This study was supported by NeuralStem, Inc., and Johns Hopkins Project RESTORE.

הערה: נהלי כל החיה אושרו על ידי ועדת הטיפול בבעלי חיים והשימוש בג&#39;ונס הופקינס. מכונות הרדמה דורשות בדיקה שנתית וכיול כנדרש. 1. הרדמה ומיצוב <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> הרדמה. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לבצע את כל הניתוחים בהרדמה עם isofluorane 2-3%. אשר רמה המתאימה של הרדמה על ידי צביטת הבוהן וקצב נשימה. בדוק שהחולדה לא נרתעה בתגובה לקמצוץ הבוהן. הערה: נרתע מצביע הרדמה שתהיינה מאוחר מדי ועשוי לדרוש הרדמה כבר לפני תחילתו או בריכוז גבוה יותר isofluorane. קצב נשימה פחות מנשימה 1 כל 2 שניות היא איטיות מדי מצביע ההרדמה הוא גבוה מדי ועשוי לדרוש הבהרה של ריכוז isofluorane. </li><li style=";text-align:right;direction:rtl"> אבטח את בעלי החיים כנדרשים כדי למנוע תנועת הראש במהלך ההליך. מניחים ירידה של לידוקאין על העין כירורגית. השתמש דמעות מלאכותיות כל 10 דקות כדי למנוע יובש ואילו בהרדמה. תןהזרקה של עצירות של 0.01 מ&quot;ג / קילוגרם כל שעות 6-8 SC מראש אופרטיבי ולאחר מכן לפי צורך. </li></ol></li><li style=";text-align:right;direction:rtl"> מיקום. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> מניחים חולדות למסגרת stereotaxic ולהתחמם עם כרית חימום. להרטיב את הפרווה הקרקפת עם טיפול נטילת אלכוהול, כדי למנוע חשיפה בעיניים. השתמש בכלים סטריליים וטכניקה סטרילית כדי למזער את הסיכון של זיהומים לאחר ניתוח. </li></ol></li></ol> 2. עין בקרה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> מניחים תפר 4-0 בלחמית הרוחב ולקשור אותו עם מספיק תפר להתיר מתיחה עדינה. </li></ol> 3. Dissection <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> Dissection הראשוני. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לעשות חתך אינץ &#39;~ 1 בעור שמעל רכס מסלולית באמצעות אזמל גודל 10 כפי שמוצג באיור 1. לחזור בו העור ואת fascia הבסיסי ולנתח בזהירות את fascia. כדי למנוע דימום יתר לניתוח, להימנע מחיתוך כלי דם בזמן לנתח את fascia. השתמש אסטרטגי plטיפים כותנה aced לספק עצירת דימום. </li></ol></li><li style=";text-align:right;direction:rtl"> Dissection עמוק יותר. הערה: עם מתיחה עדינה על לחמית משיכת העין כלפי מטה והחוצה מהשקע, השרירים מסלולית מעולים יבואו לעין. כדי לחשוף את עצב הראייה, שרירים זה יש לחתוך ולהסיר את שומן רטרו מסלולית. השומן יכול להיות מושלך ולא צריך להיות מוחלף לאחר הזרקה. מנקודה זו, fascia עצב הראייה צריך להיות גלוי כחבילה של עצב הראייה עצמו, יחד עם כלי דם עטופים בדורה (איור 1). <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לעשות חתך קטן בדורה באמצעות האזמל או מחט משופעת 31 מד לעשות פירסינג פחות טראומטי. </li></ol></li></ol> 4. פיפטה מזרק <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> משוך micropipette זכוכית לקוטר של 50-100 מיקרומטר. כדי לספק יציבות, הר micropipette על micromanipulator ולצרף מזרק המילטון מחובר למשאבת עירוי. </li><li style=";text-align:right;direction:rtl"> משוך את חרוזים (או תאי גזע)מחדש ב0.5-1.0 μl נפח לmicropipette מדרדר יחד עם 0.5 μl נפח של פתרון כחול מתיל לפני ואחרי. הערה: קצב עירוי מוגדר 0.5-2 μl לדקה מונעת טראומה לעצב הראייה. </li></ol> 5. הזרקה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> מנמיכים את קצה micropipette על עצב הראייה מעל הדורה חתוכות. הערה: תנועה זעירה, אך נמרצת של הקצה של הכוס לתוצאות עצב ראייה בנזק לפחות. כמשאבת העירוי מתחילה, הצבע הכחול מתיל צריך להדגיש את האזור של עצב הראייה המוזרק. הצבע צריך להישאר מקומי בעצב הראייה ללא דולף החוצה בחלל תת-העכבישים. </li><li style=";text-align:right;direction:rtl"> בצע את הלהקה השנייה של מתיל כחול כדי לקבוע מתי הזרקת תאי גזע היא מלאה ולכבות את משאבת העירוי כמנה השנייה של מתיל כחול מוזרקת. שמור micropipette עדיין בתוך עצב הראייה למשך 2 דקות לכל μl 1 מוזרק לתוך נפח למנוע גבוהפליטת לחץ על נסיגה של micropipette. הערה: גישה חלופית היא לחבר את קצה פיפטה למזרק מלא אוויר שניתן לטפל באופן ידני. כך או כך, למנוע בכוח מופרז כדי למזער את הנזק לעצבי ראייה. אנו ממליצים הזרקה לא יותר מ -2 μl של נפח בכל עצב ראייה. </li></ol> 6. מעקב <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> מיידי. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> כאשר ההזרקה היא מלאה, להסיר את micropipette יחד עם כל טיפים כותנה משמשים כדי לספק עצירת דימום. לתפור את העור עם 3-0 משי ולהסיר את התפר של הלחמית. </li><li style=";text-align:right;direction:rtl"> שמור חולדות חמות על כרית חימום עד שהם יוצאים מהרדמה. אל תשאיר חיה ללא השגחה עד שהוא חזר להכרה מספיק כדי לשמור על כיבה sternal. אל תחזרו לבעלי חיים שעברו ניתוח לחברה של בעלי חיים אחרים, עד שהתאושש באופן מלא. </li><li style=";text-align:right;direction:rtl"> אם בעל החיים הוא מציגים סימנים של כאב, כולל עייפות אטקסיה, נשימה מוגזמת, או מאומצתg, לנהל שיכוך כאבים מתאימים עם עצירות תוך שרירית (0.05 מ&quot;ג / קילוגרם) עד שלוש פעמים ביום. </li></ol></li><li style=";text-align:right;direction:rtl"> טווח ארוך. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לשעה 24-48 הבאה, לבחון את החולדה לסיבוכים של ניתוח כוללים נפיחות או הפרשה מהפצע או סימנים אחרים של כאב כמו בדיבור, מראה כפוף, לא טיפוח, או לא אוכל. לשקול התייעצות עם וטרינר או המתת חסד אתי כנדרש. </li></ol></li></ol>

<ol>
	<li>Dahlmann-Noor, A., Vijay, S., Jayaram, H., Limb, A., Khaw, P. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+approaches+and+future+prospects+for+stem+cell+rescue+and+regeneration+of+the+retina+and+optic+nerve.">Current approaches and future prospects for stem cell rescue and regeneration of the retina and optic nerve.</a> Canadian journal of ophthalmology Journal canadien d'ophtalmologie. 45 (4), 333-341 (2010).</li><li>Quigley, H. A., Iglesia, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stem+cells+to+replace+the+optic+nerve.">Stem cells to replace the optic nerve.</a> Eye. 18 (11), 1085-1088 (2004).</li><li>Ghaffarieh, A., Levin, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optic+nerve+disease+and+axon+pathophysiology.">Optic nerve disease and axon pathophysiology.</a> International review of neurobiology. 105, 1-17 (2012).</li><li>Fukui, Y., Hayasaka, S., Bedi, K. S., Ozaki, H. S., Takeuchi, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+study+of+the+development+of+the+optic+nerve+in+rats+reared+in+the+dark+during+early+postnatal+life.">Quantitative study of the development of the optic nerve in rats reared in the dark during early postnatal life.</a> Journal of anatomy. 174, 37-47 (1991).</li><li>Celestre, P. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimally+invasive+approaches+to+the+cervical+spine.">Minimally invasive approaches to the cervical spine.</a> The Orthopedic clinics of North America. 43 (1), 137-147 (2012).</li><li>Hallas, B. H., Wells, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Novel+Technique+for+Multiple+Injections+into+the+Mammalian+Optic+Nerve.">A Novel Technique for Multiple Injections into the Mammalian Optic Nerve.</a> Kopf Carrier. 54, (2001).</li><li>Harvey, A. R., Hellstrom, M., Rodger, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+therapy+and+transplantation+in+the+retinofugal+pathway.">Gene therapy and transplantation in the retinofugal pathway.</a> Progress in brain research. 175, 151-161 (2009).</li><li>Adachi-Usami, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optic+neuritis--from+diagnosis+to+optic+nerve+transplantation.">Optic neuritis--from diagnosis to optic nerve transplantation.</a> Nippon Ganka Gakkai zasshi. 104 (12), 841-857 (2000).</li><li>Slater, B. J., Vilson, F. L., Guo, Y., Weinreich, D., Hwang, S., Bernstein, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optic+nerve+inflammation+and+demyelination+in+a+rodent+model+of+nonarteritic+anterior+ischemic+optic+neuropathy.">Optic nerve inflammation and demyelination in a rodent model of nonarteritic anterior ischemic optic neuropathy.</a> Investigative ophthalmology &amp; visual science. 54 (13), 7952-7961 (2013).</li><li>Zarbin, M. A., Arlow, T., Ritch, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regenerative+nanomedicine+for+vision+restoration.">Regenerative nanomedicine for vision restoration.</a> Mayo Clinic proceedings. 88 (12), 1480-1490 (2013).</li></ol>

The authors have no meaningful disclosures to report.

Direct injection into the optic nerve of stem cells or other products intended to facilitate regeneration provides a convenient model compared to other means of injections into the CNS. This technique takes less time, requires less total anesthesia, avoids drilling or removing skull or bone tissue, reduces complications rates and allows for more rapid recovery following surgery. 
The most critical steps in this protocol include: 1. Adequate hemostasis in the surgical field to allow clear visua...

עצב הראייה מספק מיקום אידיאלי למערכת עצבים מרכזית (CNS) מחקר משובי לרבות תנאי ophthalmologic כגון דלקת בעצב ראיה, גלאוקומה וטראומה. זריקות של מגוון רחב של תאי גזע שגם הוכיחו את יעילות או הראו הבטחה בהחלפת המיאלין האבוד, הגדלת ספירת axonal ו / או מניעת מחלות ניווניות. 1,2 עצב הראייה האנושי מכיל כ -1.2 מיליון האקסונים מקבילים נסיעה מהרשתית אל התצלובת בקוטר של כ 3.0-3.5 מ&quot;מ. 3 מודל מחלות בבני אדם במעבדה, החולדה כבר בשימוש לעתים קרובות. עצב הראייה החולדה הבוגרת מכיל כ -100,000 האקסונים בקוטר של כ 0.5 מ&quot;מ. 4 אחת המגבלות העיקריות במערכת העצבים המרכזית מחקר משובי הוא גישה ישירה ללא עצמות. סיבוכים וסיכונים הכירורגיים לבעלי החיים הם גבוהים יותר כאשר הגולגולת או החוליות יוסרו. בדומה ליתרונות שלגישות זעירים פולשנית בעמוד השדרה, 5 זריקות עצב ראייה ישירות מבלי לפתוח את סיבוכי הצעה מופחתות גולגולת והתאוששות מהירה יותר. טכניקה זו נעשתה שימוש במחקרים קודמים. 6 בכתב היד הזה ווידאו נלווה, אנחנו מדגימים הליך מינימאלי פולשנית להזריק תאי גזע לעצב הראייה החולדה.

<table border="0" cellpadding="0" cellspacing="0" width="851">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:151px;">Company</td>
			<td style="width:151px;">Catalog Number</td>
			<td style="width:335px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Lewis rat</td>
			<td style="width:151px;">Charles River</td>
			<td align="right" style="width:151px;">4</td>
			<td>Any rat strain will work.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Anesthesia machine</td>
			<td style="width:151px;">Surgivet</td>
			<td style="width:151px;">CDS9000</td>
			<td>CDS 9000 Small Animal Anesthesia Machine - Pole Mount</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Infusion pump</td>
			<td style="width:151px;">Stoelting</td>
			<td align="right" style="width:151px;">53129</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Dissection microscope</td>
			<td style="width:151px;">National Optical</td>
			<td style="width:151px;">409-411-1105</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Fiber-optic light source</td>
			<td style="width:151px;">Fisher Scientific</td>
			<td style="width:151px;">12-562-21</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Dissection and Stereotaxic Instrument</td>
			<td style="width:151px;">Stoelting</td>
			<td align="right" style="width:151px;">51400</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Pipette Puller</td>
			<td style="width:151px;">Kopf</td>
			<td align="right" style="width:151px;">750</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Pipettes</td>
			<td style="width:151px;">World Precision Instruments</td>
			<td style="width:151px;">18150-6</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Disposable scalpel blades</td>
			<td style="width:151px;">Harvard Apparatus</td>
			<td style="width:151px;">810-15-021</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Iridectomy scissors</td>
			<td style="width:151px;">Electron Microscopy Sciences</td>
			<td style="width:151px;">Uniband LA-4XF</td>
			<td></td>
		</tr>
	</tbody>
</table>

minimally-invasive technique for injection into rat optic nerve

The rat optic nerve is a useful model for stem cell regeneration research. Direct injection into the rat optic nerve allows delivery into the central nervous system in a minimally-invasive surgery without bone removal. This technique describes an approach to visualization and direct injection of the optic nerve following minor fascial dissection from the orbital ridge, using a conjunctival traction suture to gently pull the eye down and out. Representative examples of an injected optic nerve show successful injection of dyed beads.

בסיומו של הניסוי, חולדות הוקרבו וperfused עם paraformaldehyde 4%. עצבי הראייה נותחו בזהירות ורכובים על חתך cryostat. איור 2 מראה דוגמא של עצב עכברוש כל אופטי בצריכת חשמל נמוכה שבצבע הכחול אוונס שהוזרק כדי להמחיש את האתר. החץ מזהה את המיקום המדויק של ההזרקה. נתיחה זה נעשתה תוך כ...

Watch this Scientific Journal Video about Minimally-invasive Technique for Injection into Rat Optic Nerve at JoVE.com

Minimally-invasive Technique for Injection into Rat Optic Nerve

Direct injection into the rat optic nerve is useful for regenerative research. We demonstrate a minimally-invasive technique for direct injection into a rat optic nerve that does not involve opening the skull. Using this method, surgical complications are minimized and recovery is more rapid.

טכניקה זעיר-פולשני להזרקה לתוך עצב העכברוש אופטי

The rat optic nerve is a useful model for stem cell regeneration research. Direct injection into the rat optic nerve allows delivery into the central ...

minimally-invasive-technique-for-injection-into-rat-optic-nerve

Research

JoVE Journal

Neuroscience

9.5K Views.  Johns Hopkins University. Direct injection into the rat optic nerve is useful for regenerative research. We demonstrate a minimally-invasive technique for direct injection into a rat optic nerve that does not involve opening the skull. Using this method, surgical complications are minimized and recovery is more rapid. 

Video: Minimally-invasive Technique for Injection into Rat Optic Nerve

Single-unit In vivo Recordings from the Optic Chiasm of Rat

Information about the visual world is transmitted to the brain in sequences of action potentials in retinal ganglion cell axons that make up the optic nerve. In vivo recordings of ganglion cell spike trains in several animal models have revealed much of what is known about how the early visual system processes and encodes visual information. However, such recordings have been rare in one of the most common animal models, the rat, possibly owing to difficulty in detecting spikes fired by small diameter axons. The many retinal disease models involving rats motivate a need for characterizing the functional properties of ganglion cells without disturbing the eye, as with intraocular or in vitro recordings. Here, we demonstrate a method for recording ganglion cell spike trains from the optic chiasm of the anesthetized rat. We first show how to fabricate tungsten-in-glass electrodes that can pick up electrical activity from single ganglion cell axons in rat. The electrodes outperform all commercial ones that we have tried. We then illustrate our custom-designed stereotaxic system for in vivo visual neurophysiology experiments and our procedures for animal preparation and reliable and stable electrode placement in the optic chiasm.

Single-unit In vivo Recordings from the Optic Chiasm of Rat

Retinal ganglion cells transmit visual information from the eye to the brain with sequences of action potentials. Here, we demonstrate how to record the action potentials of single ganglion cells in vivo from anesthetized rats.

יחידה יחידה ב-vivo הקלטות של התצלובת הראייה של עכברוש

Information about the visual world is transmitted to the brain in sequences of action potentials in retinal ganglion cell axons that make up the optic ...

Whole Mount Preparation of the Adult Drosophila Ventral Nerve Cord for Giant Fiber Dye Injection

To analyze the axonal and dendritic morphology of neurons, it is essential to obtain accurate labeling of neuronal structures. Preparing well labeled samples with little to no tissue damage enables us to analyze cell morphology and to compare individual samples to each other, hence allowing the identification of mutant anomalies.
In the demonstrated dissection method the nervous system remains mostly inside the adult fly. Through a dorsal incision, the abdomen and thorax are opened and most of the internal organs are removed. Only the dorsal side of the ventral nerve cord (VNC) and the cervical connective 
(CvC) containing the big axons of the giant fibers (GFs)1 are exposed, while the brain containing the GF cell body and dendrites remains2 in the 
intact head. In this preparation most nerves of the VNC should remain attached to their muscles. 
Following the dissection, the intracellular filling of the giant fiber (GF) with a fluorescent dye is demonstrated. In the CvC the GF axons are located at the dorsal surface and thus can be easily visualized under a microscope with differential interference contrast (DIC) optics. This allows the injection of the GF axons with dye at this site to label the entire GF including the axons and their terminals in the VNC. This method results in reliable and strong staining of the GFs allowing the neurons to be imaged immediately after filling with an epifluorescent microscope. Alternatively, the fluorescent signal can be enhanced using standard immunohistochemistry procedures3 suitable for high resolution confocal microscopy.

Whole Mount Preparation of the Adult Drosophila Ventral Nerve Cord for Giant Fiber Dye Injection

An in vivo dissection of the adult Drosophila ventral nerve cord (VNC) is demonstrated. This particular dissection method causes little damage to the VNC allowing the subsequent labeling of the giant fiber neurons with fluorescent dye for high resolution imaging.

הר הכנה שלמים של המבוגר תסיסנית עצבי חוט הגחון עבור הזרקת סיבים ענק דיי

To analyze the axonal and dendritic morphology of neurons, it is essential to obtain accurate labeling of neuronal structures. Preparing well labeled ...

In vivo Imaging of Optic Nerve Fiber Integrity by Contrast-Enhanced MRI in Mice

The rodent visual system encompasses retinal ganglion cells and their axons that form the optic nerve to enter thalamic and midbrain centers, and postsynaptic projections to the visual cortex. Based on its distinct anatomical structure and convenient accessibility, it has become the favored structure for studies on neuronal survival, axonal regeneration, and synaptic plasticity. Recent advancements in MR imaging have enabled the in vivo visualization of the retino-tectal part of this projection using manganese mediated contrast enhancement (MEMRI). Here, we present a MEMRI protocol for illustration of the visual projection in mice, by which resolutions of (200 &#181;m)3 can be achieved using common 3 Tesla scanners. We demonstrate how intravitreal injection of a single dosage of 15 nmol MnCl2 leads to a saturated enhancement of the intact projection within 24 hr. With exception of the retina, changes in signal intensity are independent of coincided visual stimulation or physiological aging. We further apply this technique to longitudinally monitor axonal degeneration in response to acute optic nerve injury, a paradigm by which Mn2+ transport completely arrests at the lesion site. Conversely, active Mn2+ transport is quantitatively proportionate to the viability, number, and electrical activity of axon fibers. For such an analysis, we exemplify Mn2+ transport kinetics along the visual path in a transgenic mouse model (NF-&#954;B p50KO) displaying spontaneous atrophy of sensory, including visual, projections. In these mice, MEMRI indicates reduced but not delayed Mn2+ transport as compared to wild type mice, thus revealing signs of structural and/or functional impairments by NF-&#954;B mutations.
In summary, MEMRI conveniently bridges in vivo assays and post mortem histology for the characterization of nerve fiber integrity and activity. It is highly useful for longitudinal studies on axonal degeneration and regeneration, and investigations of mutant mice for genuine or inducible phenotypes.

In vivo Imaging of Optic Nerve Fiber Integrity by Contrast-Enhanced MRI in Mice

This video illustrates a method, using a clinical 3 T scanner, for contrast-enhanced MR imaging of the na&#239;ve mouse visual projection and for repetitive and longitudinal in vivo studies of optic nerve degeneration associated with acute optic nerve crush injury and chronic optic nerve degeneration in knock-out mice (p50KO).

בהדמית vivo של יושרה סיבי עצב הראייה על ידי contrast-enhanced MRI בעכברים

The rodent visual system encompasses retinal ganglion cells and their axons that form the optic nerve to enter thalamic and midbrain centers, and ...

A Method of Nodose Ganglia Injection in Sprague-Dawley Rat

Afferent signaling via the vagus nerve transmits important general visceral information to the central nervous system from many diverse receptors located in the organs of the abdomen and thorax. The vagus nerve communicates information from stimuli such as heart rate, blood pressure, bronchopulmonary irritation, and gastrointestinal distension to the nucleus of solitary tract of the medulla. The cell bodies of the vagus nerve are located in the nodose and petrosal ganglia, of which the majority are located in the former. The nodose ganglia contain a wealth of receptors for amino acids, monoamines, neuropeptides, and other neurochemicals that can modify afferent vagus nerve activity. Modifying vagal afferents through systemic peripheral drug treatments targeted at the receptors on nodose ganglia has the potential of treating diseases such as sleep apnea, gastroesophageal reflux disease, or chronic cough. The protocol here describes a method of injection neurochemicals directly into the nodose ganglion. Injecting neurochemicals directly into the nodose ganglia allows study of effects solely on cell bodies that modulate afferent nerve activity, and prevents the complication of involving the central nervous system as seen in systemic neurochemical treatment. Using readily available and inexpensive equipment, intranodose ganglia injections are easily done in anesthetized Sprague-Dawley rats.

Afferent vagal signaling transmits important information to central nervous system from receptors located in organs of the abdomen and thorax. The nodose ganglia of vagus nerves contain many types of receptors that modulate vagal activity. This protocol describes a method of local injections of neurochemicals into the nodose ganglia.

שיטה של ​​הזרקת Nodose הגרעינים בSprague-Dawley עכברוש

Afferent signaling via the vagus nerve transmits important general visceral information to the central nervous system from many diverse receptors located ...

Intramuscular Injections Along the Motor End Plates: A Minimally Invasive Approach to Shuttle Tracers Directly into Motor Neurons

Diseases affecting the integrity of spinal cord motor neurons are amongst the most debilitating neurological conditions. Over the last decades, the development of several animal models of these neuromuscular disorders has provided the scientific community with different therapeutic scenarios aimed at delaying or reversing the progression of these conditions. By taking advantage of the retrograde machinery of neurons, one of these approaches has been to target skeletal muscles in order to shuttle therapeutic genes into corresponding spinal cord motor neurons. Although once promising, the success of such gene delivery approach has been hampered by the sub-optimal number of transduced motor neurons it has so far shown to yield. Motor end plates (MEPs) are highly specialized regions on the skeletal musculature that are in direct synaptic contact to the spinal cord &#945; motor neurons. In this regard, it is important to note that, so far, the efforts to retrogradely transfer genes into motor neurons were made without reference to the location of the MEP region in the targeted muscles. Here, we describe a simple protocol 1) to reveal the exact location of the MEPs on the surface of skeletal muscles and 2) to use this information to guide the intramuscular delivery and subsequent optimal retrograde transport of retrograde tracers into motor neurons. We hope to utilize the results from these tracing experiments in further studies into investigating retrograde transport of therapeutic genes to spinal cord motor neurons through the targeting of MEPs.

The efficacy of intramuscular uptake and retrograde transport of molecules to corresponding motor neurons depends on the location of the injection sites with respect to the motor end plates (MEPs). Here, we describe how to locate MEPs on skeletal muscles to optimise retrograde transport of tracers into motor neurons.

זריקות לשריר לאורך צלחות מוטוריות סיום: גישה פולשנית להסעות קליעים נותבים ישירות לתוך מנוע נוירונים

Diseases affecting the integrity of spinal cord motor neurons are amongst the most debilitating neurological conditions. Over the last decades, the ...

Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to monitor the disease course in the follow-up period. Changes in the VEP parameters closely correlate with pathological damage in the optic nerve. This protocol provides a detailed description about the rodent model of optic nerve microinjection, in which a partial demyelination lesion is produced in the optic nerve. VEP recording techniques are also discussed. Using skull implanted electrodes, we are able to acquire reproducible intra-session and between-session VEP traces. VEPs can be recorded on individual animals over a period of time to assess the functional changes in the optic nerve longitudinally. The optic nerve demyelination model, in conjunction with the VEP recording protocol, provides a tool to investigate the disease processes associated with demyelination and remyelination, and can potentially be employed to evaluate the effects of new remyelinating drugs or neuroprotective therapies.

Focal demyelination is induced in the optic nerve using lysolecithin microinjection. Visual evoked potentials are recorded via skull electrodes implanted over the visual cortex to examine the signal conduction along the visual pathway in vivo. This protocol details the surgical procedures underlying electrode implantation and optic nerve microinjection.

הקלטת פוטנציאל עוררה חזותית במודל עכברוש של demyelination עצב אופטי הניסויי

The visual evoked potential (VEP) recording is widely used in clinical practice to assess the severity of optic neuritis in its acute phase, and to ...

Loading a Calcium Dye into Frog Nerve Endings Through the Nerve Stump: Calcium Transient Registration in the Frog Neuromuscular Junction

One of the most feasible methods of measuring presynaptic calcium levels in presynaptic nerve terminals is optical recording. It is based on using calcium-sensitive fluorescent dyes that change their emission intensity or wavelength depending on the concentration of free calcium in the cell. There are several methods used to stain cells with calcium dyes. Most common are the processes of loading the dyes through a micropipette or pre-incubating with the acetoxymethyl ester forms of the dyes. However, these methods are not quite applicable to neuromuscular junctions (NMJs) due to methodological issues that arise. In this article, we present a method for loading a calcium-sensitive dye through the frog nerve stump of the frog nerve into the nerve endings. Since entry of external calcium into nerve terminals and the subsequent binding to the calcium dye occur within the millisecond time-scale, it is necessary to use a fast imaging system to record these interactions. Here, we describe a protocol for recording the calcium transient with a fast CCD camera.

Here, we describe a method for loading a calcium-sensitive dye through the frog nerve stump into the nerve endings. We also present a protocol for the recording and analysis of fast calcium transients in the peripheral nerve endings.

טעינת צבע סידן לתוך צפרדע עצב צפרדע דרך גדם עצב: סידן חולף הרשמה צפרדע Neuromuscular צפרדע

One of the most feasible methods of measuring presynaptic calcium levels in presynaptic nerve terminals is optical recording. It is based on using ...

Endoscopic Endonasal Trans-sphenoidal Approach: Minimally Invasive Surgery for Pituitary Adenomas

Endoscopic endonasal trans-sphenoidal surgery has become the gold standard for the surgical treatment of pituitary adenomas and many other pituitary lesions. Refinements in surgical techniques, technological advancements, and incorporation of neuronavigation have rendered this surgery minimally invasive. The complication rates of this surgery are very low while excellent results are consistently obtained through this approach.
This paper focuses on the step-by-step surgical approach to pituitary adenomas, which is based on personal experience, and details the results obtained with this minimally invasive surgery.

The aim of this paper is to describe the different steps of the endoscopic endonasal approach to the sella turcica.

בגישה אנדוסקופית דרך האף טרנס-sphenoidal: כירורגיה זעיר פולשנית עבור אדנומות בבלוטת יתרת המוח

Endoscopic endonasal trans-sphenoidal surgery has become the gold standard for the surgical treatment of pituitary adenomas and many other pituitary ...

Minimally Invasive Endoscopic Intracerebral Hemorrhage Evacuation

Intracerebral hemorrhage (ICH) is a subtype of stroke with high mortality and poor functional outcomes, largely because there are no evidence-based treatment options for this devastating disease process. In the past decade, a number of minimally invasive surgeries have emerged to address this issue, one of which is endoscopic evacuation. Stereotactic ICH Underwater Blood Aspiration (SCUBA) is a novel endoscopic evacuation technique performed in a fluid-filled cavity using an aspiration system to provide an additional degree of freedom during the procedure. The SCUBA procedure utilizes a suction device, endoscope, and sheath and is divided into two phases. The first phase involves maximal aspiration and minimal irrigation to decrease clot burden. The second phase involves increasing irrigation for visibility, decreasing aspiration strength for targeted aspiration without disturbing the cavity wall, and cauterizing any bleeding vessels. Using the endoscope and aspiration wand, this technique aims to maximize hematoma evacuation while minimizing collateral damage to the surrounding brain. Advantages of the SCUBA technique include the use of a low-profile endoscopic sheath minimizing brain disruption and improved visualization with a fluid-filled cavity rather than an air-filled one.

This paper details the surgical protocol for minimally invasive endoscopic intracerebral hemorrhage evacuation using the SCUBA technique.

פינוי דימום אנדוסקופי זעיר פולשני

Intracerebral hemorrhage (ICH) is a subtype of stroke with high mortality and poor functional outcomes, largely because there are no evidence-based ...

A Minimally Invasive Lesion Technique for Muscles Intrinsic to the Odontophore of Aplysia californica

Aplysia californica is a model system for studying the neural control of learning and behavior. This animal has a semi-open circulatory system, making it possible to access many of its internal structures without causing any significant damage. Many manipulations can be easily performed both in vivo and in vitro, making it a highly tractable model for the analysis of behavior and neural circuitry. To better understand the functions of muscles within the feeding grasper, we have developed a technique for lesioning them without opening the main body cavity of the animal or damaging the outer layers of the feeding organ (i.e., the buccal mass). In this technique, the grasper is partially everted, allowing direct access to the musculature. This procedure allows animals to recover quickly and reliably. This has made it possible to lesion the I7 muscles and sub-radular fibers, allowing us to show that both muscles significantly contribute to the opening in vivo.

A Minimally Invasive Lesion Technique for Muscles Intrinsic to the Odontophore of Aplysia californica

Here we present a protocol for minimally invasive surgical lesioning of muscles intrinsic to the feeding apparatus of the marine mollusk Aplysia californica to understand the roles of these muscles during feeding behavior.

שיטה מינימלית פולשנית לשרירי השרירים הפנימיים של השיניים של Aplysia קליפורניה

Aplysia californica is a model system for studying the neural control of learning and behavior. This animal has a semi-open circulatory system, making it ...