Journal:

Article Title: Isolation and Characterization of Cryptococcus neoformans Spores Reveal a Critical Role for Capsule Biosynthesis Genes in Spore Biogenesis ▿

doi: 10.1128/EC.00352-08

Figure Lengend Snippet: Capsule mutants have defects in sexual development and spore formation. (A) All panels show the periphery of a cross on V8 medium under a light microscope at low magnification (×200). Insets each show a single basidium under higher magnification (×400). (1) Wild-type a × α cross. (2 and 3) Wild-type a or α strain crossed with corresponding a cap67 or α cap67 strain. (4) a cap67 × α cap67 cross. The few basidia that were produced did not develop proper spore chains and yielded clumps of spores (inset in panel 4). Identical results were obtained for all additional capsule deletion strains tested (cap10Δ, cap60Δ, and cap64Δ strains). (B) Spores were stained with DAPI (blue) and DSL (green) and visualized by fluorescence micros- copy. (Left) Normal morphology of spores derived from a wild-type cross. (Right) Spores derived from a cap67 × cap67 cross forming an aggregated mass of spores. (C) Spores from wild-type and cap67 strains stained with DAPI to reveal nuclei (blue), with anti-GXM antibody (F12D2) to detect capsule (red), and with DSL to identify spores (green) were visualized by immunofluorescence microscopy. All three color channels were merged into a single image for each sample. (1) Spores derived from a wild-type a by α cross (JEC20 × JEC21). (2 and 3) Spores and yeast cells from crosses between wild-type and cap67 strains (CHY1377 × JEC21 and JEC20 × ATCC 52817, respectively). (4) Spores and yeast cells isolated from a cross between two cap67 strains (CHY1377 × ATCC 52817). Bars, 5 μm.

Article Snippet: The starting acapsular strains, cap10Δ , cap60Δ , cap64Δ , and cap67 strains (ATCC 52817-B3501 background), were all of the α mating type, so each mutant strain was crossed with JEC20 ( a ) on V8 agar ( 11 - 13 , 23 ).

Techniques: Light Microscopy, Produced, Staining, Fluorescence, Derivative Assay, Immunofluorescence, Microscopy, Isolation

Journal:

Article Title: Isolation and Characterization of Cryptococcus neoformans Spores Reveal a Critical Role for Capsule Biosynthesis Genes in Spore Biogenesis ▿

doi: 10.1128/EC.00352-08

Figure Lengend Snippet: Capsule is not efficiently transferred to the surfaces of capsule-deficient spores. Yeast cells and spores were labeled with DAPI (blue) to reveal nuclei, with DSL (green) to identify spores, and with anti-GXM antibody F12D2 (red) to highlight polysaccharide epitopes. The top row of panels shows yeast cells and spores from a wild-type cross. Note that wild-type spores and yeast cells have similar intensities of staining with anti-GXM antibody. The middle row shows yeast cells and spores from an a cap67 × α cap67 cross, in which no anti-GXM antibody staining is detectable. The bottom row of panels shows yeast cells and spores stained after incubation in capsule-conditioned medium. Anti-GXM antibody staining shows brightly stained yeast cells, with only faint staining of the spores. The final panel in each row is a merged image of all three fluorescence channels used.

Article Snippet: The starting acapsular strains, cap10Δ , cap60Δ , cap64Δ , and cap67 strains (ATCC 52817-B3501 background), were all of the α mating type, so each mutant strain was crossed with JEC20 ( a ) on V8 agar ( 11 - 13 , 23 ).

Techniques: Labeling, Staining, Incubation, Fluorescence