Overwintering of the parasitic dinoflagellate Hematodinium perezi in dredged blue crabs (Callinectes sapidus) from Wachapreague Creek, Virginia.

Parasitic dinoflagellates in the genus Hematodinium cause disease and mortality in several commercially important marine decapod crustaceans. One species, Hematodinium perezi, occurs in blue crabs, Callinectes sapidus, along the eastern seaboard and Gulf coast of the USA. The parasite infects blue crabs, other decapods, and amphipods in the high salinity waters of coastal bays. Epizootics of the parasite often reach prevalence levels of 75-80% during outbreaks with diseased crabs dying from the infection. Prevalence of the parasite is bimodal, with a minor peak in late spring or summer, and a major peak in fall, and declining rapidly to nearly zero in late November and December. The rapid decline in infections in the late fall brings up the question of whether the parasite overwinters in crabs or whether it uses an unidentified resting stage, such as a cyst. We report observations on the prevalence of the parasite from winter dredge surveys undertaken in 2011 and 2012. Crabs were examined via hemolymph smears, histology, and PCR diagnosis for the presence of H. perezi and other pathogens. Active infections were observed from January through March in 2011 and 2012, indicating the parasite can overwinter in blue crabs. However, several crabs that were positive by PCR had presumptive effete infections that were difficult to diagnose in histological slides and hemolymph smears. These infections did not appear to be active and may have been in subsidence. Dredged crabs with light and moderate active infections were held at 15°C to determine if the parasite was capable of rapid progression. In 8 cases, infections exhibited logarithmic growth progressing rapidly over 8-12days. We present evidence that overwintering of H. perezi occurs in the blue crab hosts, that infections are capable of responding rapidly to increases in temperatures, and that overwintering provides a reservoir of infected animals for transmission to occur in the spring.

Targeted deletion of AP-2alpha leads to disruption in corneal epithelial cell integrity and defects in the corneal stroma.

PURPOSE: The present study was undertaken to create a conditional knockout of AP-2alpha in the corneal epithelium. METHODS: A line of mice expressing Cre-recombinase specifically in the early lens placode was crossed with mice in which the AP-2alpha allele is flanked by two loxP sites. The resultant Le-AP-2alpha mutants exhibited a targeted deletion of AP-2alpha in lens placode derivatives, including the differentiating corneal epithelium. RESULTS: The Le-AP-2alpha mutant mice were viable and had a normal lifespan. The adult corneal epithelium exhibited a variation in the number of stratified epithelial layers, ranging from 2 to 10 cell layers. A substantial decrease in expression of the cell-cell adhesion molecule, E-cadherin, was observed in all layers of the Le-AP-2alpha mutant corneal epithelium. The basement membrane, or Bowman's layer, was thinner in the mutant cornea and in many regions was discontinuous. These defects corresponded with altered distribution of laminin and entactin, and to a lesser degree, type IV collagen. The Le-AP-2alpha mutant cornea also exhibited stromal defects, including disrupted organization of the collagen lamellae and accumulation of fibroblasts beneath the epithelium that showed increased immunoreactivity for proliferating cell nuclear antigen (PCNA), alpha-smooth muscle actin (alpha-SMA), p-Smad2, and TGF-beta2. CONCLUSIONS: In the absence of AP-2alpha, the corneal epithelium exhibits altered cell adhesion and integrity and defects in its underlying basement membrane. These defects likely caused the alterations in the corneal stroma.

Wnt1-Cre-mediated deletion of AP-2alpha causes multiple neural crest-related defects.

The AP-2alpha transcription factor is required for multiple aspects of vertebrate development and mice lacking the AP-2alpha gene (tcfap2a) die at birth from severe defects affecting the head and trunk. Several of the defects associated with the tcfap2a-null mutation affect neural crest cell (NCC) derivatives including the craniofacial skeleton, cranial ganglia, and heart outflow tract. Consequently, there is considerable interest in the role of AP-2alpha in neural crest cell function in development and evolution. In addition, the expression of the AP-2alpha gene is utilized as a marker for premigratory and migratory neural crest cells in many vertebrate species. Here, we have specifically addressed how the presence of AP-2alpha in neural crest cells affects development by creating a conditional (floxed) version of tcfap2a which has subsequently been intercrossed with mice expressing Cre recombinase under the control of Wnt1 cis-regulatory sequences. Neural crest-specific disruption of tcfap2a results in frequent perinatal lethality associated with neural tube closure defects and cleft secondary palate. A small but significant fraction of mutant mice can survive into adulthood, but have retarded craniofacial growth, abnormal middle ear development, and defects in pigmentation. The phenotypes obtained confirm that AP-2alpha directs important aspects of neural crest cell function. At the same time, we did not observe several neurocristopathies affecting the head and heart that might be expected based on the phenotype of the AP-2alpha-null mouse. These results have important implications for the evolution and function of the AP-2 gene family in both the neural crest and the vertebrate embryo.