Fresh Brain Slices in Neuroscience Research: Physiological Methods, Advantages And Application Fields

Abstract

Abstract

Neuroscience studies aiming to understand the basic properties of neurons and neuronal circuits have expanded over time to include different approaches used to examine the nervous system at different scales. These studies have gained momentum with the development of the methods used. In vivo studies and in vitro cell culture studies are frequently used in neuroscience studies. However, the limitations of in vivo studies and the out of systemic structure of in vitro culture cells are insufficient to explain neural functions. The approach known as fresh brain slices are considered to be suitable preparations for ex-perimental studies of areas out of conventional in vivo and in vitro approaches. Preserved intercellular connections and long enough vitality to allow the study of adaptive and plastic processes have led to brain slices taking an important place in neuroscience studies. Acute brain slice preparation is an ideal model for studying the details of how neurons and neuronal tissue respond to a variety of physiological conditions.
The preparation process of the brain slices differs according to the designed experimental study. In order to obtain high quality brain slices, the protocol of obtaining brain slices and the preparation of artificial cerebrospinal fluid for long-term survival are important processes. During the acquisition of electrophysiological records; to keep the cross section healthy, the preparation of the appropriate environment (pH, temperature, osmolarity), the provision of optical equipment, the provision of mechan-ical tools to stabilize the position of the microelectrodes, and the equipments of amplifying and recording electronic signals are of great importance. In this review, the use of fresh brain slices, necessary equip-ment, experimental procedures and the advantages of the technique, as well as other methods frequent-ly used in electrophysiology studies, are explained.

Keywords: Electrobiophysics, Ex-vivo recording, Field potentional, Patch-clamp

Sinirbilim Araştırmalarında Taze Beyin Kesitleri: Fizyolojik Yöntemler, Avantajları Ve Kullanım Alanları

Abstract

Nöronların ve nöronal devrelerin temel özelliklerini anlamayı hedefleyen sinirbilim çalışmaları, sinir sistemini farklı ölçeklerde incelemek için kullanılan farklı yaklaşımları içerecek şekilde zamanla genişlemiştir. Kullanılan metodların geliştirilmesiyle beraber bu çalışmalar hız kazanmıştır. Canlılığın bütünüyle devam ettiği in vivo çalışmalar ve in vitro hücre kültür çalışmaları sinirbilim çalışmalarında sıklıkla kullanılmaktadır. Ancak, in vivo çalışmaların kısıtlayıcılığı ve in vitro kültür hücrelerinin sistemik yapıdan uzak oluşu sinirsel fonksiyonların açıklanmasında yetersiz kalmaktadır. Taze beyin kesitleri olarak bilinen yaklaşım klasik in vivo ve in vitro yaklaşımların erişiminden uzakta kalan alanların deneysel çalışmaları için uygun preparatlar olarak kabul edilmektedir. Hücreler arası bağlantıların korunuyor olması ve canlılığın adaptif ve plastik süreçlerin incelenmesine imkan verecek kadar uzun olması beyin kesitlerinin sinir bilim çalışmalarında önemli yer tutmasına yol açmıştır. Akut beyin kesit hazırlığı, nöronların ve nöronal dokunun çeşitli fizyolojik koşullara nasıl tepki verdiğinin ayrıntılarını incelemek için ideal bir modeldir. Beyin kesitleri çalışmalarında kesitlerin hazırlanma süreci, tasarlanan deneysel çalışmaya göre farklılık göstermektedir. Yüksek kalitede beyin kesitlerinin eldesi için, beyin kesitlerinin elde ediliş protokolü ve canlılığın uzun süre devam edebilmesi için yapay beyin omurilik sıvısının hazırlanması önemli süreçlerdir. Elektrofizyolojik kayıtların elde edilmesi esnasında; kesiti sağlıklı tutmak için uygun ortamın hazırlanması (pH, sıcaklık, osmolarite), optik donanımın sağlanması, mikroelektrotların sabit bir şekilde konumlandırmak için mekanik araçların temini ve elektronik sinyalleri yükseltme ve kaydetme araçları büyük öneme sahiptir. Bu derlemede, elektrofizyoloji çalışmalarında sıklıkla kullanılan diğer yöntemlerin yanı sıra taze beyin kesitlerinin kullanımı, gerekli ekipmanlar, deneysel prosedürler ve tekniğin sunduğu avantajlar açıklanmaktadır.

Keywords

• Beyin kesitleri
• Ex-vivo kayıt
• Patch-clamp
• Elektrobiyofizik

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