Isolation,Culture and Long-Term Maintenance of Primary Mesencephalic Dopaminergic Neurons From Embryonic Rodent Brains

Degeneration of mesencephalic dopaminergic (mesDA) neurons is the pathological hallmark of Parkinson’s diseae. Study of the biological processes involved in physiological functions and vulnerability and death of these neurons is imparative to understanding the underlying causes and unraveling the cure for this common neurodegenerative disorder. Primary cultures of mesDA neurons provide a tool for investigation of the molecular, biochemical and electrophysiological properties, in order to understand the development, long-term survival and degeneration of these neurons during the course of disease. Here we present a detailed method for the isolation, culturing and maintenance of midbrain dopaminergic neurons from E12.5 mouse (or E14.5 rat) embryos. Optimized cell culture conditions in this protocol result in presence of axonal and dendritic projections, synaptic connections and other neuronal morphological properties, which make the cultures suitable for study of the physiological, cell biological and molecular characteristics of this neuronal population.     

Eleven polymorphic microsatellite markers in Cory's shearwater, Calonectris diomedea, and cross-species amplification on threatened Procellariiformes

For the first time, we describe 11 variable dinucleotide microsatellites and the conditions for multiplexing and simultaneous genotyping sets of loci in Cory's shearwater, Calonectris diomedea. Microsatellite variability was assessed in a colony from the Azores archipelago (Atlantic Ocean). Two to eight alleles were detected per locus, the mean gene diversity being 4.5. Cross-species amplification in three other seabirds (Diomedea exulans, Procellaria aequinoctialis and Bulweria bulwerii) revealed some variability at one, two and eight loci, respectively.     

Mathematical modelling of neuronal dendritic branching patterns in two dimensions: application to retinal ganglion cells in the cat and rat

Sholl’s analysis has been used for about 50years to study neuron branching characteristics based on a linear, semi-log or log—log method. Using the linear two- dimensional Sholl’s method, we call attention to a relationship between the number of intersections of neuronal dendrites with a circle and the numbers of branching points and terminal tips encompassed by the circle, with respect to the circle radius. For that purpose, we present a mathematical model, which incorporates a supposition that the number of dendritic intersections with a circle can be resolved into two components: the number of branching points and the number of terminal tips within the annulus of two adjoining circles. The numbers of intersections and last two sets of data are also presented as cumulative frequency plots and analysed using a logistic model (Boltzmann’s function). Such approaches give rise to several new morphometric parameters, such as, the critical, maximal and mean values of the numbers of intersections, branching points and terminal tips, as well as the abscissas of the inflection points of the corresponding sigmoid plots, with respect to the radius. We discuss these parameters as an additional tool for further morphological classification schemes of vertebrate retinal ganglion cells. To test the models, we apply them first to three groups of morphologically different cat’s retinal ganglion cells (the alpha, gamma and epsilon cells). After that, in order to quantitatively support the classification of the rat’s alpha cells into the inner and outer cells, we apply our models to two subgroups of these cells grouped according to their stratification levels in the inner plexiform layer. We show that differences between most of our parameters calculated for these subgroups are statistically significant. We believe that these models have the potential to aid in the classification of biological images.     

Stejneger's beaked whale strandings in Alaska, 1995–2020

Presented here is the first comprehensive and updated compilation of history, distribution, and findings of Stejneger's beaked whales (Mesoplodon stejnegeri) in Alaska. Stejneger's beaked whales are a poorly understood, elusive, deep-diving cetacean species found in the North Pacific Ocean. Since Stejneger's beaked whale strandings data in Alaska through 1994 were last published, 35 additional strandings have been documented. Twenty-seven animals stranded in the Aleutian Islands, seven stranded in Southcentral Alaska, and one animal stranded on St. Lawrence Island. Twenty-two carcasses were necropsied, but only four were fresh. Seventeen of the 22 died during mass stranding events and cause of death could not be definitively determined. Barotrauma was suspected in three cases and infectious disease possibly complicated by barotrauma occurred in two cases. We documented an expansion of strandings into the northern Bering Sea, characterized a sex bias, examined stomach contents that included macroplastic, and identified parasites not previously associated with Stejneger's beaked whales. Also included are data on the largest known mass stranding of Stejneger's beaked whales, which occurred on Adak Island in 2018. The history, distribution, and findings presented here are central to further our understanding of this species.