RESUMEN
Ecological disturbances caused by roadways have previously been reported, but traffic speed has not been addressed. We investigate effects of traffic speed on pollination of Centratherum punctatum (Asteraceae) along an Amazonian highway roadside. We hypothesised that frequency of flower visitors, duration of single visits and pollen deposition on stigmas will vary negatively as traffic speed increases. After measuring vehicle velocities, we classified three road sections as low-, mid- and high-velocity traffic. The main pollinator bee, Augochlora sp., visited C. punctatum inflorescences with decreasing frequency from low- to high-velocity roadside sections, whereas the nectar thief butterflies did the opposite. Duration of single visits by bees and butterflies was shorter, and arrival of pollen on C. punctatum stigmas was lower, in high- than in low-velocity roadside. Air turbulence due to passing vehicles increases with velocity and disturbed the flower visitors. Overall, results support that traffic velocity negatively affects foraging of flower visitors and the pollination of C. punctatum on roadsides.
Asunto(s)
Asteraceae/fisiología , Abejas/fisiología , Ecología , Polinización , Animales , Conducta Animal , Flores/fisiología , Néctar de las Plantas/fisiología , Polen/fisiología , ReproducciónRESUMEN
Acetate is a short-chain fatty acid secreted by Propionibacteria from the human intestine, known to induce mitochondrial apoptotic death in colorectal cancer (CRC) cells. We previously established that acetate also induces lysosome membrane permeabilization in CRC cells, associated with release of the lysosomal protease cathepsin D (CatD), which has a well-established role in the mitochondrial apoptotic cascade. Unexpectedly, we showed that CatD has an antiapoptotic role in this process, as pepstatin A (a CatD inhibitor) increased acetate-induced apoptosis. These results mimicked our previous data in the yeast system showing that acetic acid activates a mitochondria-dependent apoptosis process associated with vacuolar membrane permeabilization and release of the vacuolar protease Pep4p, ortholog of mammalian CatD. Indeed, this protease was required for cell survival in a manner dependent on its catalytic activity and for efficient mitochondrial degradation independently of autophagy. In this study, we therefore assessed the role of CatD in acetate-induced mitochondrial alterations. We found that, similar to acetic acid in yeast, acetate-induced apoptosis is not associated with autophagy induction in CRC cells. Moreover, inhibition of CatD with small interfering RNA or pepstatin A enhanced apoptosis associated with higher mitochondrial dysfunction and increased mitochondrial mass. This effect seems to be specific, as inhibition of CatB and CatL with E-64d had no effect, nor were these proteases significantly released to the cytosol during acetate-induced apoptosis. Using yeast cells, we further show that the role of Pep4p in mitochondrial degradation depends on its protease activity and is complemented by CatD, indicating that this mechanism is conserved. In summary, the clues provided by the yeast model unveiled a novel CatD function in the degradation of damaged mitochondria when autophagy is impaired, which protects CRC cells from acetate-induced apoptosis. CatD inhibitors could therefore enhance acetate-mediated cancer cell death, presenting a novel strategy for prevention or therapy of CRC.
Asunto(s)
Apoptosis/efectos de los fármacos , Autofagia/fisiología , Catepsina D/genética , Neoplasias Colorrectales/patología , Mitocondrias/patología , Acetatos/farmacología , Catepsina D/antagonistas & inhibidores , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Células HCT116 , Humanos , Mitocondrias/metabolismo , Estrés Oxidativo , Pepstatinas/farmacología , Interferencia de ARN , ARN Interferente PequeñoRESUMEN
Cathepsin D has garnered increased attention in recent years, mainly since it has been associated with several human pathologies. In particular, cathepsin D is often overexpressed and hypersecreted in cancer cells, implying it may constitute a therapeutic target. However, cathepsin D can have both anti- and pro-survival functions depending on its proteolytic activity, cellular context and stress stimulus. Therefore, a more detailed understanding of cathepsin D regulation and how to modulate its apoptotic functions is clearly needed. In this review, we provide an overview of the role of cathepsin D in physiological and pathological scenarios. We then focus on the opposing functions of cathepsin D in apoptosis, particularly relevant in cancer research. Emphasis is given to the role of the yeast protease Pep4p, the vacuolar counterpart of cathepsin D, in life and death. Finally, we discuss how insights from yeast cathepsin D and its role in regulated cell death can unveil novel functions of mammalian cathepsin D in apoptosis and cancer.
RESUMEN
Colorectal carcinoma (CRC) is one of the most common causes of cancer-related mortality. Short-chain fatty acids secreted by dietary propionibacteria from the intestine, such as acetate, induce apoptosis in CRC cells and may therefore be relevant in CRC prevention and therapy. We previously reported that acetic acid-induced apoptosis in Saccharomyces cerevisiae cells involves partial vacuole permeabilization and release of Pep4p, the yeast cathepsin D (CatD), which has a protective role in this process. In cancer cells, lysosomes have emerged as key players in apoptosis through selective lysosomal membrane permeabilization (LMP) and release of cathepsins. However, the role of CatD in CRC survival is controversial and has not been assessed in response to acetate. We aimed to ascertain whether LMP and CatD are involved in acetate-induced apoptosis in CRC cells. We showed that acetate per se inhibits proliferation and induces apoptosis. More importantly, we uncovered that acetate triggers LMP and CatD release to the cytosol. Pepstatin A (a CatD inhibitor) but not E64d (a cathepsin B and L inhibitor) increased acetate-induced apoptosis of CRC cells, suggesting that CatD has a protective role in this process. Our data indicate that acetate induces LMP and subsequent release of CatD in CRC cells undergoing apoptosis, and suggest exploiting novel strategies using acetate as a prevention/therapeutic agent in CRC, through simultaneous treatment with CatD inhibitors.
Asunto(s)
Ácido Acético/toxicidad , Apoptosis/efectos de los fármacos , Catepsina D/metabolismo , Lisosomas/metabolismo , Catepsina B/antagonistas & inhibidores , Catepsina B/metabolismo , Catepsina D/antagonistas & inhibidores , Catepsina L/antagonistas & inhibidores , Catepsina L/metabolismo , Línea Celular Tumoral , Permeabilidad de la Membrana Celular/efectos de los fármacos , Neoplasias Colorrectales/metabolismo , Neoplasias Colorrectales/patología , Humanos , Concentración de Iones de Hidrógeno , Leucina/análogos & derivados , Leucina/farmacología , Pepstatinas/farmacologíaRESUMEN
In Saccharomyces cerevisiae, Spo12p is involved in mitosis and is essential for meiosis. We found that Spo12p is imported into the nucleus by the karyopherin Kap121p. A complex containing Spo12p and Kap121p was isolated from cytosol and was also reconstituted with recombinant proteins, indicating that this interaction is direct. Spo12p was mislocalized to the cytosol in pse1-1, a temperature-sensitive strain harboring a mutation of Kap121p, at the permissive temperature, confirming an essential role for Kap121p in Spo12p import. Spo12p was also mislocalized in a pse1-1/pse1-1 homozygous strain, suggesting it is imported via the same pathway in diploid cells. Furthermore, we found that pse1-1/pse1-1 shows a sporulation defect similar to that of spo12Delta/spo12Delta. In addition, we have characterized the Spo12p nuclear localization signal, mapped it to residues 76-130, and identified residues within this region that are important for nuclear localization signal function.
Asunto(s)
Proteínas Fúngicas/metabolismo , Saccharomyces cerevisiae/metabolismo , Secuencia de Aminoácidos , Transporte Biológico , Núcleo Celular/metabolismo , Meiosis , Datos de Secuencia Molecular , Proteínas Nucleares , Saccharomyces cerevisiae/citología , Proteínas de Saccharomyces cerevisiaeRESUMEN
The possible relationship between the greater incidence of femoral hernia on the right side of the human body and anatomic differences between the right and left femoral ring were studied. The morphology and the transverse diameter of the femoral rings on both sides of 38 adult, formolized corpses were compared. There was no difference between the two sides.