.t ù ,{ ù :ó 11 l<t I I j { ( A STUDY OF THE SUBCOMMISSURAL ORGAN '1 t VULPECULA IN TIIE ADULT AND POUCH-YOUNG TRTCHOSURUS t, I i A THESIS SUBMITTED FOR TITE DEGREE Otr' DOCTOR OF PHILOSOPÏ{Y by RAì4 SrNGH TULST, D .S .M. , M. Sc. Department of Anatomy and Hístology The UniversitY of Adelaide Adelaíde South Austral-ía April,1980 11 TABLE OF CONTENTS 1X SUMMARY xt 1l DECLARATION xav ACKNOT,TLEDGEMENTS Chapter I Introduction L.2 Comnents on the Contents of the Varíous Chapters of this Thesis 1.3 REVIEI,ü OF THE LITERATT]RE r.6 Structuralfeaturesofthesubcommíssuralorgan 1.6 Development and ageíng of the subcommissural organ 1. ?5 Secretions of the subcorunlssural organ 1. 33 Reissner's fibre r.46 Nerves and related strucÈures assocíated \^líth the subcormnissural organ r.57 Functional aspects of the subcommissural organ 1. 65 Pineal recess supraependymal cells ' suPraependymal flbresandcertaínotherfeaturesofthelining' of the ventricles r.7 6 A Bríef Sunnnary of the Subcommísstrral Organ in a I'{ide Variety of Animals 1. 89 II Arl,ls, TERMINOLOGY AND ORIENTATTON Aims of the Present StudY 2.2 Factors Determíning Selectíon of Subject Species 2.3 TermínologY and Orientation 2,3 11a Chapter III THE SUBCOM}'IISSURAL ORGAN' PINEAL RECESS, AND THE AQUEDUCTAL ROOF OF THE ADI]LT ANIMAL AND THE STJ¡COMITSSUML ORGAN OF TIIE POUCH_YOIJNG STUDIED BY SO}{E SPECIFIC METHODS. Cornment related to materials and methods of all sections of the Present studY 3.1 RELATED TO MATERIALS AND METHODS OF COMMENT 3.2 CHAPTER III Scanning electron microscoPY 3.2 Transmission electron microscopy 3.2 H. & E. Stain 3.3 Periodic Acíd-Schiff reaction 3.3 3.4 Massonr s trichrome stain 3.4 AldehYde- f uchs ín me thod Cajalt s method for astrocytes 3.4 Phosphotungstic acid method for astrocytes 3.4 Marshall' s method for olígodendrocytes 3.5 Rapid Golgi method for neural elements in rhe SCO 3.5 3.5 Method for acid PhosPhatase Radioimmunoassay for melatonin in blood' SCO' 3.5 and Pineal 3.6 FINDINGS Features of the píneal and SCO observed using a díssecting mícroscoPe 3.6 Surface features of the pineal recess using SEM 3. 10 Surface features of the SCO and aqueductal roof 3. 36 viewed \^títh a SEM Líght and electron mícroscope study of the SCO 3.54 3.63 MorphologY of Reissnerts fibre InÈermediate and deep regions of the SCO and 3.70 hyPendYma Some evídence of basal secretion 3 .85 ,1 tv ,! ,t Chapter t? Labyrinths of basal lamina, perivascular space, 3.89 and perivascular neural complex I I Neural elements associated with the SCO 3.96 I I Radioimmunoassay of blood, SCO and pineal for melatonin 3.111 1 DISCUSSION 3 . 1.13 I SIJMMARY 3. 1.18 I I IV MULTIPLE METHODS OF INVESTIGATIVE APPROACH SUPPORTING PRESENCE OF EXTENSIVE SUPRAEPENDYMAL NEURAL PLEXUS ON THE VENTRICULAR SURFACE OF THE PI}IEAL/SUBCOMMISSURAL ORGAN REGION AND POSSIBLE RELATIONSHIP OF SOME OF THESE TO ADJACENT COMMISSURES AND NEUROPIL 4.r Introductíon 4.r },IATERIALS AND METIIODS 4.2 ObservaEions made wíth SEM method 4.2 ObservaÈions made with TEM method 4.s * p Observatíons made with SEl"l and TEM methods applied to the same sPecimen 4.6 Observations made with C.A. fluorescence method 4.7 Observations made with modifíed Bíelschowsky method 4.9 Observations made with methylene blue rnethod 4. 10 Observatíons made wiËh phosphotungstic acid method 4. tl DISCUSSION 4.26 SUM},IARY 4.27 V THE POUCH-YOIJNG ANIMAL: ITS PINEAL RECESS' SUBCOMIÍISSURAL ORGÆ.I, Al'lD AQUEDUCTAL ROOF 5.1 MATERIALS AI{D METIIODS 5 I Scanning electron microscope method 5 1 Transmíssion elecÈron microscope method 5 1 Aldehyde-f uchsin method 5.1 .t I ,t Chapter tr^ FINDINGS \, Introduction <t Development and maturation of the fine surface feaEures in the developing animals as seen wíth a scanning electron microscoPe: (i) Pineal recess 5 2 (ii) Subconunissural organ 5 4 (iii) Roof of the cerebral aqueduct 5 6 Fíne structural features in the developing animals as seen wÍth a light microscope and a transmíssíon electron microscope: (i) Introduction to LM and TEM study of the pouch-young animals 5.20 (ii) LM and TEM observations of the SCO in a pouch-yound with a CR length of 18 nm. 5.2t (iii¡ LM and TEM observatíons of the SCO in pouch-youngs with CR lengths ranging frorn 65 to 75 nun. 5.24 (iv) LM and TEM observations of the SCO ín a pouch-young with a CR length of 220 mrn. 5.25 DISCUSSION 5.37 SU},ÍMARY 5.38 VI SOME E)GERIMENTAL STUDIES 6.1 Introductíon 6.1 Carbon and ferritin into cerebral ventrícles 6.3 Ferritin into systemic veins 6.5 Fresh blood into cerebral ventricles 6.5 Cand'Lda a.LbLean's into cerebral ventricl-es 6.6 Ni's t ønia nøwLngì.tidi'S ir.t o c e reb r al ven t ri c le s 6.8 Mt4 xo vinu.t fJaho fj-d,írs int o cerebral ven t r ic le s 6.9 DISCUSSION 6.20 SI]MMARY 6.24 V1 I t Chapter hr 7.r VII GENERAL DISCUSSION AND CONCLUSIONS ): l !' Somestructuralfeaturesofthesubcommissural -7 ') organ 'tfi Development of the subconrnissural organ, pineal of the cerebral aqueduct 7.6 (,' recess and the roof t I basa1, and melatonín 7.L2 I Secretíons: Apical, ï Reissnerr s fibre 7.18 I I I Nervesassociatedwiththesubcommíssuralorgan 7 .2L Pineal recess' supraependymal nerves and supraePendYmal cells 7 .25 7 .33 Some exPerimental studies 8.1 VIII SU},IMARY IX (APPENDIX I) 9.1 DETAILS OF Æ{IMALS USED Conrnent related to rnaterials and meÈhods of all sections of the Present studY 9.1 I summary table of TtuLchoÃÚLu^ vu.b¡tøcu.tA" used in A 9.2 the Present study For scanning electron microscope method: - Adult 9.3 Pouch-Young 9.4 For transmission electron microscope method: - Adult 9.4 Pouch-Young 9.5 For llaen:atoxylínand Eosin (adult and pouch-young) 9.6 Tor Períodic Acid Schiff reactlon (adult and pouch-Young 9.6 For l"fassonts Trichrome SËain (adult) 9.6 For aldehyde-fuchsín method (adult and pouch-young) 9.7 For Cajalts method for astrocytes (adult and pouch- young) 9.7 For Phosphotungstic acid method for astrocytes (adul-t) 9.8 i vt1 1 Chapter For Marshall's method for oligodendrocytes (adult) 9.8 ¡ l I For rapid Golgi method (adult) 9.8 t t For acid phosphatase method (adult and pouch- young) 9.9 { I t For radioimmunoassay for melatonin in blood, ! SCO and píneal (adult) 9.9 For modified Bielschowsky meÈhod (adult and pouch-young) 9. r0 For methylene blue rnethod (adult) 9.10 For C.A. fluorescence method (adult and pouch- Young) 9.10 For carbon and ferrítin method (adult) 9.11 For ferrltin I.V. method (adult and pouch-young) 9. 1r Eor Candida a.LbLcavrs method (adult) 9. l1 For lllqxovinu yto,ttotlúi's method (adult) 9. 11 I I Fox Neis'se¡ia m¿ningibLdi^ method (adult) 9.L2 For blood into the ventricle of the brain nethod (adult and pouch-young) 9.L2 X (APPENDIX II) EETAILS OF METHODS USED 10. 1 Scanning electron microscope method 10.1 Method of counting cíliated cells L0.2 Transmission elecÈron microscope method r0.2 Method of counting dark and light cells of rhe sco 10.4 SEM and TEM examinations carried out on the same specimen 10. 5 Haematoxylfn and eosin method 10. 5 Periodic acid-Schiff reaction 10. 6 Massonr s Trichrome stain method 10. 6 va]-l- Chapter Aldehyde-fuchsin method 10. 7 I Cajal's method for astrocytes 10. I Phosphotungstic acid method for: astrocytes 10. 9 Marshallrs method for oligodendrocytes 10.9 Rapid Golgí rnethod for neural elements of SCO 10. 10 MeÈhod for acid phosPhatase 10.11 Radíoinununoassay method for melatonin in blood, SCO and pineal 10. 11 Modified Bielschowsky method IO.I2 Methylene blue method r0. 13 C.A. fluorescence method using glyoxylic acid 10. 14 Details of stereoÈaxic technique for intraventrícular ínjection of various substances 10. 15 XI (APPENDIX III) 11. 1 Thís chapEer contains abstracts/stnrunaries of papers read at meetings of Èhe Anatomical Society of Australia and New Zealand and papers published ín a number of journals. BIBLIOGRAPHY B.l lx SUMMARY Ànextensivemorphologicinvestigationofthesubcommissrrralorgan' marsupial pineal recess and the aqueductal roof of the Australian Trí'ehas?l]iusDulpeeuLawascarriedoutinnormalanimalsandothers sr-rbjected to several experimental procedl;res' developmental stages In addition an ontogenetic study of various , of the pouch-yor:ng animats was carried out' electron Light microscopic techniques, transmission- and scanning mlcroscopyh'ereusedtoanalysetheeperrdlnna]surfacesoftheseareas. fibres' Particular attention I¡Ias paid to (a) supraependymal cell-s and (b) cilia, and (c) secretory processes of the subcommissural organ' (1- The following results were obtained to 6): 1,. The subcommissural organ ot T¡iChO^urLtA Vu'LytøUú^ has resembles basically those in other mammafs' It two longitudinally-n:nning paramedi-an ridges separated (parasub- by a median groove (gutter), and two grooves commissural organ gutters) lateral to the paramedian ridges. The secretory ependl'rnal cells of the which subcommissural- organ produce an apical secretion coalesces to form Reissnerrs fibre' The secretory activity and the size of Reissnerrs fibre did not pouch- appear to be influenced by sex or season' In the young, indications of basaf secretion were observed' in Although some basal secretory materiaf was observed X the adult, capillaries with fenestrated endothelium as found in other endocrine organs were not present. Perivascular space associated with labyrinthine basal l-amina and nerve terminals, which resemble nerve endings in the neurophypophysisr were al-so observed.