DARWIN Digitale Dissertationen German Version Strich

FU Berlin
Digitale Dissertation

Dirk Eulitz :
Heterogeneous distribution of Kir3 potassium channel proteins within dopaminergic neurons in the mesencephalon of the rat brain
Dopaminerge Neurone im motorischen und limbischen Mesencephalon der Ratte unterscheiden sich in ihrer Ausstattung mit Kir3-Kanälen

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|Abstract| |Table of Contents| |More Information|

Abstract

Dopaminergic neurons in the substantia nigra pars compacta (SNc) and the ventral tegmental area (VTA) of the ventral mesencephalon play an important role in the regulation of the parallel basal ganglia loops. Their dysfunction can result in significant locomotor (SNc) and neuropsychiatric (VTA) disorders. Due to the high similarity between the closely related loops, a selective treatment without any side-effect is not possible. Ion channels could be potential targets for a more selective therapy.
The aim of the current study was therefore to investigate the distribution of the Kir3?channel proteins in the dopaminergic neurons in the mesencephalon of the rat brain.
For this reason, specific antibodies against each of the four members of the Kir3-family were produced and affinity-purified. Immunocytochemical labelling with the purified antibodies at the light and electron microscopical level was subsequently used to investigate the distribution of these Kir3-channel proteins within the dopaminergic neurons of the ventral mesencephalon.
All four members of the Kir3-family were present in this region. Their distribution was heterogeneous, with differences in quantity as well in ultrastructural distribution being detected. The Kir3.2 channel protein distribution was especially striking. The four channel proteins could be found in the cytoplasm of dopaminergic neurons (frequently close to the ER) but only Kir3.2 was also located at the plasma membrane of these neurons. In addition, the distribution of Kir3.2 protein was shown to be heterogeneous. A significant increase of the protein concentration from the medially situated VTA to the laterally situated SNc could be observed.
Thus it seems possible that the newly characterised heterogeneous distribution of Kir3.2 channel protein in dopaminergic neurons could perhaps offer a new chance for a more selective therapy of locomotor and neuropsychiatric disorders.
 

Table of Contents

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1 Einleitung
8
1.1 Ionenkanäle
8
1.2 Kaliumkanäle
8
1.3 Einwärtsgleichrichtende Kaliumkanäle (Kir-Kanäle)
11
1.4 Kir3-Kanäle
14
1.5 Basalganglien und ventrales Mesencephalon
18
1.6 Ziel der Arbeit
23
2 Material und Methoden
24
2.1 Allgemein verwendete Materialien
24
2.2 Klonierung
25
2.3 Gewinnung der Antikörper
39
2.4. Aufreinigung der Antikörper
42
2.5 Gewinnung der Rattenhirngewebe für die Immuncytochemie
46
2.6 Gewinnung von Rattenhirnhomogenaten für Western-Blots
47
2.7 Immuncytochemie
48
3 Ergebnisse
59
3.1 Generierung und Charakterisierung der Antikörper
59
3.2 Reinigung der Antikörper am Beispiel von Kir3.1
64
3.3 Charakterisierung der gereinigten und eingeengten Antikörper
68
3.4 Ergebnisse der Lichtmikroskopie
78
3.5 Ergebnisse der Elektronenmikroskopie
89
4 Diskussion
97
4.1 Alle vier Kir3-Kanalproteine sind im ventralen Mesencephalon vorhanden
97
4.2 Bewertung des Kir3.2 Gradienten
102
4.3 Ausblick
104
5 Anhang
105
5.1 Zusammenfassung
105
5.2 Abkürzungen
106
5.3 Literaturverzeichnis
109
5.4 Publikationen
117
5.5 Lebenslauf
119
5.6 Danksagung
120

More Information:

Online available: http://www.diss.fu-berlin.de/2001/41/indexe.html
Language of PhDThesis: german
Keywords: dopamine potassium channel VTA SNc mesencephalon Kir3 Girk
DNB-Sachgruppe: 32 Biologie
Date of disputation: 29-Jan-2001
PhDThesis from: Fachbereich Biologie, Chemie, Pharmazie, Freie Universität Berlin
First Referee: Prof. Dr. Rüdiger W. Veh
Second Referee: Prof. Dr. Randolf Menzel
Contact (Author): dirk.eulitz@gmx.de
Contact (Advisor): ruediger.veh@charite.de
Date created:26-Mar-2001
Date available:28-Mar-2001

 

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