Introduction/Background

The plan in approaching the problem of dissecting the sheep's brain is the following: First, an examination of the external features of the specimen will be undertaken. Second, the brain will be bisected along the longitudinal midline, and the several features of the medial face of the hemisphere considered. Third, the brain will be sectioned in a coronal manner, and the deep structures of the brain studied from that point of view. (For your dissection you will use only the right hemisphere of the brain for your coronal sections).
Directional Terminology
This guide will use a number of terms that refer to position and direction in describing the brain. In quadrupeds (virtually everyone but us) directional terms are based on four basic reference points: head - up, tail - behind, back - top, and belly - down. In bipeds (e.g., humans), the neuraxis bends through a 90ø angle in the skull and human directional references shift with this bend. The directional terminology used here is largely appropriate for the anatomy of quadruped brains.
Most users of this software do not have an intrinsic interest in this species. Rather, the sheep brain serves as a less expensive alternative to the human brain. For their benefit, some poetic license has been taken in labeling certain structures and human terminology has been inaccurately incorporated. Our apologies to the purists. (You can appear sage by pointing out our errors.)

Directional Terms

Rostral, Anterior       Head or front end
Caudal, Posterior      Tail or hind end
Dorsal                     Back or top side
Ventral                    Belly or bottom side
 
Lateral                     Away from the midline
Medial                     Toward the midline
Proximal                  Closer
Distal                      Farther away


Three directional planes exist in the brain: rostral/caudal, dorsal/ventral, and medial/lateral. When sectioning (cutting) the brain, which planes are visible is determined by the type of section and two of these are used in this guide. In the sagittal section (which is made parallel to the midline, dorsal to ventral) the rostral/caudal and dorsal/ventral planes can be seen. In the coronal or cross section (made perpendicular to the midline, dorsal to ventral) the medial/lateral and dorsal/ventral planes can be seen.
Subdivisions of the Brain
Forebrain
Telencephalon: cerebral cortex, corpora striata (caudate nucleus, internal capsule, putamen), and rhinencephalon (olfactory brain, e.g., olfactory bulb, hippocampus, amygdala, septal region, and cingulate cortex)
Diencephalon: thalamus, hypothalamus, pineal gland, and posterior lobe of the pituitary (a migrated portion of the hypothalamus)
Midbrain
Mesencephalon: corpora quadrigemia (tectum--inferior and superior colliculi), tegmentum
Hindbrain
Metencephalon: cerebellum, pons
Myelencephalon: medulla oblongata
The Brain Stem is composed of the thalamus, hypothalamus, midbrain, pons and medulla.

Dissection Instructions:

On the ventral side of the brain, about midway between rostral and caudal ends, locate the pituitary gland. It is a midline mass of tissue lying caudal to the optic chiasm. If your brain still has all of its membranes intact, a number of types of tissue should be visible. For example, the grayish, semi-transparent, tough membrane is the dura mater, while the dark brown, spongy material on either side of the pituitary are capillary beds. Rostrally, you may also find large globs of yellowish fat that are associated with the eyes. Buried in all this is the bulbous pituitary itself.

 

Dissection Instructions:

Carefully dissect connective tissues from the caudal aspect of the pituitary and gently lift the pituitary mass from its caudal end. You should be able to see the IIIrd (oculomotor) cranial nerve pair attached to the ventral surface of the brain, on either side of the midline. These nerves are fairly broad, but quite flat, and may be difficult to see if they are lying down directly on the brain. Directly on the midline, anterior to the oculomotor nerves, you may find the thin stalk of the pituitary (the infundibulum) which connects the body of the pituitary to the base of the brain. Keeping the pituitary lifted away from the ventral surface of the brain, use a pair of small scissors to sever the two nerves (III) and the infundibulum as far away from the brain as you can. Carefully interrupt any other connective tissue present, lift the pituitary away, mark the caudal or rostral aspect of the pituitary (you'll forget), and set it aside. You can examine it later to see the difference between its anterior (rostral) and posterior (caudal) lobes. Continuing with the ventral aspect of your specimen, at its very rostral limit, locate the two light colored pad-like flaps of tissue which are the olfactory bulbs. Caudal to the olfactory bulbs, with a little removal of dura and fat, you should be able to find the cut stumps of the optic nerves (II). Follow these back and you'll see that they blend into an "X" on the midline. The fused part of the X is the optic chiasm. Caudal to the chiasm are the optic tracts, which are part of the ventral surface of the brain.

Dissection Instructions:
Stay on the Ventral Surface--

Just caudal to the optic tracts, two very large bundles of fibers form the ventral surface of the brain. These are the cerebral peduncles. On the medial part of each you should find the oculomotor nerve (III) that we saw earlier. While carefully removing the membranes between the cerebral and cerebellar cortices, look up between them to find to see the fine spaghetti-like trochlear nerve (IV) as it proceeds ventralwards from its exit point on the dorsal aspect of the brain stem. The large trigeminal nerve (V) runs rostrally through this region. It is usually quite difficult to remove its protective membrane, consequently, it is often severed fairly close to the brain stem, leaving only a stump on the side of the pons.

 

Stay on the Ventral Surface--

Several cranial nerves can be seen exiting from the medulla. At its rostral end, you may be able to see the thin (and easily ripped off, see "Other View") abducens nerve (VI). The abducens exits from the trapezoid body (a thin transverse band of fibers on the surface of the brain just caudal to the pons, see Plate 8) at a point just lateral to the longitudinally coursing pyramidal tracts (see Plate 8). Lateral to the abducens is the more substantial facial nerve (VII). Lateral to this, tucked up just under the cerebellum, you may see the tuft that forms the vestibulocochlear nerve (VIII). Caudal to the vestibulocochlear nerve you may see another light tuft of nerves that are made up of the glossopharyngeal nerve(IX) blending into the more caudal vagus nerve (X). Caudal to this you may see the spinal accessory nerve (XI) which appears as a line of connected filaments running in a rostral-caudal direction (they may take on the appearance of a broad yard rake). The hypoglossal nerve (XII) exits in several branches from the lateral edge of the caudal medulla (see "Other View").

Dissection Instructions: A New Perspective--

Place the brain on its ventral surface. Look down from the top at the most ventrocaudal point of the cerebellum. If your thin arachnoid membrane is still in place (it isn't in Plate 6), note how the arachnoid forms the roof of a space, the cisterna magna, as it sweeps caudally from the cerebellum to the medulla. Carefully separate the caudal part of the cerebellum from the medulla; as you lift the cerebellum the arachnoid will rupture, and you should be able to see yet another membrane (or fragments of it); this is the tela chorioidea, forming the posterior roof of the 4th ventricle. Separate the cerebellum from the medulla until that membrane ruptures; the internal space revealed by this maneuver is the 4th ventricle. The caudal point at which the two sides of the tela choroidea come together is called the obex. This can be seen on the dorsal surface of the medulla and forms the caudal boundary of the 4th ventricle. Looking into the 4th ventricle, you may see some dark spongy tufts; these are pieces of choroid plexus.

Another Perspective--

Leaving the brain on its ventral surface, we will now look down over the rostral end of the cerebellum. Looking down from the top, careful bending of the cerebellum in a caudal direction will reveal a second cistern, the superior cistern, under the rostral surface of the cerebellum (not visible in this Plate). Looking further down the brain stem, you may be able to see the white membrane forming the rostral roof of the 4th ventricle, the anterior medullary velum. Rostrally, this membrane attaches to the caudal inferior colliculus. Caudally, it continues under the cerebellum.

Back to the Ventral Surface--

After you have located the cranial nerves, and the cisterns just described, carefully strip away any remaining dura and arachnoid from the medulla. On the ventral aspect of the medulla a number of surface features can be readily seen: Note the longitudinal ridges coursing immediately on either side of the midline (marked by the ventral median sulcus); these are the pyramidal tracts. At the rostral end of the medulla, locate the band of transverse fibers paralleling the pons that form the trapezoid body.

Use Ventral and Lateral Perspectives--

The very prominent swelling just anterior to the trapezoid body; this is the pons (L. bridge). Three cerebellar peduncles attach the Cerebellum to the brain stem. The large middle cerebellar peduncle (brachium pontis) can be seen from the lateral perspective.

Dissection Instructions: Another look between Cerebral and Cerebellar Cortices--

Looking down from the top, once again, gently pull back on the cerebellum to reveal the dorsal surface of the brain stem. Here you should be able to easily see the corpora quadrigemina, that is, the superior (rostral) and inferior (caudal) colliculi. The superior colliculi are considerably larger than the inferior. Collectively, this area is also called the tectum.

Ventral-Rostral Features--

Just rostral to the pons, the ventral surface of the brain is formed by the cerebral peduncles, which run up to the optic tracts (seen earlier). The IIIrd cranial nerves can be seen exiting from the peduncles. The interpeduncular cistern lies in the "V" on the midline where the caudal-medial aspects of the peduncles meet.

Ventral-Rostral Features--

Just anterior to the interpeduncular cistern, locate the small, but distinct, protuberance lying on the midline; this is the mammillary body, and it marks the caudal limit of the hypothalamus, as seen from the ventral approach. The rostral border of the hypothalamus is marked by the optic chiasm, and the lateral boundaries are the medial edges of the cerebral peduncles. The general outline of the hypothalamus from the ventral aspect, takes on something of a diamond configuration. The remainder of the diencephalon (i.e., the thalamus) cannot be seen without sectioning the brain.

Ventral-Rostral Features--

Still viewing the brain from the ventral aspect, notice the fairly large, relative smooth masses of cortical tissue just lateral to the cerebral peduncles, extending, at the caudal limit, from the lateral-most part of the pons rostrally to include the olfactory bulbs. This mass of tissue is the rhinencephalon. The large rhinal fissure marks the lateral boundary of this region. The larger part, beginning at about the rostral/caudal level of the optic chiasm and proceeding caudally is the hippocampal gyrus. Within this gyrus resides the amygdala and part of the hippocampus (not visible without sectioning the brain). Rostral to the hippocampal gyrus you will find the lateral olfactory gyrus. Running along the medial surface of the lateral olfactory gyrus, locate the lateral olfactory stria, a band of fibers originating in the olfactory bulbs and coursing caudally. The medial olfactory stria is seen as a fiber bundle coursing medially, disappearing into the longitudinal fissure just anterior to the optic chiasm.

Dorsal-Rostral Perspective--

The two cerebral hemispheres are separated by the longitudinal fissure. Each hemisphere is divided into four major lobes. The frontal lobes are limited caudally by the cruciate fissure. The area caudal to the cruciate fissure is the parietal lobe whose line of separation from the more posteriorly placed occipital lobe is ill-defined. The temporal lobe in the sheep is very little developed in contrast to primates. In the sheep, it is represented by a slight bulge superior to the hippocampal gyrus. The cruciate fissure is somewhat variable in the sheep, and sometimes difficult to locate. However, the superior frontal sulcus is easily located. Parallelling the longitudinal fissure, the superior frontal sulcus divides the frontal poles into approximately equal left- and right-halves, and, if traced caudally, it is seen to "T-end" into the cruciate fissure (see "Other View").

The Medial Face--

If you are very fortunate, you may be able to see the central canal of the caudal medulla and spinal cord as it moves rostrally and opens up under the cerebellum, becoming the 4th ventricle (cerebrospinal fluid actually tends to run caudally in the ventricular system). Under the rostral cerebellum, the relatively thick anterior medullary velum should be easily located, It forms the anterior roof of the 4th ventricle and the rostral limit of the 4th ventricle is marked by the attachment of the anterior medullary velum to the caudal inferior colliculus. The 4th ventricle is continuous with the cerebral aqueduct of the midbrain. Tissue dorsal to the middle of the aqueduct is the tectum, and that ventral to its middle, excluding the cerebral peduncles, is the tegmentum.
The tectum consists of the corpora quadrigemina and the underlying lamina quadrigemina; the latter can be seen as the cut surface immediately superior to the aqueduct, appearing somewhat like the end of a piece of plywood (which is a laminated structure). The cerebral aqueduct opens up into the 3rd ventricle (discussed further in Plate 16) which in turn is continuous with the two lateral ventricles that run out into each cerebral hemisphere.

Rostral Medial Face--

The relationship between the 3rd ventricle and the thalamus is somewhat unusual. Most of the medial portions of the two thalami are fused in a structure called the massa intermedia (appropriately named, if you remember your latin roots). Where this fusion exists, of course, there can be no ventricular space. Consequently, the 3rd ventricle must run around the massa intermedia. In the vicinity of the massa intermedia, the lateral walls of the 3rd ventricle are formed by the unfused medial nuclei of the two thalami. Ventrally the lateral walls of the 3rd ventricle are formed by the medial nuclei of the hypothalami. At about 2 o'clock from the massa intermedia (assuming the brain is horizontal with the ventral side up), locate the small piece of tissue that is just outside the 3rd ventricle. This is the pineal body. Looking at the most ventral part of the 3rd ventricle, you should be able to appreciate how it extends down into the hollow stalk attaching the pituitary and hypothalamus (the infundibulum). Just caudal to this region, you will find the cut surface of the mammillary body. Just rostral to this region, you will find the cut surface of the optic chiasm. The corpus callosum is the very prominent collection of axons that extends for some distance along the medial face of the cerebral hemispheres. At the rostral end, it curves ventrally and caudally, virtually making a 180ø turn. The area of turning is called the genu, and if you look carefully (with a perfectly cut brain), you will see that the caudal extending portion of this bend comes to a point and ends; this is the rostrum. At the caudal end of the corpus callosum it can be seen that a similar 180ø turn is made; this bend is the splenium. The main "body" of the corpus callosum runs between these two turns.

Callosal Sulcus: The sulcus on the medial surface of the cerebral hemisphere which separates the corpus callosum from the cingulate gyrus.

Rostral Medial Face--

Immediately dorsal to the body of the corpus callosum (i.e., that part between the splenium and genu) locate the crease that forms the callosal sulcus. The cortical outfolding just dorsal to the callosal sulcus is the cingulate gyrus, which, in turn, is bounded dorsally by a crease, the cingulate sulcus. Follow the cingulate gyrus caudally, and you will find that it courses ventrally and laterally, eventually becoming continuous with the hippocampal gyrus that we saw on the ventral surface of the rostral brain. Just inferior to the genu and rostrum of the corpus callosum is a region of medial face cortex called the septal area. As you will see in cross-sections, this region does not extend very deeply from the midline. At the caudal edge of the septal area, you should be able to locate the small, light-colored dot which is a cross-section of the anterior commissure. Just inferior to the attachment point of the pineal body, there is a much smaller (you won't be able to see it here, but it may appear in cross-section) bundle of decussating fibers called the posterior commissure. If the bisection of the brain was exactly on-center, there will be a membrane extending ventralwards from the body of the corpus callosum, the septum pellucidum, which separates the two lateral ventricles from each other. The fornix is a bundle of fibers that parallels the corpus callosum through much of its course. You can find the body of the fornix lying about midway between the massa intermedia and the body of the corpus callosum. You'll get a better look at the fornix in Plate 18.

Time for a Little Digging--

You are about to scrape away part of the medial face of one of your brain halves. Choose your half wisely. If your cut is off center, be certain that the structures we are interested in are still in the half you choose to scrape, they don't lie far from the midline. The most caudal part of the fornix is made up, in part, of decussating fibers called the hippocampal commissure (also known as the commissure of the fornix), which interconnect the two hippocampi. The more rostral part of the structure is made up of fibers largely originating in the hippocampi, paralleling the midline, and these are called the body of the fornix. Just dorsal to the anterior commissure, the fibers of the body of the fornix separate into two distinct bundles of fibers which move just lateral to the midline, forming the columns of the fornix. Some cell bodies in the mammillary body send their axons to the dorsal anterior thalamus through the mammillothalamic tract, which lies just beneath the medial face of the hemisphere. Use a scalpel, and, holding the blade at a right angle to the medial face of the hemisphere, very carefully scrape away the tissue between the anterior commissure and the mammillary body in order to uncover the column of the fornix, which will appear as a distinct white line arcing ventrocaudally from the body of the fornix. Next, do the same thing in the area between the mammillary body and the dorsal anterior part of the massa intermedia, uncovering the mammillothalamic tract.

The Tree of Life (Internal Structure of the Cerebellum)--

Note the internal structure of the cerebellum. The thick core of myelinated axons branching out into separate folia, reminiscent of a tree (arbor vitae), frosty window pane (for those of you in northern latitudes), or cauliflower florets (any analogy to food is a good analogy)

 

Now you get to do some serious slicing. You will be making a series of coronal sections using one of your hemi-brains. Typically, you should use the half that you haven't scraped to see the mammillothalamic tract and columns of the fornix . Once again, you should minimize cutting artifacts by making one clean pass through the tissue, rather than sawing through it. You might also consider the following: As you are asked to make sections of the brain, rather than attempting to make the section at the exact level in one single cut, make a number of thinner cuts until the cut-face of your specimen resembles the relevant figure. If you are skilled at cutting you can end up with a deck of brain slices each about as thick as a robust slice of bologna (don't shuffle this deck). Plate 20 shows the results of a cut that passes through the genu of the corpus callosum. Try to have the angle of your cuts match that of the sagittal section that is shown prior to each coronal section. Substantial deviations from this angle will make it somewhat challenging for you to match our plates with your sections. Looking at the cut surface of your first section, the darker material is mostly cell bodies, and the lighter tissue is mostly axons. Working from the ventral aspect of your section, look laterally until you find the first infolding, this is the rhinal fissure. To appreciate the full extent of this fissure, either look at the ventral lateral surface of your uncut hemi- brain or look back at Plate 13. Obviously, you should be able to see the rhinal fissure in many of your coronal sections, we will only label it here, however. Impress your friends by finding it elsewhere. Just medial to the rhinal fissure you may be able to locate the lateral olfactory stria. Moving to the medial face of the ventral part of your section, the gray matter here is the septal region. You should have no trouble locating the corpus callosum lying under the ventral surface of the medial cerebral cortex. As you did for the rhinal fissure, either look at the uncut hemi- brain (medial face) or look back at Plate 16 to appreciate the full extent of the corpus callosum. Obviously, the corpus callosum will show up in many of your coronal sections, we have only labeled it here. You may also see part of the rostrum of the corpus callosum as it extends ventrally and caudally from the genu. The thin space above the corpus callosum is the callosal sulcus, the cortex above that is the cingulate gyrus , and the space above that is the cingulate sulcus. We have only labeled them here, but, as you would expect, these structures will occur in many of your crossections (Examine either the medial face of the uncut hemi-brain or Plate 17). Off the midline, the space below the corpus callosum is lateral ventricle. The caudate nucleus forms the ventral wall of the 3th ventricle. Just lateral and ventral to the caudate nucleus, find the stippled band of white and gray matter. The white matter is the internal capsule. Ventral and lateral to the internal capsule you should be able to see the putamen (a nuclear group). This arrangement of nuclei and fibers (caudate nucleus, internal capsule, and putamen) gives the area a striated appearance, and thus led to terming this group the corpus striatum. Immediately lateral to the putamen, you should be able to see a solid streak of white matter, the external capsule. Just lateral to this, typically, you can see the triangularly shaped claustrum. The caudate, internal capsule, putamen, external capsule, claustrum and septal area will appear in subsequent sections of this guide, however they will not always be labelled.

 

Notice how the septal area has become larger on the ventral medial face of the brain. The caudate is still prominent and the internal capsule should be easier to see. You may be able to see the beginning of the globus pallidus, which is contiguous with the putamen on its ventral medial face. The globus pallidus has a lighter and more homogeneous coloration than the putamen. If you had trouble seeing the claustrum in the last plate, it should be clear here.