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Related Experiment Video

Updated: Jul 15, 2026

Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting
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NKCC1 does not accumulate chloride in developing retinal neurons.

Ling-Li Zhang1, Eric Delpire, Noga Vardi

  • 1Dept of Neuroscience, University of Pennsylvania, Philadelphia, PA 19104-6058, USA.

Journal of Neurophysiology
|May 12, 2007
PubMed
Summary

The chloride transporter NKCC1 does not accumulate chloride in immature retinal neurons, contrary to common belief. Instead, it may help Müller cells buffer extracellular chloride levels.

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Area of Science:

  • Neuroscience
  • Developmental Biology
  • Cellular Physiology

Background:

  • Gamma-aminobutyric acid (GABA) typically inhibits mature neurons but excites immature ones due to high intracellular chloride.
  • The chloride cotransporter NKCC1 is widely believed to be responsible for this high chloride accumulation in developing neurons.

Purpose of the Study:

  • To investigate the role of NKCC1 in chloride accumulation in immature retinal amacrine and ganglion cells.
  • To determine if NKCC1 is the primary transporter responsible for GABAergic excitation in these developing retinal neurons.

Main Methods:

  • Western blotting and immunocytochemistry to detect and localize NKCC1 expression in the developing retina.
  • Calcium imaging and intracellular chloride measurements in wild-type and NKCC1-null retinas.

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  • Functional assays using bumetanide (NKCC1 blocker) and manipulation of extracellular sodium.

    • Main Results:

    • NKCC1 expression in retinal neurons peaked early and decreased with age, while KCC2 expression increased.
    • GABA-evoked calcium rises in immature retinal neurons were not abolished in NKCC1-null retinas or with bumetanide treatment.
    • Intracellular chloride concentrations remained high in NKCC1-null retinas and were unaffected by bumetanide or altered sodium levels.
    • NKCC1 was primarily localized to Müller cells at early developmental stages.

    Conclusions:

    • NKCC1 does not accumulate chloride in immature retinal amacrine and ganglion cells.
    • The mechanism of chloride accumulation and GABAergic excitation in these cells requires further investigation.
    • NKCC1 in Müller cells may play a role in buffering extracellular chloride concentrations.