We would like to thank the members of the Montgomery and Madison labs for helpful discussion. We acknowledge the funding received from the following sources in this research: NFNZ, AMRF, Marsden Fund, HRC, and NIH.

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The authors have nothing to disclose

כאן יש לנו תיארנו את הדרישות להקמת הקלטות תא מוצלחות לזווג כל בתרבויות פרוסה בהיפוקמפוס organotypic. יכולות גם להתבצע הקלטות מותאמים בהכנות מרובות, כוללים פרוסות חריפות ומערכות תרבות ניתקו 26,27. למרות ההתמקדות כאן הייתה על הגיוס של צורות ארוכות יותר של פלסטיות הסינפ?...

קולטני גלוטמט לתווך רוב שידור סינפטי מעורר בסינפסות במערכת עצבים מרכזית. שני תתי הסוגים העיקריים של ionotropic קולטני גלוטמט המקומי בראש עמוד השדרה של קרום postsynaptic הם N-methyl-D-aspartate (NMDA) וα-אמינו-3-הידרוקסי-5-methylisoxazole-4-proprionic חומצת קולטנים (AMPA). בנחים פוטנציאלי קרום, קולטני AMPA לשאת רוב הנוכחי postsynaptic במהלך שידור סינפטי. בהיפוקמפוס, רצפטור NMDA ממלא תפקיד מרכזי בגרימת שינויים במספר הקולטנים AMPA בממברנה postsynaptic: על ידי מתנהג כמו &quot;גלאי צירוף מקרים&quot; 1 ליזום שינויים בכוח הסינפטי 1, רצפטור NMDA משתתף במנגנונים סינפטיים כי הם חשבו כדי לחזק את למידה וזיכרון ברמת subcellular. בתגובה לשלילת קוטביות של הנוירון postsynaptic במקביל לשחרור משדר presynaptic, סידן נכנס דרך NMDAקולט ליזום הכנסת קולטן AMPA או הסרת 2. דינמיקת קולט אלה עומדת בבסיס פלסטיות סינפסה: עלייה בכח הסינפטי היא 2,3 הגברה לטווח ארוך (LTP), ואילו ירידה בכוח הסינפטי היא דיכאון לטווח ארוך 4 (בע&quot;מ). לכן הוא חשב תנועת קולטן AMPA שיהיה אחראי לביטוי פלסטיות הסינפטית, ואילו קולטני NMDA הם חשבו לשלוט האינדוקציה שלה. קביעת המנגנון המדויק שבבסיס שידור סינפטי ופלסטיות דורשת לימוד אוכלוסיות קטנות של סינפסות, באופן אידיאלי סינפסות בודדות. בעוד כמה סינפסות הן מאוד מתאימות למחקר ברמה זו, למשל, Calyx של הלד 5, לאוכלוסיות הסינפטי ביותר זה קשה מאוד בשל האופי הקטן ומפוזר של הקשרים סינפטיים. שתי טכניקות עיקריות אלקטרופיזיולוגיה פותחו כדי לבחון קשרים סינפטיים אחת: הראשון הוא גירוי מינימאלי, wherדואר סיב אחד presynaptic הוא בחזקת מגורה extracellularly. הטכניקה השנייה היא זיווג הקלטות, שבו שתי הקלטות תא כל בו זמנית מתא העצב מחובר synaptically מתבצעת. יתרון עיקרי של גירוי מינימאלי הוא שזה מהיר ופשוט יחסית לביצוע, הכולל מיקום של האלקטרודה מגרה תאית לתוך מערכת axonal בעת הקלטה בו זמנית מתא עצב postsynaptic. הדאגה העיקרית בעת השימוש בטכניקה זו היא שגירוי אמין של תא בודד יכול לעתים רחוקות להיות מובטח משפט אחרי המשפט. במהלך חמישה עשר השנים האחרונות יש לנו בשימוש שיגרתי לזווג כל תא הקלטות משני נוירונים פירמידה מחוברים synaptically 6-17. היתרון העיקרי של שיטה זו הוא כי רק אחד נוירון presynaptic מגורה באופן עקבי ואמין. זה גם מאפשר לא רק אפיון אלקטרו אלא גם מניפולציה תרופתית של נוירון presynaptic 6,18 &lt;/ Sup&gt;. עם זאת, ההסתברות של קישוריות הסינפטית בין תאי העצב היא נמוכה, מה שהופך את זוגות מחוברים קשה להשיג 19. השימוש בתרבויות פרוסה מוח organotypic עוקפת את המכשול הזה כקישוריות הסינפטית יכול להקים מחדש במבחנה ויותר מכך את טבעו של הקישוריות וכתוצאה מכך הוא דומה לזה ברקמת מוח ילידי 20. בנוסף, תרבויות organotypic להביע LTP, בע&quot;מ 7-10,12-15,21 וצורות נוספות של פלסטיות הסינפטית לטווח קצר כולל הנחיית לזווג דופק (PPF) ודיכאון (PPD) 6,22,23, המאפשרים למנגנוני פלסטיות ל יילמד בזוגות של תאי עצב. כאן אנו מתארים את המתודולוגיה מפורטת הכרוכה בהשגת הצלחת הקלטות לזווג במערכת זו במבחנה. מידע זה בקלות יכול להיות מותאם למערכות ניסיוניות אחרות, כוללים פרוסות חריפות ואזורים אחרים במוח.

<table><tbody><tr><td>Minimum Essential Medium</td><td></td><td></td><td>Stable motorized micromanipulators&amp;nbsp;</td></tr><tr><td>Penicillin-Streptomycin solution&amp;nbsp;</td><td></td><td></td><td>Shallow tissue bath</td></tr><tr><td>HEPES buffer solution</td><td></td><td></td><td>DIC camera</td></tr><tr><td>1 M Tris stock solution</td><td></td><td></td><td>Amplifier</td></tr><tr><td>Hank&amp;rsquo;s Balanced Salt Solution</td><td></td><td></td><td>Computer</td></tr><tr><td>Horse Serum&amp;nbsp;</td><td></td><td></td><td>Vibration isolation table</td></tr><tr><td>Plastic-coated miniature spatulas</td><td></td><td></td><td>Upright microscope</td></tr><tr><td>Soft paintbrush</td><td></td><td></td><td>Data acquistion and analysis software</td></tr><tr><td>Manual tissue chopper</td><td></td><td></td><td>Electrode puller</td></tr><tr><td>#2 Filter paper</td><td></td><td></td><td>Faraday cage</td></tr><tr><td>#5 Forceps</td><td></td><td></td><td></td></tr><tr><td>Membrane inserts</td><td></td><td></td><td></td></tr><tr><td>CO&lt;sub&gt;2&lt;/sub&gt; incubator</td><td></td><td></td><td></td></tr><tr><td>Dissection hood</td><td></td><td></td><td></td></tr><tr><td>Class II hood</td><td></td><td></td><td></td></tr></tbody></table>

paired whole cell recordings in organotypic hippocampal slices

Pair recordings involve simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling not only direct electrophysiological characterization of the synaptic connections between individual neurons, but also pharmacological manipulation of either the presynaptic or the postsynaptic neuron. When carried out in organotypic hippocampal slice cultures, the probability that two neurons are synaptically connected is significantly increased. This preparation readily enables identification of cell types, and the neurons maintain their morphology and properties of synaptic function similar to that in native brain tissue. A major advantage of paired whole cell recordings is the highly precise information it can provide on the properties of synaptic transmission and plasticity that are not possible with other more crude techniques utilizing extracellular axonal stimulation. Paired whole cell recordings are often perceived as too challenging to perform. While there are challenging aspects to this technique, paired recordings can be performed by anyone trained in whole cell patch clamping provided specific hardware and methodological criteria are followed. The probability of attaining synaptically connected paired recordings significantly increases with healthy organotypic slices and stable micromanipulation allowing independent attainment of pre- and postsynaptic whole cell recordings. While CA3-CA3 pyramidal cell pairs are most widely used in the organotypic slice hippocampal preparation, this technique has also been successful in CA3-CA1 pairs and can be adapted to any neurons that are synaptically connected in the same slice preparation. In this manuscript we provide the detailed methodology and requirements for establishing this technique in any laboratory equipped for electrophysiology.

קישוריות הסינפטית ניכרה על ידי גירוי תא עצב presynaptic לירות פוטנציאל פעולה על ידי העברת דופק depolarizing נוכחי (בדרך כלל 20-50 הרשות הפלסטינית ל20 אלפית שני) באמצעות אלקטרודה ההקלטה. העקבות הנוכחיות postsynaptic לאחר מכן נבדקו לנוכחות של EPSC monosynaptic עוררו ב( &lt;אלפיות 5) קצרים, וזמן התגוב?...

Watch this Scientific Journal Video about Paired Whole Cell Recordings in Organotypic Hippocampal Slices at JoVE.com

Paired Whole Cell Recordings in Organotypic Hippocampal Slices

Pair recordings are simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling precise electrophysiological and pharmacological characterization of the synapses between individual neurons. Here we describe the detailed methodology and requirements for establishing this technique in organotypic hippocampal slice cultures in any laboratory equipped for electrophysiology.

הקלטות תא כל לזווג בפרוסות בהיפוקמפוס Organotypic

Pair recordings involve simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling not only direct ...

paired-whole-cell-recordings-in-organotypic-hippocampal-slices

Research

JoVE Journal

Neuroscience

14.6K Views.  University of Auckland. Pair recordings are simultaneous whole cell patch clamp recordings from two synaptically connected neurons, enabling precise electrophysiological and pharmacological characterization of the synapses between individual neurons. Here we describe the detailed methodology and requirements for establishing this technique in organotypic hippocampal slice cultures in any laboratory equipped for electrophysiology.

Video: Paired Whole Cell Recordings in Organotypic Hippocampal Slices

Whole Cell Recording from an Organotypic Slice Preparation of Neocortex

We have been studying the expression and functional roles of voltage-gated potassium channels in pyramidal neurons from rat neocortex. Because of the lack of specific pharmacological agents for these channels, we have taken a genetic approach to manipulating channel expression. We use an organotypic culture preparation (16) in order to maintain cell morphology and the laminar pattern of cortex. We typically isolate acute neocortical slices at postnatal days 8-10 and maintain the slices in culture for 3-7 days. This allows us to study neurons at a similar age to those in our work with acute slices and minimizes the development of exuberant excitatory connections in the slice. We record from visually-identified pyramidal neurons in layers II/III or V using infrared illumination (IR-) and differential interference contrast microscopy (DIC) with whole cell patch clamp in current- or voltage-clamp. We use biolistic (Gene gun) transfection of wild type or mutant potassium channel DNA to manipulate expression of the channels to study their function. The transfected cells are easily identified by epifluorescence microscopy after co-transfection with cDNA for green fluorescent protein (GFP). We compare recordings of transfected cells to adjacent, untransfected neurons in the same layer from the same slice.

This is a protocol to prepare and maintain a neocortical slice preparation in organotypic culture for the purpose of making electrical recordings from pyramidal neurons.

הקלטה נייד שלמים מן ההכנה פורסים Organotypic של הניאוקורטקס

We have been studying the expression and functional roles of voltage-gated potassium channels in pyramidal neurons from rat neocortex.  Because of the ...

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs through activation of their ion channels and neurotransmitter receptors, and process information in part through activity-dependent release of gliotransmitters. Furthermore, astrocytes constitute the main uptake system for glutamate, contribute to potassium spatial buffering, as well as to GABA clearance. These cells therefore constantly monitor synaptic activity, and are thereby sensitive indicators for alterations in synaptically-released glutamate, GABA and extracellular potassium levels. Additionally, alterations in astroglial uptake activity or buffering capacity can have severe effects on neuronal functions, and might be overlooked when characterizing physiopathological situations or knockout mice. Dual recording of neuronal and astroglial activities is therefore an important method to study alterations in synaptic strength associated to concomitant changes in astroglial uptake and buffering capacities. Here we describe how to prepare hippocampal slices, how to identify stratum radiatum astrocytes, and how to record simultaneously neuronal and astroglial electrophysiological responses. Furthermore, we describe how to isolate pharmacologically the synaptically-evoked astroglial currents.

The preparation of acute brain slices from isolated hippocampi, as well as the simultaneous electrophysiological recordings of astrocytes and neurons in stratum radiatum during stimulation of schaffer collaterals is described. The pharmacological isolation of astroglial potassium and glutamate transporter currents is demonstrated.

קלטות אלקטרו כפולות של תגובות Astroglial ועצביות עוררו-synaptically בפרוסות ביפוקמפוס חריפות

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

Simultaneous Whole-cell Recordings from Photoreceptors and Second-order Neurons in an Amphibian Retinal Slice Preparation

One of the central tasks in retinal neuroscience is to understand the circuitry of retinal neurons and how those connections are responsible for shaping the signals transmitted to the brain. Photons are detected in the retina by rod and cone photoreceptors, which convert that energy into an electrical signal, transmitting it to other retinal neurons, where it is processed and communicated to central targets in the brain via the optic nerve. Important early insights into retinal circuitry and visual processing came from the histological studies of Cajal1,2 and, later, from electrophysiological recordings of the spiking activity of retinal ganglion cells - the output cells of the retina3,4.
A detailed understanding of visual processing in the retina requires an understanding of the signaling at each step in the pathway from photoreceptor to retinal ganglion cell. However, many retinal cell types are buried deep in the tissue and therefore relatively inaccessible for electrophysiological recording. This limitation can be overcome by working with vertical slices, in which cells residing within each of the retinal layers are clearly visible and accessible for electrophysiological recording.
Here, we describe a method for making vertical sections of retinas from larval tiger salamanders (Ambystoma tigrinum). While this preparation was originally developed for recordings with sharp microelectrodes5,6, we describe a method for dual whole-cell voltage clamp recordings from photoreceptors and second-order horizontal and bipolar cells in which we manipulate the photoreceptor's membrane potential while simultaneously recording post-synaptic responses in horizontal or bipolar cells. The photoreceptors of the tiger salamander are considerably larger than those of mammalian species, making this an ideal preparation in which to undertake this technically challenging experimental approach. These experiments are described with an eye toward probing the signaling properties of the synaptic ribbon - a specialized synaptic structure found in a only a handful of neurons, including rod and cone photoreceptors, that is well suited for maintaining a high rate of tonic neurotransmitter release7,8 - and how it contributes to the unique signaling properties of this first retinal synapse.

We describe the preparation of thin retinal slices from aquatic tiger salamanders (Ambystoma tigrinum) and explain how we use these slices to study synaptic processing in the retina by obtaining dual whole-cell voltage clamp recordings from photoreceptors and second-order horizontal and bipolar cells.

כל תא הקלטות בו זמנית מקולטני אור ועל נוירונים מסדר שני בהכנה פורסים רשתית אמפיבי

One of the central tasks in retinal neuroscience  is to understand the circuitry of retinal neurons and how those connections are  responsible for shaping ...

Whole-cell Patch-clamp Recordings from Morphologically- and Neurochemically-identified Hippocampal Interneurons

GABAergic inhibitory interneurons play a central role within neuronal circuits of the brain. Interneurons comprise a small subset of the neuronal population (10-20%), but show a high level of physiological, morphological, and neurochemical heterogeneity, reflecting their diverse functions. Therefore, investigation of interneurons provides important insights into the organization principles and function of neuronal circuits. This, however, requires an integrated physiological and neuroanatomical approach for the selection and identification of individual interneuron types. Whole-cell patch-clamp recording from acute brain slices of transgenic animals, expressing fluorescent proteins under the promoters of interneuron-specific markers, provides an efficient method to target and electrophysiologically characterize intrinsic and synaptic properties of specific interneuron types. Combined with intracellular dye labeling, this approach can be extended with post-hoc morphological and immunocytochemical analysis, enabling systematic identification of recorded neurons. These methods can be tailored to suit a broad range of scientific questions regarding functional properties of diverse types of cortical neurons.

Cortical networks are controlled by a small, but diverse set of inhibitory interneurons. Functional investigation of interneurons therefore requires targeted recording and rigorous identification. Described here is a combined approach involving whole-cell recordings from single or synaptically-coupled pairs of neurons with intracellular labeling, post-hoc morphological and immunocytochemical analysis.

הקלטות תיקון מהדק כל התא מMorphologically- וinterneurons היפוקמפוס זיהה Neurochemically

GABAergic inhibitory interneurons play a central role within neuronal circuits of the brain. Interneurons comprise a small subset of the neuronal ...

Whole-cell Patch-clamp Recordings in Brain Slices

Whole-cell patch-clamp recording is an electrophysiological technique that allows the study of the&#160;electrical properties of a substantial part of the neuron. In this configuration, the micropipette is in tight contact with the cell membrane, which prevents current leakage and thereby provides more accurate ionic current measurements than the previously used intracellular sharp electrode recording method. Classically, whole-cell recording can be performed on neurons in various types of preparations, including cell culture models, dissociated neurons, neurons in brain slices, and in intact anesthetized or awake animals. In summary, this technique has immensely contributed to the understanding of passive and active biophysical properties of excitable cells. A major advantage of this technique is that it provides information on how specific manipulations (e.g., pharmacological, experimenter-induced plasticity) may alter specific neuronal functions or channels in real-time. Additionally, significant opening of the plasma membrane allows the internal pipette solution to freely diffuse into the cytoplasm, providing means for introducing drugs, e.g., agonists or antagonists of specific intracellular proteins, and manipulating these targets without altering their functions in neighboring cells. This article will focus on whole-cell recording performed on neurons in brain slices,&#160;a preparation that has the advantage of recording neurons in relatively well preserved brain circuits, i.e., in a physiologically relevant context. In particular,&#160;when combined with appropriate pharmacology, this technique is a powerful tool allowing identification of specific neuroadaptations that occurred following any type of experiences, such as learning, exposure to drugs of abuse, and stress. In summary, whole-cell patch-clamp recordings in brain slices provide means to measure in ex vivo preparation long-lasting changes in neuronal functions that have developed in intact awake animals.

This protocol describes basic procedural steps for performing whole-cell patch-clamp recordings. This technique allows the study of&#160;the electrical behavior of neurons, and when performed in brain slices, allows the assessment of various neuronal functions from neurons that are still integrated in relatively well preserved brain circuits.

הקלטות תיקון מהדק כל תא פרוסות המוח

Whole-cell patch-clamp recording is an electrophysiological technique that allows the study of the&#160;electrical properties of a substantial part of the ...

A Model of Epileptogenesis in Rhinal Cortex-Hippocampus Organotypic Slice Cultures

Organotypic slice cultures have been widely used to model brain disorders and are considered excellent platforms for evaluating a drug&#8217;s neuroprotective and therapeutic potential. Organotypic slices are prepared from explanted tissue and represent a complex multicellular ex vivo environment. They preserve the three-dimensional cytoarchitecture and local environment of brain cells, maintain the neuronal connectivity and the neuron-glia reciprocal interaction. Hippocampal organotypic slices are considered suitable to explore the basic mechanisms of epileptogenesis, but clinical research and animal models of epilepsy have suggested that the rhinal cortex, composed of perirhinal and entorhinal cortices, play a relevant role in seizure generation.
Here, we describe the preparation of rhinal cortex-hippocampus organotypic slices. Recordings of spontaneous activity from the CA3 area under perfusion with complete growth medium, at physiological temperature and in the absence of pharmacological manipulations, showed that these slices depict evolving epileptic-like events throughout time in culture. Increased cell death, through propidium iodide uptake assay, and gliosis, assessed with fluorescence-coupled immunohistochemistry, was also observed. The experimental approach presented highlights the value of rhinal cortex-hippocampus organotypic slice cultures as a platform to study the dynamics and progression of epileptogenesis and to screen potential therapeutic targets for this brain pathology.

Here, we describe the preparation of rhinal cortex-hippocampus organotypic slices. Under a gradual and controlled deprivation of serum, these slices depict evolving epileptic-like events and can be considered an ex vivo model of epileptogenesis. This system represents an excellent tool for monitoring the dynamics of spontaneous activity, as well as for assessing the progression of neuroinflammatory features throughout the course of epileptogenesis.

מודל של אפילפטוגנזה בתרבויות פרוסות קליפת המוח-היפוקמפוס של רינאל קליפת המוח

Organotypic slice cultures have been widely used to model brain disorders and are considered excellent platforms for evaluating a drug&#8217;s ...

Preparing Acute Brain Slices from the Dorsal Pole of the Hippocampus from Adult Rodents

Whole-cell patch-clamp recordings from acute rodent brain slices are a mainstay of modern neurophysiological research, allowing precise measurement of cellular and synaptic properties. Nevertheless, there is an ever increasing need to perform correlated analyses between different experimental modes in addition to slice electrophysiology, for example: immunohistochemistry, molecular biology, in vivo imaging or electrophysiological recording; to answer evermore complex questions of brain function. However, making meaningful conclusions from these various experimental approaches is not straightforward, as even within relatively well described brain structures, a high degree of sub-regional variation of cellular function exists. Nowhere is this better exemplified than in the CA1 of the hippocampus, which has well-defined dorso-ventral properties, based on cellular and molecular properties. Nevertheless, many published studies examine protein expression patterns or behaviorally correlated in vivo activity in the dorsal extent of the hippocampus; and explain findings mechanistically with cellular electrophysiology from the ventro-medial region. This is further confounded by the fact that many acute slice electrophysiological experiments are performed in juvenile animals, when other experimental modes are performed in more mature animals. To address these issues, this method incorporates transcardial perfusion of mature (&#62;60 day old rodents) with artificial cerebrospinal fluid followed by preparation of modified coronal slices including the septal pole of the dorsal hippocampus to record from CA1 pyramidal cells. This process leads to the generation of healthy acute slices of dorsal hippocampus allowing for slice-based cellular electrophysiological interrogation matched to other measures.

The purpose of this protocol is to describe a method to produce slices of the dorsal hippocampus for electrophysiological examination. This procedure employs perfusion with chilled ACSF prior to slice preparation with a near-coronal slicing angle which allows for preservation of healthy principal neurons.

Whole-cell patch-clamp recordings from acute rodent brain slices are a mainstay of modern neurophysiological research, allowing precise measurement of ...

Acute Mouse Brain Slicing to Investigate Spontaneous Hippocampal Network Activity

Acute rodent brain slicing offers a tractable experimental approach to gain insight into the organization and function of neural circuits with single-cell resolution using electrophysiology, microscopy, and pharmacology. However, a major consideration in the design of in vitro experiments is the extent to which different slice preparations recapitulate naturalistic patterns of neural activity as observed in vivo. In the intact brain, the hippocampal network generates highly synchronized population activity reflective of the behavioral state of the animal, as exemplified by the sharp-wave ripple complexes (SWRs) that occur during waking consummatory states or non-REM sleep. SWRs and other forms of network activity can emerge spontaneously in isolated hippocampal slices under appropriate conditions. In order to apply the powerful brain slice toolkit to the investigation of hippocampal network activity, it is necessary to utilize an approach that optimizes tissue health and the preservation of functional connectivity within the hippocampal network. Mice are transcardially perfused with cold sucrose-based artificial cerebrospinal fluid. Horizontal slices containing the hippocampus are cut at a thickness of 450 &#956;m to preserve synaptic connectivity. Slices recover in an interface-style chamber and are transferred to a submerged chamber for recordings. The recording chamber is designed for dual surface superfusion of artificial cerebrospinal fluid at a high flow rate to improve oxygenation of the slice. This protocol yields healthy tissue suitable for the investigation of complex and spontaneous network activity in vitro.

This protocol describes the preparation of horizontal hippocampal-entorhinal cortex (HEC) slices from mice exhibiting spontaneous sharp-wave ripple activity. Slices are incubated in a simplified interface holding chamber and recordings are performed under submerged conditions with fast-flowing artificial cerebrospinal fluid to promote tissue oxygenation and the spontaneous emergence of network-level activity.

חריף המוח עכבר חותכת לחקור פעילות רשת היפוקמפוס ספונטנית

Acute rodent brain slicing offers a tractable experimental approach to gain insight into the organization and function of neural circuits with single-cell ...

Preparation of Acute Slices from Dorsal Hippocampus for Whole-Cell Recording and Neuronal Reconstruction in the Dentate Gyrus of Adult Mice

Although the general architecture of the hippocampus is similar along its longitudinal axis, recent studies have revealed prominent differences in molecular, anatomical and functional criteria suggesting a division into different sub-circuits along its rostro-caudal extent. Owing to differential connectivity and function the most fundamental distinction is made between the dorsal and the ventral hippocampus, which are preferentially involved in spatial and emotional processing, respectively. Accordingly, in vivo work regarding spatial memory formation has focused on the dorsal hippocampus.
In contrast, electro-physiological in vitro recordings have been preferentially performed on intermediate-ventral hippocampus, largely motivated by factors like slice viability and circuit integrity. To allow for direct correlation of in vivo data on spatial processing with in vitro data we have adapted previous sectioning methods to obtain highly viable transverse brain slices from the dorsal-intermediate hippocampus for long-term recordings of principal cells and interneurons in the dentate gyrus. As spatial behavior is routinely analyzed in adult mice, we have combined this transversal slicing procedure with the use of protective solutions to enhance viability of brain tissue from mature animals. We use this approach for mice of about 3 months of age. The method offers a good alternative to the coronal preparation which is frequently used for in vitro studies on dorsal hippocampus. We compare these two preparations in terms of quality of recordings and preservation of morphological features of recorded neurons.

We present a protocol to prepare acute-slices from the dorsal-intermediate hippocampus of mice. We compare this transversal preparation with coronal slicing in terms of quality of recordings and preservation of morphological features of recorded neurons.

הכנת פרוסות חריפות מהיפוקמפוס הגבי להקלטה של תא שלם ושחזור עצבי בGyrus דנטאטה של עכברים בוגרים

Although the general architecture of the hippocampus is similar along its longitudinal axis, recent studies have revealed prominent differences in ...

Photostimulation and Whole-Cell Patch Clamp Recordings of Neurons in Mouse Hippocampal Slices

Source: Richevaux, L., et al. <a href="https://app.jove.com/v/59534">In Vivo Intracerebral Stereotaxic Injections for Optogenetic Stimulation of Long-Range Inputs in Mouse Brain Slices.</a> J. Vis. Exp. (2019)
The video demonstrates a protocol to study synaptic interactions and neuronal responses in a transfected mouse hippocampal brain slice. It involves illuminating long-range sensory neurons to trigger action potentials and recording the resulting excitatory post-synaptic potentials in the recorded neuron using the whole-cell patch-clamp technique.

פוטו-טימולציה והקלטות מהדק טלאי של תא שלם של תאי עצב בפרוסות היפוקמפוס עכבר

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.
1. Solutions for acute slice ...