Cytoplasmic bulk flow propels nuclei in mature hyphae of Neurospora crassa

Silvia L. Ramos-García, Robert Roberson, Michael Freitage, Salomón Bartnicki-García, Rosa R. Mouriño-Pérez

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48 Scopus citations

Abstract

We used confocal microscopy to evaluate nuclear dynamics in mature, growing hyphae of Neurospora crassa whose nuclei expressed histone H1-tagged green fluorescent protein (GFP). In addition to the H1-GFP wild-type (WT) strain, we examined nuclear displacement (passive transport) in four mutants deficient in microtubule-related motor proteins (ro-1, ro-3, kin-1, and a ro-1 kin-1 double mutant). We also treated the WT strain with benomy1 and cytochalasin A to disrupt microtubules and actin microfilaments, respectively. We found that the degree of nuclear displacement in the subapical regions of all strains correlated with hyphal elongation rate. The WT strain and that the ro-1 kin-1 double mutant showed the highest correlation between nuclear movement and hyphal elongation. Although most nuclei seemed to move forward passively, presumably carried by the cytoplasmic bulk flow, a small proportion of the movement detected was either retrograde or accelerated anterograde. The absence of a specific microtubule motor in the mutants ro-1, ro-3, or kin-1 did not prevent the anterograde and retrograde migration of nuclei; however, in the ro-1 kin-1 double mutant retrograde migration was absent. In the WT strain, almost all nuclei were elongated, whereas in all other strains a majority of nuclei were nearly spherical. With only one exception, a sizable exclusion zone was maintained between the apex and the leading nucleus. The ro-1 mutant showed the largest nucleus exclusion zone; only the treatment with cytochalasin A abolished the exclusion zone. In conclusion, the movement and distribution of nuclei in mature hyphae appear to be determined by a combination of forces, with cytoplasmic bulk flow being a major determinant. Motor proteins probably play an active role in powering the retrograde or accelerated anterograde migrations of nuclei and may also contribute to passive anterograde displacement by binding nuclei to microtubules.

Original languageEnglish (US)
Pages (from-to)1880-1890
Number of pages11
JournalEukaryotic Cell
Volume8
Issue number12
DOIs
StatePublished - Dec 1 2009

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

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    Ramos-García, S. L., Roberson, R., Freitage, M., Bartnicki-García, S., & Mouriño-Pérez, R. R. (2009). Cytoplasmic bulk flow propels nuclei in mature hyphae of Neurospora crassa. Eukaryotic Cell, 8(12), 1880-1890. https://doi.org/10.1128/EC.00062-09