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Updated: Apr 1, 2026

Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting

Study of the Functions and Activities of Neuronal K-Cl Co-Transporter KCC2 Using Western Blotting

Published on: December 9, 2022

Developmental Expression Patterns of KCC2 and Functionally Associated Molecules in the Human Brain.

Goran Sedmak¹, Nataša Jovanov-Milošević¹, Martin Puskarjov²

¹Croatian Institute for Brain Research.

Cerebral Cortex (New York, N.Y. : 1991)

|October 3, 2015

Summary

The cation-chloride cotransporter (CCC) family, specifically KCC2, is expressed early in the human fetal brain. This finding refutes the idea that KCC2 absence causes anticonvulsant inefficacy in neonates.

Keywords:

KCC2 KCC3 NKCC1 Na-K ATPase SLC12 TrkB calpain calpastatin cation-chloride cotransporters human brain

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Area of Science:

Neuroscience
Developmental Biology
Molecular Biology

Background:

Rodent studies show KCC2 upregulation around birth establishes hyperpolarizing GABAergic responses.
The cation-chloride cotransporter (CCC) family includes KCC2, a key neuronal chloride extruder.
Understanding CCC family expression in the human brain is crucial for developmental neuroscience.

Purpose of the Study:

To map the spatiotemporal expression of the entire CCC family in the developing human brain.
To investigate the early expression of KCC2 and associated proteins (Na-K ATPase, TrkB) in human fetal brain development.
To re-evaluate the role of KCC2 in neonatal seizure susceptibility and anticonvulsant efficacy.

Main Methods:

Analysis of KCC2 mRNA expression in human brain tissue from 10th postconceptional week (PCW).
Immunohistochemical detection of KCC2-immunoreactive (KCC2-ir) neurons in the developing human brain (18-25 PCW).
Examination of mRNA expression for Na-K ATPase and TrkB in relation to KCC2 expression patterns.

Main Results:

KCC2 mRNA detected as early as 10 PCW in amygdala, cerebellum, and thalamus.
KCC2-ir neurons were abundant in the subplate by 18 PCW and in both subplate and cortical plate by 25 PCW.
Expression patterns of Na-K ATPase and TrkB mRNA align with known interactions promoting KCC2 upregulation, supporting conserved mechanisms.

Conclusions:

KCC2 and functionally associated proteins are expressed early in the human fetal brain, starting in the first trimester.
These findings facilitate the translation of rodent CCC research to human development.
The study refutes the notion that lack of KCC2 causes poor anticonvulsant efficacy in neonates; instead, perinatal calpain activation may downregulate KCC2 during trauma.