A homozygous mutation in the tight-junction protein JAM3 causes hemorrhagic destruction of the brain, subependymal calcification, and congenital cataracts

The tight junction, or zonula occludens, is a specialized cell-cell junction that regulates epithelial and endothelial permeability, and it is an essential component of the blood-brain barrier in the cerebrovascular endothelium. In addition to functioning as a diffusion barrier, tight junctions are also involved in signal transduction. In this study, we identified a homozygous mutation in the tight-junction protein gene JAM3 in a large consanguineous family from the United Arab Emirates. Some members of this family had a rare autosomal-recessive syndrome characterized by severe hemorrhagic destruction of the brain, subependymal calcification, and congenital cataracts. Their clinical presentation overlaps with some reported cases of pseudo-TORCH syndrome as well as with cases involving mutations in occludin, another component of the tight-junction complex. However, massive intracranial hemorrhage distinguishes these patients from others. Homozygosity mapping identified the disease locus in this family on chromosome 11q25 with a maximum multipoint LOD score of 6.15. Sequence analysis of genes in the candidate interval uncovered a mutation in the canonical splice-donor site of intron 5 of JAM3. RT-PCR analysis of a patient lymphoblast cell line confirmed abnormal splicing, leading to a frameshift mutation with early termination. JAM3 is known to be present in vascular endothelium, although its roles in cerebral vasculature have not been implicated. Our results suggest that JAM3 is essential for maintaining the integrity of the cerebrovascular endothelium as well as for normal lens development in humans.     

Long-term variation in the winter resident bird community of Guánica Forest, Puerto Rico: lessons for measuring and monitoring species richness

ABSTRACT.   Because the winter season is potentially limiting for migratory birds, understanding their nonbreeding distributional patterns is essential. At a given site, patterns of species occurrence and abundance may vary over time and, within a species, wintering strategies may vary with regard to the degree that individuals are site-faithful both within and between winters. We examined long-term patterns in the composition of a winter resident bird community to determine how long a site must be studied to understand the wintering community. Over a 34-yr period of constant-effort mist netting at a site in Guánica, Puerto Rico, we captured 21 species of winter resident birds, with mean total captures varying from 8.3 to 18.9 individuals per net line and 6–14 species captured per year. Species richness capture/recapture models generated numbers similar to actual capture rates. Capture and recapture data allowed us to categorize winter residents into three groups: sporadic winter residents (14 species), regular winter residents (four species captured nearly every year), and dominant winter residents (three species captured each year with high rates of recapture). Our results suggest that sampling for at least three consecutive winters is needed to accurately characterize the bird community at a site. However, sampling for 5 yr is better, and 10-yr samples generate patterns similar to those based on our entire 34-yr sample. A 1-yr sample provides minimal information about the composition and characteristics of a winter resident bird community.     

Effects of the herbivorous minnow,southern redbelly dace (<Emphasis Type="Italic">Phoxinus erythrogaster</Emphasis>), on stream productivity and ecosystem structure

We used field and mesocosm experiments to measure effects of southern redbelly dace (Phoxinus erythrogaster), a grazing minnow, on stream ecosystem structure and function. Ecosystem structure was quantified as algal filament length, algal biomass, size distribution of particulate organic matter (POM), algal assemblage structure, and invertebrate assemblage structure, whereas ecosystem function was based on gross and net primary productivity. Our experiments showed that moderate densities of Phoxinus temporarily reduced mean algal filament length and mean size of POM relative to fishless controls. However, there was no detectable effect on algal biomass or ecosystem primary productivity. Several factors could explain the lack of effect of Phoxinus on primary productivity including increased algal production efficiency in grazed treatments or increased grazing by other organisms in fishless treatments. The inability of Phoxinus to reduce algal biomass and system productivity contrasts with experimental results based on other grazing minnows, such as the central stoneroller (Campostoma anomalum), and questions the generality of grazer effects in stream ecosystems. However, environmental venue and the spatial and temporal scale of ecosystem measurements can greatly influence the outcome of these experiments. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users. The experiments described herein comply with the current laws of the country in which the experiments were performed.     

Paternal, maternal, and biparental inheritance of the chloroplast genome in Passiflora (Passifloraceae): implications for phylogenetic studies

Patterns of inheritance of the chloroplast genome in Passiflora were analyzed by examining the progeny from both interspecific and intraspecific crosses. Artificial crosses of field-collected material were performed in greenhouses at The University of Texas at Austin. DNA from fresh leaf material was analyzed by Southern blot techniques to identify the donor of the chloroplast genome. Initially, single progeny were analyzed for 11 crosses; two intraspecific crosses demonstrated maternal inheritance, whereas the nine interspecific crosses had paternal inheritance. Subsequently, the donor of the chloroplast genome was determined for multiple progeny in seven crosses. Passiflora oerstedii × P. retipetala showed strict paternal inheritance in all of 17 progeny. A series of five crosses and backcrosses between P. oerstedii and P. menispermifolia demonstrated strictly paternal inheritance. Finally, when 15 progeny were analyzed for the P. costaricensis × P. costaricensis cross, 12 of the 15 showed maternal inheritance, whereas the remaining three were biparental. Interestingly, all interspecific crosses had primarily paternal inheritance, whereas all intraspecific crosses had primarily maternal inheritance. The implications of heteroplasmy on phylogenetic analyses of chloroplast DNA are discussed.     

Treatment with CRH-1 antagonist antalarmin reduces behavioral and endocrine responses to social stressors in marmosets (Callithrix kuhlii)

Corticotropin-releasing hormone (CRH) has multiple roles in coordinating the behavioral and endocrine responses to a host of environmental challenges, including social stressors. In the present study we evaluated the role of CRH in mediating responses to a moderate social stressor in Wied's black tufted-eared marmosets (Callithrix kuhlii). Male and female marmosets (n=14) were administered antalarmin (a selective CRH-1 receptor antagonist; 50 microg/kg, p.o.) or vehicle in a blind, counterbalanced, crossover design. One hr after treatment, marmosets were separated from long-term pairmates and then housed alone in a novel enclosure for 7 hr. Behavior was recorded during separation and upon reunion with the partner, and urine samples for cortisol assay collected before, during, and after the intervention. Separation from partners elevated urinary cortisol concentrations over baseline for both conditions, but antalarmin treatment reduced the magnitude of the elevation. Antalarmin also lowered rates of behavioral patterns associated with arousal (alarm and "e-e" vocalizations, object manipulate/chew), but had no effect on contact calls, locomotory activity or alertness. Although most patterns of social behavior upon reunion with the partner were not affected by antalarmin, antalarmin-treated marmosets displayed more sexual behavior (mounts and copulations) upon reunion. These data indicate that antagonism of the CRH-1 receptor acts to reduce the magnitude of both endocrine and behavioral responses to a moderate social stressor without causing any overall reduction in alertness or general activity. This supports the hypothesis that CRH, acting through its type 1 receptor, is involved in coordinating the responses to anxiety-producing events. These results further suggest that the marmoset is a useful model for exploration of the role of CRH in mediating the behavioral and neuroendocrine responses to psychosocial stressors, particularly in the context of heterosexual social relationships.     

AICAR induces phosphorylation of AMPK in an ATM-dependent, LKB1-independent manner

AMPK is an AMP-activated protein kinase that plays an important role in regulating cellular energy homeostasis. Metabolic stress, such as heat shock and glucose starvation, causes an energy deficiency in the cell and leads to elevated levels of intracellular AMP. This results in the phosphorylation and activation of AMPK. LKB1, a tumor suppressor, has been identified as an upstream kinase of AMPK. We found that in response to treatment with 5-aminoimidazole-4-carboxamide-1-β-4-ribofuranoside (AICAR), the LKB1 deficient cancer cell line, HeLa, exhibited AMPK-α phosphorylation. This indicates the existence of an LKB1-independent AMPK-α phosphorylation pathway. ATM is a protein that is deficient in the disease ataxia telangiectasia (A-T). We measured the activation of AMPK by AICAR in the normal mouse embryo fibroblast cell line, A29, and the mouse cell line lacking the ATM protein, A38. In A38 cells, the level of AICAR-induced AMPK-α phosphorylation was significantly lower than that found in A29 cells. Furthermore, phosphorylation of AMPK in HeLa and A29 cells was inhibited by an ATM specific inhibitor, KU-55933. Our results demonstrate that AICAR treatment could lead to phosphorylation of AMPK in an ATM-dependent and LKB1-independent manner. Thus, ATM may function as a potential AMPK kinase in response to AICAR treatment.     

The use of chemical and visual cues in female choice in the butterfly Bicyclus anynana

Investigating the relative importance of multiple cues for mate choice within a species may highlight possible mechanisms that led to the diversification of closely related species in the past. Here, we investigate the importance of close-range pheromones produced by male Bicyclus anynana butterflies and determine the relative importance of these chemical cues versus visual cues in sexual selection by female choice. We first blocked putative androconial organs on the fore- and hindwings of males, while also manipulating the ability of females to perceive chemical signals via their antenna. We found that male chemical signals were emitted by both fore- and hindwing pairs and that they play an important role in female choice. We subsequently tested the relative importance of these chemical cues versus visual cues, previously identified for this species, and found that they play an equally important role in female choice in our laboratory setting. In addition, females will mate with males with only one signal present and blocking both androconial organs on males seems to interfere with male to male recognition. We discuss the possible functions of these signals and how this bimodal system may be used in intra- and interspecific mate evaluation.     

Extra-renal 25-hydroxyvitamin D3-1α-hydroxylase in human health and disease

Although ectopic expression of 25-hydroxyvitamin D₃-1α-hydroxylase (1α-OHase) has been recognized for many years, the precise function of this enzyme outside the kidney remains open to debate. Three specific aspects of extra-renal 1α-OHase have attracted most attention: (i) expression and regulation in non-classical tissues during normal physiology; (ii) effects on the immune system and inflammatory disease; (iii) expression and function in tumors. The most well-recognized manifestation of extra-renal 1α-OHase activity remains that found in some patients with granulomatous diseases where locally synthesized 1α,25(OH)₂D₃ has the potential to spill-over into the general circulation. However, immunohistochemistry and mRNA analyses suggest that 1α-OHase is also expressed by a variety of normal human tissues including the gastrointestinal tract, skin, vasculature and placenta. This has promoted the idea that autocrine/paracrine synthesis of 1,25(OH)₂D₃ contributes to normal physiology, particularly in mediating the potent effects of vitamin D on innate (macrophage) and acquired (dendritic cell) immunity. We have assessed the capacity for synthesis of 1,25(OH)₂D₃ in these cells and the functional significance of autocrine responses to 1α-hydroxylase. Data suggest that local synthesis of 1,25(OH)₂D₃ may be a preferred mode of response to antigenic challenge in many tissues.