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All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Dissection of Palate Tissue from Adult Mouse
<ol>
	<li>Sacrifice an adult C57BL/6J mouse (either male or female) by cervical dislocation in compliance with the facility&#39;s regulations relating to animal welfare.
	<ol>
		<li>Remove the hair around the mouth with a shaver. Using scissors, cut from the cheek toward the jaw, on both sides.&#160;&#160;&#160; 
		NOTE: The mouse needs to be anesthetized before sacrifice. An anesthetic mixture of medetomidine, midazolam, and butorphanol is used (see&#160;Table of Materials).</li>
	</ol>
	</li>
	<li>Use forceps to open the mouth wide and absorb any blood using a cotton swab. To disinfect the palate, wipe the inside of the mouth with a cotton swab containing 10% povidone-iodine.</li>
	<li>To harvest the mouse palate, first, use a surgical scalpel blade to make a full-thickness marginal incision along the palate side of the maxillary teeth. Then, carefully dissect the entire palate using a raspatorium. 
	NOTE: A raspatorium is a tool used to elevate a mucoperiosteal flap (see&#160;Table of Materials) (Figure 1B).</li>
	<li>Quickly transfer the palate tissue to a 15 mL tube containing 4 mL of complete medium + antibiotic-antimycotic solution. Keep the tissues on ice until ready for incubation. 
	NOTE: For collection from multiple mice, palate tissues may be kept on ice for up to 4 h.</li>
</ol>

<table><tbody><tr><td>Iris Scissors</td><td>Muromachi Kikai</td><td>14090-09</td><td></td></tr><tr><td>Medetomidine</td><td>Nippon Zenyaku Kogyo</td><td></td><td>Domitor</td></tr><tr><td>Midazolam</td><td>Astellas Pharma</td><td></td><td>Dormicum</td></tr><tr><td>No.15 Disposable scalpel</td><td>Feather</td><td>219AABZX00136000</td><td></td></tr><tr><td>Butorphanol</td><td>Meiji Seika Pharma</td><td></td><td>Vetorphale</td></tr><tr><td>Half-curved forceps</td><td>BRC Bio Research Center</td><td>PRI13-3376</td><td></td></tr><tr><td>Povidone-iodine</td><td>Y&#039;s Square</td><td>872612</td><td></td></tr><tr><td>Raspatorium #14</td><td>AS ONE</td><td>8-4599-01</td><td></td></tr><tr><td>Fine forceps</td><td>BRC Bio Research Center</td><td>PRI13-3374</td><td></td></tr><tr><td>Antibiotic-Antimycotic 100x</td><td>Gibco</td><td>15240062</td><td></td></tr><tr><td>15 mL sterile conical tube</td><td>Falcon</td><td>352096</td><td></td></tr><tr><td>Cotton</td><td>AS ONE</td><td>63-1452-97</td><td></td></tr><tr><td>EpiLife Defined Growth Supplement (EDGS)&amp;nbsp;</td><td>Gibco&amp;nbsp;</td><td>S0120</td><td></td></tr></tbody></table>

dissection of palate tissue from adult murine model: a surgical procedure to harvest hard palate and soft palate from oral cavity of adult mouse

Source: Xuan Ngo, Y. et al. <a href="https://www.jove.com/video/62820" target="_blank">Isolation and Culture of Primary Oral Keratinocytes from the Adult Mouse Palate</a>.&#160;J. Vis. Exp.&#160;(2021)
This video demonstrates a surgical procedure to dissect the palate tissue from a murine model.

<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/20740/20740_Figure1.jpg" /> 
Figure 1: Overview of the dissection procedure and isolation of mouse oral keratinocytes.&#160;(A) Schematic representation of the mouse oral cavity. (B) Instruments used to dissect the palate and isolate mouse oral keratinocytes. (C) Brightfield image of the mouse palate. Scale bar: 100 &#181;m. (D) Summary of th...

Dissection of Palate Tissue from Adult Murine Model: A Surgical Procedure to Harvest Hard Palate and Soft Palate from Oral Cavity of Adult Mouse

Source: Xuan Ngo, Y. et al. Isolation and Culture of Primary Oral Keratinocytes from the Adult Mouse Palate.&#160;J. Vis. Exp.&#160;(2021)
This video ...

In mammals, the palate, the roof of the mouth, is divided into two parts - hard palate and soft palate. The soft palate, the posterior muscular part of the palate, is continuous with the hard palate, the anterior bony part of the palate. To harvest the palate tissue, begin with a euthanized mouse in the supine position. Next, shave the area around the animal&#8217;s mouth.
Now, make surgical incisions from the cheek towards the jaw on both sides of the mouse. Open its mouth wide and sterilize the palate area. Further, make a marginal incision along the palate side between the two sides of the maxillary dental arch.
Carefully lift the portion of the hard palate mucoperiosteum - connective tissue covering the bones of the hard palate formed by the union of mucous membrane with the dense fibrous connective tissue, the periosteum. This step facilitates the detachment of the hard palate from the underlying bony structures and further releases the soft palate from the oral cavity.
Immediately transfer the harvested palate into a tube containing suitable media supplemented with antibiotics and antimycotics to prevent bacterial and fungal contamination within the tissue. Store the tissue at lower temperatures until further processing.

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1.7K Views. Source: Xuan Ngo, Y. et al. Isolation and Culture of Primary Oral Keratinocytes from the Adult Mouse Palate. J. Vis. Exp. (2021) This video demonstrates a surgical procedure to dissect the palate tissue from a murine model. 

Video: Dissection of Palate Tissue from Adult Murine Model: A Surgical Procedure to Harvest Hard Palate and Soft Palate from Oral Cavity of Adult Mouse - Concept

Method of Studying Palatal Fusion using Static Organ Culture

Cleft lip and palate are among the most common of all birth defects. The secondary palate forms from mesenchymal shelves covered with epithelium that adheres to form the midline epithelial seam (MES). The theories suggest that MES cells follow an epithelial to mesenchymal transition (EMT), apoptosis and migration, making a fused palate 1. Complete disintegration of the MES is the final essential phase of palatal confluence with surrounding mesenchymal cells. We provide a method for palate organ culture. The developed in vitro protocol allows the study of the biological and molecular processes during fusion. The applications of this technique are numerous, including evaluating responses to exogenous chemical agents, effects of regulatory and growth factors and specific proteins. Palatal organ culture has a number of advantages including manipulation at different stages of development that is not possible using in vivo studies.

Studies of palate development are motivated by the incidence of cleft palate, a birth defect that imposes a tremendous health burden and can leave lasting disfigurement. We demonstrate here a technique to culture palatal shelves that can be used to study different signaling pathways involved in palatal development and fusion.

Cleft lip and palate are among the most common of all birth defects. The secondary palate forms from mesenchymal shelves covered with epithelium that ...

JoVE Journal

Developmental Biology

Isolation and Time-Lapse Imaging of Primary Mouse Embryonic Palatal Mesenchyme Cells to Analyze Collective Movement Attributes

Development of the palate is a dynamic process, which involves vertical growth of bilateral palatal shelves next to the tongue followed by elevation and fusion above the tongue. Defects in this process lead to cleft palate, a common birth defect. Recent studies have shown that palatal shelf elevation involves a remodeling process that transforms the orientation of the shelf from a vertical to a horizontal one. The role of the palatal shelf mesenchymal cells in this dynamic remodeling has been difficult to study. Time-lapse-imaging-based quantitative analysis has been recently used to show that primary mouse embryonic palatal mesenchymal (MEPM) cells can self-organize into a collective movement. Quantitative analyses could identify differences in mutant MEPM cells from a mouse model with palate elevation defects. This paper describes methods to isolate and culture MEPM cells from E13.5 embryos-specifically for time-lapse imaging-and to determine various cellular attributes of collective movement, including measures for stream formation, shape alignment, and persistence of direction. It posits that MEPM cells can serve as a proxy model for studying the role of palatal shelf mesenchyme during the dynamic process of elevation. These quantitative methods will allow investigators in the craniofacial field to assess and compare collective movement attributes in control and mutant cells, which will augment the understanding of mesenchymal remodeling during palatal shelf elevation. Furthermore, MEPM cells provide a rare mesenchymal cell model for investigation of collective cell movement in general.

We present a protocol for isolation and culture of primary mouse embryonic palatal mesenchymal cells for time-lapse imaging of two-dimensional (2D) growth and wound-repair assays. We also provide the methodology for analysis of the time-lapse imaging data to determine cell-stream formation and directional motility.

Development of the palate is a dynamic process, which involves vertical growth of bilateral palatal shelves next to the tongue followed by elevation and ...

Whole-Mount Staining, Visualization, and Analysis of Fungiform, Circumvallate, and Palate Taste Buds

Taste buds are collections of taste-transducing cells specialized to detect subsets of chemical stimuli in the oral cavity. These transducing cells communicate with nerve fibers that carry this information to the brain. Because taste-transducing cells continuously die and are replaced throughout adulthood, the taste-bud environment is both complex and dynamic, requiring detailed analyses of its cell types, their locations, and any physical relationships between them. Detailed analyses have been limited by tongue-tissue heterogeneity and density that have significantly reduced antibody permeability. These obstacles require sectioning protocols that result in splitting taste buds across sections so that measurements are only approximated, and cell relationships are lost. To overcome these challenges, the methods described herein involve collecting, imaging, and analyzing whole taste buds and individual terminal arbors from three taste regions: fungiform papillae, circumvallate papillae, and the palate. Collecting whole taste buds reduces bias and technical variability and can be used to report absolute numbers for features including taste-bud volume, total taste-bud innervation, transducing-cell counts, and the morphology of individual terminal arbors. To demonstrate the advantages of this method, this paper provides comparisons of taste bud and innervation volumes between fungiform and circumvallate taste buds using a general taste-bud marker and a label for all taste fibers. A workflow for the use of sparse-cell genetic labeling of taste neurons (with labeled subsets of taste-transducing cells) is also provided. This workflow analyzes the structures of individual taste-nerve arbors, cell type numbers, and the physical relationships between cells using image analysis software. Together, these workflows provide a novel approach for tissue preparation and analysis of both whole taste buds and the complete morphology of their innervating arbors.

This paper describes methods for tissue preparation, staining, and analysis of whole fungiform, circumvallate, and palate taste buds that consistently yield whole and intact taste buds (including the nerve fibers that innervate them) and maintain the relationships between structures within taste buds and the surrounding papilla.

Taste buds are collections of taste-transducing cells specialized to detect subsets of chemical stimuli in the oral cavity. These transducing cells ...

Dissection of Palate Tissue from Adult Murine Model: A Surgical Procedure to Harvest Hard Palate and Soft Palate from Oral Cavity of Adult Mouse

Transcript

Dissection of Palate Tissue from Adult Murine Model: A Surgical Procedure to Harvest Hard Palate and Soft Palate from Oral Cavity of Adult Mouse

Method of Studying Palatal Fusion using Static Organ Culture

Isolation and Time-Lapse Imaging of Primary Mouse Embryonic Palatal Mesenchyme Cells to Analyze Collective Movement Attributes

Whole-Mount Staining, Visualization, and Analysis of Fungiform, Circumvallate, and Palate Taste Buds