Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.1 Lab #4 - Biological Sciences 102 – Animal Biology This lab is designed to introduce you to multicellular animals and their basic tissue structure with particular reference to the simplest of the metazoans–the sponges of the Phylum Porifera. Metazoans The diversity seen among the unicellular protozoa is a product of their various subcellular structures and organelles. In contrast, the complexity of metazoans or multicellular animals is due to the evolution of cells working together to form larger units each dedicated to a specific function necessary for the survival of the entire animal. Metazoan cells cannot survive on their own outside of the whole organism (except in cell culture with lots of feeding and maintenance by biologists). The simplest metazoans such as the sponges are collections of cells that show some division of labor, but less distinction in the tasks of specific cells seen in more complex metazoans. Animals that have true tissues possess cells that work together as a highly coordinated unit. In most animals, these tissues are further organized into organs. Metazoans are part of the opisthokont lineage (the eukaryotic clade that has a single posterior flagellum, if a flagellum is present) along with the fungi, choanoflagellates and microsporidians. Microsporidians are protozoan parasites living inside the cells of many animals including humans. Choanoflagellates are solitary and colonial protozoans in which each cell has a flagellum surrounded by a collar of microvilli. Choanoflagellates closely resemble the feeding cells (choanocytes) seen in sponges. Sponges are basically aggregations of cells held together by an extracellular matrix. Biologists are currently studying the cell-to-cell interactions and signaling mechanisms that lead to colony formation in choanoflagellates in order to better understand the evolution of the first simple multicellular animals. See and study the textbook regarding the syncytical ciliate hypothesis and the colonial flagellate hypothesis (look them up in the textbook index). Phylum Porifera: The Sponges While lacking true tissues, sponges have a cellular level of organization. There is division of labor among their cells, but there are no organs, no systems, no mouth or digestive tract, and only the hints of nervous integration. There are no germ layers (ectoderm, mesoderm or endoderm). Adult sponges are all sessile in form. Some have no regular form or symmetry; others have a characteristic shape and radial symmetry. Sponges may be either solitary or colonial. See lab exercise #7 on sponges in the Hickman lab manual for diagrams and photos. Important characteristics of sponges are: Sponges do not possess true tissue organization possess pores and canal systems flagellated sponge feeding cells, called choanocytes, which line their cavities and create currents of water internal skeletons of spicules and/or protein fibers (spongin). internal cavity (spongocoel) that opens to the outside by an osculum. most sponges are marine, though there are a few freshwater species; freshwater forms are found in small, slimy masses attached to sticks, leaves, or other objects in quiet ponds and streams Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.2 Lab #4 - Biological Sciences 102 – Animal Biology Classification of the Phylum Porifera Class Calcarea Cal-ca're-a (Gr. calc, limy). About 700 species; sponges with spicules of calcium carbonate, needle-shaped or three-rayed or four-rayed; canal systems asconoid, syconoid, or leuconoid; all marine. Examples: Sycon, Leucosolenia Class Hexactinellida (hex-ak-tin-el'i-da) (Gr. hex) six, + aktis, ray). About 500 species; sponges with three-dimensional, six-rayed siliceous spicules; spicules often united to form network; body often cylindrical or funnel-shaped; canal systems syconoid or leuconoid; all marine, mostly deep water. Examples: Euplectella (Venus' flower basket), Hyalonema Class Demospongiae (de-mo-spun'je-e) (Gr. demos, people, + spongos, sponge). About 7000 species; sponges with siliceous spicules (not six-rayed), spongin, or both; canal systems leuconoid; one family freshwater, all others marine. Most sponges belong to this class. Examples: Spongilla (freshwater sponge), Spongia, (commercial bath sponge), Cliona (a boring sponge) Your instructor will review the basic structure of sponges including asconoid, syconoid and leuconoid canal systems, cell types found in sponges such as choanocytes, their skeletons and basic reproduction. Class Calcarea Class Demospongiae Class Hexactinellida Types of Animal Tissues & Histology (the study of tissues) See lab exercise #4 on animal tissues in the Hickman lab manual for diagrams of tissues. As we proceed through the various animal phyla, we will be discussing most of these tissues in lab and lecture. You are not responsible for knowing all of this material now, but you must know the four basic types of tissues, their basic characteristics and an example of each. You will want to use these pages as reference material during the course. There are four basic types of tissues that are recognized in animals. Some animals show less distinction between the four separate types. connective tissue epithelial tissue muscular tissue nervous tissue CONNECTIVE TISSUES In general, connective tissue is quite diverse and is not as cellular as other types of tissue. Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.3 Lab #4 - Biological Sciences 102 – Animal Biology All connective tissues arise from the mesoderm layer (mesenchyme) of the developing embryo. Connective tissue provides a stiffness to the animal body (bone) and helps the body withstand tension, stress and shearing (twisting) forces. Basically, connective tissue is composed of: 1. An intercellular matrix (in the intercellular space between connective tissue cells) This matrix can be fluid as in blood or it can be more solid as in bone. The matrix is a mixture of proteins that are secreted by the cells in the connective The proteins in the matrix are usually fibrous (long strands). tissue. 2. Connective tissue cells These cells secrete the intercellular matrix and can have a variety of functions. For instance, the red blood cells (erythrocytes) & white blood cells (leukocytes) in blood, the osteocytes of bone, or the adipocytes in fat tissue. Often some of these cells make new connective tissue (blast cells) while other cells degrade the connective tissue (clast cells). CONNECTIVE TISSUE EXAMPLES: Dense Connective Tissue: function = tightly packed fibers that are aligned with the forces applied to the tissue; makes this a strong, somewhat flexible tissue found in tendons, aponeuroses, elastic tissues and ligaments Tendons = cords of dense regular connective tissue that attach skeletal muscles to bones Aponeuroses = collagenous sheets or ribbons that resemble flat tendons; these often cover the surface of some muscles Elastic tissue = tolerates expansion and contraction; found in the walls of blood vessels and respiratory passageways; often underlies transitional epithelia Ligaments = resemble tendons, but they connect bone to bone; ligaments have significant amounts of elastic as well as collagenous fibers TYPES AND CHARACTERISTICS OF CONNECTIVE TISSUE There are two different types of Dense Connective Tissue: 1. Dense Regular Connective Tissue = has the collagen fibers aligned tightly in a parallel fashion There is a grain or orientation to dense regular connective tissue (such as tendon) 2. Dense Irregular Connective Tissue = has the collagen fibers that run in many directions and there is no particular orientation to the fibers which makes this tissue strong in several directions (such as skin) Structure Mostly collagenous fibers (collagen) bundled like fibers in a rope = strength Histology = mostly fibers, may be “wavy” in appearance (regular) or “cracked” in appearance (irregular) Cells = fibroblasts which secrete the fibers and become fibrocytes Fibers = collagen fibers Loose (Areolar) Connective Tissue: function = wraps organs, occurs between some muscles in sheets, occurs at the boundaries of tissues such as between fat and muscle Histology = large, pink stained collagen fibers; smaller & darker elastic fibers and a collection of cells in a relatively random collection; looks like handfuls of straw that have been thrown into the air and allowed to fall in a random arrangement; loosely packed Cells = fibroblasts, fibrocytes, mast cells, macrophages (the last 2 are types of immune cells) Fibers = large collagen fibers with smaller elastin fibers Elastic Connective Tissue: function = occurs in the walls of arteries Histology = looks like dark squiqqly lines in the walls of arteries Cells = fibroblasts & fibrocytes Fibers = elastic fibers made of the protein elastin Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.4 Lab #4 - Biological Sciences 102 – Animal Biology Reticular Connective Tissue: function = found in internal organs such as the liver, spleen and lymph nodes; reticular connective tissue provides an internal framework for the organ Histology = if the slide is stained with silver, the reticular fibers will appear black, if the slide is stained with Masson stain, the reticular fibers will appear blue; look for branched fibers among small round cells Cells = fibroblasts & fibrocytes Fibers = reticular fibers Adipose Tissue (fat): function = very cellular; store large amounts of lipids (fats); the nucleus of the cell and the cytoplasm remain on the outer part of the cell near the cell membrane Histology = in prepared slides, the fat has typically been dissolved during preparation. You should see large, empty cells with some cell nuclei near the edge of some cells; may look like a honeycomb Cells = adipocytes (fat cells) Fibers = few fibers if any at all Osseous Tissue (vertebrate bone): function = support, rigidity, protection, attachment point of muscles for movement, cavity for the formation of blood cells (erythrocytes & leukocytes). The exoskeleton of arthropods is comprised of proteins and the polysaccharide chitin. Structure In vertebrates, the matrix of bone consists of: about 30% collagen (protein) = flexibility and tensile strength about 70% calcium and phosphate crystallized into bone salts = hydroxyapatite = makes bone rigid Osteocytes produce the cartilaginous matrix of bone, bone is a physiologically active, living tissue with its own blood supply that allows bones to heal relatively quickly; bone is constantly reworked and reformed according to your body’s needs Histology = dense bone looks like tree rings while trabecular (spongy) bone looks like scaffolding with spaces in it Cells = osteocytes = cells found in mature bone that arose from osteoblast cells osteoblasts = bone forming cells that produce new bone tissue osteoclasts = bone degrading cells (“bone eating” cells); large, multi-nucleate cells that make the medullary cavity (hollow inside of long bones) and degrade bone to release calcium and/or phosphate as needed Fibers = collagen fibers Microscopic View of Vertebrate Bone - Osseous Tissue formed of Osteons - Cartilage (and Perichondrium) = a relatively simple, structural connective that is nonvascular (without a direct blood supply) and grows relatively slowly tissue Structure Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.5 Lab #4 - Biological Sciences 102 – Animal Biology Consists primarily of intercellular matrix with cells chondrocytes (chondro = from the Greek for grain = cartilage) interspersed in the matrix within small pockets called lacunae. Cartilage is a pliable material that allows for movement. The intercellular matrix is made up of protein and carbohydrate that forms a gel in which fibers and cells are found. The perichondrium is dense irregular connective tissue that is found on the surface of cartilage. Blood vessels run through the perichondrium and nutrients diffuse into the chondrocytes, so if cartilage becomes damaged, it heals slowly as there is no direct blood supply to each chondrocyte. There are three types of cartilage in the human body as described below. Function of cartilage Cartilage in the joints provides a smooth surface through the full range of movement during which it is continuously subjected to severe stresses such as the pull of gravity in weight bearing joints and muscle tension in all joints Hyaline Cartilage: Function = most common type in the body; found at the apex of the nose, the ends of many long bones within joints, between the ribs and sternum, the larynx Structure Glassy and clear in appearance in fresh tissue; hyaline cartilage is like crunchy gristle Histology = often appears clearish with a pinkish tint in prepared slides; fibers are not obvious as they blend into the matrix; chondrocytes are often found in pairs; relatively large lacunae Cells = chondrocytes are the cells that secrete the proteins that make up cartilage Fibers = collagenous fibers (fibers made of collagen). Chondrin is a major protein component of collagen Elastic Cartilage: Function = flexible with resilience, it can be flexed but it will “bounce back”; forms the pinna of the ear (earlobe), epiglottis, parts of the nose Structure Made from chondrocytes with proportionally less chondrin protein than hyaline cartilage; contains a lot of the protein elastin that is a coiled protein at the molecular level. Elastin has fibers that run in random directions to help provide elasticity. Histology = if stained with silver then the elastic fibers appear black Cells = chondrocytes Fibers = mainly elastin fibers Fibrocartilage: Function = similar to hyaline cartilage with more collagenous fibers (collagen); fibrocartilage can withstand much tension, but it still has some flexibility; located in areas with greater stress such as the intervertebral disks, pubic symphysis, and the menisci of each knee Structure lots of collagen; collagen can be bent, but it resists stretching (as the fibers tend to run parallel and are densely interwoven = durable and tough) Histology = collagen fibers are relatively easy to see; chondrocytes are often found in rows of four or five; lacunae are less obvious Cells = chondrocytes Fibers = mainly collagen fibers Blood (vertebrates)/Endolymph (many invertebrates) = is composed of four major portions: erythrocytes, leukocytes, thrombocytes (platelets) and plasma (noncellular fluid portion of blood); blood will be covered in much greater detail in later lectures Structure Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.6 Lab #4 - Biological Sciences 102 – Animal Biology Matrix is fluid (mostly water) with no fibers present Cells = erythrocytes (red blood cells) & leukocytes (white blood cells) Histology = pink discs = RBCs; darker purple = nuclei of leukocytes Erythrocytes (Red Blood Cells) of various vertebrate classes Note that mammals have red blood cells which lack a cell nucleus while all other vertebrates (fish, amphibians, reptiles and birds) have erythrocytes that possess a cell nucleus EPITHELIAL TISSUE Epithelial tissue is highly cellular tissue that forms coverings and linings of an animal’s body. Epithelial tissue attaches to underlying layers by way of a basement membrane that is a non-cellular adhesive layer. For instance, in the skin, the epidermis is made of epithelial tissue that is attached to the underlying dermis by a basement membrane. The basement membrane consists of glycoproteins and both fine and coarse protein filament which are produced by connective tissue cells. Epithelial tissue is classified according to the shape of the cells and the number of cell layers present. Epithelial cell shapes = squamous (flattened), cuboidal, or columnar Simple epithelium is only one cell layer thick Stratified epithelium is more than one cell layer thick Simple Squamous Epithelium Location in the animal body: air sacs of the lungs, lining of blood vessels, many body membranes, part of the kidney tubules and nephridia Structure: one layer of flat cells that lie on the basement membrane Function: provides a smooth, slippery surface to reduce friction and allows for diffusion of substances during absorption or secretion Simple Cuboidal Epithelium Location in the animal body: forms many of the major glands and ducts, lines tubules (such as sweat ducts) and tubules of the kidney and nephridia Structure: one layer of tube-like or pie-like shaped cells (sort of like squashed cubes) that are fairly uniform in diameter attached to the basement membrane; often involved in the secretion of fluids (sweat, oil) or in filtration (as in the kidney) Simple Columnar Epithelium Location in the animal body: ciliated type lines the oviducts; non-ciliated type lines the digestive tract Structure: one layer of tall column-like cells attached to the basement membrane; the cells may be ciliated or non-ciliated Function: assists in the movement of substances through tubes in the body and plays vital roles in the process of absorption in the intestines; protection, absorption, and secretion cilium (= singular; cilia is plural) = a slender organelle that extends above the free surface of an epithelial cell, and usually undergoes coordinated cycles of movement. Cilia possess the same 9 + 2 arrangement of microtubules seen in flagella, but cilia are shorter and more numerous. Cilia are not a type of tissue, but a cell organelle seen in the cells of some tissues. Pseudostratified Ciliated Columnar Epithelium Location in the animal body: lines portions of the respiratory tract and the male reproductive tract Structure: one layer of ciliated columnar cells attached to the basement membrane, although it may look like it is formed from multiple cell layers Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.7 Lab #4 - Biological Sciences 102 – Animal Biology Function: produces a mucus and traps small particles which are carried out of the lungs via coordinated movements of the cilia Stratified Squamous Epithelium Location in the animal body: surface of the skin, lines the female reproductive system, mouth, throat and anus Structure: multiple layers of flat cells attached to the basement membrane; may have cuboidal cells at the basement layer, but the cells along the free edge are flattened; these flattened cells may be keratinized or non-keratinized; keratin is a tough protein that hardens cells in the outer layers of the skin, hair and nails Function: multiple cell layers protect underlying layers from abrasive, pathogenic, or chemical damage Stratified Cuboidal Epithelium (multiple layers of cuboidal cells) is relatively rare, but provides for some protection, secretion and absorption in some ducts (e.g. sweat gland ducts) Stratified Columnar Epithelium (multiple layers of columnar cells) is relatively rare, but provides for some protection in salivary gland ducts, the epiglottis, mammary ducts and the urethra Transitional Epithelium Location in the animal body: lines the renal pelvis, urinary bladder and ureters in vertebrates Structure: multiple layers of irregularly shaped cells lie on top of the bladder, but the cells along the free edge are not flattened (as in stratified squamous epithelium); when the bladder fills, the cells along the free edge flatten out Function: the cell structure and multiple layers have an unusual amount of stretching capacity that allow for significant changes in the volume of portions of the urinary tract MUSCLE TISSUE Muscle tissue is a highly cellular tissue characterized by its ability to contract in response to an appropriate excitatory stimulus. Muscle fibers are individual muscle cells, each of which is contractile and can shorten in length due to the sliding of protein filaments (actin, myosin & other proteins) along one another within the cell. Skeletal Muscle Fibers Note that skeletal muscle fibers are striated (“striped”) and multinucleate (there are multiple cell nuclei per cell membrane) Three Types of Muscle Tissue = Skeletal, Cardiac, and Smooth Skeletal Muscle (striated, voluntary, multinucleate) Location in the animal body: all the muscles which are used voluntarily to control movement; this is the type of muscle that moves bones (in both arthropods and vertebrates) and is found near all the joints of the skeletal system Structure: striated due to the organization of the protein filaments in the muscle fibers; skeletal muscle cells form a syncitium, that is, they are multinucleate (more than one nucleus within a single cell membrane) Function: modulate all voluntary muscle action & movement Cardiac Muscle (striated, involuntary, uninucleate) Location in the animal body: only found in the heart and heart-like structures; makes up most of the heart; involuntary Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.8 Lab #4 - Biological Sciences 102 – Animal Biology Structure: striated, but the striations are less regular and obvious as compared to skeletal muscle; cardiac muscle cells are usually uninucleate (one nucleus per cell); individual cells are connected to each other by intercalated discs that hold the cells together and allow for electrical communication between certain cells of the heart Function: provides for the involuntary pumping of blood by the heart Smooth Muscle (nonstriated, involuntary, uninucleate) Location in the animal body: found in the digestive tract, reproductive tracts and other internal organ systems; involuntary Structure: nonstriated, spindle-shaped uninucleate cells with a centrally located fusiform shaped cell nucleus when the cells are relaxed; when the cells are contracted, the cell nucleus has a corkscrew shape Function: provides for the involuntary movement (peristalsis) of food products through the digestive tract, the movement of sperm and egg in the female reproductive tract, the movement of excretory products, the activation of pili muscles in the skin, movements of sensory organs, the contractions of the uterus at parturition (labor), etc NERVOUS TISSUE Nervous tissue is a highly cellular tissue characterized by an ability to conduct electrochemical signals. The individual cells are termed neurons. Each neuron is composed of dendrites (which receive the incoming signal), a cell body (or soma where the cell nucleus is found), an axon (a long cytoplasmic extension, “like a wire”), and a terminal (or synaptic knob) near the synapse where one neuron conveys the impulse to another neuron. There are many different shapes of neurons found in the animal kingdom. Location in the body: the nervous system or nerve net of an animal Structure: composed of neurons and also contains special support and nursing cells called glial cells (or neuroglia) or similar cell types. Function: conduct electrochemical impulses that allow the nerve net, cerebral ganglia or brain and spinal cord to communicate with other parts of the animal’s body A typical multipolar nerve cell (neuron) Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.9 Lab #4 - Biological Sciences 102 – Animal Biology LAB PROCEDURE NAME: LAB SCORE: Organization of a Syconoid Sponge Using your textbook and/or the Internet, label the diagram below with the following terms: osculum (surrounded by long protruding spicules) flagellated canal (radial canal) incurrent canal spongocoel ostium With a colored pen or pencil, indicate (with arrows) the path of water flowing into and out of the sponge. Grantia sp. as seen under a dissecting scope Examine a prepared slide of a Grantia sp. cross-section under the compound scope and compare it to the diagram below. Be sure you can identify the basic parts indicated on page 4.10. Metazoans, Animal Tissues & Sponges: Phylum Porifera Lab #4 - Biological Sciences 102 – Animal Biology What are the evolutionary and functional advantages of being multicellular? 4.10 Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.11 Lab #4 - Biological Sciences 102 – Animal Biology Using your textbook and/or the Internet, label the diagram below with the following terms: ostium flagellated canal (radial canal) prosopyle apopyle mesohyl (mesenchyme) Sponge Skeletons: Spicules incurrent canal amoebocyte choanocyte spicule Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.12 Lab #4 - Biological Sciences 102 – Animal Biology Various Types of Sponge Spicules Note that spicules can have one, two, three, four, five, six rays or more Spicules can be made of protein, calcium carbonate or silica Prepare and examine some Grantia sp. spicules by doing the following: 1. 2. 3. 4. 5. Place a small piece of the sponge in a depression slide and add a drop or two of bleach. Wait about five minutes for the bleach to take effect. Add a cover slip. Observe them under the compound scope. Note that there are no common intact sponge spicule types that exhibit square ends. If features such as these are observed, they represent fragments of damaged spicules and should not be sketched. Look around for undamaged spicules. 6. In the space below, draw a few representative examples of the types of spicules you observe. Note how many rays (points) are found on each spicule. 7. After you have drawn the spicules, add a drop or two of acid (HCl) to the spicules. Do these spicules dissolve in acid? To which Class of sponges does Grantia sp. belong? Draw a sketch of the Grantia sponge spicules below. Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.13 Lab #4 - Biological Sciences 102 – Animal Biology Prepare and examine some spicules from the local sponge, Tethya aurantia. by removing a small fragment from the cortex of the sponge and following the same procedure as above. 1. In the space below, draw a few representative examples of the types of spicules you observe from Tethya aurantia. Note how many rays (points) are found on each spicule? 2. After you have drawn the spicules, add a drop or two of acid (HCl) to the spicules. Do these spicules dissolve in acid? What compound are these spicules made of? Draw a sketch of the Tethya sponge spicules below. The axial filament of the strongyle megasclere spicules is a protein fiber. This fiber is the substrate on to which silica is deposited during spicule biosynthesis. Sponges of the Class Calcarea lack this organic core within their spicules as control of calcification is mediated by proteins occluded within the various CaC03 phases (amorphous or crystalline). Live Sponge Specimens Carefully observe all of the live sponge specimens displayed in the lab noting the similarities and differences in their basic body plans, color, etc. Fill in the table below with the requested information for any of the live sponge species. The canal system may not be obvious by visual inspection of the sponge alone. Often canal systems must be determined by microscopic examination. You should refer to the Internet for sources to determine the type of canal system for each species. Genus 1. 2. 3. 4. 5. species Class Type of canal system Metazoans, Animal Tissues & Sponges: Phylum Porifera 4.14 Lab #4 - Biological Sciences 102 – Animal Biology Classification of an Unknown Sponge Working with your instructor, attempt to key out and identify at least the class of the unknown sponge displayed in the lab. Note that the following characteristics will be useful in determining the classification of the unknown sponge: General shape of specimen Size, quantity and location of the ostia (these may be difficult to see without a dissecting scope) Size, quantity and location of the oscula (these may be difficult to see without a dissecting scope) General dimensions of the specimen Color of the specimen Type of spicules – calcium, silica, and/or protein based Number of spicule rays Class of the unknown sponge: Genus and species of the unknown sponge: Important Terms Related to Animal Tissues homeostasis = relative to an animal’s physiology, the maintenance of the relatively stable internal environment that is necessary for the animal to survive in a changing external environment. Homeostasis is a state of equilibrium characterized by a dynamic interplay between outside forces that tend to change an organism's internal environment and the internal control mechanisms that oppose such changes. Homeostasis is essential for the survival of each cell, and each cell, through its specialized activities, contributes as part of a body system to the maintenance of the internal environment shared by all cells tissue = a cooperative unit of many similar cells that perform a specific function organ = a structure consisting of at least two of the basic tissue types adapted as a group to perform a specific function. organ system = a collection of organs the work together as a unit for a specific function necessary to maintain homeostasis for an animal (eg. the circulatory system, the digestive system, the respiratory system, etc.) parenchyma = the chief functional cells that comprise an organ (eg. cardiac muscle cells in a heart) stroma = supportive tissues in an organ (eg. the pericardial sac made of connective tissue that surrounds and lubricates the beating heart) LABORATORY NOTES: Metazoans, Animal Tissues & Sponges: Phylum Porifera Lab #4 - Biological Sciences 102 – Animal Biology LABORATORY NOTES: 4.15