Mammal dung-dung beetle trophic networks: an improved method based on gut-content DNA.

Background: Dung beetles provide many important ecosystem services, including dung decomposition, pathogen control, soil aeration, and secondary seed dispersal. Yet, the biology of most dung beetles remains unknown. Natural diets are poorly studied, partly because previous research has focused on choice or attraction experiments using few, easily accessible dung types from zoo animals, farm animals, or humans. This way, many links within natural food webs have certainly been missed. In this work, we aimed to establish a protocol to analyze the natural diets of dung beetles using DNA gut barcoding. Methods: First, the feasibility of gut-content DNA extraction and amplification of 12s rDNA from six different mammal dung types was tested in the laboratory. We then applied the method to beetles caught in pitfall traps in Ecuador and Germany by using 12s rDNA primers. For a subset of the dung beetles caught in the Ecuador sampling, we also used 16s rDNA primers to see if these would improve the number of species we could identify. We predicted the likelihood of amplifying DNA using gut fullness, DNA concentration, PCR primer, collection method, and beetle species as predictor variables in a dominance analysis. Based on the gut barcodes, we generated a dung beetle-mammal network for both field sites (Ecuador and Germany) and analyzed the levels of network specificity. Results: We successfully amplified mammal DNA from dung beetle gut contents for 128 specimens, which included such prominent species as Panthera onca (jaguar) and Puma concolor (puma). The overall success rate of DNA amplification was 53%. The best predictors for amplification success were gut fullness and DNA concentration, suggesting the success rate can be increased by focusing on beetles with a full gut. The mammal dung-dung beetle networks differed from purely random network models and showed a moderate degree of network specialization (H2': Ecuador = 0.49; Germany = 0.41). Conclusion: We here present a reliable method of extracting and amplifying gut-content DNA from dung beetles. Identifying mammal dung via DNA reference libraries, we created mammal dung-dung beetle trophic networks. This has benefits over previous methods because we inventoried the natural mammal dung resources of dung beetles instead of using artificial mammal baits. Our results revealed higher levels of specialization than expected and more rodent DNA than expected in Germany, suggesting that the presented method provides more detailed insights into mammal dung-dung beetle networks. In addition, the method could have applications for mammal monitoring in many ecosystems.

Morphometric changes on dung beetle Dichotomius problematicus (Coleoptera: Scarabaeidae: Scarabaeinae) related to conversion of forest into grassland: A case of study in the Ecuadorian Amazonia.

The conversion of forest into grassland can induce differentiation in the functional morphology of resilient species. To assess this effect, we have chosen a dung beetle Dichotomius problematicus, as a model species. We established 20 sampling points distributed along a transect for a forest and grassland located in the Podocarpus National Park in Ecuador. Four pit-fall traps were baited with pig feces per sample point and were left open for 48 h. We sexed and measured 13 morphological traits of 269 individuals. Nonmetric multidimensional scaling was carried out to evaluate the influence of habitat and sexual dimorphism on the traits. We applied a principal component analysis to evaluate the morphological features that best explain the differences between land use and sexual dimorphism. We used generalized linear models to evaluate the explanatory variables: habitat and sexual dimorphism with respect to morphological traits. Five traits contributed over 70% body thickness, Pronotum width, Pronotum length, Head width and Elytra length, following the results of a principal component analysis. Both habitat and sex influence traits. In the forest, the individuals are larger than grassland likely due to available resources, but in grassland, the structures in charge of the burial process head, protibia are larger, displaying a strong pronotum and possible a greater reproductive capacity given by spherecity. These patterns of changes in the size of beetles and their structures could reflect the conservation state of an ecosystem.

Different motifs regulate trafficking of SorCS1 isoforms.

The type I transmembrane protein SorCS1 is a member of the Vps10p-domain receptor family comprised of Sortilin, SorLA and SorCS1, -2 and -3. Current information indicates that Sortilin and SorLA mediate intracellular protein trafficking and sorting, but little is known about the cellular functions of the SorCS subgroup. SorCS1 binds platelet-derived growth factor-BB (PDGF-BB) and is expressed in isoforms differing only in their cytoplasmic domains. Here, we identify two novel isoforms of mouse SorCS1 designated m-SorCS1c and -d. In situ hybridization revealed a combinatorial expression pattern of the variants in brain and embryonic tissues. We demonstrate that among the mouse variants, only SorCS1c mediates internalization and that the highly conserved SorCS1c is internalized through a canonical tyrosine-based motif. In contrast, human SorCS1a, whose cytoplasmic domain is completely different from mouse SorCS1a, is internalized through a DXXLL motif. We report that the human SorCS1a cytoplasmic domain interacts with the alphaC/sigma2 subunits of the adaptor protein (AP)-2 complex, and internalization of human SorCS1a and -c is mediated by AP-2. Our results suggest that the endocytic isoforms target internalized cargo to lysosomes but are not engaged in Golgi-endosomal transport to a significant degree.

Fkbp8: novel isoforms, genomic organization, and characterization of a forebrain promoter in transgenic mice.

The immunophilin homolog FKBP8 has been implicated in the regulation of apoptosis. Here we show that the 38-kDa form of FKBP8 (FKBP38) derives from a truncated ORF. The extended FKBP8 ORFs are 46 and 44 kDa in mouse and 45 kDa in human. Although the genomic organization of mouse and human FKBP8 is evolutionarily conserved, additional first exons are encoded by the murine locus. A 4.4-kb murine Fkbp8 gene fragment, containing a GC-rich potential promoter, directed expression of a LacZ reporter gene to forebrain neurons in transgenic mice. Expression of the transgene was observed in CA1 pyramidal neurons of the hippocampus in transgenic mice from three lines. One transgenic founder mouse exhibited widespread forebrain expression of the LacZ transgene that resembles the pattern for the endogenous Fkbp8 gene. Thus promoter/enhancer elements for forebrain expression are located around the first exons of the mouse Fkbp8 gene.

Increased mean arterial pressure and aldosterone-to-renin ratio in Persian cats with polycystic kidney disease.

Polycystic kidney disease (PKD) in Persian cats has been increasingly reported and compared to human autosomal dominant polycystic kidney disease (ADPKD) in the last decade. In cats, however, few studies have dealt with the occurrence and hormonal determinants of hypertension, one of the most common extrarenal manifestations of ADPKD in humans. The purpose of this study was to compare Persian cats >4 years old with PKD to unaffected control cats with regard to blood pressure (BP), plasma renin activity (PRA), serum aldosterone concentration, plasma atrial natriuretic peptide (ANP) concentration, and aldosterone-to-renin ratio (ARR). Three gender- and age-matched groups were studied, each consisting of 7 cats: (1) a control group without cysts, (2) a group with mild PKD, and (3) a group with severe PKD (multiple cysts and renal enlargement). Mild renal insufficiency was found in only 1 of 14 cats with PKD. Cats with PKD had a higher mean arterial pressure (P = .04) and more often had a high ARR (P = .047) than did control cats. Tendencies toward higher diastolic and systolic arterial pressures (DAPs and SAPs, respectively) and lower PRAs were observed in cats with PKD compared to controls (.05 < P < or = .1). No significant differences were found between the groups in serum aldosterone and plasma ANP concentrations. None of the cats had echocardiographic evidence of cardiac hypertrophy. In conclusion, cats with PKD had a minor increase in mean arterial pressure compared to control cats, and half of the cats had a high ARR.

Astroglial c-Myc overexpression predisposes mice to primary malignant gliomas.

Malignant astrocytomas are common human primary brain tumors that result from neoplastic transformation of astroglia or their progenitors. Here we show that deregulation of the c-Myc pathway in developing astroglia predisposes mice to malignant astrocytomas within 2-3 weeks of age. The genetically engineered murine (GEM) gliomas harbor a molecular signature resembling that of human primary glioblastoma multiforme, including up-regulation of epidermal growth factor receptor and Mdm2. The GEM gliomas seem to originate in an abnormal population of glial fibrillary acidic protein-expressing cells in the ventricular zone and, analogous to human glioblastomas, exhibit molecular and morphological heterogeneity. Levels of connexin 43 in the majority of the tumors are unaltered from normal tissue, indicating that GEM tumors have retained the capacity to establish syncytial networks. In line with this, individual glioma foci are composed of a mixture of actively proliferating cells expressing c-Myc and proliferating cell nuclear antigen and less dividing bystander cells that express glial fibrillary acidic protein and the broad complex tramtrack bric-a-brac/poxvirus and zinc finger domain protein HOF. A subset of the transgenic mice harbored, in addition to brain tumors, vestigial cerebellums in which granule cell migration and radial Bergman glial cell differentiation were disturbed. These observations argue for a window of vulnerability during astrocyte development where c-Myc overexpression is sufficient to trigger the neoplastic process, presumably by inducing the sustained growth of early astroglial cells. This is in contrast to most other transgenic studies in which c-Myc overexpression requires co-operating transgenes for rapid tumor induction.

Mouse Atf5: molecular cloning of two novel mRNAs, genomic organization, and odorant sensory neuron localization.

The activating transcription factor (ATF) family comprises a group of basic region-leucine zipper (bZIP) proteins, which have roles in the development of species as diverse as insects and mammals. Here we describe two novel mRNAs encoding a single, 30-kDa mouse polypeptide, designated mouse ATF5, which is 58% identical to mouse ATF4 in the carboxy-terminal bZIP region. Both transcripts harbor highly complex 5' untranslated regions that impede translation of the ATF5 open reading frame. The mouse and human ATF5 loci consist of at least four exons contained within 5 kb of genomic sequence. During mouse embryonic development, expression of Atf5 is pronounced at the late gestational period and appears to be confined to cells of the neuronal layers of the olfactory epithelium and vomeronasal organ. This suggests a role for ATF5 in odorant sensory neuron differentiation.

Evaluation of an oscillometric blood pressure monitor for use in anesthetized cats.

OBJECTIVE: To determine accuracy of an oscillometric blood pressure monitor used over a wide range of pressures in anesthetized cats. DESIGN: Prospective study. ANIMALS: 6 healthy cats. PROCEDURE: 4 female cats and 2 male cats that weighed 2.7 to 4.5 kg (5.9 to 9.9 lb) and were 2 to 8 years old were anesthetized. Blood pressure was measured directly with an arterial catheter placed in the right femoral artery and indirectly from the left antebrachium by use of an oscillometric monitor. A series of diastolic arterial pressure (DAP), mean arterial pressure (MAP), and systolic arterial pressure (SAP) measurements were obtained during hypotension, normotension, and hypertension. Values obtained indirectly and directly were compared. RESULTS: The oscillometric monitor was accurate for DAP and MAP throughout the entire pressure range and met the standards of the Association for the Advancement of Medical Instrumentation (mean +/- SD difference from values obtained directly, < or = 5 +/- 8 mm Hg). The SAP was increasingly underestimated with increasing overall pressure; mean differences from direct measurements were -5.2, -12.1, and -17.7 mm Hg during hypo-, normo-, and hypertension, respectively. Standard deviations for SAP were all < or = 8 mm Hg. The monitor gave readings during all attempts. The direct blood pressure recording system appeared to perform well with neither under- nor overdamping. CONCLUSIONS AND CLINICAL RELEVANCE: Except for a minor underestimation of SAP during normo- and hypertension, the oscillometric monitor yielded reliable and easily obtainable blood pressure measurements in anesthetized cats.

Characterization of two novel nuclear BTB/POZ domain zinc finger isoforms. Association with differentiation of hippocampal neurons, cerebellar granule cells, and macroglia.

BTB/POZ (broad complex tramtrack bric-a-brac/poxvirus and zinc finger) zinc finger factors are a class of nuclear DNA-binding proteins involved in development, chromatin remodeling, and cancer. However, BTB/POZ domain zinc finger factors linked to development of the mammalian cerebral cortex, cerebellum, and macroglia have not been described previously. We report here the isolation and characterization of two novel nuclear BTB/POZ domain zinc finger isoforms, designated HOF(L) and HOF(S), that are specifically expressed in early hippocampal neurons, cerebellar granule cells, and gliogenic progenitors as well as in differentiated glia. During embryonic development of the murine cerebral cortex, HOF expression is restricted to the hippocampal subdivision. Expression coincides with early differentiation of presumptive CA1 and CA3 pyramidal neurons and dentate gyrus granule cells, with a sharp decline in expression at the CA1/subicular border. By using bromodeoxyuridine labeling and immunohistochemistry, we show that HOF expression coincides with immature non-dividing cells and is down-regulated in differentiated cells, suggesting a role for HOF in hippocampal neurogenesis. Consistent with the postulated role of the POZ domain as a site for protein-protein interactions, both HOF isoforms are able to dimerize. The HOF zinc fingers bind specifically to the binding site for the related promyelocytic leukemia zinc finger protein as well as to a newly identified DNA sequence.