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The centrosomal protein pericentrin identified at the basal body complex of the connecting cilium in mouse photoreceptors.

Mühlhans J, Brandstätter JH, Giessl A - PLoS ONE (2011)

Bottom Line: Here, Pcnt colocalized with the whole protein machinery responsible for transport processes between the two compartments.Surprisingly, photoreceptors expressed a small Pcnt splice transcript - most likely a modified variant of Pcnt S - which was not present in receptor neurons of the olfactory epithelium.The individual patchwork of different Pcnt splice transcripts seems to reflect the complexity of Pcnt function, an assumption corroborated by the heterogeneous clinical manifestations associated with mutations in the Pcnt gene.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Animal Physiology, University of Erlangen-Nuremberg, Erlangen, Germany.

ABSTRACT

Background: Pericentrin (Pcnt), a conserved protein of the pericentriolar material, serves as a multifunctional scaffold for numerous proteins and plays an important role in microtubule organization. Recent studies indicate that Pcnt mutations are associated with a range of diseases including primordial dwarfism and ciliopathies. To date, three Pcnt splice variants from orthologous genes in mice and humans are known.

Principal findings: We generated a specific Pcnt antiserum detecting all known Pcnt splice variants and examined the cellular and subcellular distribution of Pcnt in ciliated tissues of the mouse, the olfactory epithelium and the retina. For the first time, we identified Pcnt and its centrosomal interaction partners at the basal body complex of mouse retinal photoreceptors. Photoreceptors are morphologically and functionally subdivided into the light sensitive outer segment and the inner segment comprising the metabolic function of the cell. The two compartments are linked via a modified, specialized, non-motile cilium, the connecting cilium. Here, Pcnt colocalized with the whole protein machinery responsible for transport processes between the two compartments. Surprisingly, photoreceptors expressed a small Pcnt splice transcript - most likely a modified variant of Pcnt S - which was not present in receptor neurons of the olfactory epithelium.

Conclusions: Our findings suggest distinct functional roles of several Pcnt variants in different ciliated tissues and sensory neurons, like the olfactory epithelium and the retina of the mouse. The individual patchwork of different Pcnt splice transcripts seems to reflect the complexity of Pcnt function, an assumption corroborated by the heterogeneous clinical manifestations associated with mutations in the Pcnt gene.

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Localization of Pericentrin during the cell cycle in NIH 3T3 mouse fibroblasts.(A–D) Triple staining of endogenous Pcnt (A, green), γ-tubulin (B, red) as a centrosomal marker, and DAPI (D, blue) as a nuclear marker in NIH 3T3 mouse fibroblasts. (C–D) The merge of the stainings shows the localization of Pcnt at the centrosomes in dividing (arrowheads) and in non-dividing cells (arrow). (E–H) NIH 3T3 mouse fibroblast cells stained for endogenous Pcnt (E, green), α-tubulin (F, red) as a marker for microtubules, and DAPI (H, blue). (G–H) In the merge pictures Pcnt is localized at the centrosomes. Scale bars: 5 µm (H, D).
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pone-0026496-g002: Localization of Pericentrin during the cell cycle in NIH 3T3 mouse fibroblasts.(A–D) Triple staining of endogenous Pcnt (A, green), γ-tubulin (B, red) as a centrosomal marker, and DAPI (D, blue) as a nuclear marker in NIH 3T3 mouse fibroblasts. (C–D) The merge of the stainings shows the localization of Pcnt at the centrosomes in dividing (arrowheads) and in non-dividing cells (arrow). (E–H) NIH 3T3 mouse fibroblast cells stained for endogenous Pcnt (E, green), α-tubulin (F, red) as a marker for microtubules, and DAPI (H, blue). (G–H) In the merge pictures Pcnt is localized at the centrosomes. Scale bars: 5 µm (H, D).

Mentions: Additional immunocytochemical stainings of active NIH 3T3 mouse fibroblasts with the MmPeriC1 antiserum revealed that Pcnt overlapped with γ-tubulin, a protein associated with the mother and daughter centriole of the centrosome in all phases of the cell cycle (Fig. 2A–D). Centrosomal structures also colocalize with α-tubulin, a marker for microtubules (Fig. 2E–H). These results indicate the presence of Pcnt at centrosomal or BBC structures in every stage of the cell cycle, regardless of whether the cell is dividing or resting, and support the assumption that Pcnt may also play a role in resting, and thus often ciliated cells.


The centrosomal protein pericentrin identified at the basal body complex of the connecting cilium in mouse photoreceptors.

Mühlhans J, Brandstätter JH, Giessl A - PLoS ONE (2011)

Localization of Pericentrin during the cell cycle in NIH 3T3 mouse fibroblasts.(A–D) Triple staining of endogenous Pcnt (A, green), γ-tubulin (B, red) as a centrosomal marker, and DAPI (D, blue) as a nuclear marker in NIH 3T3 mouse fibroblasts. (C–D) The merge of the stainings shows the localization of Pcnt at the centrosomes in dividing (arrowheads) and in non-dividing cells (arrow). (E–H) NIH 3T3 mouse fibroblast cells stained for endogenous Pcnt (E, green), α-tubulin (F, red) as a marker for microtubules, and DAPI (H, blue). (G–H) In the merge pictures Pcnt is localized at the centrosomes. Scale bars: 5 µm (H, D).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3198765&req=5

pone-0026496-g002: Localization of Pericentrin during the cell cycle in NIH 3T3 mouse fibroblasts.(A–D) Triple staining of endogenous Pcnt (A, green), γ-tubulin (B, red) as a centrosomal marker, and DAPI (D, blue) as a nuclear marker in NIH 3T3 mouse fibroblasts. (C–D) The merge of the stainings shows the localization of Pcnt at the centrosomes in dividing (arrowheads) and in non-dividing cells (arrow). (E–H) NIH 3T3 mouse fibroblast cells stained for endogenous Pcnt (E, green), α-tubulin (F, red) as a marker for microtubules, and DAPI (H, blue). (G–H) In the merge pictures Pcnt is localized at the centrosomes. Scale bars: 5 µm (H, D).
Mentions: Additional immunocytochemical stainings of active NIH 3T3 mouse fibroblasts with the MmPeriC1 antiserum revealed that Pcnt overlapped with γ-tubulin, a protein associated with the mother and daughter centriole of the centrosome in all phases of the cell cycle (Fig. 2A–D). Centrosomal structures also colocalize with α-tubulin, a marker for microtubules (Fig. 2E–H). These results indicate the presence of Pcnt at centrosomal or BBC structures in every stage of the cell cycle, regardless of whether the cell is dividing or resting, and support the assumption that Pcnt may also play a role in resting, and thus often ciliated cells.

Bottom Line: Here, Pcnt colocalized with the whole protein machinery responsible for transport processes between the two compartments.Surprisingly, photoreceptors expressed a small Pcnt splice transcript - most likely a modified variant of Pcnt S - which was not present in receptor neurons of the olfactory epithelium.The individual patchwork of different Pcnt splice transcripts seems to reflect the complexity of Pcnt function, an assumption corroborated by the heterogeneous clinical manifestations associated with mutations in the Pcnt gene.

View Article: PubMed Central - PubMed

Affiliation: Department of Biology, Animal Physiology, University of Erlangen-Nuremberg, Erlangen, Germany.

ABSTRACT

Background: Pericentrin (Pcnt), a conserved protein of the pericentriolar material, serves as a multifunctional scaffold for numerous proteins and plays an important role in microtubule organization. Recent studies indicate that Pcnt mutations are associated with a range of diseases including primordial dwarfism and ciliopathies. To date, three Pcnt splice variants from orthologous genes in mice and humans are known.

Principal findings: We generated a specific Pcnt antiserum detecting all known Pcnt splice variants and examined the cellular and subcellular distribution of Pcnt in ciliated tissues of the mouse, the olfactory epithelium and the retina. For the first time, we identified Pcnt and its centrosomal interaction partners at the basal body complex of mouse retinal photoreceptors. Photoreceptors are morphologically and functionally subdivided into the light sensitive outer segment and the inner segment comprising the metabolic function of the cell. The two compartments are linked via a modified, specialized, non-motile cilium, the connecting cilium. Here, Pcnt colocalized with the whole protein machinery responsible for transport processes between the two compartments. Surprisingly, photoreceptors expressed a small Pcnt splice transcript - most likely a modified variant of Pcnt S - which was not present in receptor neurons of the olfactory epithelium.

Conclusions: Our findings suggest distinct functional roles of several Pcnt variants in different ciliated tissues and sensory neurons, like the olfactory epithelium and the retina of the mouse. The individual patchwork of different Pcnt splice transcripts seems to reflect the complexity of Pcnt function, an assumption corroborated by the heterogeneous clinical manifestations associated with mutations in the Pcnt gene.

Show MeSH
Related in: MedlinePlus