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LABORATORY 9 Nervous System Histology Objectives 1. Identify the following portions of a multipolar neuron using a diagram, a model or a prepared slide: cell body (soma, perikaryon) nucleus Schwann cell myelin sheath axon collateral axon dendrite Nissl bodies axon hillock node of Ranvier neurofibril neurilemma telodendria (terminal branches) axonal terminals (synaptic knobs, boutons) 2. Differentiate between pseudounipolar neurons, bipolar neurons and multipolar neurons using diagrams, models and prepared slides. 3. Identify the following components of a cross sectioned nerve using diagrams and prepared slides: myelin sheath, nerve fibers, fascicles, endoneurium, perineurium and epineurium Introduction The functional unit of the nervous system is the neuron, a cell that is capable of generating and propagating electrical signals in the form of action potentials. Neurons can be found in the central nervous system (brain and spinal cord) and in the nerves of the peripheral nervous system. All neurons have three essential components: a cell body (soma), one or more dendrites and a single axon. Neurons can be structurally classified as unipolar (having a single projection from the cell body), bipolar (having two projections from the cell body) of multipolar (having many projections from the cell body). Neurons are supported structurally and functionally by cells called neuroglia (glial cells). An example of a glial cell is a Schwann cell which produces an insulating myelin sheath for the neurons of the peripheral nervous system. A nerve is a bundle of axons that is found in the peripheral nervous system. These axons are bundled into fascicles and are supported by a number of connective tissue wrappings. The endoneurium is a delicate layer of loose connective tissue that surrounds individual axons within a fascicle and electrically insulates each axon. The 105 perineurium, a coarser connective tissue wrapping, surrounds the fascicles within the nerve. The entire nerve is surrounded by an epineurium, which is a strong, dense layer of fibrous connective tissue. Materials 1. Each student should have a compound microscope. 2. Each pair of students should have: Lens paper Immersion oil Lens cleaner Box of prepared slides Colored pencils 3. Class materials to be shared by students: Three dimensional models of neurons Charts of nervous system histology Activity 1: Resources: The Structure of Multipolar Neurons Textbook: Photographic Atlas: pages 140, 390 – 393 page 27 (Figures 3.41) page 28 (Figures 3.44, 3.45) page 102 (Figures 9.2, 9.3) 1. Identify the following components of a multipolar neuron on a three dimensional model and on diagrams: cell body (soma, perikaryon) nucleus Schwann cell myelin sheath axon collateral axon dendrite Nissl bodies axon hillock node of Ranvier neurofibril neurilemma telodendria (terminal branches) axonal terminals (synaptic knobs, boutons) 1. Observe a prepared slide of an ox spinal cord smear. Select one neuron to study and identify the cell body, nucleus, nucleolus and Nissl bodies. Try to differentiate between the axon and dendrites. Sketch the neuron on the laboratory worksheet. Note that the cytoplasm is coarsely textured with dark staining Nissl bodies (rough endoplasmic reticulum). A large nucleus contains a prominent “Owl’s eye” nucleolus. A thin axon emerges from the cell body at a conical shaped axon hillock. Both the axon and axon hillock lack ribosomes and may be recognizable because of their pale staining cytoplasm. It 106 is also possible that the axon is attached to the top or the bottom of the cell body and is therefore not in your field of view. 2. Observe a prepared slide of myelinated nerve fibers. Identify the nodes of Ranvier, axon (axis cylinder), Schwann cell nuclei and myelin sheath. Sketch the nerve fiber on the laboratory worksheet. Activity 2: Resources: Structural Classification of Neurons Textbook: Photographic Atlas: pages 393-395; 550 page 117 (Figure 11.4) page 118 (Figure 11.6) 1. Observe prepared slides of pyramidal cells (cerebral cortex) and purkinje cells (cerebellum), and make sketches on your lab worksheet. Both are examples of multipolar neurons of the central nervous system. Pyramidal cells were named because of the shape of their cell body in specimens’ sectioned perpendicular to the brain surface. The apex of the pyramid points to the brain surface. Multiple dendrites emerge from the apex and the corners of the pyramid. A single axon emerges from the base and travels deep into brain tissue. Purkinje cells are located in the cortex of the cerebellum between its granular and molecular layers. Purkinje cells have large cell bodies, a massive array of finely branching dendrites that extend towards the surface, and a single small axon that extends into deeper portions of the cerebellum. 2. Observe a prepared slide of the retina (eye) and locate the bipolar cells. Make a sketch on your lab worksheet. Note: It is fairly easy to identify the bipolar cell nuclei but more difficult to distinctly see the axons and dendrites attached to the cell bodies. 3. Observe a prepared slide of a spinal cord section and locate the dorsal root ganglion. Located within the dorsal root ganglion are the cell bodies of pseudounipolar neurons. Make a labeled sketch on your laboratory worksheet. Note: The cell bodies of these neurons are large, rounded with pale cytoplasm and prominent nuclei and nucleoli. The cell bodies are surrounded by a layer of satellite cells. Activity 3: Resources: Nerve Histology Textbook: Photographic Atlas: pages 490-491 page 27 (Figures 3.40, 3.41,3.43) Observe a prepared slide of a nerve in cross section and identify the following: axon, myelin sheath (if visible), endoneurium, fascicle, perineurium and epineurium. Make a labeled sketch on your lab worksheet. 107 Checklist: Neuron Structures Cell body (soma, perikaryon) Model/Diagram ____________ Prepared Slide ____________ Nissl bodies ____________ ____________ Nucleus ____________ ____________ Nucleolus ____________ ____________ Dendrites ____________ ____________ Axon Hillock ____________ ____________ Axon ____________ ____________ Schwann cell ____________ ____________ Myelin sheath ____________ ____________ Node of Ranvier ____________ ____________ Axon collateral ____________ NA Neurofibril ____________ NA Neurilemma ____________ NA Telodendria (Terminal branches) ____________ NA Axon terminals (synaptic knobs, boutons) ____________ NA A. Structural Classes of Neurons ____ Multipolar neuron (Pyramidal cell) ____ Multipolar neuron (Purkinje cell) ____ Bipolar neuron (retina) ____ Pseudounipolar neuron (dorsal root ganglia) B. Peripheral Nerve Anatomy ____ axon ____ endoneurium ____ fascicle ____ perineurium ____ epineurium 108 Lab 9 Worksheet Name: Score: Neuron Total Magnification: ___________________ ___________________ Nerve Fiber ____________ Total Magnification: Label the nucleus, soma, nucleolus, and, if possible, axon and dendrites ________ Label the nodes of Ranvier, axon, and neurilemma Pyramidal Cell (mulitpolar neuron) Purkinje Cell (mulitpolar neuron) Total magnification: ___________ Label the soma, nucleus, processes Total magnification: _________ Label the soma, nucleus, processes 109 Retina (Bipolar neuron) Dorsal Root Ganglion (Unipolar neuron) Total magnification: _________ Label the bipolar cell layer Total magnification: __________ Label the soma and nucleus Nerve Cross Section Total magnification: ____________ Label the nerve fiber, endoneurium, fascicle, perineurium, and epineurium 110 Post lab worksheet lab 9 1. Label the structures of the neuron 2. Match the term with the description: a. b. c. d. e. neurofibril Schwann cell axon hillock dendrite Nissl bodies f. axon g. telodendria h. node of Ranvier i. axon terminal j. soma ____ rough endoplasmic reticulum found in the cell body; also called chromatophilic substance because of its affinity for basic dyes ____ distal knoblike endings of terminal branches that store neurotransmitter ____ neuronal process that acts as a receptive area for incoming stimuli ____ neuroglial cell that surrounds and forms myelin around larger nerve fibers in the peripheral nervous system ____ biosynthetic center of a neuron; contains the nucleus, nucleolus and ribosomes ____ terminal branches of an axon ____ gaps in a myelin sheath ____ bundles of intermediate filaments that, along with microtubules, help to maintain the shape of a neuron ____ the cone shaped area of the cell body that gives rise to an axon ____ neuronal process that generates action potentials and propagates them, typically away from the cell body 111 3. What is a nerve? _________________________________________________________ _________________________________________________________ _________________________________________________________ _________________________________________________________ 4. Match the term with its description. Each term can be used more than once. a. mulitpolar neuron b. bipolar neuron c. unipolar neuron (pseudounipolar neuron) ____ sensory neurons of the peripheral nervous system in which a single process is attached to the cell body; the process divides to form a peripheral process and a central process ____ major type of neuron in the central nervous system ____ has two processes attached to the cell body, a single axon and a single dendrite ____ has a single axon and two or more dendrites attached to the cell body ____ act as receptor cells for special sense organs, such as the eye and nose ____ act as motor (efferent) neurons, transmitting impulses away from the central nervous system to effectors 5. What is the location of each nerve connective tissue layer listed below? Endoneurium: _________________________________________________ Epineurium: _________________________________________________ Perineurium: _________________________________________________ 112