Allelic variation of polymorphic locus lytB, encoding a choline-binding protein, from streptococci of the mitis group

The choline-binding protein LytB, an N-acetylglucosaminidase of Streptococcus pneumoniae, is the key enzyme for daughter cell separation and is believed to play a critical pathogenic role, facilitating bacterial spreading during infection. Because of these peculiarities LytB is a putative vaccine target. To determine the extent of LytB polymorphism, the lytB alleles from seven typical, clinical pneumococcal isolates of various serotypes and from 13 additional streptococci of the mitis group (12 atypical pneumococci and the Streptococcus mitis type strain) were sequenced. Sequence alignment showed that the main differences among alleles were differences in the number of repeats (range, 12 to 18) characteristic of choline-binding proteins. These differences were located in the region corresponding to repeats 11 to 17. Typical pneumococcal strains contained either 14, 16, or 18 repeats, whereas all of the atypical isolates except strains 1283 and 782 (which had 14 and 16 repeats, respectively) and the S. mitis type strain had only 12 repeats; atypical isolate 10546 turned out to be a DeltalytB mutant. We also found that there are two major types of alternating repeats in lytB, which encode 21 and 23 amino acids. Choline-binding proteins are linked to the choline-containing cell wall substrate through choline residues at the interface of two consecutive choline-binding repeats that create a choline-binding site. The observation that all strains contained an even number of repeats suggests that the duplication events that gave rise to the choline-binding repeats of LytB involved two repeats simultaneously, an observation that is in keeping with previous crystallographic data. Typical pneumococcal isolates usually grew as diplococci, indicating that an active LytB enzyme was present. In contrast, most atypical isolates formed long chains of cells that did not disperse after addition of purified LytB, suggesting that in these strains chains were produced through mechanisms unrelated to LytB.     

Autoradiographic demonstration of specific binding sites for E. coli enterotoxin in various epithelia of the North American opossum

Summary In the North American opossum, heat-stable specific binding sites for E. coli enterotoxin are observed (i) in epithelial cells lining the small intestine, colon, gall bladder, cystic duct, common bile duct and trachea, and (ii) in epithelial cells forming the duodenal (Brunner's) glands, liver, kidneys (metanephros, mesonephros) and testis, as demonstrated by autoradiography. Enterotoxin-specific binding sites in the intestinal tract are only found in intestinal epithelial cells with the highest concentration in the microvillus border. Enterotoxin-specific binding sites also occur in epithelial cells comprising the secretory tubules and ducts of the duodenal glands. In the kidneys (metanephros and mesonephros), enterotoxin-specific binding sites are confirmed primarily to the proximal tubules, whereas in the testis they are localized in seminiferous tubules. In the liver, enterotoxin-specific binding sites are confined primarily to hepatocytes. E. coli enterotoxin caused a 7-fold increase of cGMP in the liver and a 30-fold increase in the duodenal glands. The liver responded in about half of the animals studied, whereas the duodenal glands gave a consistent response in each case. Likewise, the duodenal glands consistently showed strong labelling for ¹²⁵I-enterotoxin, whereas receptor labelling of hepatocytes was inconsistent in nearly half the incubations and corresponds to the observed cGMP measurements.Supported by a Weldon Springs Grant, University of Missouri and by funds from the Medical Research Service, Department of Veteran's Affairs     

Transcriptomic analysis of skin pigmentation variation in the Virginia opossum (Didelphis virginiana)

Skin pigmentation and coat pigmentation are two of the best‐studied examples of traits under natural selection given their quantifiable fitness interactions with the environment (e.g., camouflage) and signalling with other organisms (e.g., warning coloration). Previous morphological studies have found that skin pigmentation variation in the Virginia opossum (Didelphis virginiana) is associated with variation in precipitation and temperatures across its distribution range following Gloger's rule (lighter pigmentation in temperate environments). To investigate the molecular mechanism associated with skin pigmentation variation, we used RNA‐Seq and quantified gene expression of wild opossums from tropical and temperate populations. Using differential expression analysis and a co‐expression network approach, we found that expression variation in genes with melanocytic and immune functions is significantly associated with the degree of skin pigmentation variation and may be underlying this phenotypic difference. We also found evidence suggesting that the Wnt/β‐catenin signalling pathway might be regulating the depigmentation observed in temperate populations. Based on our study results, we present several alternative hypotheses that may explain Gloger's rule pattern of skin pigmentation variation in opossum, including changes in pathogen diversity supporting a pathogen‐resistant hypothesis, thermal stress associated with temperate environments, and pleiotropic and epistatic interactions between melanocytic and immune genes.     

Placentation in the opossum, Didelphis virginiana

For the first 9 days of gestation, opossum embryos float in uterine secretions, separated from maternal tissues by a shell membrane. Each embryo is part of the wall of its hollow embryonic sphere. By the 10th day of development, the embryo becomes enveloped by both the amnion and yolk-sac. The yolk-sac consists of vascular and non-vascular portions and, together with the surrounding trophectoderm (trophoblast), forms the yolk-sac placenta of the opossum: the allantois does not contribute to formation of the placenta. The vascular portion of the yolk-sac placenta establishes an intimate relationship with the uterine epithelium soon after loss of the shell membrane. The yolk-sac placenta is non-invasive. Cells of the trophoblast exhibit numerous microvilli, an apical endocytic complex and the lateral and basal cell membrane are elaborately folded. These features suggest a cell that is active in the transport of materials. Junctional complexes between cells of the trophoblast and uterine epithelium were not observed. The uterine epithelium changes from ciliated pseudostratified columnar with few infoldings of lateral and basal cell membranes, to non-ciliated simple columnar in which these membranes show elaborate infoldings. The cells show numerous inclusions and mitochondria are polarized to the basal half of the cell. These features suggest a cell that also is active in the transport of materials.     

Natural variation in phytochrome signaling

The phytochromes, photoreceptors sensitive to red and far-red light, are critical for sensing foliage shade, canopy breaks, and neighbor proximity. A combination of molecular genetic, evolutionary, and ecological techniques are being used to understand how phytochromes function in the natural environment. We discuss studies on the adaptive value of phytochrome mediated plasticity, as well as the role that variation in phytochrome expression and function might play in allowing plants to adapt to unique light environments. Continued study of phytochrome signaling variation may reveal how natural selection acts at the molecular level.     

The extensor assembly in two species of opossum,Philander opossum and Didelphis marsupialis

In considering primate and hominoid phylogeny, the fundamental position assigned to opossums is explained partially by the characteristic morphology of their hands and feet. One of the main functional features of the human hand is the ability to make a stabilized arch of the finger. Because the extensor assembly plays a key role in establishing an arched finger, the extensor systems of the digits of both the hands and feet were studied in two species of opossum, Philander opossum and Didelphis marsupialis. In the foot, two extensor tendons join in each toe to form one tendinous plate, which inserts onto the base of the second phalanx. Lumbricals join this plate along the tibial side, and interosseus insertions are found, although a true interosseus wing is lacking. At the proximal interphalangeal level, a terminal tendon takes its origin from this tendinous plate. This terminal tendon is oval in cross-section and contains elastic structures. Oblique bands arise from this terminal tendon and run proximally along the proximal interphalangeal joint inserting onto the base of the first phalanx. There are elastic structures in the flexor tendon on the dorsal side near its site of insertion. In the hand, the main extensor tendons are arranged differently and the interossei contribute substantially to the extensor assembly. Otherwise, the extensor assembly of the hands and feet are quite similar. The function of the so-called paratendinous intravaginal flexors is discussed as are evolutionary aspects of the extensor assembly.     

Natural genetic variation in plant photosynthesis

Natural genetic variation in plant photosynthesis is a largely unexplored and as a result an underused genetic resource for crop improvement. Numerous studies show genetic variation in photosynthetic traits in both crop and wild species, and there is an increasingly detailed knowledge base concerning the interaction of photosynthetic phenotypes with their environment. The genetic factors that cause this variation remain largely unknown. Investigations into natural genetic variation in photosynthesis will provide insights into the genetic regulation of this complex trait. Such insights can be used to understand evolutionary processes that affect primary production, allow greater understanding of the genetic regulation of photosynthesis and ultimately increase the productivity of our crops.     

The pineal region in the opossum,Didelphis virginiana

Summary In the pineal region of the opossum, Didelphis virginiana, two types of cells predominate: 1) pinealocytes, and 2) fibrous astrocytes. Pinealocytes are characterized by the presence of prominent Golgi bodies, numerous clear and dense-cored vesicles, sensory cilia (9+0), vesicle-crowned rods, and condensation of a material that was always associated with the rough endoplasmic reticulum. In addition, two other cell types are occasionally seen. These include 1) neuron-like cells, and 2) darker staining cells of unknown identity. The endoplasmic reticulum of the darker staining cells is typically expanded and filled with an amorphous substance. Although the pineal region is small in size, the present findings suggest that pinealocytes in this species are metabolically active cells displaying a secretory function. Moreover, the presence of sensory cilia (9+0) and vesicle-crowned rods indicates that pinealocytes of the opossum are phylogenetically related to the photoreceptor cells found in the pineal organ of lower vertebrates.