The Morphology of the Nephridia and Genital Ducts of Cirriformia Spirobrancha Thomas T. Gilbert Hopkins Marine Station of Stanford University Pacific Grove, California 1. Permanent Address: C The Morphology of the Nephridia and Genital Ducts of Cirriformia Spirobrancha I. Introduction The object of this study was the polychaete annelid Cirriformia spirobrancha (Moore, 1904). A member of the family Cirratulidae, C. spirobrancha is a sedentary worm, up to six inches long and 1 inch in diameter, living in black mud in intertidal and shallow subtidal mud flats and sheltered tide pools. It lives burrowed with its anterior end just below the substrate surface and its tentacles extended above the surface. The literature of the nephridial anatomy of the annelidae is scanty at best with the exceptions of the Nereidae (Jones, 1957) and Lumbricus, an oligochaete. Goodrich's excellent summary (1945) covers nearly all the known literature to date. Thus there is a limited back- ground of well-described species with which to compare new findings. Goodrich cites the work of E. Meyer (1887) describing the family Cirratulidae in general terms. He found two types of nephridia, an anterior pair of mixonephridia with very long excretory canal and numerous posterior mixo- nephridia in the genital region composed almost entirely of large coelom- ostomes. Goodrich also mentions Caullery and Mesnil's findings (1898) that in Dodecaria concharum the anterior organs are as expected, but in epitokus forms large coelomostomes develop late from the coelomic epithelium on the anterior face of the posterior septa, fuze with the epidermis, and form pores through which issue the genital products. No nephridial canal element was found contributing to these genital ducts and possibly they represent coelomostomes only. It had been observed by students and faculty of this course prior to this study that a pair of large anterior nephridia seemed to be present in only about one-fifth of the C. spirobrancha individuals observed. Further, no smaller nephridia or coelomoducts were seen posteriorly. The objectives 98 of this study were thus to describe the nephridial anatomy of C. spiro- brancha and to determine the extent and possible explanation of the observed dimorphism. This study determined that all individuals of C. spirobrancha possess a single pair of large anterior mixonephridia and segmental nephridia or coelomoducts in the genital region presumably serving as gonoducts. The anterior organs have a unique sac-like morphology and an extensive blood supply, while the genital organ's anatomy is not entirely clear. II. Materials and Methods Worms were collected from intertidal mud flats by municipal pier fl, Monterey, California. Both live dissection and fixed and stained serial sections in transverse, medial, and sagittal planes were used. In dissection, the worms were anesthetized by addition of crystals of magnesium chloride to sea water in a dissecting pan to a concentration of approximately 7.5%. The body wall was opened by a lateral incision along the dorsolateral row of setae. For prepared sections, worms were relaxed in magnesium chloride, fixed in Bouin's fluid, embedded in paraffin, cut at 10 and 14 microns, and stained according to Masson's trichrome connective tissue technique as modified by Foot and by Coldner. A series of camera lucida sketches on index cards was prepared to aid in visualizing the internal anatomy of the nephridia. III. Description of the Nephridia and Gonoducts The nephridia are a pair of sac-like organs located in the first two segments posterior to the prostomium ventrolateral to the pharynx. They were not consistently observed early in the course because dissections as a rule were made with dorsal or ventral incisions either missing or destroying the nephridia. The nephrostome is a large heavily ciliated funnel opening inte the prostomial coelomic cavity in close proximity to the lateral body wall and the extensive prostomial capillary beds. The funnel opening faces dorsally and measures about 100 microns in diameter in fixed and stained preparations. The whole funnel structure is 150 to 200 microns long. The funnel narrows posteriorly as it passes through the septum and leads into the cavity of the nephridium. The body of the nephridium is about 6mm. long by 3 mm. high by 2 mm. wide and shows some involution in its outer wall. Its color varies from light yellow to black over its surface; the darker colors are due to blood capillaries in the outer wall. The wall is fragile and easily torn in dissection. When opened, the interior surface of the nephridial sac appears convoluted; in fixed and stained material subject to some shrinkage and contraction, the interior of the sac is filled with these convolutions. In dissection, the convolutions follow a characteristic pattern. The dark color of blood vessels close to the surface is especially evident on the convolutions. In fixed and stained preparations, the inter- ior wall appears lightly ciliated, the cilia ocurring in tufts uniformly distributed over the entire interior surface. The wall appears to be two to three cell layers thick. The cell boundaries are indistinct; the outer layer seems to be a continuation of the lining of the coelom. The wall shows a lack of connective tissue or muscle fibers and is heavily supplied with blood capillaries. The nephridiopores are located posterior to the first septum and lie on either side of the ventral nerve cord posterior to the point of junction of the two circumpharyngeal connectives. The tubule leading to the pore is 25-30 microns in diameter. The duct, unlike the body of the nephridium, does not appear ciliated. The region of ocurrence of the presumed gonoducts corresponds very /8 closely with the region of production and storage of the gametes. The ducts were not seen in fixed and stained material due to tearing of the sections by grit in the gut. In dissection they appear white in color and lie just posterior to each septum in this region one pair to a segment. They are about O.3 mm. long and about 50 microns in outside diameter. They appear to open to the outside at the level of the ventrolateral ros of setae. No obvious funnel was present at the inner end of the duct and the duct was not seen to pierce the septum. There appeared to be little or no blood supply. The inner end lies close to the venteral nerve cord and ventral blood vessel at the base of the coelomic cavity. IV. Discussion The nephridia do not have the convoluted tubule morphology described for the Nereidae (Jones, 1957). However, the nephridial anatomy described for C. spirobrancha is reasonable. The worm is sedentary, yet does not irrigate its burrow as do some other burrowing polychaetes. Thus it must excrete its waste products at its anterior end. The worm does not appear to move enough to be able to leave its wastes behind it as it burrows. Waste products presumably are carried in the blood from the posterior parts of the body to the capillary nets in the nephridia or to those lining the coelomic cavity of the prostomium into which the nephrostome extends. These capillary nets are the locus of exchange of solutes between blood and coelomic fluid. The nephridia then pump out the dissolved wastes in the urine. The cilia in the nephrostome provide the current of fluid through the nephridium. The cilia and the convolutions in the wall of the organ help direct the flow and keep the fluid near the walls in constant motion facilitating exchange. Reabsorbtion of substances from the coelomic fluid to the blood is similarly accomplished. Contractions of the extensive musculature associated with the pharynx and body wall in the region of the 101 nephridia may also help provide a fluid flow. It would be interesting to follow the embryological development of these organs to facilitate comparison with the tubular nephridia of other polychaetes. Physiological work on the functions of the organs is also necessary. In line with the preceeding discussion, it is presumed that the posterior gonoducts do not function as nephridia. Their location posterior to the anterior septum in each segment suggests they may consist of nephridial tissue and may function as simple nephridia. Caullery and Mesnil (1898) noted in Dodecaria concharum that coelomostomes with short ducts developed at the time of spawning. The present study was carried out when most worms were spawning and the ducts observed could have been in the process of development. They may be absent or vestigial during the rest of the yaer or the rest of the worms' life cycle. V. Conclusions 1. The nephridia of Cirriformia spirobrancha have been described to be a pair of sac-like organs with a large blood supply and extensive internal surface area located in the first two post-prostomial segments. 2. The presumed gonoducts have been described to be simple tubules located in the genital region. 3. Possible functions for both nephridia and gonoducts have been postulated. 08 Cb MC A FIGURE Mc FIGURE 2 11 N cb MC 0 L (2 60 FIGURE s Nb S J FIGURE 4 — SLe FIGURE 5 - Figure Captions Figure 1. Camera lucida sketch of a transverse section at the level of the first septum. Note the extensive capillary beds and the large size of the nephrostome. Figure 2. Camera lucida sketch of a transverse section of the body of the nephridium showing the large size of the cavity. Figure 3. Camera lucida sketch of a sagittal section. The ventral body wall is shown at the bottom; anterior is to the right. Figure 4. Diagram of ventral view showing the location of the nephridia in the body. Figure 5. View from dorsally of the interior ventrolateral body wall in the genital region showing the presumed gonoducts. Mc Ne Nd Np Se Si Vnb Key to Figures Body wall Blood vessel Coelomic cavity Capillary bed Circumpharyngeal connective Nephridial cavity Prostomial coelom Gut musculature Gonoduct Longitudinal muscle Circular muscle Nephridial wall tissue: For clarity wall blood vessels have been omitted from all figures except figure 2 where they were not as extensive as usual. Body of Nephridium Ventral nerve cord Duct from nephridium to nephridiopore Nephridiopore Nepkrostome Prostomium Septum Setae and setal musculature Ventrolateral row of setae Ventral blood vessel and nerve cord Acknowledgements: I would like fo thank Roger Szal and Bruce Belman for teaching me,with great patience, all the techniques used in this work and for much good advice. This work was supported in part by the undergraduate research participation program of the National Science Foundation, grant+ GY-4369. Footnote: 1. Jones, R. MeC., MeClung's Handbook of Microscopical Technique, p. 249-50. Literature Cited 1. Goodrich, E. S. 1945, The Study of Nephridia and Genital Ducts since 1895, Quart. J. Microscop. Sci. 86:113-392. 2. Jones, M. L. 1957, On the Morphology of the Nephridium of Nereis Vexillosa Grube, Biol. Bull. 113:407-13. 3. Jones, R. McC., ed., MeClung's Handbook of Microscopical Technique, ded., Haffner Publishing Co., New York, 1950. C C