Inter-strain differences in nitrogen use by the coccolithophore Emiliania huxleyi, and consequences for predation by a planktonic ciliate

Four strains of the coccolithophore Emiliania huxleyi (CCMP strains 370, 373, 374, 379) were tested for their ability to grow on various nitrogen sources. All strains grew on ammonium, nitrate, and urea, although growth of CCMP379 on urea was low. Responses to other dissolved organic nitrogen (DON) sources varied. CCMP379 did not grow on any DON source other than urea. All other strains grew on one of the two tested amino acids: CCMP370 and CCMP373 on glutamine, and CCMP374 on alanine. All three of these strains also grew on hypoxanthine; in addition, two grew well on acetamide and one on ethanolamine. E. huxleyi strains also differed in their susceptibility to predation by the ciliate Strobilidium sp. CCMP374 was ingested at substantially higher rates than CCMP373 regardless of E. huxleyi growth condition. Ciliate feeding rates also depended on E. huxleyi growth condition. For CCMP374, feeding rates were 2× higher on growing E. huxleyi cells than on non-growing cells (average 27.5 versus 15.6 cells ciliate⁻¹ h⁻¹, respectively). For CCMP373, a relationship between E. huxleyi growth rate and ciliate feeding rate was not evident, but E. huxleyi grown on some N sources (ammonium, nitrate, urea) were ingested at consistently higher rates than E. huxleyi grown on other sources (ethanolamine, glutamine). Interstrain differences in the ability to utilize DON and resist predation may contribute to maintenance of high genetic diversity within this cosmopolitan, bloom-forming species.     

Grazing by the calanoid copepod Neocalanus cristatus on the microbial food web in the coastal Gulf of Alaska

Neocalanus cristatus feeding on phytoplankton and microzooplanktonwas measured in the coastal Gulf of Alaska during spring andearly summer of 2001 and 2003. Neocalanus cristatus CV fed primarilyon particles >20 µm. Particles in the 5- to 20-µmsize range were ingested in some experiments under nonbloomconditions but not under bloom conditions. Particles <5 µmwere not ingested but increased during incubations because N.cristatus consumed their microzooplanktonic predators. Neocalanuscristatus are sufficiently abundant in nature to induce sucha cascade effect in situ. Microzooplankton provided >70%of the carbon ingested by N. cristatus under nonbloom conditionsbut only     

Murine mesenchymal cells that express elevated levels of the CDK inhibitor p16(Ink4a) in vivo are not necessarily senescent

Age-related health decline has been attributed to the accumulation of senescent cells recognized in vivo by p16(Ink4a) expression. The pharmacological elimination of p16(Ink4a)-positive cells from the tissues of mice was shown to extend a healthy lifespan. Here, we describe a population of mesenchymal cells isolated from mice that are highly p16(INK4a)-positive are proficient in proliferation but lack other properties of cellular senescence. These data, along with earlier reports on p16(Ink4a)-positive macrophages, indicate that p16(Ink4a)-positive and senescent cell populations only partially intersect, therefore, extending the list of potential cellular targets for anti- aging therapies.     

Antigen-induced unresponsiveness results in altered T cell signaling

Pretransplant exposure to allogeneic lymphocytes can result in donor-specific unresponsiveness and prolonged allograft survival. Intracellular signaling events have been described in anergic T cell clones, but the biochemical events underlying in vivo induced unresponsiveness have not been studied in detail. We employed a TCR transgenic mouse, bearing the 2C TCR, providing adequate numbers of homogenous peripheral T cells to study biochemical aspects of T cell unresponsiveness in vivo. 2C mice exposed to semiallogeneic lymphocytes (H-2b x H-2d) experienced prolonged H-2d cardiac allograft survival, and cells from these mice did not proliferate or make IL-2 in response to alloantigen (H-2d). Importantly, there were marked differences in TCR-associated tyrosine phosphorylation activation patterns. The targets for the unresponsive state appear to be diminished Lck activation and absent ZAP-70 and LAT (linker for activation of T cells) phosphorylation. Our study demonstrates that Ag-induced tolerance in vivo is accompanied by altered early TCR-mediated signaling events.     

Expression of duplicate msa genes in the salmonid pathogen Renibacterium salmoninarum

Renibacterium salmoninarum is a gram-positive bacterium responsible for bacterial kidney disease of salmon and trout. R. salmoninarum has two identical copies of the gene encoding major soluble antigen (MSA), an immunodominant, extracellular protein. To determine whether one or both copies of msa are expressed, reporter plasmids encoding a fusion of MSA and green fluorescent protein controlled by 0.6 kb of promoter region from msa1 or msa2 were constructed and introduced into R. salmoninarum. Single copies of the reporter plasmids integrated into the chromosome by homologous recombination. Expression of mRNA and protein from the integrated plasmids was detected, and transformed cells were fluorescent, demonstrating that both msa1 and msa2 are expressed under in vitro conditions. This is the first report of successful transformation and homologous recombination in R. salmoninarum.     

A family of Rab27-binding proteins. Melanophilin links Rab27a and myosin Va function in melanosome transport

The Rab27a GTPase regulates diverse processes involving lysosome-related organelles, including melanosome motility in melanocytes, and lytic granule release in cytotoxic T lymphocytes. Toward an understanding of Rab27a function, we searched for proteins that interact with Rab27a(GTP) using the yeast two-hybrid system and identified JFC1/Slp1, a protein of unknown function. JFC1/Slp1 and related proteins, including melanophilin, contain a conserved amino-terminal domain similar to the Rab3a-binding domain of Rabphilin-3. We used several methods to demonstrate that this conserved amino-terminal domain is a Rab27-binding domain. We show that this domain interacts directly, and in a GTP-dependent manner with Rab27a. Furthermore, overexpression of this domain in melanocytes results in perinuclear clustering of melanosomes, suggesting that this region is sufficient for interaction with, and perturbation of function of, Rab27a in a physiological context. Thus, we identified a novel family of Rab27-binding proteins. We also show that melanophilin associates with Rab27a and myosin Va on melanosomes in melanocytes, and present evidence that a domain within the carboxyl-terminal region of melanophilin interacts with the carboxyl-terminal tail of the melanocyte-specific splice isoform of myosin Va. Thus, melanophilin can associate simultaneously with activated Rab27a and myosin Va via distinct regions, and serve as a linker between these proteins.