What is the clinical evidence from this study?

Intervention: Isoproterenol. Primary outcome: Membrane-proximal acidic granular organelle dynamics and behavior.

What question did this study set out to answer?

To establish an isolation and culture protocol for primary juxtaglomerular cells and evaluate the dynamic behavior of renin-containing secretory granules using high-resolution live-cell microscopy.

November 10, 2016AJP Renal PhysiologyOpen Access

Bimodal dynamics of granular organelles in primary renin-expressing cells revealed using TIRF microscopy

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Key result

Isoproterenol stimulation of primary renin-expressing cells revealed two populations of granular organelles with distinct functional responses.

Population

Primary renin-expressing cells isolated from RenGFP+ transgenic mice

Design

Preclinical

Authors

CBCharlotte BuckleyUniversidad Mayor ADAlison R. DunHeriot-Watt University APAudrey PeterBritish Heart Foundation Centre for Cardiovascular Science

Discussion

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Implication

Hypothesis-generating in primary cells; leaves open whether granule subpopulations differentially regulate renin release in vivo.

Key Points

To establish an isolation and culture protocol for primary juxtaglomerular cells and evaluate the dynamic behavior of renin-containing secretory granules using high-resolution live-cell microscopy.
Purified primary renin-expressing cells from RenGFP+ transgenic mice using Percoll density gradient centrifugation and cultured them on fibronectin-coated glass coverslips.
Labeled acidic granule and lysosome populations with Lysotracker Red and stimulated cells with 100 nM isoproterenol.
Recorded real-time, membrane-proximal organelle dynamics at the plasma membrane using total internal reflection fluorescence microscopy (TIRFM).
Fibronectin-coated coverslips optimized cell adhesion and enabled high-speed, high-resolution TIRF imaging of primary renin-expressing cells.
Isoproterenol stimulation revealed two functionally distinct populations of membrane-proximal acidic granular organelles with bimodal dynamic responses.

Structured PICO

Population

Primary renin-expressing cells isolated from RenGFP+ transgenic mice

Intervention

Stimulation with 100 nM isoproterenol

Outcome

Membrane-proximal acidic granular organelle dynamics and behaviorsurrogate

High-resolution TIRF microscopy reveals two distinct populations of granular organelles with different functional responses to isoproterenol in primary renin-expressing cells.

Cite This Study

Buckley et al. (2016) studied this question. Isoproterenol was evaluated on Membrane-proximal acidic granular organelle dynamics and behavior. Isoproterenol stimulation of primary renin-expressing cells revealed two populations of granular organelles with distinct functional responses.

synapsesocial.com/papers/6a846c7ec7e37ebe3168dd6a https://doi.org/10.1152/ajprenal.00384.2016

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Bimodal dynamics of granular organelles in primary renin-expressing cells revealed using TIRF microscopy

Discussion

Member takes

Implication

Key Points

Structured PICO

Cite This Study

Bimodal dynamics of granular organelles in primary renin-expressing cells revealed using TIRF microscopy

Discussion

Member takes

Implication

Key Points

Structured PICO

Cite This Study

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