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A morphological, enzyme-cytochemical, and physiological study of the blood-gonad barrier in the hermaphroditic snail Lymnaea stagnalis

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In the hermaphroditic pulmonate snail Lymnaea stagnalis a blood-gonad (blood-testis) barrier appears to exist. Septate junctions between Sertoli cells and epithelial cells of the neck areas of the gonadal acini constitute this barrier; they separate the male from the female compartment. Experiments with tracer substances (colloidal gold particles, lanthanum nitrate, tannic acid) showed that the basal lamina around the acini hardly forms a barrier; only the larger colloidal gold particles do not pass this lamina.

Physiologically, the blood-gonad barrier is apparent in studies on the composition of gonadal fluid, which differs considerably from that of haemolymph. The osmolarity and the concentration of protein and amino acids in gonadal fluid exceed those of haemolymph. As to the major ions, in the gonadal fluid Na+ is partly replaced by K+, and HCO 3 - is almost totally replaced by Cl-. Such a distribution of HCO 3 - and Cl- is indicative of metabolic acidosis. The cytochemical localization of carbonic anhydrase activity in cells lining the acinar lumen (Sertoli cells, epithelial cells) suggests that these cells are involved in the process of ion exchange. The metabolic acidosis in the gonad might result from the anaerobic production of lactate and succinate by Sertoli cells; these cells lack the enzymes cytochrome oxidase, lactate dehydrogenase, and succinate dehydrogenase. Spermatogenic cells, on the other hand, do possess these enzymes. This probably indicates that these cells metabolize lactate and succinate secreted by Sertoli cells.

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  1. Abraham M, Rahamin E, Tibika H, Golenser E, Kieselstein M (1980) The blood-testis barrier in Aphanius dispar (Teleostei). Cell Tissue Res 211:207–214

  2. Anderson WA, Personne P, André J (1968) Chemical compartimentalization in Helix spermatozoa. J Microsc 7:367–390

  3. Boer HH, Witteveen J (1980) Ultrastructural localization of carbonic anhydrase in tissues involved in shell formation and ionic regulation in the pond snail Lymnaea stagnalis. Cell Tissue Res 209:383–390

  4. Brown AC, Brown RJ (1965) The fate of thorium dioxide injected into the pedal sinus of Bullia (Gastropoda: Prosobranchiata). J Exp Biol 42:509–519

  5. DeWolfe MS, Baskurt S, Cockrane WA (1967) Automatic amino acid analysis of blood serum and plasma. Clin Biochem 1:75–81

  6. Dogterom AA (1979) The effect of the growth hormone of the freshwater snail Lymnaea stagnalis on biochemical composition and nitrogeneous wastes. Comp Biochem Physiol 65B:163–167

  7. Dongen WJ van, Kort EJM de (1959) Spermiation in the common frog (Rana temporaria) I. Experiments on the isolated testis. Proc Kon Ned Akad Wet C62:320–326

  8. Dongen WJ van, Draisma JR, Kort EJM de (1959) Spermiation in the common frog (Rana temporaria) II. Experiments in vivo. Proc Kon Ned Akad Wet C62:327–332

  9. Dongen WJ van, Ballieux RE, Geursen HJ, Offermans T (1960) Spermiation in the common frog (Rana temporaria) III. Histochemical and chemical investigations. Proc Kon Ned Akad Wet C63:257–263

  10. Dym M (1973) The fine structure of the monkey Sertoli cell and its role in maintaining the blood-testis barrier. Anat Rec 173:639–656

  11. Dym M, Cavicchia J (1977) Further observations on the bloodtestis barrier in monkeys. Biol Reprod 17:390–403

  12. Eppig JJ (1977) Mouse oocyte development in vitro with various culture systems. Dev Biol 60:371–388

  13. Gilula NB, Fawcett DW, Aoki A (1976) The Sertoli cell occluding junctions and gap junctions in mature and developing mammalian testis. Dev Biol 50:142–168

  14. Guraya SS (1980) Recent progress in the morphology, histochemistry, biochemistry, and physiology of developing and maturing mammalian testis. Int Rev Cytol 62:188–309

  15. Hanker JS (1975) Oxidoreductases. In: Hayat MA (ed) Electron microscopy of enxymes, Vol 4. Principles and methods. Van Nostrand Reinhold Company New York Cincinnati Toronto London Melbourne, pp 1–139

  16. Hogarth PJ (1978) Biology of reproduction. Tertiary level biology. Blackie, Glasgow and London

  17. Johnson MH (1970a) Changes in the blood-testis barrier of the guinea pig in relation to histological damage following isoimmunization with testis. J Reprod Fertil 22:119–127

  18. Johnson MH (1970b) An immunological barrier in the guinea pig testis. J Pathol 101:129–139

  19. Johnson MH, Setchell BP (1968) Protein and immunoglobin content of rete testis fluid of rams. J Reprod Fertil 17:403–406

  20. Jones RTh (1978) The blood/germ cell barrier in male Schistocerca gregaria: the time of its establishment and factors affecting its formation. J Cell Sci 31:145–163

  21. Jong-Brink M de, Wit A de, Kraal G, Boer HH (1976) A light and electron microscope study on oogenesis in the freshwater pulmonate snail Biomphalaria glabrata. Cell Tissue Res 171:195–219

  22. Jong-Brink M de, Boer HH, Hommes TG, Kodde A (1977) Spermatogenesis and the role of Sertoli cells in the freshwater snail Bimophalaria glabrata. Cell Tissue Res 181:37–58

  23. Jong-Brink M de, Schot LPC, Schoenmakers HJN, Bergamin-Sassen MJM (1981) A biochemical and quantitative electron microscope study on steroidogenesis in ovotestis and digestive gland of the pulmonate snail Lymnaea stagnalis. Gen Comp Endocrinol 45:30–38

  24. Jong-Brink M de, Boer HH, Joosse J (1983) Mollusca. In: Adiyodi KG, Adiyodi RG (eds) Reproductive Biology of Invertebrates Vol I. Oogenesis, oviposition, and oosorption. John Wiley & Sons Ltd, New York, pp 297–355

  25. Joosse J, Reitz D (1969) Functional anatomical aspects of the ovotestis of Lymnaea stagnalis. Malacologia 9:101–109

  26. Jutte NHPM, Grootegoed JA, Rommerts FFG, Molen JH van der (1981) Exogenous lactate is essential for metabolic activities in isolated spermatocytes and spermatids. J Reprod Fertil 62:399–405

  27. Knaap WPW van der, Sminia T, Edelenbosch P (1981) The role of serum factors in phagocytosis of foreign particles by blood cells of the freshwater snail Lymnaea stagnalis. Dev Comp Immunol 3:37–44

  28. Knaap WPW van der, Doderer A, Boerrigter-Barendsen LH, Sminia T (1982) Some properties of an agglutinin in the haemolymph of the pond snail Lymnaea stagnalis. Biol Bull 162:404–412

  29. Koskimies AJ, Kormano M (1973) The proteins in fluids from the seminiferous tubules and rete testis of the rat. J Reprod Fertil 34:433–444

  30. Lacroix M, Smith FE, Fritz JB (1977) Secretion of plasminogen activator by Sertoli cells enriched cultures. Mol Cell Endocrinol 9:227–236

  31. Le Gac F, de Kretser DM (1982) Inhibin production by Sertoli cell cultures. Mol Cell Endocrinol 28:487–498

  32. Livingstone DR (1982) Energy production in the muscle tissues of different kinds of molluscs. In: Addink ADF, Spronk N (eds) Exogenous and endogenous influences on metabolic and neural control. Pergamon Press Oxford New York Toronto Sidney Paris Frankfurt, pp 257–274

  33. Louis BG, Fritz B (1977) Stimulation by androgens of the production of androgen binding protein by cultured Sertoli cells. Mol Cell Endocrinol 7:9–16

  34. Luchtel D (1972a) Gonadal development and sex determination in pulmonate molluscs. I. Arion circumscriptus. Z Zellforsch 130:279–301

  35. Luchtel D (1972b) Gonadal development and sex determination in pulmonate molluscs. II. Arion ater rufus and Deroceras reticulatum. Z Zellforsch 130–302–311

  36. Marcaillou C, Szöllösi A (1975) Variations de perméabilité du follicule testiculaire chez le criquet Locusta migratorioïdes (Orthoptère) au cours du dernier stade larvaire. CR Acad Sci 281:2001–2004

  37. Marcaillou C, Szöllösi A (1980) A blood-testis barrier in a trematode and a fish. Generalizable concept. J Ultrastruct Res 70:128–136

  38. Meistrich ML, Trostle PK, Frapart M, Erickson RP (1977) Biosynthesis and localization of lactate dehydrogenase-X in pachytene spermatocytes and spermatids of mouse testes. Dev Biol 60:428–441

  39. Morisawa M, Suzuki K (1980) Osmolarity and potassium ion: their roles in initiation of sperm motility in teleosts. Science 210:1145–1147

  40. Noorden CJF van, Vogels IMC, James J, Tas J (1982) A sensitive cytochemical staining method for glucose-6-phosphate dehydrogenase activity in individual erythrocytes. I. Optimalization of the staining procedure. Histochemistry 75:493–506

  41. Pearse AGE (1968) Histochemistry. Theoretical and applied. London: I & A Churchill, Ltd. Vol 1

  42. Pelletier RM, Nenirovsky MS, Calvert R, Hugon JS (1981) Effects of immunization with Freund's complete adjuvant and isologous spermatozoa on the seminiferous epithelium and bloodtestis barrier in guinea pigs. Anat Rec 199:197–211

  43. Ramsay JA, Brown RHJ (1955) Simplified apparatus and procedure for freezing-point determinations upon small volumes of fluid. J Sc Instrum 32:372–375

  44. Rieder H, Teutsch HF, Sasse D (1978) NADP-dependent dehydrogenases in rat liver parenchyma I. Methodological studies on the qualitative histochemistry of G6PDH, 6PGDH, malic enzyme and ICDH. Histochemistry 56:283–298

  45. Rommerts FFG, Krüger-Sewnarain BC, Woerkom-Blik A van, Grootegoed JA, Molen HJ van der (1978) Secretion of proteins by Sertoli cells enriched cultures: effects of follicle stimulating hormone, dibutyryl cAMP and testosterone and correlation with secretion of oestradiol and androgen binding protein. Mol Cell Endocrinol 10:39–55

  46. Seligman AM, Karnovsky MJ, Wasserkrug HL, Hanker JS (1968) Nondroplet ultrastructural demonstration of cytochrome oxidase activity with a polymerizing osmiophilic reagent, diaminobenzidine (DAB). J Cell Biol 38:1–14

  47. Setchell BP (1974) Secretions of the testis and epididymis. J Reprod Fertil 37:165–177

  48. Setchell BP (1980) The functional significance of the blood-testis barrier. J Androl 1:3–10

  49. Setchell BB, Waites GMH (1975) The blood-testis barrier. In: Hamilton DW, Greep RO (eds) Male reproductive system Section 7. Handbook of physiology Vol V. Washington DC American Physiology Society, pp 143–172

  50. Sminia T (1972) Structure and function of blood and connective tissue cells of the freshwater pulmonate Lymnaea stagnalis studied by electron microscopy and enzyme histochemistry. Z Zellforsch 130:497–526

  51. Steen WJ van der, Hoven NP van den, Jager JC (1969) A method for breeding and studying freshwater snails under continuous water change with some remarks on growth and reproduction in Lymnaea stagnalis L. Neth J Zool 19:131–139

  52. Szöllösi A, Rieman J, Marcaillou C (1980) Localization of the blood-testis barrier in the testis of the moth, Anagasta kuehniella. J Ultrastruct Res 72(2):189–199

  53. Tuck RR, Setchell BP, Waites GMH, Young JA (1970) The composition of fluid collected by micropuncture and catheterization from the seminiferous tubules and rete testis of rats. Pfluegers Arch 318:225–243

  54. Verhoef HA (1981) Water balance in Collembola and its relation to habitat selection: water content, haemolymph osmotic pressure and transpiration during an instar. J Insect Physiol 27:755–760

  55. Vitale-Calpe R, Burgos MH (1970) The mechanism of spermiation in the hamster. I. Ultrastructure of spontaneous spermiation. J Ultrastruct Res 31:381–393

  56. Wendelaar Bonga SE (1970) Ultrastructure and histochemistry of neurosecretory cells and neurohaemal areas in the pond snail Lymnaea stagnalis (L). Z Zellforsch 108:190–224

  57. With ND de (1978) The effects of starvation and feeding on the ionic composition of the haemolymph in the freshwater snail Lymnaea stagnalis. Proc Kon Ned Akad Wet C81:241–248

  58. With ND de, Sminia T (1980) The effects of the nutritional state and the external calcium concentration on the ionic composition of the haemolymph and on the calcium cells in the pulmonate freshwater snail Lymnaea stagnalis. Proc Kon Ned Akad Wet C83:217–227

  59. Zachariae F, Jensen CE (1958) Studies on the mechanism of ovulation. Histochemical and physico-chemical investigations on genuine follicular fluids. Acta Endocrinol (Kbh) 27:343–355

  60. Zwaan A de, Mohamed AM, Geraerts WPM (1976) Anaerobiosis in Lymnaea stagnalis. Neth J Zool 26:549–557

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Correspondence to Dr. Marijke de Jong-Brink.

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The authors are greatly indebted to Prof. Dr. H.H. Boer for stimulating comments during this study, to Dr. C.J.F. van Noorden and Miss Ilse M.C. Vogels for localization of dehydrogenase activity; to Mr. R. van Elk for determining amino acids, and to Dr. W.P.W. van der Knaap for performing the agglutinin assay

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de Jong-Brink, M., de With, N.D., Hurkmans, P.J.M. et al. A morphological, enzyme-cytochemical, and physiological study of the blood-gonad barrier in the hermaphroditic snail Lymnaea stagnalis . Cell Tissue Res. 235, 593–600 (1984).

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Key words

  • Blood-gonad barrier
  • Septate junctions
  • Basal lamina
  • Composition of gonadal fluid
  • Enzyme cytochemistry