Influence of Extracellular Matrix Components on the Expression of Integrins and Regeneration of Adult Retinal Ganglion Cells

Purpose Retinal ganglion cells (RGCs) are exposed to injury in a variety of optic nerve diseases including glaucoma. However, not all cells respond in the same way to damage and the capacity of individual RGCs to survive or regenerate is variable. In order to elucidate factors that may be important for RGC survival and regeneration we have focussed on the extracellular matrix (ECM) and RGC integrin expression. Our specific questions were: (1) Do adult RGCs express particular sets of integrins in vitro and in vivo? (2) Can the nature of the ECM influence the expression of different integrins? (3) Can the nature of the ECM affect the survival of the cells and the length or branching complexity of their neurites? Methods Primary RGC cultures from adult rat retina were placed on glass coverslips treated with different substrates: Poly-L-Lysine (PL), or PL plus laminin (L), collagen I (CI), collagen IV (CIV) or fibronectin (F). After 10 days in culture, we performed double immunostaining with an antibody against beta III-Tubulin to identify the RGCs, and antibodies against the integrin subunits: alpha V, alpha 1, alpha 3, alpha 5, beta 1 or beta 3. The number of adhering and surviving cells, the number and length of the neurites and the expression of the integrin subunits on the different substrates were analysed. Results PL and L were associated with the greatest survival of RGCs while CI provided the least favourable conditions. The type of substrate affected the number and length of neurites. L stimulated the longest growth. We found at least three different types of RGCs in terms of their capacity to regenerate and extend neurites. The different combinations of integrins expressed by the cells growing on different substrata suggest that RGCs expressed predominantly alpha 1 beta 1 or alpha 3 beta 1 on L, alpha 1 beta 1 on CI and CIV, and alpha 5 beta 3 on F. The activity of the integrins was demonstrated by the phosphorylation of focal adhesion kinase (FAK). Conclusions Adult rat RGCs can survive and grow in the presence of different ECM tested. Further studies should be done to elucidate the different molecular characteristics of the RGCs subtypes in order to understand the possible different sensitivity of different RGCs to damage in diseases like glaucoma in which not all RGCs die at the same time.

Distribution and biology of prowfish (Zaprora silenus) in the northeast Pacific

The prowfish (Zaprora silenus) is an infrequent component of bottom trawl catches collected on stock assessment surveys. Based on presence or absence in over 40,000 trawl catches taken throughout Alaskan waters southward to southern California, prowfish are most frequently encountered in the Gulf of Alaska and the Aleutian Islands at the edge of the continental shelf. Based on data from two trawl surveys, relative abundance indicated by catch per swept area reaches a maximum between 100 m and 200 m depth and is much higher in the Aleutian Islands than in the Gulf of Alaska. Females weigh 3.7% more than males of the same length. Weight-length functions are W (g) = 0.0164 L2.92 (males) and W = 0.0170 L2.92 (females). Length at age does not differ between sexes and is described by L = 89.3(1 – e–0.181(t+0.554)), where L is total length in cm and t is age in years. Females reached 50% maturity at a length of 57.0 cm and an age of 5.1 years. Prowfish diet is almost entirely composed of gelatinous zooplankton, primarily scyphozoa and salps.

Abundance of cetaceans in the oceanic northern Gulf of Mexico from 2003 and 2004 ship surveys

The Gulf of Mexico (GMx) is a subtropical marginal sea of the western North Atlantic Ocean with a diverse cetacean community. Ship-based, line-transect abundance surveys were conducted in oceanic waters (>200 m deep) of the northern GMx within U.S. waters (380,432 square km) during summer 2003 and spring 2004. Data from these surveys were pooled and minimum abundance estimates were based on 10,933 km of effort and 433 sightings of at least 17 species.The most commonly sighted species (number of groups) were pantropical spotted dolphin, Stenella attenuata (115); sperm whale, Physeter macrocephalus (85); dwarf/pygmy sperm whale, Kogia sima/breviceps (27); Risso’s dolphin, Grampus griseus (26); and bottlenose dolphin, Tursiops truncatus (26). The most abundant species (number of individuals; coefficient of variation) were S. attenuata (34,067; 0.18); Clymene dolphin, S. clymene (6,575; 0.36); T. truncatus (3,708; 0.42); and striped dolphin, S. coeruleoalba (3,325; 0.48). The only large whales sighted were P. macrocephalus (1,665; 0.20) and Bryde’s whale, Balaenoptera edeni (15; 1.98). Abundances for other species or genera ranged from 57 to 2,283 animals. Cetaceanswere sighted throughout the oceanic northern GMx, and whereas many species were widely distributed, some had more regional distributions. Compared to abundance estimates for this area based on 1996-2001 surveys, the estimate for S. attenuata was significantly smaller (P <0.05) and that for the spinner dolphin, S. longirostris, appeared much smaller. Also, P. macrocephalus estimates were based on less negatively biased estimates of group-size using 90-minute counts during 2003 and 2004.

Description and growth of larval and pelagic juvenile pygmy rockfish (Sebastes wilsoni) (family Sebastidae)

A developmental series of larval and pelagic juvenile pygmy rockfish (Sebastes wilsoni) from central California is illustrated and described. Sebastes wilsoni is a non- commercially, but ecologically, important rockfish, and the ability to differentiate its young stages will aid researchers in population abundance studies. Pigment patterns, meristic characters, morphometric measurements, and head spination were recorded from specimens that ranged from 8.1 to 34.4 mm in standard length. Larvae were identified initially by meristic characters and the absence of ventral and lateral midline pigment. Pelagic juveniles developed a prominent pigment pattern of three body bars that did not extend to the ventral surface. Species identification was confirmed subsequently by using mitochondrial sequence data of four representative specimens of various sizes. As determined from the examination of otoliths, the growth rate of larval and pelagic juvenile pygmy rockfish was 0.28 mm/day, which is relatively slow in comparison to the growth rate of other species of Sebastes. These data will aid researchers in determining species abundance.

Abundance of cetatceans in the southern U.S. North Atlantic Ocean during summer 1998

The U.S. Marine Mammal Protection Act requires that the abundance of marine mammals in U.S. waters be assessed. Because this requirement had not been met for a large portion of the North Atlantic Ocean (U.S. waters south of Maryland), a ship-based, line-transect survey was conducted with a 68 m research ship between Maryland (38.00°N) and central Florida (28.00°N) from the 10-m isobath to the boundary of the U.S. Exclusive Economic Zone. The study area (573,000 km2) was surveyed between 8 July and 17 August 1998. Minimum abundance estimates were based on 4163 km of effort and 217 sightings of at least 13 cetacean species and other taxonomic categories. The most commonly sighted species (number of groups) were bottlenose dolphins, Tursiops truncatus (38); sperm whales, Physeter macrocephalus (29); Atlantic spotted dolphins, Stenella frontalis (28); and Risso’s dolphins, Grampus griseus (22). The most abundant species (abundance; coeffi cient of variation) were Atlantic spotted dolphins (14,438; 0.63); bottlenose dolphins (13,085; 0.40); pantropical spotted dolphins, S. attenuate (12,747; 0.56); striped dolphins, S. coeruleoalba (10,225; 0.91); and Risso’s dolphins (9533; 0.50). The abundance estimate for the Clymene dolphin, S. clymene (6086; 0.93), is the first for the U.S. Atlantic Ocean. Sperm whales were the most abundant large whale (1181; 0.51). Abundances for other species or taxonomic categories ranged from 20 to 5109. There were an estimated 77,139 (0.23) cetaceans in the study area. Bottlenose dolphins and Atlantic spotted dolphins were encountered primarily in continental shelf (<200 m) and continental slope waters (200−2000 m). All other species were generally sighted in oceanic waters (>200 m). The distribution of some species varied north to south. Striped dolphins, Clymene dolphins, and sperm whales were sighted primarily in the northern part of the study area; whereas pantropical spotted dolphins were sighted primarily in the southern portion.