Name: full-circle cauterization of limbal vascular plexus for surgically induced glaucoma in rodents
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Description: Watch this Scientific Journal Video about Full-Circle Cauterization of Limbal Vascular Plexus for Surgically Induced Glaucoma in Rodents at JoVE.com

हम अपने प्रयोगशाला तकनीशियन जोस को स्वीकार करते हैं; निल्सन डॉस सैंटोस, डायने मंडेरिनो टोरेस, जोस फ्रांसिस्को टिबुर्सियो, गिल्डो ब्रिटो डी सूजा, और लुसियानो कैवलकेंट फरेरा। इस शोध को FAPERJ, CNPQ और CAPES द्वारा वित्त पोषित किया गया था।

सभी प्रक्रियाओं को एसोसिएशन फॉर रिसर्च इन विजन एंड ओप्थाल्मोलॉजी (एआरवीओ) से नेत्र और दृश्य अनुसंधान में जानवरों के उपयोग के लिए बयान के अनुपालन में किया गया था और स्वास्थ्य विज्ञान केंद्र, रियो डी जन?...

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लिम्बल प्लेक्सस कॉटराइजेशन (एलपीसी) एक नया पोस्ट-ट्रैब्युलर मॉडल है जिसका लाभ यह है कि यह आसानी से सुलभ संवहनी संरचनाओं को लक्षित करता है जिन्हें नेत्रश्लेष्मला या टेनोन विच्छेदन17,28<sup class="xr...

चिकित्सा साहित्य ग्लूकोमा को ऑप्टिक न्यूरोपैथियों के एक विषम समूह के रूप में समझता है जो रेटिना गैंग्लियन कोशिकाओं (आरजीसी), डेंड्राइट, सोमा और अक्षतंतु के प्रगतिशील अध: पतन की विशेषता है, जिसके परिणामस्वरूप ऑप्टिक डिस्क की संरचनात्मक कपिंग (उत्खनन) और ऑप्टिक तंत्रिका की कार्यात्मक गिरावट होती है, जिससे आंखसे मस्तिष्क तक दृश्य जानकारी के संचरण को बाधित करके अनियंत्रित मामलों में एमॉरोसिस होता है।. ग्लूकोमा वर्तमान में दुनिया भर में अपरिवर्तनीय अंधापन का सबसे आम कारण है, 20402 में लगभग 111.8 मिलियन लोगों तक पहुंचने की भविष्यवाणी की गई है, इस प्रकार रोगियों के जीवन की गुणवत्ता (क्यूओएल) को गहराई से प्रभावित करता है और महत्वपूर्ण सामाजिकआर्थिक चिंताओं को जन्म देता है।
ऊंचा इंट्राओकुलर दबाव (आईओपी) ग्लूकोमा के विकास और प्रगति के लिए सबसे महत्वपूर्ण और एकमात्र परिवर्तनीय जोखिम कारक में से एक है। ग्लूकोमा के कई प्रकारों में, सामान्य तनाव ग्लूकोमा (एनटीजी) को छोड़कर सभी, रोग के नैदानिक इतिहास में किसी समय ऊंचा आईओपी से जुड़े होते हैं। आईओपी को लक्षित करने और रोग की प्रगति को धीमा करने या रोकने के लिए उल्लेखनीय नैदानिक और शल्य चिकित्सा प्रगति के बावजूद, ग्लूकोमा 4,5 के कारण रोगी अभी भी दृष्टि खो देते हैं। इसलिए, इस बीमारी के जटिल और बहुक्रियाशील पैथोफिज़ियोलॉजी की पूरी तरह से समझ अधिक प्रभावी उपचार के विकास के लिए अनिवार्य है, खासकर आरजीसी को न्यूरोप्रोटेक्शन प्रदान करने के लिए।
रोग तंत्र की समझ के लिए विभिन्न प्रयोगात्मक दृष्टिकोणों के बीच, ओकुलर हाइपरटेंशन (ओएचटी) पर आधारित पशु मॉडल मानव ग्लूकोमा से सबसे अधिक मिलते-जुलते हैं। कृंतक मॉडल विशेष रूप से उपयोगी होते हैं क्योंकि वे कम लागत वाले होते हैं, संभालना आसान होता है, आनुवंशिक रूप से हेरफेर किया जा सकता है, एक छोटा जीवनकाल होता है, और मनुष्यों की तुलना में ओकुलर शारीरिक और शारीरिक विशेषताएं पेश करते हैं, जैसे जलीय हास्य उत्पादन और जल निकासी 6,7,8,9,10,11,12,13।. वर्तमान में उपयोग किए जाने वाले मॉडलों में एपिस्क्लरल नसों 14 में हाइपरटोनिक सलाइन के इंजेक्शन के बाद ट्रैब्युलर मेशवर्क का स्क्लेरोसिस, माइक्रोबीड्स 15 या विस्कोस्टिक पदार्थों का इंट्राकैस्ट्रियल इंजेक्शन 16, भंवर नसों का संयोजन 17, आर्गन लेजर 18 के साथ ट्रेब्युलर जाल का फोटोकॉएक्शन, सर्कमलिम्बल सीवन 19, और उम्र से संबंधित ओएचटी (डीबीए / 2 जे चूहों) के ट्रांसजेनिक मॉडल का उपयोग शामिल है।. हालांकि, इनवेसिवनेस, कॉर्निया के पोस्ट-ऑपरेटिव ओपेसिफिकेशन, एंटीरियर सेगमेंट व्यवधान, व्यापक सीखने के मोड़, महंगे उपकरण, और अत्यधिक परिवर्तनीय पोस्टऑपरेटिव आईओपी, वर्तमान मॉडल से जुड़े कुछ रिपोर्ट किए गए नुकसान ों में से हैं, जिससे ओएचटी के वैकल्पिक मॉडल का विकासइन समस्याओं को दूर करने की मांग करता है।
वर्तमान प्रोटोकॉल कृन्तकों में लिम्बल प्लेक्सस कॉटराइजेशन (एलपीसी) के आधार पर ग्लूकोमा के प्रॉक्सी के रूप में ओएचटी को प्रेरितकरने के लिए एक नई शल्य चिकित्सा प्रक्रिया को औपचारिक रूप देता है। यह एक आसान, प्रतिलिपि प्रस्तुत करने योग्य, सुलभ और गैर-इनवेसिव मॉडल है जो आईओपी ऊंचाई की उच्च दक्षता और कम परिवर्तनशीलता प्रदान करता है, जो पूर्ण नैदानिक वसूली की विशिष्ट उच्च दर से जुड़ा हुआ है, इसलिए प्रत्येक प्रयोग में उपयोग किए जाने वाले जानवरों की कम संख्या में विवो कार्यात्मक मूल्यांकन प्रदान करता है। सर्जरी तकनीक कुछ दिनों में बेसलाइन स्तरों पर धीरे-धीरे वापसी के साथ सबस्यूट ओएचटी को प्रेरित करती है, जो तीव्र कोण-बंद ग्लूकोमा में देखे गए उच्च रक्तचाप से ग्रस्त हमले को मॉडल करती है। इसके अलावा, मॉडल में आईओपी रिकवरी के बाद निरंतर ग्लूकोमाटस न्यूरोडीजेनेरेशन होता है, जो आरजीसी के माध्यमिक अध: पतन के भविष्य के यांत्रिक अध्ययनों के लिए उपयोगी है, जो आईओपी के पर्याप्त नियंत्रण के बावजूद मानव ग्लूकोमा के कई मामलों में होता है।

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Acetone</td>
			<td>Isofar</td>
			<td>201</td>
			<td>Used for electron microscopy tissue preparation (step 5)</td>
		</tr>
		<tr>
			<td>Active electrode for electroretinography</td>
			<td>Hansol Medical Co</td>
			<td>-</td>
			<td>Stainless steel needle 0.25 mm &#215; 15 mm</td>
		</tr>
		<tr>
			<td>Anestalcon</td>
			<td>Novartis Bioci&#234;ncias S/A</td>
			<td>MS-1.0068.1087</td>
			<td>Proxymetacaine hydrochloride 0.5%</td>
		</tr>
		<tr>
			<td>Calcium chloride</td>
			<td>Vetec</td>
			<td>560</td>
			<td>Used for electron microscopy tissue preparation (step 5)</td>
		</tr>
		<tr>
			<td>Cautery Low Temp Fine Tip 10/bx</td>
			<td>Bovie Medical Corporation</td>
			<td>AA00</td>
			<td>Low-temperature ophthalmic cautery</td>
		</tr>
		<tr>
			<td>Cetamin</td>
			<td>Syntec do Brasil Ltda</td>
			<td>000200-3-000003</td>
			<td>Ketamine hydrochloride 10%</td>
		</tr>
		<tr>
			<td>DAKO</td>
			<td>Dako North America</td>
			<td>S3023</td>
			<td>Antifade mounting medium</td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Thermo Fisher Scientific</td>
			<td>28718-90-3</td>
			<td>diamidino-2-phenylindole; blue fluorescent nuclear counterstain; emission at 452&#177;3 nm</td>
		</tr>
		<tr>
			<td>Ecofilm</td>
			<td>Crist&#225;lia Produtos Qu&#237;micos Farmac&#234;uticos Ltda</td>
			<td>MS-1.0298.0487</td>
			<td>Carmellose sodium 0.5%</td>
		</tr>
		<tr>
			<td>EPON Resin</td>
			<td>Polysciences, Inc.</td>
			<td>-</td>
			<td>Epoxy resin used for electron microscopy, composed of a mixture of four reagents: Poly/Bed 812 Resin (CAT#08791); DDSA - Dodecenylsuccinic Anhydride (CAT#00563); NMA - Nadic Methyl Anhydride (CAT#00886); DMP-30 - 2,4,6-tris(dimethylaminomethyl)phenol (CAT#00553)</td>
		</tr>
		<tr>
			<td>Glutaraldehyde</td>
			<td>Electron Microscopy Sciences</td>
			<td>16110</td>
			<td>Used for electron microscopy tissue preparation (step 5)</td>
		</tr>
		<tr>
			<td>Hyabak</td>
			<td>Uni&#227;o Qu&#237;mica Farmac&#234;utica Nacional S/A</td>
			<td>MS-8042140002</td>
			<td>Sodium hyaluronate 0.15%</td>
		</tr>
		<tr>
			<td>Icare Tonolab</td>
			<td>Icare Finland Oy</td>
			<td>TV02 (model number)</td>
			<td>Rebound handheld tonometer</td>
		</tr>
		<tr>
			<td>IgG donkey anti-mouse antibody + Alexa Fluor 555</td>
			<td>Thermo Fisher Scientific</td>
			<td>A31570</td>
			<td>Secondary antibody solution</td>
		</tr>
		<tr>
			<td>LCD monitor 23 inches</td>
			<td>Samsung Electronics Co. Ltd.</td>
			<td>S23B550</td>
			<td>Model LS23B550, for electroretinogram recording</td>
		</tr>
		<tr>
			<td>LSM 510 Meta</td>
			<td>Carl Zeiss</td>
			<td>-</td>
			<td>Confocal epifluorescence microscope</td>
		</tr>
		<tr>
			<td>Maxiflox</td>
			<td>Crist&#225;lia Produtos Qu&#237;micos Farmac&#234;uticos Ltda</td>
			<td>MS-1.0298.0489</td>
			<td>Ciprofloxacin 3.5 mg/g</td>
		</tr>
		<tr>
			<td>MEB-9400K</td>
			<td>Nihon Kohden Corporation</td>
			<td>-</td>
			<td>System for electroretinogram recording</td>
		</tr>
		<tr>
			<td>monoclonal IgG1 mouse anti-Brn3a</td>
			<td>MilliporeSigma</td>
			<td>MAB-1585</td>
			<td>Brn3a primary antibody solution</td>
		</tr>
		<tr>
			<td>Neuropack Manager v08.33</td>
			<td>Nihon Kohden Corporation</td>
			<td>-</td>
			<td>Software for electroretinogram signal processing</td>
		</tr>
		<tr>
			<td>Optomotry</td>
			<td>CerebralMechanics</td>
			<td>-</td>
			<td>System for optomotor response analysis</td>
		</tr>
		<tr>
			<td>Osmium tetroxide</td>
			<td>Electron Microscopy Sciences</td>
			<td>19100</td>
			<td>Used for electron microscopy tissue preparation (step 5)</td>
		</tr>
		<tr>
			<td>Potassium ferrocyanide</td>
			<td>Electron Microscopy Sciences</td>
			<td>20150</td>
			<td>Used for electron microscopy tissue preparation (step 5)</td>
		</tr>
		<tr>
			<td>Reference and ground electrodes for electroretinography</td>
			<td>Chalgren Enterprises</td>
			<td>110-63</td>
			<td>Stainless steel needles 0.4 mm &#215; 37 mm</td>
		</tr>
		<tr>
			<td>Sodium cacodylate buffer</td>
			<td>Electron Microscopy Sciences</td>
			<td>12300</td>
			<td>Used for electron microscopy tissue preparation (step 5)</td>
		</tr>
		<tr>
			<td>Ster MD</td>
			<td>Uni&#227;o Qu&#237;mica Farmac&#234;utica Nacional S/A</td>
			<td>MS-1.0497.1287</td>
			<td>Prednisolone acetate 0.12%</td>
		</tr>
		<tr>
			<td>Terolac</td>
			<td>Crist&#225;lia Produtos Qu&#237;micos Farmac&#234;uticos Ltda</td>
			<td>MS-1.0497.1286</td>
			<td>Ketorolac trometamol 0.5%</td>
		</tr>
		<tr>
			<td>Terramicina</td>
			<td>Laborat&#243;rios Pfizer Ltda</td>
			<td>MS-1.0216.0024</td>
			<td>Oxytetracycline hydrochloride 30 mg/g + polymyxin B 10,000 U/g</td>
		</tr>
		<tr>
			<td>Tono-Pen XL</td>
			<td>Reichert Technologies</td>
			<td>230635</td>
			<td>Digital applanation handheld tonometer</td>
		</tr>
		<tr>
			<td>TO-PRO-3</td>
			<td>Thermo Fisher Scientific</td>
			<td>T3605</td>
			<td>Far red-fluorescent nuclear counterstain; emission at 661 nm</td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma-Aldrich</td>
			<td>9036-19-5</td>
			<td>Non-ionic surfactant</td>
		</tr>
		<tr>
			<td>Uranyl acetate</td>
			<td>Electron Microscopy Sciences</td>
			<td>22400</td>
			<td>Used for electron microscopy tissue preparation (step 5)</td>
		</tr>
		<tr>
			<td>Xilazin</td>
			<td>Syntec do Brasil Ltda</td>
			<td>7899</td>
			<td>Xylazine hydrochloride 2%</td>
		</tr>
		<tr>
			<td></td>
			<td>Carl Zeiss</td>
			<td>-</td>
			<td>Stereo microscope for surgery and retinal dissection</td>
		</tr>
	</tbody>
</table>

full-circle cauterization of limbal vascular plexus for surgically induced glaucoma in rodents

ग्लूकोमा, दुनिया भर में अंधापन का दूसरा प्रमुख कारण, ओकुलर विकारों का एक विषम समूह है जो ऑप्टिक तंत्रिका और रेटिना गैंग्लियन सेल (आरजीसी) अध: पतन को संरचनात्मक क्षति की विशेषता है, जिसके परिणामस्वरूप आंख से मस्तिष्क तक दृश्य जानकारी के संचरण को बाधित करके दृश्य शिथिलता होती है। ऊंचा इंट्राओकुलर दबाव सबसे महत्वपूर्ण जोखिम कारक है; इस प्रकार, रोग के कारणों और प्रभावों की जांच के लिए आनुवंशिक या प्रयोगात्मक दृष्टिकोण द्वारा कृन्तकों में ओकुलर उच्च रक्तचाप के कई मॉडल विकसित किए गए हैं। उनमें से, कुछ सीमाओं की सूचना दी गई है जैसे कि सर्जिकल इनवेसिवनेस, अपर्याप्त कार्यात्मक मूल्यांकन, व्यापक प्रशिक्षण की आवश्यकता, और रेटिना क्षति का अत्यधिक परिवर्तनीय विस्तार। वर्तमान कार्य कृन्तकों में ओकुलर उच्च रक्तचाप को प्रेरित करने के लिए एक सरल, कम लागत वाली और कुशल विधि की विशेषता है, जो जलीय हास्य जल निकासी के एक प्रमुख घटक, लिम्बल संवहनी जाल के कम तापमान, पूर्ण-सर्कल कॉटराइजेशन पर आधारित है। नया मॉडल एक तकनीकी रूप से आसान, गैर-आक्रामक, और प्रतिलिपि प्रस्तुत करने योग्य सबस्यूट ओकुलर उच्च रक्तचाप प्रदान करता है, जो प्रगतिशील आरजीसी और ऑप्टिक तंत्रिका अध: पतन से जुड़ा हुआ है, और एक अद्वितीय पोस्ट-ऑपरेटिव नैदानिक वसूली दर है जो इलेक्ट्रोफिजियोलॉजिकल और व्यवहार दोनों तरीकों से विवो कार्यात्मक अध्ययन में अनुमति देता है।

मात्रात्मक चर को माध्य (एसईएम) के माध्य ± मानक त्रुटि के रूप में व्यक्त किया जाता है। ओएचटी और नियंत्रण समूहों (चित्रा 1 एफ) के बीच आईओपी गतिशीलता की तुलना को छोड़कर, सांख्यिकीय विश्लेषण दो-तरफ...

Watch this Scientific Journal Video about Full-Circle Cauterization of Limbal Vascular Plexus for Surgically Induced Glaucoma in Rodents at JoVE.com

Full-Circle Cauterization of Limbal Vascular Plexus for Surgically Induced Glaucoma in Rodents

इस प्रोटोकॉल का लक्ष्य लिम्बल वैस्कुलर प्लेक्सस के 360 डिग्री थर्मिक कॉटराइजेशन के आधार पर ग्लूकोमाटस न्यूरोडीजेनेरेशन के एक नए मॉडल को चिह्नित करना है, जो सबस्यूट ओकुलर हाइपरटेंशन को प्रेरित करता है।

कृन्तकों में शल्य चिकित्सा से प्रेरित ग्लूकोमा के लिए लिम्बल वैस्कुलर प्लेक्सस का फुल-सर्कल कॉटराइजेशन

Glaucoma, the second leading cause of blindness worldwide, is a heterogeneous group of ocular disorders characterized by structural damage to the optic ...

full-circle-cauterization-limbal-vascular-plexus-for-surgically

Research

JoVE Journal

Neuroscience

Preparing Undercut Model of Posttraumatic Epileptogenesis in Rodents

Partially isolated cortex ("undercut") is an animal model of posttraumatic epileptogenesis. The surgical procedure involves cutting through the sensorimotor cortex and the underneath white matter (undercut) so that a specific region of the cerebral cortex is largely isolated from the neighboring cortex and subcortical regions1-3. After a latency of two or more weeks following the surgery, epileptiform discharges can be recorded in brain slices from rodents1; and electrical or behavior seizures can be observed in vivo from other species such as cat and monkey4-6. This well established animal model is efficient to generate and mimics several important characteristics of traumatic brain injury. However, it is technically challenging attempting to make precise cortical lesions in the small rodent brain with a free hand. Based on the procedure initially established in Dr. David Prince's lab at the Stanford University1, here we present an improved technique to perform a surgery for the preparation of this model in mice and rats. We demonstrate how to make a simple surgical device and use it to gain a better control of cutting depth and angle to generate more precise and consistent results. The device is easy to make, and the procedure is quick to learn. The generation of this animal model provides an efficient system for study on the mechanisms of posttraumatic epileptogenesis.

Partially isolated cortex (&#x201C;undercut&#x201D;) is an efficient animal model of posttraumatic epileptogenesis. Here we demonstrate how to make a novel surgical device and use it to make more precise and consistent lesions to generate this model.

कृन्तकों में Posttraumatic Epileptogenesis की काटकर अलग कर देना मॉडल की तैयारी

Partially isolated cortex ("undercut") is an animal model of posttraumatic epileptogenesis. The surgical procedure involves cutting through the ...

Whole Animal Perfusion Fixation for Rodents

The goal of fixation is to rapidly and uniformly preserve tissue in a life-like state. While placing tissue directly in fixative works well for small pieces of tissue, larger specimens like the intact brain pose a problem for immersion fixation because the fixative does not reach all regions of the tissue at the same rate 5,7. Often, changes in response to hypoxia begin before the tissue can be preserved 12. The advantage of directly perfusing fixative through the circulatory system is that the chemical can quickly reach every corner of the organism using the natural vascular network. In order to utilize the circulatory system most effectively, care must be taken to match physiological pressures 3. It is important to note that physiological pressures are dependent on the species used. Techniques for perfusion fixation vary depending on the tissue to be fixed and how the tissue will be processed following fixation. In this video, we describe a low-cost, rapid, controlled and uniform fixation procedure using 4% paraformaldehyde perfused via the vascular system: through the heart of the rat to obtain the best possible preservation of the brain for immunohistochemistry. The main advantage of this technique (vs. gravity-fed systems) is that the circulatory system is utilized most effectively.

Here we describe a low-cost, rapid, controlled and uniform fixation procedure using 4% paraformaldehyde perfused via the vascular system: through the heart of the rat to obtain the best possible preservation of the brain.

कृन्तकों के लिए पूरे पशु परफ्यूज़न फिक्सेशन

The goal of fixation is to rapidly and uniformly preserve tissue in a life-like state. While placing tissue directly in fixative works well for small ...

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects1-11. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design.
Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (<a href="http://www.neuronexustech.com/" >http://www.neuronexustech.com/</a>; <a href="http://www.sbmicrosystems.com/">http://www.sbmicrosystems.com/</a>; <a href="http://www.acreo.se/">http://www.acreo.se/</a>) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process.

We describe methods for large-scale recording of multiple single units and local field potential in behaving rodents with silicon probes. Drive fabrication, probe attachment to the drive and probe implantation processes are illustrated in sufficient details for easy replication.

कृन्तकों व्यवहार में बड़े पैमाने पर चल सिलिकॉन जांच से न्यूरॉन्स की रिकॉर्डिंग

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

A Low Cost Setup for Behavioral Audiometry in Rodents

In auditory animal research it is crucial to have precise information about basic hearing parameters of the animal subjects that are involved in the experiments. Such parameters may be physiological response characteristics of the auditory pathway, e.g. via brainstem audiometry (BERA). But these methods allow only indirect and uncertain extrapolations about the auditory percept that corresponds to these physiological parameters. To assess the perceptual level of hearing, behavioral methods have to be used. A potential problem with the use of behavioral methods for the description of perception in animal models is the fact that most of these methods involve some kind of learning paradigm before the subjects can be behaviorally tested, e.g. animals may have to learn to press a lever in response to a sound. As these learning paradigms change perception itself 1,2 they consequently will influence any result about perception obtained with these methods and therefore have to be interpreted with caution. Exceptions are paradigms that make use of reflex responses, because here no learning paradigms have to be carried out prior to perceptual testing. One such reflex response is the acoustic startle response (ASR) that can highly reproducibly be elicited with unexpected loud sounds in na&iuml;ve animals. This ASR in turn can be influenced by preceding sounds depending on the perceptibility of this preceding stimulus: Sounds well above hearing threshold will completely inhibit the amplitude of the ASR; sounds close to threshold will only slightly inhibit the ASR. This phenomenon is called pre-pulse inhibition (PPI) 3,4, and the amount of PPI on the ASR gradually depends on the perceptibility of the pre-pulse. PPI of the ASR is therefore well suited to determine behavioral audiograms in na&iuml;ve, non-trained animals, to determine hearing impairments or even to detect possible subjective tinnitus percepts in these animals. In this paper we demonstrate the use of this method in a rodent model (cf. also ref. 5), the Mongolian gerbil (Meriones unguiculatus), which is a well know model species for startle response research within the normal human hearing range (e.g. 6).

A fast and inexpensive method for the behavioral determination of hearing parameters like hearing thresholds, hearing impairments or phantom perceptions (subjective tinnitus) is described. It uses pre-pulse inhibition of the acoustic startle response and can be easily implemented in a personal computer using a programmable AD/DA-converter and a piezo sensor.

कृन्तकों में व्यवहार ऑडियोमेट्री के लिए कम लागत सेटअप

In auditory animal research it is crucial to have precise information about basic hearing parameters of the animal subjects that are involved in the ...

Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, including rats, hamsters, and mice, are active during the night and relatively inactive during the day. Many other behavioral and physiological measures also exhibit daily rhythms, but in rodents, running-wheel activity serves as a particularly reliable and convenient measure of the output of the master circadian clock, the suprachiasmatic nucleus (SCN) of the hypothalamus. In general, through a process called entrainment, the daily pattern of running-wheel activity will naturally align with the environmental light-dark cycle (LD cycle; e.g. 12 hr-light:12 hr-dark). However circadian rhythms are endogenously generated patterns in behavior that exhibit a ~24 hr period, and persist in constant darkness. Thus, in the absence of an LD cycle, the recording and analysis of running-wheel activity can be used to determine the subjective time-of-day. Because these rhythms are directed by the circadian clock the subjective time-of-day is referred to as the circadian time (CT). In contrast, when an LD cycle is present, the time-of-day that is determined by the environmental LD cycle is called the zeitgeber time (ZT).
Although circadian rhythms in running-wheel activity are typically linked to the SCN clock6-8, circadian oscillators in many other regions of the brain and body9-14 could also be involved in the regulation of daily activity rhythms. For instance, daily rhythms in food-anticipatory activity do not require the SCN15,16 and instead, are correlated with changes in the activity of extra-SCN oscillators17-20. Thus, running-wheel activity recordings can provide important behavioral information not only about the output of the master SCN clock, but also on the activity of extra-SCN oscillators. Below we describe the equipment and methods used to record, analyze and display circadian locomotor activity rhythms in laboratory rodents.

Circadian rhythms in voluntary wheel-running activity in mammals are tightly coupled to the molecular oscillations of a master clock in the brain. As such, these daily rhythms in behavior can be used to study the influence of genetic, pharmacological, and environmental factors on the functioning of this circadian clock.

और कृन्तकों में गतिविधि पहिया चल रहा है में circadian लय का विश्लेषण रिकॉर्डिंग

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for using blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) to image rodents with simultaneous DBS. DBS fMRI presents a number of technical challenges, including accuracy of electrode implantation, MR artifacts created by the electrode, choice of anesthesia and paralytic to minimize any neuronal effects while simultaneously eliminating animal motion, and maintenance of physiological parameters, deviation from which can confound the BOLD signal.&#160;Our laboratory has developed a set of procedures that are capable of overcoming most of these possible issues. For electrical stimulation, a homemade tungsten bipolar microelectrode is used, inserted stereotactically at the stimulation site in the anesthetized subject. In preparation for imaging, rodents are fixed on a plastic headpiece and transferred to the magnet bore. For sedation and paralysis during scanning, a cocktail of dexmedetomidine and pancuronium is continuously infused, along with a minimal dose of isoflurane; this preparation minimizes the BOLD ceiling effect of volatile anesthetics. In this example experiment, stimulation of the subthalamic nucleus (STN) produces BOLD responses which are observed primarily in ipsilateral cortical regions, centered in motor cortex. Simultaneous DBS and fMRI allows the unambiguous modulation of neural circuits dependent on stimulation location and stimulation parameters, and permits observation of neuronal modulations free of regional bias. This technique may be used to explore the downstream effects of modulating neural circuitry at nearly any brain region, with implications for both experimental and clinical DBS.

This protocol describes a standard method for simultaneous functional magnetic resonance imaging and deep brain stimulation in the rodent. The combined use of these experimental tools allows for the exploration of global downstream activity in response to electrical stimulation at virtually any brain target.

कृंतक में एक साथ fMRI साथ डीप ब्रेन उसकाव

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for ...

Quantification of Cerebral Vascular Architecture using Two-photon Microscopy in a Mouse Model of HIV-induced Neuroinflammation

Human Immunodeficiency Virus 1 (HIV-1) infection frequently results in HIV-1 Associated Neurocognitive Disorders (HAND), and is characterized by a chronic neuroinflammatory state within the central nervous system (CNS), thought to be driven principally by virally-mediated activation of microglia and brain resident macrophages. HIV-1 infection is also accompanied by changes in cerebrovascular blood flow (CBF), raising the possibility that HIV-associated chronic neuroinflammation may lead to changes in CBF and/or in cerebral vascular architecture. To address this question, we have used a mouse model for HIV-induced neuroinflammation, and we have tested whether long-term exposure to this inflammatory environment may damage brain vasculature and result in rarefaction of capillary networks. In this paper we describe a method to quantify changes in cortical capillary density in a mouse model of neuroinflammatory disease (HIV-1 Tat transgenic mice). This generalizable approach employs in vivo two-photon imaging of cortical capillaries through a thin-skull cortical window, as well as ex vivo two-photon imaging of cortical capillaries in mouse brain sections. These procedures produce images and z-stack files of capillary networks, respectively, which can be then subjected to quantitative analysis in order to assess changes in cerebral vascular architecture.

This paper describes a method by which the vascular architecture in the brain can be quantified using in vivo and ex vivo two-photon microscopy.

एचआईवी प्रेरित neuroinflammation के एक माउस मॉडल में दो photon माइक्रोस्कोपी का उपयोग मस्तिष्क संवहनी वास्तुकला की मात्रा

Human Immunodeficiency Virus 1 (HIV-1) infection frequently results in HIV-1 Associated Neurocognitive Disorders (HAND), and is characterized by a chronic ...

Cannula Implantation into the Cisterna Magna of Rodents

Cisterna magna cannulation (CMc) is a straightforward procedure that enables direct access to the cerebrospinal fluid (CSF) without operative damage to the skull or the brain parenchyma. In anesthetized rodents, the exposure of the dura mater by blunt dissection of the neck muscles allows the insertion of a cannula into the cisterna magna (CM). The cannula, composed either by a fine beveled needle or borosilicate capillary, is attached via a polyethylene (PE) tube to a syringe. Using a syringe pump, molecules can then be injected at controlled rates directly into the CM, which is continuous with the subarachnoid space. From the subarachnoid space, we can trace CSF fluxes by convective flow into the perivascular space around penetrating arterioles, where solute exchange with the interstitial fluid (ISF) occurs. CMc can be performed for acute injections immediately following the surgery, or for chronic implantation, with later injection in anesthetized or awake, freely moving rodents. Quantitation of tracer distribution in the brain parenchyma can be performed by epifluorescence, 2-photon microscopy, and magnetic resonance imaging (MRI), depending on the physico-chemical properties of the injected molecules. Thus, CMc in conjunction with various imaging techniques offers a powerful tool for assessment of the glymphatic system and CSF dynamics and function. Furthermore, CMc can be utilized as a conduit for fast, brain-wide delivery of signaling molecules and metabolic substrates that could not otherwise cross the blood brain barrier (BBB).

Here we describe a protocol to perform cisterna magna cannulation (CMc), a minimally invasive way to deliver tracers, substrates and signaling molecules into the cerebrospinal fluid (CSF). Combined with different imaging modalities, CMc enables glymphatic system and CSF dynamics assessment, as well as brain-wide delivery of various compounds.

कुतर के Cisterna Magna में प्रवेशनी आरोपण

Cisterna magna cannulation (CMc) is a straightforward procedure that enables direct access to the cerebrospinal fluid (CSF) without operative damage to ...

Intracranial Pharmacotherapy and Pain Assays in Rodents

Pain is a salient sensory experience with affective and cognitive dimensions. However, central mechanisms for pain remain poorly understood, hindering the development of effective therapeutics. Intracranial pharmacology presents an important tool for understanding the molecular and cellular mechanisms of pain in the brain, as well as for novel treatments. Here we present a protocol that integrates intracranial pharmacology with pain behavior testing. Specifically, we show how to infuse analgesic drugs into a select brain region, which may be responsible for pain modulation. Furthermore, to determine the effect of the candidate drug in the central nerve system, pain assays are performed after intracranial treatment. Our results demonstrate that intracranial administration of analgesic drugs in a targeted region can provide relief of pain in rodents. Thus, our protocol successfully demonstrates that intracranial pharmacology, combined with pain behavior testing, can be a powerful tool for the study of pain mechanisms in the brain.

Here we present a protocol to perform intracranial pharmacological experiments followed by pain behavior assays in rodents. This protocol allows researchers to deliver molecular and cellular targets in the brain, for pharmacologic agents in the treatment of pain.

Intracranial औषधीय और दर्द के साथ-साथ Rodents में Assays

Pain is a salient sensory experience with affective and cognitive dimensions. However, central mechanisms for pain remain poorly understood, hindering the ...

The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in neurocognitive disorders. Despite the popularization of prepulse inhibition (PPI) in recent years, many current protocols promote using a percent of control measure, thereby precluding the assessment of temporal processing. The present study used cross-modal PPI and gap prepulse inhibition (gap-PPI) to demonstrate the benefits of employing a range of interstimulus intervals (ISIs) to delineate effects of sensory modality, psychostimulant exposure, and age. Assessment of sensory modality, psychostimulant exposure, and age reveals the utility of an approach varying the interstimulus interval (ISI) to establish the shape of the ISI function, including increases (sharper curve inflections) or decreases (flattening of the response amplitude curve) in startle amplitude. Additionally, shifts in peak response inhibition, suggestive of a differential sensitivity to the manipulation of ISI, are often revealed. Thus, the systematic manipulation of ISI affords a critical opportunity to evaluate temporal processing, which may reveal the underlying neural mechanisms involved in neurocognitive disorders.

Temporal processing, a preattentive process, may underlie deficits in higher-level cognitive processes, including attention, commonly observed in neurocognitive disorders. Using prepulse inhibition as an exemplar paradigm, we present a protocol for manipulating interstimulus interval (ISI) to establish the shape of the ISI function to provide an assessment of temporal processing.