Comparisons of allozymes in individuals representing three mother-to-daughter generations raised in the laboratory suggest that they reproduce by parthenogenetic cloning. In addition to two previously described morphotypes (pattern classes A and B) that occur in southern Texas, we report the existence of three all-female clonal lineages based on allozymes. Individuals of at least one of these lineages occasionally hybridize in nature with males of A. gularis, producing viable and healthy triploid offspring that can grow to adulthood, one of which herself produced an offspring in the laboratory and could have represented a new, clonal triploid species. The possibility exists that cloned offspring of triploid hybrids are present in South Texas and/or northern Mexico, awaiting discovery. These would represent a new species that would appear to be very similar to A. laredoensis

POPDC1 scaffolds a complex of adenylyl cyclase 9 and the potassium channel TREK-1 in heart

The establishment of macromolecular complexes by scaffolding proteins is key to the local production of cAMP by anchored adenylyl cyclase (AC) and the subsequent cAMP signaling necessary for cardiac functions. We identify a novel AC scaffold, the Popeye domain-containing (POPDC) protein. The POPDC family of proteins is important for cardiac pacemaking and conduction, due in part to their cAMP-dependent binding and regulation of TREK-1 potassium channels. We show that TREK-1 binds the AC9:POPDC1 complex and copurifies in a POPDC1-dependent manner with AC9 activity in heart. Although the AC9:POPDC1 interaction is cAMP-independent, TREK-1 association with AC9 and POPDC1 is reduced upon stimulation of the β-adrenergic receptor (βAR). AC9 activity is required for βAR reduction of TREK-1 complex formation with AC9:POPDC1 and in reversing POPDC1 enhancement of TREK-1 currents. Finally, deletion of the gene-encoding AC9 (Adcy9) gives rise to bradycardia at rest and stress-induced heart rate variability, a milder phenotype than the loss of Popdc1 but similar to the loss of Kcnk2 (TREK-1). Thus, POPDC1 represents a novel adaptor for AC9 interactions with TREK-1 to regulate heart rate control

Studies of the dose-effect relation

Dose-effect relations and, specifically, cell survival curves are surveyed with emphasis on the interplay of the random factors — biological variability, stochastic reaction of the cell, and the statistics of energy deposition —that co-determine their shape. The global parameters mean inactivation dose, , and coefficient of variance, V, represent this interplay better than conventional parameters. Mechanisms such as lesion interaction, misrepair, repair overload, or repair depletion have been invoked to explain sigmoid dose dependencies, but these notions are partly synonymous and are largely undistinguishable on the basis of observed dose dependencies. All dose dependencies reflect, to varying degree, the microdosimetric fluctuations of energy deposition, and these have certain implications, e.g. the linearity of the dose dependence at small doses, that apply regardless of unresolved molecular mechanisms of cellular radiation action

Structural basis of adenylyl cyclase 9 activation

Adenylyl cyclase 9 (AC9) is a membrane-bound enzyme that converts ATP into cAMP. The enzyme is weakly activated by forskolin, fully activated by the G protein Gαs subunit and is autoinhibited by the AC9 C-terminus. Although our recent structural studies of the AC9-Gαs complex provided the framework for understanding AC9 autoinhibition, the conformational changes that AC9 undergoes in response to activator binding remains poorly understood. Here, we present the cryo-EM structures of AC9 in several distinct states: (i) AC9 bound to a nucleotide inhibitor MANT-GTP, (ii) bound to an artificial activator (DARPin C4) and MANT-GTP, (iii) bound to DARPin C4 and a nucleotide analogue ATPαS, (iv) bound to Gαs and MANT-GTP. The artificial activator DARPin C4 partially activates AC9 by binding at a site that overlaps with the Gαs binding site. Together with the previously observed occluded and forskolin-bound conformations, structural comparisons of AC9 in the four conformations described here show that secondary structure rearrangements in the region surrounding the forskolin binding site are essential for AC9 activation

Hybridization among whiptail lizards.

43 p. : ill. (some col.) ; 26 cm. "October 11, 2010."The natural origin of diploid parthenogenesis in whiptail lizards has been through interspecific hybridization. Genomes of the parthenogens indicate that they originated in one generation, as the lizards clone the F₁ hybrid state. In addition, hybridization between diploid parthenogens and males of bisexual species has resulted in triploid parthenogenetic clones in nature. Consequently, the genus Aspidoscelis contains numerous gonochoristic (= bisexual) species and numerous unisexual species whose closest relatives are bisexual, and from whom they originated through instantaneous sympatric speciation and an abrupt and dramatic switch in reproductive biology. In order to study this phenomenon more closely, with hopes (unfulfilled) to witness the origin of parthenogenetic cloning in one generation, we maintained whiptail lizards in captivity. For more than 29 years, we caged males of bisexual species with females of bisexual and of unisexual species in attempts to obtain laboratory hybrids. Hybrids were raised to adulthood to see whether they would reproduce, but none did. The hybrid status of suspected laboratory hybrids was confirmed by karyotypic, allozyme, and morphological analyses, and histological studies were made on reproductive tissues of the hybrids, which were apparently sterile. The present paper focuses on the laboratory hybrids of two bisexual species, A. inornata arizonae ([female]) x A. tigris marmorata ([male]). These three individuals from one clutch of eggs were the only hybrids between two bisexual species that we obtained. The hybrids had a karyotype, allozymes (21 loci tested), and external morphology that were similar to those of A. neomexicana, which is a diploid parthenogen that had a hybrid origin in nature that was the reciprocal cross: A. t. marmorata ([female]) x A. inornata ([male]). Histological study showed that the largest and oldest laboratory hybrid raised, which appeared to be a female with inherited X chromosome of A. t. marmorata, was an intersex with an enormous adrenal. The other hybrid that reached adult size, a male, was also apparently sterile. Later, we review and summarize the information on the other laboratory hybrids we obtained over the years. These include two different combinations of hybrids between a male of a bisexual species and females of unisexual species (one diploid, one triploid), producing triploid and tetraploid hybrids, respectively, as a haploid genome from the male was added to the cloned egg. Considering only those specimens whose hybrid status was confirmed with genetic analyses, a total of only five hybrids from three crosses were obtained over 29 years. The effort involved having a total of 74 males of four species caged with 156 females of nine species, where individuals were caged together for at least six months (or less, if mating behavior was observed). Despite our extensive efforts to provide for their comfort and best health and captive environment, the lizards at times experienced health problems such as metabolic bone disease and a Salmonella infection. These definitely had a negative effect on reproduction, the full extent of which is unknown. Nevertheless, we estimate that successful hybridization among whiptail lizards (i.e., which results in healthy offspring capable of reproduction) is much more rare than we previously thought, although, paradoxically, it is far more common among Aspidoscelis than among nearly all other genera of lizards in the world, with the possible exception of lacertids

Structural basis of adenylyl cyclase 9 activation

Adenylyl cyclase 9 (AC9) is a membrane-bound enzyme that converts ATP into cAMP. The enzyme is weakly activated by forskolin, fully activated by the G protein Gαs subunit and is autoinhibited by the AC9 C-terminus. Although our recent structural studies of the AC9-Gαs complex provided the framework for understanding AC9 autoinhibition, the conformational changes that AC9 undergoes in response to activator binding remains poorly understood. Here, we present the cryo-EM structures of AC9 in several distinct states: (i) AC9 bound to a nucleotide inhibitor MANT-GTP, (ii) bound to an artificial activator (DARPin C4) and MANT-GTP, (iii) bound to DARPin C4 and a nucleotide analogue ATPαS, (iv) bound to Gαs and MANT-GTP. The artificial activator DARPin C4 partially activates AC9 by binding at a site that overlaps with the Gαs binding site. Together with the previously observed occluded and forskolin-bound conformations, structural comparisons of AC9 in the four conformations described here show that secondary structure rearrangements in the region surrounding the forskolin binding site are essential for AC9 activation

Morphological characterization of the blood cells in the endangered Sicilian endemic pond turtle,Emys trinacris(Testudines: Emydidae)

In this study, measurements of morphological parameters, sizes and frequencies of peripheral blood cells (erythrocytes, leukocytes, thrombocytes) on blood smear preparation devices stained with May-Grünwald stain were evaluated for both sexes in 20 Emys trinacris (Testudines: Emydidae) specimens. Erythrocytes were higher in male than in female specimens. The leukocyte of E. trinacris contains eosinophil, basophil, monocyte, heterophil and lymphocyte. The eosinophil was higher in males than in females whereas lymphocytes were higher in females than in males. The erythrocyte morphological parameters (EL [erythrocyte length], EW [erythrocyte width], L/W [length/width], ES [erythrocyte size]) were compared with the same data from Emys orbicularis s.l, and from species belonging to other chelonian genera. The erythrocyte size did not vary within the studied Palearctic Emys taxa, whereas it proved to differ from that observed in other chelonians

Accurate automated quantitative imaging of tortoise erythrocytes using the NIS image analysis system

The standard method for assessing blood cell characteristics using an ocular micrometer is time-consuming and limited. We used the Nikon NIS Elements imaging software and May-Grünwald-Giemsa staining to determine whether automated image analysis is suitable for rapid and accurate quantitative morphometry of erythrocytes. Blood was collected during four seasons from 126 geometric tortoises and the blood smears were evaluated for cell (C) and nuclear (N) characteristics of the erythrocytes. We measured area, length (L), width (W), perimeter, elongation and pixelation intensity, and calculated L/W and N/C areas. Erythrocyte size differed among cohorts; females, the larger sex, had smaller erythrocytes than either males or juveniles. Males had more elongated erythrocytes than females and erythrocytes of adults were more elongated than those of juveniles. Erythrocyte size and shape influence the efficiency of gas exchange owing to surface area to volume ratios, which are greater for small, elongated cells than for large, round cells. The high N/C ratio and low pixelation intensities of males and juveniles indicate that they may have had more immature erythrocytes in their circulation than females. The use of pixelation intensity to indicate the presence of immature erythrocytes was validated by seasonal differences that corresponded to the biology of the tortoises. Pixelation intensity was lowest in winter. We found that automated image analysis is a rapid and reliable method for determining cell size and shape, and it offers the potential for distinguishing among developmental stages that differ in staining intensity. The method should be useful for rapid health assessments, particularly of threatened species, and for comparative studies among different vertebrates.Web of Scienc

The Circadian Neuropeptide PDF Signals Preferentially through a Specific Adenylate Cyclase Isoform AC3 in M Pacemakers of Drosophila

To synchronize a network of pacemakers in the Drosophila brain, a neuropeptide receptor specifically associates with adenylate cyclase 3 to create a “circadian signalosome.