Name: a model of glaucoma induced by circumlimbal suture in rats and mice
Uploaded: 2018-10-05T14:00:00
Description: Watch this Scientific Journal Video about A Model of Glaucoma Induced by Circumlimbal Suture in Rats and Mice at JoVE.com

This work is funded by National Health and Medical Research Council of Australia project grant (1046203), Australian Research Council Future Fellowship (FT130100338).

All experimental procedures were conducted according to the Australian Code of Practice for the Care and Use of Animals for Scientific Purposes, set by the National Health and Medical Research Council in Australia. Ethics approval was obtained from the Howard Florey Institute Animal Ethics Committee (approval number 13-044-UM and 13-068-UM for rats and mice, respectively).
1. Intraocular Pressure Measurement in Conscious Rats
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	<li>Set the laboratory rebound tonometer to the rat setting....

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The circumlimbal suture is a new model of chronic ocular hypertension. In addition to the studies from which the representative results are sourced16,18, this animal model has been utilized in a number of recent studies15,23,24,25,26. Comparison across these previous reports shows that the method produ...

Animal models provide an important platform for laboratory investigation of cellular processes underlying glaucoma pathogenesis, as well as to evaluate potential therapeutic interventions. Several inducible models have been developed to produce sustained intraocular pressure (IOP) elevation, the most important risk factor for glaucoma. Methods that have been applied to elevate IOP include: hypertonic saline injection in episcleral veins1, laser photocoagulation of the trabecular meshwork2 or of the limbal veins3, and intracameral injection of substances such as ghost red blood cells4, microbeads5,6 and viscoelastic agents7. Each approach has its advantages and limitations.
A good model for glaucoma should mimic the disease process, with minimal complication such as trauma, inflammation and media opacities. These complications are frequently associated with the procedures used to induce IOP elevation, and can confound interpretation of outcomes. For example, paracentesis of the anterior chamber, even when foreign substances are not introduced, has been shown to cause trauma and inflammation that is not representative of typical glaucomatous change8,9. In addition to the importance of avoiding inflammation, maintaining optical clarity facilitates in vivo imaging and electrophysiology to monitor disease progression. Although it is unclear to what extent these complications may affect disease investigations, it may be better to avoid penetrating the eye during model induction. The circumlimbal suture approach avoids penetration of the globe and facilitates in vivo longitudinal assessment of retinal structure and function. More importantly, this model differs from previous ones in its capacity to return IOP to baseline values by removal of the suture when required. IOP normalization may be useful for studying the cellular and molecular correlates of reversible and irreversible ganglion cell injury10,11,12,13,14.
This article focuses on the technique for model induction. Characterization of retinal injury&#160;induced by this model in rats and mice can be found in greater detail elsewhere15,16,17,18,19.

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	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:157px;">normal saline</td>
			<td style="width:219px;">Baxter International Inc</td>
			<td style="width:135px;">AHB1323</td>
			<td style="width:365px;">Maintain corneal hydration during surgery</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:157px;">Chlorhexadine 0.5%</td>
			<td style="width:219px;">Orion Laboratories</td>
			<td style="width:135px;">27411, 80085</td>
			<td style="width:365px;">Disinfection of surgical instrument</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:157px;">Isoflurane 99.9%</td>
			<td style="width:219px;">Abbott Australasia Pty Ltd</td>
			<td style="width:135px;">CAS 26675-46-7</td>
			<td style="width:365px;">Proprietory Name: Isoflo(TM) Inhalation anaaesthetic. Pharmaceutical-grade inhalation anesthetic mixed with oxygen gas for suture procedure</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:157px;">ocular lubricant</td>
			<td style="width:219px;">Alcon Laboratories&#160;</td>
			<td style="width:135px;">1618611</td>
			<td style="width:365px;">Proprietory Name: Genteal, ocular lubricant to keep the other eye moist</td>
		</tr>
		<tr height="32">
			<td height="32" style="height:32px;width:157px;">Needle holder (microsurgery)</td>
			<td style="width:219px;">World Precision Instruments</td>
			<td style="width:135px;">555419NT</td>
			<td style="width:365px;">To hold needle during ocular surgery</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:157px;">Proxymetacaine 0.5%</td>
			<td style="width:219px;">Alcon Laboratories&#160;</td>
			<td style="width:135px;">CAS 5875-06-9</td>
			<td style="width:365px;">Topical ocular analgesia</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:157px;">Scissors (microsurgery)</td>
			<td style="width:219px;">World Precision Instruments</td>
			<td style="width:135px;">501232</td>
			<td style="width:365px;">To cut excessive suture stump during ligation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:157px;">Surgical drape</td>
			<td style="width:219px;">Vital Medical Supplies</td>
			<td style="width:135px;">GM29-612EE</td>
			<td style="width:365px;">Ensure sterile enviornment during surgery</td>
		</tr>
		<tr height="32">
			<td height="32" style="height:32px;width:157px;">Suture needle for rats (microsurgery)</td>
			<td style="width:219px;">Ninbo medical needles</td>
			<td style="width:135px;">151109</td>
			<td style="width:365px;">8-0 nylon suture attached with round needle, cutting edge 3/8, dual-needle, suture length 30cm</td>
		</tr>
		<tr height="32">
			<td height="32" style="height:32px;width:157px;">Suture needle for mice (microsurgery)</td>
			<td style="width:219px;">Ninbo medical needles</td>
			<td style="width:135px;">160905</td>
			<td style="width:365px;">10-0 nylon suture attached with round needle, cutting edge 3/8, dual-needle, suture length 30cm</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:157px;">Tweezers (microsurgery)</td>
			<td style="width:219px;">World Precision Instruments</td>
			<td style="width:135px;">500342</td>
			<td style="width:365px;">Manipulate tissues during ocular surgery</td>
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		<tr height="20">
			<td height="20" style="height:20px;width:157px;">rebound tonometer</td>
			<td style="width:219px;">TONOLAB, iCare, Helsinki, Finland</td>
			<td style="width:135px;">TV02</td>
			<td style="width:365px;">for intraocular pressure monitoring</td>
		</tr>
	</tbody>
</table>

a model of glaucoma induced by circumlimbal suture in rats and mice

The circumlimbal suture is a technique for inducing experimental glaucoma in rodents by chronically elevating intraocular pressure (IOP), a well-known risk factor for glaucoma. This protocol demonstrates a step-by-step guide on this technique in Long Evans rats and C57BL/6 mice. Under general anesthesia, a "purse-string" suture is applied on the conjunctiva, around the equator and behind the limbus of the eye. The fellow eye serves as an untreated control. Over the duration of our study, which was a period of 8 weeks for rats and 12 weeks for mice, IOP remained elevated, as measured regularly by rebound tonometry in conscious animals without topical anesthesia. In both species, the sutured eyes showed electroretinogram features consistent with preferential inner retinal dysfunction. Optical coherence tomography showed selective thinning of the retinal nerve fiber layer. Histology of the rat retina in cross-section found reduced cell density in the ganglion cell layer, but no change in other cellular layers. Staining of flat-mounted mouse retinae with a ganglion cell specific marker (RBPMS) confirmed ganglion cell loss. The circumlimbal suture is a simple, minimally invasive and cost-effective way to induce ocular hypertension that leads to ganglion cell injury in both rats and mice.

The following results in rats18 and mice16 have been previously reported and are summarized here. The circumlimbal suture produced a similar pattern of IOP elevation in rats and mice (Figure 2). A brief IOP spike, up to 58.1 &#177; 2.7 mmHg in rats and 38.7 &#177; 2.2 mmHg in mice, was found immediately after the suture procedure. In rats, IOP magnitude gradually reduced over time to be 44 &#177; 6 mmHg and 32 &...

Watch this Scientific Journal Video about A Model of Glaucoma Induced by Circumlimbal Suture in Rats and Mice at JoVE.com

A Model of Glaucoma Induced by Circumlimbal Suture in Rats and Mice

Chronic ocular hypertension is induced by applying a circumlimbal suture in rats and mice, leading to functional and structural deterioration of the retinal ganglion cells consistent with glaucoma.

The circumlimbal suture is a technique for inducing experimental glaucoma in rodents by chronically elevating intraocular pressure (IOP), a well-known ...

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