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Determination of Mother Centriole Maturation in CPAP-Depleted Cells Using the Ninein Antibody.

Lee M, Rhee K - Endocrinol Metab (Seoul) (2014)

Bottom Line: We immunostained CPAP-depleted cells using the ninein antibody, which selectively detects subdistal appendages in mature mother centrioles.Ninein signals were significantly impaired in CPAP-depleted cells.The selective absence of centriolar appendages in young mother centrioles may be responsible for asymmetric spindle pole formation in CPAP-depleted cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Seoul National University, Seoul, Korea.

ABSTRACT

Background: Mutations in centrosomal protein genes have been identified in a number of genetic diseases in brain development, including microcephaly. Centrosomal P4.1-associated protein (CPAP) is one of the causal genes implicated in primary microcephaly. We previously proposed that CPAP is essential for mother centriole maturation during mitosis.

Methods: We immunostained CPAP-depleted cells using the ninein antibody, which selectively detects subdistal appendages in mature mother centrioles.

Results: Ninein signals were significantly impaired in CPAP-depleted cells.

Conclusion: The results suggest that CPAP is required for mother centriole maturation in mammalian cells. The selective absence of centriolar appendages in young mother centrioles may be responsible for asymmetric spindle pole formation in CPAP-depleted cells.

No MeSH data available.


Related in: MedlinePlus

Ninein distribution within the centriole. Asynchronous HeLa cells were coimmunostained with centrin-2 (green) and ninein (red) antibodies. DNA was stained with 4',6-diamidino-2-phenylindole. Scale bar=10 µm. Insets are magnified views of the centrosomes. The staining patterns of ninein on centrioles were taken from the right side.
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Figure 3: Ninein distribution within the centriole. Asynchronous HeLa cells were coimmunostained with centrin-2 (green) and ninein (red) antibodies. DNA was stained with 4',6-diamidino-2-phenylindole. Scale bar=10 µm. Insets are magnified views of the centrosomes. The staining patterns of ninein on centrioles were taken from the right side.

Mentions: To determine the precise localization of ninein within the centriole, we coimmunostained HeLa cells with ninein and centrin-2 antibodies. Ninein was localized at only one of the two unduplicated centrioles in G1 phase cells (Fig. 3). Ninein was detected in both the proximal and distal regions of the mother centriole, as either a ring or three dots (Fig. 3). A second ninein signal appeared as a dot at the daughter centriole during late-stage G1 (Fig. 3). Strong and weak ninein signals were located at the old and young mother centrioles, respectively (Fig. 3). This staining pattern was maintained until the young mother centriole reached maturity during late-stage G2 (data not shown).


Determination of Mother Centriole Maturation in CPAP-Depleted Cells Using the Ninein Antibody.

Lee M, Rhee K - Endocrinol Metab (Seoul) (2014)

Ninein distribution within the centriole. Asynchronous HeLa cells were coimmunostained with centrin-2 (green) and ninein (red) antibodies. DNA was stained with 4',6-diamidino-2-phenylindole. Scale bar=10 µm. Insets are magnified views of the centrosomes. The staining patterns of ninein on centrioles were taken from the right side.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
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getmorefigures.php?uid=PMC4384682&req=5

Figure 3: Ninein distribution within the centriole. Asynchronous HeLa cells were coimmunostained with centrin-2 (green) and ninein (red) antibodies. DNA was stained with 4',6-diamidino-2-phenylindole. Scale bar=10 µm. Insets are magnified views of the centrosomes. The staining patterns of ninein on centrioles were taken from the right side.
Mentions: To determine the precise localization of ninein within the centriole, we coimmunostained HeLa cells with ninein and centrin-2 antibodies. Ninein was localized at only one of the two unduplicated centrioles in G1 phase cells (Fig. 3). Ninein was detected in both the proximal and distal regions of the mother centriole, as either a ring or three dots (Fig. 3). A second ninein signal appeared as a dot at the daughter centriole during late-stage G1 (Fig. 3). Strong and weak ninein signals were located at the old and young mother centrioles, respectively (Fig. 3). This staining pattern was maintained until the young mother centriole reached maturity during late-stage G2 (data not shown).

Bottom Line: We immunostained CPAP-depleted cells using the ninein antibody, which selectively detects subdistal appendages in mature mother centrioles.Ninein signals were significantly impaired in CPAP-depleted cells.The selective absence of centriolar appendages in young mother centrioles may be responsible for asymmetric spindle pole formation in CPAP-depleted cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Sciences, Seoul National University, Seoul, Korea.

ABSTRACT

Background: Mutations in centrosomal protein genes have been identified in a number of genetic diseases in brain development, including microcephaly. Centrosomal P4.1-associated protein (CPAP) is one of the causal genes implicated in primary microcephaly. We previously proposed that CPAP is essential for mother centriole maturation during mitosis.

Methods: We immunostained CPAP-depleted cells using the ninein antibody, which selectively detects subdistal appendages in mature mother centrioles.

Results: Ninein signals were significantly impaired in CPAP-depleted cells.

Conclusion: The results suggest that CPAP is required for mother centriole maturation in mammalian cells. The selective absence of centriolar appendages in young mother centrioles may be responsible for asymmetric spindle pole formation in CPAP-depleted cells.

No MeSH data available.


Related in: MedlinePlus