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ISSN 0073-6767 Proceedings of the Academy Indiana of Science Volume 106 Number 3-4 ThePROCEEDINGSOFTHEINDIANAACADEMYOFSCIENCEisaquarterlyjournaldedicatedtopro- moting scientific research and the diffusion ofscientific information; to encouraging communication and cooperationamongscientists; andtoimprovingeducationinthesciences. EDITOR GaryE.Dolph IndianaUniversityKokomo 2300S.WashingtonSt. Kokomo,Indiana46904-9003 [email protected] 765-455-9303 EDITORIALBOARD HansO.Andersen WiltonN.Melhorn IndianaUniversity PurdueUniversity RitaBan- PaulRothrock PurdueUniversity TaylorUniversity ErnestE. Campaigne AlfredR. Shmidt IndianaUniversity RoseHulmanInstituteofTechnology WilliamR.Clark ThomasP. Simon Ball StateUniversity UnitedStatesEnvironmentalProtectionAgency DonaldR.Cochran PaulM. Stewart BallStateUniversity IndianaUniversity-PurdueUniversityFortWayne RobertF.Dale MichaelTansey PurdueUniversity IndianaUniversity KaraW.Eberly RobertD.Waltz St.Mary'sCollege IndianaDepartmentofNaturalResources UweJ.Hansen J. DanWebster IndianaStateUniversity HanoverCollege DarlyR. Karns HarmonWeeks HanoverCollege PurdueUniversity N.GaryLane JohnO.Whitaker,Jr. IndianaUniversity IndianaStateUniversity PaulC. MacMillan HanoverCollege EXECUTIVECOMMITTEE JamesD.Haddock President IndianaUniversity-PurdueUniv. FortWayne RebeccaDolan President-Elect ButlerUniversity SusanM.Johnson Secretary BallStateUniversity EdwardL. Frazier Treasurer 5007W. 14thStreet, Speedway JamesW. Berry DirectorofPublicRelations ButlerUniversity GaryE. Dolph EditoroftheProceedings IndianaUniversity Kokomo JamesGammon ImmediatePastPresident DePauwUniversity Nelson R. Shaffer ExecutiveOfficer IndianaGeologicalSurvey EXCHANGEITEMS: ItemssentinexchangefortheProceedingsandcorrespondenceconcerningexchange arrangements should be sent to the Indiana Academy ofScience, John S. Wright Memorial Library, 140 North SenateAvenue,Indianapolis, Indiana46204. REPRINTS AND PERMISSIONS: Write to the Executive Officer, School of Civil Engineering, Purdue University,WestLafayette,IN47907. EDITORIALCORRESPONDENCE:AllmatterforpublicationshouldbeaddressedtotheEditor. PROCEEDINGS OF THE INDIANA ACADEMY OF SCIENCE Founded 29 December 1885 Volume 106 Number 3-4 1997 Published at Bloomington, Indiana May 12, 2000 Copyright © 1999 by the IndianaAcademy ofScience PROCEEDINGS OF THE INDIANAACADEMY OF SCIENCE Volume 106, No. 3-4(1997) CONTENTS BIOMEDICAL M.S. Jarial. SCANNINGAND TRANSMISSION ELECTRON MICROSCOPIC STUDY OFTHEARACHNOID VILLI IN SQUIRRELMONKEYIN RELATIONTO CEREBROSPINAL FLUIDABSORPTION 161 C.E. Mays, J.J. DeJongh, and E.A. Hellmann. GENETIC AND ENVIRONMENTALEFFECTS OF SIDESTREAM SMOKE ON PUPSURVIVORSHIPOFTHREE INBRED STRAINS OF MICE 175 j BOTANY D.G. Ruch, K. Nurtjahja, and K.S. Badger. THE DIFFERENCE BETWEEN MALATE SYNTHASE SPECIFICACTIVITY OF LIGHTAND DARK SPOREDAGARICS IS NOT DUETO PHENOLIC CONTAMINATION 191 ECOLOGY S.J. Burgdorfand H.P. Weeks, Jr. AERIALCENSUSING OF WHITE-TAILED DEERAND COMPARISON TO SEX-AGE-KILLPOPULATION ESTIMATES IN NORTHERN INDIANA 201 C.R. Webster and G.R. Parker. THE EFFECTS OF WHITE-TAILED DEER ON PLANT COMMUNITIES WITHIN INDIANASTATE PARKS 213 ENVIRONMENTALQUALITY M. Liberti and J. Pichtel. SPATIAL DISTRIBUTION OFTRACE METALS IN DELAWARE COUNTY, INDIANA, SURFACE SOILS 233 GEOLOGY J.B. Droste andA.S. Horowitz. SOME HIGHLIGHTS OFTHE CARBONDALE GROUP (PENNSYLVANIAN) IN THE SUBSURFACE IN INDIANA 247 HISTORYOF SCIENCE J.W. Delleur andW.N. Melhorn. DAN WIERSMA, PIONEER AGRONOMIST 253 N.I. Johansen. NEW HARMONY, INDIANA, ACENTURYAND AHALF OF SCIENCEAND ENGINEERING 257 PLANT SYSTEMATICS AND BIODIVERSITY PE. Rothrock. THE VASCULAR FLORAOF FOGWELLFOREST NATURE PRESERVE, ALLEN COUNTY, INDIANA 267 D.E. Wujek and G.A. Bechtel. SILICA-SCALED CHRYSOPHYTES FROM INDIANA. II 291 PSYCHOLOGY R.E. Osborne, J. Penticuff, J. Norman, and M. Robinson. I AM! THEREFORE, I VOTE! SELF-MONITORINGAND 1996 PRESIDENTIALVOTING CHOICES 299 SOILANDATMOSPHERIC SCIENCES R.F. Dale and K.L. Scheeringa. DO INDIANAPOLIS AIRPORT TEMPERATURES REPRESENT INDIANA'S ENERGY NEEDS? 307 ZOOLOGY J.O. Whitaker, Jr. NOTES ONAWINTER COLONYOF BIG BROWN BATS ATWILLIAMSPORT, WARREN COUNTY, INDIANA 319 RESEARCH NOTES E.M. Shull. THE MASSASAUGAIN INDIANA 327 N.D. Simons and E.P. Ellingson. THE REDISCOVERYOF LATHYRUS OCHROLEUCUS (LEGUMINOSAE) IN INDIANA 329 MANUSCRIPT REVIEWERS 333 INDEX 335 ProceedingsoftheIndianaAcademyofScience 161 (1997)Volume 106p. 161-173 SCANNING AND TRANSMISSION ELECTRON MICROSCOPIC STUDY OF THE ARACHNOID VILLI IN SQUIRREL MONKEY IN RELATION TO CEREBROSPINAL FLUID ABSORPTION Mohinder S. Jarial Muncie Center for Medical Education Ball State University Muncie, Indiana47306 ABSTRACT: The surfacefeatures andfine structureofarachnoidvilliwere examinedusingscanning(SEM)andtransmissionelectronmicroscopy(TEM). With SEM, the endotheliumofthe superiorsagittal sinus was seenextend- ingoverthe surfaceofthevilliasacontinuouslayerofendothelialcellsthat exhibitedfoldsandcrypts.Thebulgingluminalsurfaceoftheendothelialcells displayedslenderprocesses,shortmicrovilli,andporesofvarioussizes.Using TEM,theuninterruptedendothelialcellsdisplayedslenderinterdigitatingcyto- plasmic processes that werejoinedby desmosomes. The cytoplasmofthe endothelialcellscontainednumerousmicropinocytoticvesiclesandgiantvac- uoles.Thegiantvacuolescommunicatedwithbasalpinocytotic vesicles and surfacepores, apparentlycreatingtranscellularchannelsintheendothelium. Theendothelialcoveringseparatedthesubendothelialspacefromthevenous sinus. The cores contained arachnoidcells, fibroblasts, macrophages, and a network ofanastomosing channels. The slender, overlapping cytoplasmic processes ofthe arachnoidcells linedthechannels. The villi were devoidof endothelium-lined tubes andblood vessels. The shallow endothelial crypts seenin some villi were closed offfromthe subendothelial space by desmosomes.Thevilliwereinnervatedbymyelinatedaxons.Theultrastructuralfea- turesofthe arachnoidvilliofthe squirrelmonkeyrevealedbythis study are consistent with theirfunction ofCSFabsorption by transcellularbulkflow andstreamingthroughthe surfacepores intothedural venous sinuses. KEYWORDS:Arachnoidcells,channels,desmosomes,endothelialcells,giant vacuoles,pores, vesicles. INTRODUCTION Thearachnoidvilli, orgranulations, are small, bluntherniations ofthe arachnoid membrane which project into the cerebral veins and dural sinuses through smalldeficienciesintheduramater.The villiplay anessentialrole inthe drainage and absorption ofcerebrospinal fluid (CSF) into the venous sinuses. However, themechanismby whichCSFis transportedacrossthe villi intothe venous blood is unclear. The histology ofthe arachnoid villi was described by Weed (1914), Welch and Friedman (1960), Turner (1961), Millen and Woolam (1962), Jayatilaka (1965a), and Potts, et al. (1972). A number ofultrastructural studies were carried out on the arachnoid villi ofvarious mammals (Jayatilaka, 1965b; Alksne 162 Biomedical: Jarial Vol. 106 (1997) and White, 1965; Shabo and Maxwell, 1968a, b; Alksne and Lovings, 1972a, b; Gomez, etal., 1973; Gomez and Potts, 1974; Gomez, etal, 1974; Peters, et al, 1976; Tripathi, 1973). These studies demonstrated that the arachnoid villi were invested by endothelial cells, that internally the villi were composed of arachnoid tissue and collagen bundles, and that the villi were traversed by a labyrinth ofintercellular channels which communicated with the subarachnoid spacearound the brain. Some investigators asserted thatthe endotheliumofthe dural venous sinuses that invested the villi was continuous andjoined by tight junctions, supportingWeed's (1923) "closed" systemhypothesis whichimplied that CSF absorption took place across the endothelial covering ofthe arachnoid villi (Shabo andMaxwell, 1968a, b;Alksne andLovings, 1972a, b). Other researchers showed that the villus core contained tubular channels lined by endothelium which was continuous with the lining ofthe dural sinuses, allow- ing direct flow ofCSF from the subarachnoid space into the venous sinuses. Thus, an "open" system was proposed (Welch and Friedman, 1960; Welch and Pollay, 1961; Jayatilaka, 1965a, b; Hayes, etal, 1971; Potts, etal, 1972; Gomez andPotts, 1974; Gomez, etal, 1974). Furthermore,Tripathi (1973) andTripathi and Tripathi (1974) demonstrated that the giant vacuoles formed within the endotheliallining ofmonkey arachnoidvilli allowedmovementofCSFintothe venous system by bulk flow. Kida, etal. (1988) reported that the entire luminal surface ofhuman arach- noidvilliwas investedbyendothelialcells. However, otherinvestigators showed that only a small portion ofthe human arachnoid villus is covered by endothelium, therestismainlyinvestedwithalayerofarachnoidcells (UptonandWeller, 1985; Yamashima, 1986). Studies by d'Avella, etal. (1980, 1983) on human arachnoid villi revealed the presence ofgiant intracellularvacuoles, pinocytot- ic vesicles, andlargegapsbetweenthemintheendothelialcells, supportingboth the "closed" and "open" mechanisms ofCSF absorption. However, Upton and Weller (1985) did not observe any pores in the endothelium covering human arachnoid villi. Thus, the mechanismofCSFabsorption throughthe arachnoidvilli intothe venous sinuses stillremainscontroversial. The aimofthepresentstudyistoelu- cidate the surface features and fine structure ofthe arachnoid villi in squirrel monkey and to relate them to the mechanism ofCSF absorption. MATERIALS AND METHODS The squirrel monkey, Saimirisciureus, usedinthepresentstudywas obtained from the monkey colony maintained at the Yerkes Regional Primate Research Center, Emory University,Atlanta, Georgia.Anormal adultsquirrel monkey was anesthetized at the Histochemical Laboratory at the Yerkes Primate Research Center using an appropriate dose of sodium nembutal given intraperitoneally and was perfused with 2.5% glutaraldehyde in 0.1M phosphate buffer at pH 7.4 for 30 minutes. Aftercraniotomy, twelve arachnoid villi were excised from the inner wall ofthe superior sagittal sinus under a dissecting microscope and Vol. 106 (1997) IndianaAcademy of Science 163 Figures 1 and 2. Figure 1.Ascanning electron micrograph (SEM) ofan arachnoid villus (AV) projecting into the lumen ofthe superior sagittal sinus. Note the continuity of the endothelial lining (END) ofthe sinus with that ofthe villus (675X). Figure 2.An SEM ofthe same villus shown in Figure 1 (1,500X). Note the somewhat roundedpro- files ofthe endothelial cells (EC) with cytoplasmic processes (CP), crypts (CT), folds (FO), andpores (P). 164 Biomedical: Jarial Vol. 106 (1997) fixed in fresh 2.5% glutaraldehyde in 0.1M phosphate buffer (pH 7.4) for two hours at room temperature. The samples were rinsed in phosphate buffer and post-fixed in 1% osmium tetroxide in the same buffer. The material was dehy- dratedinanethanol series, transferredtopropyleneoxide, andembeddedinEpon 812 (Luft, 1961). Polymerization was carried out overnight at 60° C. The sec- MT tions werecutonaPorter-Blum ultramicrotome, stainedwithuranylacetate 2 and lead citrate, and examined with an RCA-3C and a Hitachi HU-11Atrans- mission electron microscope (TEM). Similarly fixed material was dried using the liquid C0 critical point method, coated with gold/palladium, and exam- 2 ined in an ETEC autoscan scanning electron microscope (SEM). Two (2) u,m thick sections were cut using glass knives, stained with azure II, and examined under a light microscope (LM). RESULTS Scanning Electron Microscopy. After opening the dorsal wall of the superior sagittal sinus, many arachnoid villi measuring 125-200 u,m in diameter were seen protruding into the sinus lumen. Apanoramic view ofthe inner wall ofthe sinus revealedthatthe intactendothelial lining ofthe superiorsagittal sinus extended over the villi to form their endothelial covering (Figure 1). Theendothelialcoveringdisplayedfolds andcrypts.Theendothelialcellsappeared somewhatrounded in contour, possibly due to the presence ofunderlying giant vacuoles and/or nuclei (Figure 2). The cells had slender cytoplasmic processes and displayed short, club-shaped microvilli on their luminal surface (Figures 3-5). A prominent surface feature ofthe endothelial cells was the presence of pores measuring about 0.4 u,m in diameter with somewhat thickened margins. More pores were found at the attenuatedperiphery than in the central region of the endothelial cells (Figures 3-5). Longitudinal sections through the middle of the villi revealed that the central core contained arachnoid cells with stout cytoplasmic processes and channels containing collagen bundles (Figure 6). Light and Transmission Electron Microscopy. In semi-thin sections, the arachnoidvilli werecomposedofacontinuous, thinendothelialcoveringandan underlying core ofarachnoid cells and interconnecting channels (Figures 7 and 8).Thearachnoidvilliweredevoidofbloodvessels, andtheylackedtheendothe- lium-lined tubes which have been reported in the arachnoid villi ofother ani- mals (Figures 7 and 8: Jayatilaka, 1965a; Potts, etal, 1972).Apictorial summary ofthegeneralorganizationofthearachnoidvilliofthesquirrelmonkeyasrevealed by electron microscopy is presented in Figure 9. Transmission electron micrographs of the arachnoid villi revealed that theircontinuous endothelialcovering wascomposedoffusiformendothelialcells displaying short microvilli on their luminal surface. The cells were separated fromthe underlyingcoreby asubendothelial space (Figure 10). Thecentralcore ofa villus contained loosely packed arachnoid cells, fibroblasts, macrophages, and bundles of collagen. The core was traversed by a network of channels (0.6-1.5 jxm in width) that were in continuity with the subendothelial space