RESUMEN
Protein secretion in filamentous fungi has been shown to be restricted to actively growing hyphal tips. To determine whether an increase in the amount f growing surface area of a fungus can lead to an increase in the amount of protein secretion, we examined secretion in a temperature-sensitive Neurospora crassa mcb mutant that shows a loss of growth polarity when incubated at restrictive temperature. Incubation of the mcb mutant at restrictive temperature results in a three- to fivefold increase in the level of extracellular protein and a 20-fold increase in carboxymethyl cellulase activity relative to a wild-type strain. A mutation in the cr-1 gene has been shown previously to suppress the apolar growth phenotype of the mcb mutant, and we find that the level of extracellular protein produced by a mcb; cr-1 double mutant was reduced to that of the wild-type control. Immunolocalization of a secreted endoglucanase revealed that proteins are secreted mainly at hyphal tips in hyphae exhibiting polar growth and over the entire surface area of bulbous regions of hyphae that are produced after a shift of the mcb mutant to restrictive temperature. These results support the hypothesis that secretion of extracellular protein by a filamentous fungus can be significantly increased by mutations that alter growth polarity.
Asunto(s)
Celulasa , Proteínas Fúngicas/metabolismo , Neurospora crassa/crecimiento & desarrollo , Neurospora crassa/metabolismo , Adenilil Ciclasas/genética , Glicósido Hidrolasas/metabolismo , Neurospora crassa/genéticaRESUMEN
This study was conducted to determine whether the rat suprachiasmatic nucleus (SCN) is characterized by circadian expression of brain-derived neurotrophic factor (BDNF). In constant darkness, SCN content of both BDNF mRNA and protein oscillated in a circadian fashion. BDNF mRNA and protein levels in the SCN reached peak values during the early subjective day and during the subjective night, respectively. In contrast, the hippocampus showed no sign of circadian rhythmicity in its expression of BDNF mRNA and protein. Since BDNF enhances synaptic transmission in other brain regions, the coincidence between peak expression of BDNF protein in the SCN and the known interval of circadian pacemaker sensitivity to the phase-shifting effects of light may have some implications for the role of BDNF in the circadian regulation of the SCN pacemaker by photic signals from the retinohypothalamic tract.