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Cell proliferation, movement and differentiation during maintenance of the adult mouse adrenal cortex.

Chang SP, Morrison HD, Nilsson F, Kenyon CJ, West JD, Morley SD - PLoS ONE (2013)

Bottom Line: Pulse-chase-labelling coupled with phenotypic immunostaining showed that, unlike cells in the inner cortex, most BrdU-positive outer cortical cells did not express steroidogenic markers, while co-staining for BrdU and Ki67 revealed that some outer cortical BrdU-positive cells were induced to proliferate following acute adrenocorticotropic hormone (ACTH) treatment.Together, these observations are consistent with the location of both slow-cycling stem/progenitor and transiently amplifying cell populations in the outer cortex.Understanding the relationships between these distinct adrenocortical cell populations will be crucial to clarify mechanisms underpinning adrenocortical maintenance and long-term adaptation to pathophysiological states.

View Article: PubMed Central - PubMed

Affiliation: Centre for Integrative Physiology, University of Edinburgh, Edinburgh, United Kingdom.

ABSTRACT
Appropriate maintenance and regeneration of adult endocrine organs is important in both normal physiology and disease. We investigated cell proliferation, movement and differentiation in the adult mouse adrenal cortex, using different 5-bromo-2'-deoxyuridine (BrdU) labelling regimens and immunostaining for phenotypic steroidogenic cell markers. Pulse-labelling showed that cell division was largely confined to the outer cortex, with most cells moving inwards towards the medulla at around 13-20 µm per day, though a distinct labelled cell population remained in the outer 10% of the cortex. Pulse-chase-labelling coupled with phenotypic immunostaining showed that, unlike cells in the inner cortex, most BrdU-positive outer cortical cells did not express steroidogenic markers, while co-staining for BrdU and Ki67 revealed that some outer cortical BrdU-positive cells were induced to proliferate following acute adrenocorticotropic hormone (ACTH) treatment. Extended pulse-chase-labelling identified cells in the outer cortex which retained BrdU label for up to 18-23 weeks. Together, these observations are consistent with the location of both slow-cycling stem/progenitor and transiently amplifying cell populations in the outer cortex. Understanding the relationships between these distinct adrenocortical cell populations will be crucial to clarify mechanisms underpinning adrenocortical maintenance and long-term adaptation to pathophysiological states.

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Effect of acute ACTH treatment on BrdU-pulse-chase-labelled cells in the adult mouse adrenal cortex.Adrenal sections from female F1 mice treated acutely with ACTH 6 weeks after 1-week BrdU infusion, immunostained for (A,B) BrdU, (C,D) Ki67 and (E,F) BrdU/Ki67. Mice (n =6/group) were injected with saline vehicle (A,C,E) or ACTH (B,D,F), as described in the Materials and Methods section. Ki67-positive cells are indicated by brown nuclear staining while BrdU-labelled cells are identified by blue nuclear staining. Typical examples of BrdU/Ki67 dual-labelled cells are shown by arrows in F. Sections C and D were lightly counterstained with haematoxylin but sections A,B,E & F were mounted without counterstaining. Scale = 100 µm.
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pone-0081865-g004: Effect of acute ACTH treatment on BrdU-pulse-chase-labelled cells in the adult mouse adrenal cortex.Adrenal sections from female F1 mice treated acutely with ACTH 6 weeks after 1-week BrdU infusion, immunostained for (A,B) BrdU, (C,D) Ki67 and (E,F) BrdU/Ki67. Mice (n =6/group) were injected with saline vehicle (A,C,E) or ACTH (B,D,F), as described in the Materials and Methods section. Ki67-positive cells are indicated by brown nuclear staining while BrdU-labelled cells are identified by blue nuclear staining. Typical examples of BrdU/Ki67 dual-labelled cells are shown by arrows in F. Sections C and D were lightly counterstained with haematoxylin but sections A,B,E & F were mounted without counterstaining. Scale = 100 µm.

Mentions: To distinguish between slow-cycling and terminally differentiated cells in specific adrenocortical zones, BrdU pulse-chase-labelled cells were assessed for their ability to proliferate following acute ACTH stimulation, by dual-staining for BrdU and Ki67. This showed that 4 hours after ACTH treatment, BrdU-labelled cells remained widely distributed throughout the adrenal cortex, but nuclei positive for the Ki67 cell proliferation marker were found mostly in the outer cortex (Figure 4). Quantitative analysis showed that BrdU-labelled cell numbers showed a moderate increase in the ZG and outer and inner ZF following ACTH treatment, while Ki67-positive cells were increased significantly in the outer ZF (Figure 5A-B). Crucially, BrdU/Ki67 dual-labelled cell numbers showed a statistically significant increase in the ZG, with a similar, but non-significant (P = 0.14), increase in the outer ZF (Figure 5C). Cell proliferation in response to ACTH was not affected by prior BrdU labelling, since the proportions of Ki67-positive cells did not differ significantly between BrdU-labelled and unlabelled cells in any of the zones before and after ACTH treatment (P > 0.1), while acute ACTH-treatment had no significant effect on cell size (average cross-sectional area) in any zone (data not shown).


Cell proliferation, movement and differentiation during maintenance of the adult mouse adrenal cortex.

Chang SP, Morrison HD, Nilsson F, Kenyon CJ, West JD, Morley SD - PLoS ONE (2013)

Effect of acute ACTH treatment on BrdU-pulse-chase-labelled cells in the adult mouse adrenal cortex.Adrenal sections from female F1 mice treated acutely with ACTH 6 weeks after 1-week BrdU infusion, immunostained for (A,B) BrdU, (C,D) Ki67 and (E,F) BrdU/Ki67. Mice (n =6/group) were injected with saline vehicle (A,C,E) or ACTH (B,D,F), as described in the Materials and Methods section. Ki67-positive cells are indicated by brown nuclear staining while BrdU-labelled cells are identified by blue nuclear staining. Typical examples of BrdU/Ki67 dual-labelled cells are shown by arrows in F. Sections C and D were lightly counterstained with haematoxylin but sections A,B,E & F were mounted without counterstaining. Scale = 100 µm.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC3852665&req=5

pone-0081865-g004: Effect of acute ACTH treatment on BrdU-pulse-chase-labelled cells in the adult mouse adrenal cortex.Adrenal sections from female F1 mice treated acutely with ACTH 6 weeks after 1-week BrdU infusion, immunostained for (A,B) BrdU, (C,D) Ki67 and (E,F) BrdU/Ki67. Mice (n =6/group) were injected with saline vehicle (A,C,E) or ACTH (B,D,F), as described in the Materials and Methods section. Ki67-positive cells are indicated by brown nuclear staining while BrdU-labelled cells are identified by blue nuclear staining. Typical examples of BrdU/Ki67 dual-labelled cells are shown by arrows in F. Sections C and D were lightly counterstained with haematoxylin but sections A,B,E & F were mounted without counterstaining. Scale = 100 µm.
Mentions: To distinguish between slow-cycling and terminally differentiated cells in specific adrenocortical zones, BrdU pulse-chase-labelled cells were assessed for their ability to proliferate following acute ACTH stimulation, by dual-staining for BrdU and Ki67. This showed that 4 hours after ACTH treatment, BrdU-labelled cells remained widely distributed throughout the adrenal cortex, but nuclei positive for the Ki67 cell proliferation marker were found mostly in the outer cortex (Figure 4). Quantitative analysis showed that BrdU-labelled cell numbers showed a moderate increase in the ZG and outer and inner ZF following ACTH treatment, while Ki67-positive cells were increased significantly in the outer ZF (Figure 5A-B). Crucially, BrdU/Ki67 dual-labelled cell numbers showed a statistically significant increase in the ZG, with a similar, but non-significant (P = 0.14), increase in the outer ZF (Figure 5C). Cell proliferation in response to ACTH was not affected by prior BrdU labelling, since the proportions of Ki67-positive cells did not differ significantly between BrdU-labelled and unlabelled cells in any of the zones before and after ACTH treatment (P > 0.1), while acute ACTH-treatment had no significant effect on cell size (average cross-sectional area) in any zone (data not shown).

Bottom Line: Pulse-chase-labelling coupled with phenotypic immunostaining showed that, unlike cells in the inner cortex, most BrdU-positive outer cortical cells did not express steroidogenic markers, while co-staining for BrdU and Ki67 revealed that some outer cortical BrdU-positive cells were induced to proliferate following acute adrenocorticotropic hormone (ACTH) treatment.Together, these observations are consistent with the location of both slow-cycling stem/progenitor and transiently amplifying cell populations in the outer cortex.Understanding the relationships between these distinct adrenocortical cell populations will be crucial to clarify mechanisms underpinning adrenocortical maintenance and long-term adaptation to pathophysiological states.

View Article: PubMed Central - PubMed

Affiliation: Centre for Integrative Physiology, University of Edinburgh, Edinburgh, United Kingdom.

ABSTRACT
Appropriate maintenance and regeneration of adult endocrine organs is important in both normal physiology and disease. We investigated cell proliferation, movement and differentiation in the adult mouse adrenal cortex, using different 5-bromo-2'-deoxyuridine (BrdU) labelling regimens and immunostaining for phenotypic steroidogenic cell markers. Pulse-labelling showed that cell division was largely confined to the outer cortex, with most cells moving inwards towards the medulla at around 13-20 µm per day, though a distinct labelled cell population remained in the outer 10% of the cortex. Pulse-chase-labelling coupled with phenotypic immunostaining showed that, unlike cells in the inner cortex, most BrdU-positive outer cortical cells did not express steroidogenic markers, while co-staining for BrdU and Ki67 revealed that some outer cortical BrdU-positive cells were induced to proliferate following acute adrenocorticotropic hormone (ACTH) treatment. Extended pulse-chase-labelling identified cells in the outer cortex which retained BrdU label for up to 18-23 weeks. Together, these observations are consistent with the location of both slow-cycling stem/progenitor and transiently amplifying cell populations in the outer cortex. Understanding the relationships between these distinct adrenocortical cell populations will be crucial to clarify mechanisms underpinning adrenocortical maintenance and long-term adaptation to pathophysiological states.

Show MeSH
Related in: MedlinePlus