Name: organotypic slice cultures of embryonic ventral midbrain: a system to study dopaminergic neuronal development in vitro
Uploaded: 2012-01-31T12:00:00
Description: Watch this Scientific Journal Video about Organotypic Slice Cultures of Embryonic Ventral Midbrain: A System to Study Dopaminergic Neuronal Development in vitro at JoVE.com

हम Martine Emond और इसाबेल Brachmann organotypic टुकड़ा संस्कृति प्रणाली और वोल्फगैंग Hübner और Liviu गेब्रियल Bodea पांडुलिपि के महत्वपूर्ण पढ़ने के लिए स्थापित करने में उनकी मदद के लिए धन्यवाद. हम R26 संवाददाता चूहों श्श्श CreER चूहों के लिए और क्लिफ Tabin के लिए फ्रैंक COSTANTINI धन्यवाद देना चाहूंगा. उत्तर राइन वेस्टफेलिया (Programm zur Förderung der Rückkehr डेस wissenschaftlichen Spitzennachwuchses aus dem Ausland) के विज्ञान और अनुसंधान मंत्रालय से इस अध्ययन में एक अनुसंधान पुरस्कार द्वारा वित्त पोषित किया गया था.

इस प्रोटोकॉल के पार्ट्स Daza एट अल से संशोधित कर रहे हैं. 7 2007. 1. तैयारी <ol><li> एक दिन अग्रिम में तैयार किया जा सकता है <ol><li> 1X क्रेब्स बफर (1.5 एल) तैयार: 126 मिमी NaCl, 2.5, 1.2 मिमी मिमी KCl नाह 2 4 प?...

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organotypic टुकड़ा संस्कृति विधि यहाँ प्रस्तुत और भ्रूण वेंट्रल midbrain में डोपामिनर्जिक न्यूरॉन्स उनके प्रवासी और प्रक्षेपण मार्गों के विकास के इन विट्रो विश्लेषण में अल्पावधि के लिए एक प्रणाली उपलब्?...

विशिष्ट अभिकर्मकों और उपकरणों की तालिका <table border="1"><tr><th> अभिकर्मक के नाम </th><th> कंपनी </th><th> सूचीपत्र संख्या </th><th> टिप्पणियां (वैकल्पिक) </th></tr><tr><td> DMEM </td><td> सिग्मा Aldrich </td><td> D6429 </td><td></td></tr><tr><td> 30% ग्लूकोज </td><td> सिग्मा Aldrich </td><td> G7528-250 </td><td></td></tr><tr><td> हार्स सीरम </td><td> Invitrogen </td><td> 26050-088 </td><td></td></tr><tr><td> DMEM (4,5 छ / एल GLC, एल GLN के साथ, ना Pyr, NaHCO3) </td><td> सिग्मा Aldrich </td><td> D6429-500 </td><td></td></tr><tr><td> पेनिसिलीन / 100x स्ट्रेप्टोमाइसिन </td><td> सिग्मा Aldrich </td><td> P4333 - 20 </td><td></td></tr><tr><td> एल ascorbic एसिड </td><td> सिग्मा Aldrich </td><td> A4403 </td><td> -20 डिग्री सेल्सियस पर 200mm शेयर और दुकान को तैयार </td></tr><tr><td> Ultrapure एलएमपी agarose </td><td> Invitrogen </td><td> 15517-022 </td><td></td></tr><tr><td> Millicel आवेषण </td><td> Millipore </td><td> PICMORG50 </td><td></td></tr><tr><td> μ पकवान 35 मिमी, कम </td><td> Ibidi </td><td> 80136 </td><td></td></tr><tr><td> Vibratome </td><td> Microm </td><td> एचएम 650V </td><td></td></tr><tr><td> धार </td><td> Plano, GmbH </td><td> 121-6 </td><td></td></tr><tr><td> Histoacryl गोंद </td><td> BRAU9381104 </td><td> ब्रौन Aesculap </td><td></td></tr><tr><td> छिद्रित चम्मच व्यास व्यास 15 मिमी </td><td> ललित विज्ञान उपकरण </td><td> 10,370 -18 </td><td></td></tr><tr><td> संदंश 5 Dumoxel </td><td> ललित विज्ञान उपकरण </td><td> 11252 - 30 </td><td></td></tr></table> Immunostainings के लिए इस्तेमाल किया एंटीबॉडी: <table border="1"><tr><tघ&gt; नाम एंटीबॉडी के </td><td> कंपनी </td><td > सूचीपत्र संख्या </td><td> टिप्पणियां (वैकल्पिक) </td></tr><tr><td> खरगोश hydroxylase विरोधी-tyrosine </td><td> Millipore </td><td> AB152 </td><td> 1:500 मन्दन </td></tr><tr><td> विरोधी-tyrosine hydroxylase माउस </td><td> Millipore </td><td> MAB318 </td><td> 1:500 मन्दन </td></tr><tr><td> विरोधी BrdU माउस </td><td> बी.डी. Pharmingen </td><td> 555627 </td><td> कमजोर पड़ने 1:200 </td></tr><tr><td> खरगोश विरोधी cleaved 3 कस्पासे </td><td> संकेतन सेल प्रौद्योगिकी </td><td> 9661 </td><td> कमजोर पड़ने 1:200 </td></tr><tr><td> विरोधी खरगोश गधा आईजीजी Alexa 488 </td><td> Invitrogen </td><td> A21206 </td><td> 1:500 मन्दन </td></tr><tr><td> गधा आईजीजी Cy3 विरोधी माउस </td><td> जैक्सन ImmunoResearch </td><td> 715-165-150 </td><td> कमजोर पड़ने 1:200 </td></tr></table>

organotypic slice cultures of embryonic ventral midbrain: a system to study dopaminergic neuronal development in vitro

The mouse is an excellent model organism to study mammalian brain development due to the abundance of molecular and genetic data. However, the developing mouse brain is not suitable for easy manipulation and imaging in vivo since the mouse embryo is inaccessible and opaque. Organotypic slice cultures of embryonic brains are therefore widely used to study murine brain development in vitro. Ex-vivo manipulation or the use of transgenic mice allows the modification of gene expression so that subpopulations of neuronal or glial cells can be labeled with fluorescent proteins. The behavior of labeled cells can then be observed using time-lapse imaging. Time-lapse imaging has been particularly successful for studying cell behaviors that underlie the development of the cerebral cortex at late embryonic stages 1-2. Embryonic organotypic slice culture systems in brain regions outside of the forebrain are less well established. Therefore, the wealth of time-lapse imaging data describing neuronal cell migration is restricted to the forebrain 3,4. It is still not known, whether the principles discovered for the dorsal brain hold true for ventral brain areas. In the ventral brain, neurons are organized in neuronal clusters rather than layers and they often have to undergo complicated migratory trajectories to reach their final position. The ventral midbrain is not only a good model system for ventral brain development, but also contains neuronal populations such as dopaminergic neurons that are relevant in disease processes. While the function and degeneration of dopaminergic neurons has been investigated in great detail in the adult and ageing brain, little is known about the behavior of these neurons during their differentiation and migration phase 5. We describe here the generation of slice cultures from the embryonic day (E) 12.5 mouse ventral midbrain. These slice cultures are potentially suitable for monitoring dopaminergic neuron development over several days in vitro. We highlight the critical steps in generating brain slices at these early stages of embryonic development and discuss the conditions necessary for maintaining normal development of dopaminergic neurons in vitro. We also present results from time lapse imaging experiments. In these experiments, ventral midbrain precursors (including dopaminergic precursors) and their descendants were labeled in a mosaic manner using a Cre/loxP based inducible fate mapping system 6.

Watch this Scientific Journal Video about Organotypic Slice Cultures of Embryonic Ventral Midbrain: A System to Study Dopaminergic Neuronal Development in vitro at JoVE.com

Organotypic Slice Cultures of Embryonic Ventral Midbrain: A System to Study Dopaminergic Neuronal Development in vitro

E12.5 murine भ्रूण midbrain से organotypic स्लाइस उत्पन्न विधि वर्णित है. organotypic टुकड़ा संस्कृतियों डोपामिनर्जिक न्यूरॉन्स या अन्य वेंट्रल मध्यमस्तिष्क न्यूरॉन्स के व्यवहार का निरीक्षण करने के लिए इस्तेमाल किया जा सकता है.

भ्रूणीय ventral midbrain के Organotypic स्लाइस संस्कृति: एक डोपामिनर्जिक neuronal विकास अध्ययन प्रणाली इन विट्रो में</em

The  mouse is an excellent model organism to study mammalian brain development due  to the abundance of molecular and genetic data. However, the ...

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Organotypic Slice Cultures of Embryonic Ventral Midbrain: A System to Study Dopaminergic Neuronal Development in vitro

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