Functional aspects of early brain development are preserved in tuberous sclerosis complex (TSC) epileptogenic lesions

Gabriele Ruffolo, Anand Iyer, Pierangelo Cifelli, Cristina Roseti, Angelika Mühlebner, Jackelien van Scheppingen, Theresa Scholl, Johannes A. Hainfellner, Martha Feucht, Pavel Krsek, Josef Zamecnik, Floor E. Jansen, Wim G M Spliet, Cristina Limatola, Eleonora Aronica, Eleonora Palma

Research output: Contribution to journalArticle

Abstract

Tuberous sclerosis complex (TSC) is a rare multi-system genetic disease characterized by several neurological disorders, the most common of which is the refractory epilepsy caused by highly epileptogenic cortical lesions. Previous studies suggest an alteration of GABAergic and glutamatergic transmission in TSC brain indicating an unbalance of excitation/inhibition that can explain, at least in part, the high incidence of epilepsy in these patients. Here we investigate whether TSC cortical tissues could retain GABAA and AMPA receptors at early stages of human brain development thus contributing to the generation and recurrence of seizures. Given the limited availability of pediatric human brain specimens, we used the microtransplantation method of injecting Xenopus oocytes with membranes from TSC cortical tubers and control brain tissues. Moreover, qPCR was performed to investigate the expression of GABAA and AMPA receptor subunits (GABAA α1–5, β3, γ2, δ; GluA1, GluA2) and cation chloride co-transporters NKCC1 and KCC2. The evaluation of nine human cortical brain samples, from 15 gestation weeks to 15 years old, showed a progressive shift towards more hyperpolarized GABAA reversal potential (EGABA). This shift was associated with a differential expression of the chloride cotransporters NKCC1 and KCC2. Furthermore, the GluA1/GluA2 mRNA ratio of expression paralleled the development process. On the contrary, in oocytes micro-transplanted with epileptic TSC tuber tissue from seven patients, neither the GABAA reversal potential nor the GluA1/GluA2 expression showed similar developmental changes. Our data indicate for the first time, that in the same cohort of TSC patients, the pattern of both GABAAR and GluA1/GluA2 functions retains features that are typical of an immature brain. These observations support the potential contribution of altered receptor function to the epileptic disorder of TSC and may suggest novel therapeutic approaches. Furthermore, our findings strengthen the novel hypothesis that other developmental brain diseases can share the same hallmarks of immaturity leading to intractable seizures.

Original languageEnglish
Pages (from-to)93-101
Number of pages9
JournalNeurobiology of Disease
Volume95
DOIs
Publication statusPublished - Nov 1 2016

Fingerprint

Tuberous Sclerosis
Brain
AMPA Receptors
GABA-A Receptors
Oocytes
Chlorides
Epilepsy
Member 2 Solute Carrier Family 12
Seizures
Symporters
Inborn Genetic Diseases
Brain Diseases
Human Development
Xenopus
Nervous System Diseases
Cations
Pediatrics
Recurrence
Pregnancy
Messenger RNA

Keywords

  • Brain development
  • Epilepsy
  • GABA receptor
  • Oocytes
  • Tuberous sclerosis complex

ASJC Scopus subject areas

  • Neurology

Cite this

Functional aspects of early brain development are preserved in tuberous sclerosis complex (TSC) epileptogenic lesions. / Ruffolo, Gabriele; Iyer, Anand; Cifelli, Pierangelo; Roseti, Cristina; Mühlebner, Angelika; van Scheppingen, Jackelien; Scholl, Theresa; Hainfellner, Johannes A.; Feucht, Martha; Krsek, Pavel; Zamecnik, Josef; Jansen, Floor E.; Spliet, Wim G M; Limatola, Cristina; Aronica, Eleonora; Palma, Eleonora.

In: Neurobiology of Disease, Vol. 95, 01.11.2016, p. 93-101.

Research output: Contribution to journalArticle

Ruffolo, G, Iyer, A, Cifelli, P, Roseti, C, Mühlebner, A, van Scheppingen, J, Scholl, T, Hainfellner, JA, Feucht, M, Krsek, P, Zamecnik, J, Jansen, FE, Spliet, WGM, Limatola, C, Aronica, E & Palma, E 2016, 'Functional aspects of early brain development are preserved in tuberous sclerosis complex (TSC) epileptogenic lesions', Neurobiology of Disease, vol. 95, pp. 93-101. https://doi.org/10.1016/j.nbd.2016.07.014
Ruffolo, Gabriele ; Iyer, Anand ; Cifelli, Pierangelo ; Roseti, Cristina ; Mühlebner, Angelika ; van Scheppingen, Jackelien ; Scholl, Theresa ; Hainfellner, Johannes A. ; Feucht, Martha ; Krsek, Pavel ; Zamecnik, Josef ; Jansen, Floor E. ; Spliet, Wim G M ; Limatola, Cristina ; Aronica, Eleonora ; Palma, Eleonora. / Functional aspects of early brain development are preserved in tuberous sclerosis complex (TSC) epileptogenic lesions. In: Neurobiology of Disease. 2016 ; Vol. 95. pp. 93-101.
@article{db27cbfeceef4928b688cf4bbe0c40b2,
title = "Functional aspects of early brain development are preserved in tuberous sclerosis complex (TSC) epileptogenic lesions",
abstract = "Tuberous sclerosis complex (TSC) is a rare multi-system genetic disease characterized by several neurological disorders, the most common of which is the refractory epilepsy caused by highly epileptogenic cortical lesions. Previous studies suggest an alteration of GABAergic and glutamatergic transmission in TSC brain indicating an unbalance of excitation/inhibition that can explain, at least in part, the high incidence of epilepsy in these patients. Here we investigate whether TSC cortical tissues could retain GABAA and AMPA receptors at early stages of human brain development thus contributing to the generation and recurrence of seizures. Given the limited availability of pediatric human brain specimens, we used the microtransplantation method of injecting Xenopus oocytes with membranes from TSC cortical tubers and control brain tissues. Moreover, qPCR was performed to investigate the expression of GABAA and AMPA receptor subunits (GABAA α1–5, β3, γ2, δ; GluA1, GluA2) and cation chloride co-transporters NKCC1 and KCC2. The evaluation of nine human cortical brain samples, from 15 gestation weeks to 15 years old, showed a progressive shift towards more hyperpolarized GABAA reversal potential (EGABA). This shift was associated with a differential expression of the chloride cotransporters NKCC1 and KCC2. Furthermore, the GluA1/GluA2 mRNA ratio of expression paralleled the development process. On the contrary, in oocytes micro-transplanted with epileptic TSC tuber tissue from seven patients, neither the GABAA reversal potential nor the GluA1/GluA2 expression showed similar developmental changes. Our data indicate for the first time, that in the same cohort of TSC patients, the pattern of both GABAAR and GluA1/GluA2 functions retains features that are typical of an immature brain. These observations support the potential contribution of altered receptor function to the epileptic disorder of TSC and may suggest novel therapeutic approaches. Furthermore, our findings strengthen the novel hypothesis that other developmental brain diseases can share the same hallmarks of immaturity leading to intractable seizures.",
keywords = "Brain development, Epilepsy, GABA receptor, Oocytes, Tuberous sclerosis complex",
author = "Gabriele Ruffolo and Anand Iyer and Pierangelo Cifelli and Cristina Roseti and Angelika M{\"u}hlebner and {van Scheppingen}, Jackelien and Theresa Scholl and Hainfellner, {Johannes A.} and Martha Feucht and Pavel Krsek and Josef Zamecnik and Jansen, {Floor E.} and Spliet, {Wim G M} and Cristina Limatola and Eleonora Aronica and Eleonora Palma",
year = "2016",
month = "11",
day = "1",
doi = "10.1016/j.nbd.2016.07.014",
language = "English",
volume = "95",
pages = "93--101",
journal = "Neurobiology of Disease",
issn = "0969-9961",
publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - Functional aspects of early brain development are preserved in tuberous sclerosis complex (TSC) epileptogenic lesions

AU - Ruffolo, Gabriele

AU - Iyer, Anand

AU - Cifelli, Pierangelo

AU - Roseti, Cristina

AU - Mühlebner, Angelika

AU - van Scheppingen, Jackelien

AU - Scholl, Theresa

AU - Hainfellner, Johannes A.

AU - Feucht, Martha

AU - Krsek, Pavel

AU - Zamecnik, Josef

AU - Jansen, Floor E.

AU - Spliet, Wim G M

AU - Limatola, Cristina

AU - Aronica, Eleonora

AU - Palma, Eleonora

PY - 2016/11/1

Y1 - 2016/11/1

N2 - Tuberous sclerosis complex (TSC) is a rare multi-system genetic disease characterized by several neurological disorders, the most common of which is the refractory epilepsy caused by highly epileptogenic cortical lesions. Previous studies suggest an alteration of GABAergic and glutamatergic transmission in TSC brain indicating an unbalance of excitation/inhibition that can explain, at least in part, the high incidence of epilepsy in these patients. Here we investigate whether TSC cortical tissues could retain GABAA and AMPA receptors at early stages of human brain development thus contributing to the generation and recurrence of seizures. Given the limited availability of pediatric human brain specimens, we used the microtransplantation method of injecting Xenopus oocytes with membranes from TSC cortical tubers and control brain tissues. Moreover, qPCR was performed to investigate the expression of GABAA and AMPA receptor subunits (GABAA α1–5, β3, γ2, δ; GluA1, GluA2) and cation chloride co-transporters NKCC1 and KCC2. The evaluation of nine human cortical brain samples, from 15 gestation weeks to 15 years old, showed a progressive shift towards more hyperpolarized GABAA reversal potential (EGABA). This shift was associated with a differential expression of the chloride cotransporters NKCC1 and KCC2. Furthermore, the GluA1/GluA2 mRNA ratio of expression paralleled the development process. On the contrary, in oocytes micro-transplanted with epileptic TSC tuber tissue from seven patients, neither the GABAA reversal potential nor the GluA1/GluA2 expression showed similar developmental changes. Our data indicate for the first time, that in the same cohort of TSC patients, the pattern of both GABAAR and GluA1/GluA2 functions retains features that are typical of an immature brain. These observations support the potential contribution of altered receptor function to the epileptic disorder of TSC and may suggest novel therapeutic approaches. Furthermore, our findings strengthen the novel hypothesis that other developmental brain diseases can share the same hallmarks of immaturity leading to intractable seizures.

AB - Tuberous sclerosis complex (TSC) is a rare multi-system genetic disease characterized by several neurological disorders, the most common of which is the refractory epilepsy caused by highly epileptogenic cortical lesions. Previous studies suggest an alteration of GABAergic and glutamatergic transmission in TSC brain indicating an unbalance of excitation/inhibition that can explain, at least in part, the high incidence of epilepsy in these patients. Here we investigate whether TSC cortical tissues could retain GABAA and AMPA receptors at early stages of human brain development thus contributing to the generation and recurrence of seizures. Given the limited availability of pediatric human brain specimens, we used the microtransplantation method of injecting Xenopus oocytes with membranes from TSC cortical tubers and control brain tissues. Moreover, qPCR was performed to investigate the expression of GABAA and AMPA receptor subunits (GABAA α1–5, β3, γ2, δ; GluA1, GluA2) and cation chloride co-transporters NKCC1 and KCC2. The evaluation of nine human cortical brain samples, from 15 gestation weeks to 15 years old, showed a progressive shift towards more hyperpolarized GABAA reversal potential (EGABA). This shift was associated with a differential expression of the chloride cotransporters NKCC1 and KCC2. Furthermore, the GluA1/GluA2 mRNA ratio of expression paralleled the development process. On the contrary, in oocytes micro-transplanted with epileptic TSC tuber tissue from seven patients, neither the GABAA reversal potential nor the GluA1/GluA2 expression showed similar developmental changes. Our data indicate for the first time, that in the same cohort of TSC patients, the pattern of both GABAAR and GluA1/GluA2 functions retains features that are typical of an immature brain. These observations support the potential contribution of altered receptor function to the epileptic disorder of TSC and may suggest novel therapeutic approaches. Furthermore, our findings strengthen the novel hypothesis that other developmental brain diseases can share the same hallmarks of immaturity leading to intractable seizures.

KW - Brain development

KW - Epilepsy

KW - GABA receptor

KW - Oocytes

KW - Tuberous sclerosis complex

UR - http://www.scopus.com/inward/record.url?scp=84978733864&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84978733864&partnerID=8YFLogxK

U2 - 10.1016/j.nbd.2016.07.014

DO - 10.1016/j.nbd.2016.07.014

M3 - Article

AN - SCOPUS:84978733864

VL - 95

SP - 93

EP - 101

JO - Neurobiology of Disease

JF - Neurobiology of Disease

SN - 0969-9961

ER -