image Exploring the Pre-Bötzinger complex through 3D Modeling

1Mariah Geritano, 3Sharon Lam, 1Janine Tirone, 2 Irene Solomon and 1Marvin H. O’Neal III

1Undergraduate Biology, Stony Brook University; 2Physiology and Biophysics, Stony Brook University; 3Syosset High School, Syosset, New York

Previous work from our laboratory has used histological analyses to confirm functionally identified sites in the cat pre-Botzinger complex (pre-BotC). These sites have been represented in a 2-D fashion based on our histological sections in conjunction with plates from Berman’s stereotactic atlas (1968). To gain further insight into the anatomical distribution of these sites, this project used rapid prototyping technology to generate multiple 3-D physical models representing the relevant regions/nuclei of the cat brainstem (based on Berman’s atlas) and the histological/stereotaxic coordinates associated with our microinjection sites. To accomplish this, plates from Berman’s atlas were scanned and compiled into a 3-D configuration slide box, the stereotaxic coordinates of 348 pre-BötC microinjection sites were entered into an x/y/z matrix based on histologically confirmed coordinates with the origin (0, 0, 0) referenced to the caudal pole of the retrofacial nucleus, midline, and dorsal surface of the medulla, and a 3-D digital representation was generated and printed. This approach compliments both the 2-D representations (previously used) and the digital 3-D representation, and provides additional insight into the anatomical specificity of the functionally identified and histologically confirmed microinjection sites.

Figure 1: Construction of Blender Slide  The Blender default cube in the viewport was modified using the scale tool.  A single slide was created, showing thickness along the x, y, z plane. The default cube in the Blender viewport was modified to create the base of the rectangular slide box. The tool loop cut was used to create lines along the border. Loop cut was used to create nine evenly-spaced grooves.
Figure 1: Construction of Blender Slide
The Blender default cube in the viewport was modified using the scale tool. A single slide was created, showing thickness along the x, y, z plane. The default cube in the Blender viewport was modified to create the base of the rectangular slide box. The tool loop cut was used to create lines along the border. Loop cut was used to create nine evenly-spaced grooves.
Figure 2: UV Mapping Images on Slides The top left image shows the histology of cat brainstem at 14.8 mm. The top right image shows the transparency of the cat brainstem at 14.8 mm outlining respiratory nuclei of interest. The edges of the slide were marked as seams and unwrapped in the UV layout window to the right. A rectangle automatically appears, which highlights area to be shown on the slide.
Figure 2: UV Mapping Images on Slides
The top left image shows the histology of cat brainstem at 14.8 mm. The top right image shows the transparency of the cat brainstem at 14.8 mm outlining respiratory nuclei of interest. The edges of the slide were marked as seams and unwrapped in the UV layout window to the right. A rectangle automatically appears, which highlights area to be shown on the slide.
Figure 3:  Identification of Relative Location of Slides in Brainstem Slides were lined up and plates shown in the Berman’s atlas, indicated by a certain coordinate, were moved to the right. The rectangular slide box was laid in juxtaposition to the slides of interest so that grooves could be made at the specific place.
Figure 3: Identification of Relative Location of Slides in Brainstem
Slides were lined up and plates shown in the Berman’s atlas, indicated by a certain coordinate, were moved to the right. The rectangular slide box was laid in juxtaposition to the slides of interest so that grooves could be made at the specific place.
Figure 4: Brain Stem Model Top halves of slides were modeled to resemble planes of brain stem
Figure 4: Brain Stem Model
Top halves of slides were modeled to resemble planes of brain stem
Figure 5: Extruded Nuclei Model Top right:  Side View of Extruded Nuclei Model Bottom right: Frontal View Top left: Slide 9 Extrusion View 1 Bottom left:  Slide 9 Extrusion View 2
Figure 5: Extruded Nuclei Model
Top right: Side View of Extruded Nuclei Model
Bottom right: Frontal View
Top left: Slide 9 Extrusion View 1
Bottom left: Slide 9 Extrusion View 2
Figure 6: Full Slide Box Frontal view of the 3D printed rectangular slide box showing the scaled rostral-caudal position of each slide depicting Berman’s atlas.
Figure 6: Full Slide Box
Frontal view of the 3D printed rectangular slide box showing the scaled rostral-caudal position of each slide depicting Berman’s atlas.
Figure 7: (L) 3D View of the location of Microinjections and types of responses in the brainstems (Solomon et al., 1999) (R) 3D Visualization of Fisher Linear Discriminant of microinjections and corresponding respiratory responses.  (Solomon et al., 1999)
Figure 7:
(L) 3D View of the location of Microinjections and types of responses in the brainstems (Solomon et al., 1999)
(R) 3D Visualization of Fisher Linear Discriminant of microinjections and corresponding respiratory responses. “Blob” 3D model was created based off of these data points.
(Solomon et al., 1999)

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