Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1979 Jul 1;82(1):264–277. doi: 10.1083/jcb.82.1.264

Luteinizing hormone-accelerated redistribution of lysosome-like organelles preceding dissolution of the nuclear envelope in rat oocytes maturing in vitro

PMCID: PMC2110411  PMID: 573271

Abstract

Maturation of the mammalian oocyte is characterized in part by dissolution of the nuclear envelope, or germinal vesicle breakdown (GVB). By fluorescence microscopy after vital uptake of acridine orange (AO), redistribution and perinuclear accumulation of organelles corresponding to lysosomes occur before GVB in rat oocytes undergoing meiotic maturation in vitro. In follicle-enclosed oocytes explanted during the preovulatory gonadotropin surge (GS) and individually cultured as such in chemically defined medium at approximately 22 degrees C, lysosomes aggregated into disperse clusters after 30 min; by 60 min, perinuclear concentration of lysosomes and their essential disappearance from the cortical ooplasm were observed. GVB occurred within 120 min. In contrast, follicle-enclosed oocytes explanted before the GS displayed a generally homogeneous distribution of lysosomes and intact GV for up to 5 h in culture. In oocytes aspirated from follicles before the GS, partially denuded of granulosa cells, and cultivated without added hormone, most lysosomes concentrated around the GV within 60 min, with GVB occurring generally by 120 min. Luteinizing hormone (LH) added in vitro to the isolated preparation at 3 or 30 x 10(-8) M sharply accelerated these events. The effects of LH, not seen with 1.5 x 10(-8) M hormone, were blocked by anti-LH IgG. Up to 60 x 10(-8) M follicle-stimulating hormone or 80 x 10(-8) M prolactin were ineffective in accelerating lysosome redistribution or GVB. After GVB, lysosomes became once again uniformly dispersed and unresponsive, even to 60 x 10(-8) M added LH, a finding consistent with tachyphylaxis of target cells by independent criteria. The present data, all statistically significant at P less than 0.05, demonstrate that mobilization of lysosomes before GVB is a specific response to factors that promote resumption of meiotic maturation of rat oocytes.

Full Text

The Full Text of this article is available as a PDF (1.2 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. ALLISON A. C., YOUNG M. R. UPTAKE OF DYES AND DRUGS BY LIVING CELLS IN CULTURE. Life Sci. 1964 Dec;3:1407–1414. doi: 10.1016/0024-3205(64)90082-7. [DOI] [PubMed] [Google Scholar]
  2. Anderson E., Albertini D. F. Gap junctions between the oocyte and companion follicle cells in the mammalian ovary. J Cell Biol. 1976 Nov;71(2):680–686. doi: 10.1083/jcb.71.2.680. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ascoli M., Puett D. Intracellular uptake and catabolism of lutropin by testicular tissue in vivo. FEBS Lett. 1977 Mar 15;75(1):77–82. doi: 10.1016/0014-5793(77)80057-4. [DOI] [PubMed] [Google Scholar]
  4. Ayalon D., Tsafriri A., Lindner H. R., Cordova T., Harell A. Serum gonadotrophin levels in pro-oestrous rats in relation to the resumption of meiosis by the oocytes. J Reprod Fertil. 1972 Oct;31(1):51–58. doi: 10.1530/jrf.0.0310051. [DOI] [PubMed] [Google Scholar]
  5. Balakier H., Czolowska R. Cytoplasmic control of nuclear maturation in mouse oocytes. Exp Cell Res. 1977 Dec;110(2):466–469. doi: 10.1016/0014-4827(77)90314-7. [DOI] [PubMed] [Google Scholar]
  6. Brachet J., Hanocq F., Van Gansen P. A cytochemical and ultrastructural analysis of in vitro maturation in amphibian oocytes. Dev Biol. 1970 Feb;21(1):157–195. doi: 10.1016/0012-1606(70)90067-9. [DOI] [PubMed] [Google Scholar]
  7. Calarco P. G., Donahue R. P., Szollosi D. Germinal vesicle breakdown in the mouse oocyte. J Cell Sci. 1972 Mar;10(2):369–385. doi: 10.1242/jcs.10.2.369. [DOI] [PubMed] [Google Scholar]
  8. Carrillo A. J., Sawyer C. H. Observations on successive pro-oestrous LH surges in individual 4-day cycling and post-pseudopregnant rats. Acta Endocrinol (Copenh) 1978 Jun;88(2):274–284. doi: 10.1530/acta.0.0880274. [DOI] [PubMed] [Google Scholar]
  9. Chen T. T., Abel J. H., Jr, McClellan M. C., Sawyer H. R., Diekman M. A., Niswender G. D. Localization of gonadotropic hormones in lysosomes of ovine luteal cells. Cytobiologie. 1977 Apr;14(3):412–420. [PubMed] [Google Scholar]
  10. De Duve C., Wattiaux R. Functions of lysosomes. Annu Rev Physiol. 1966;28:435–492. doi: 10.1146/annurev.ph.28.030166.002251. [DOI] [PubMed] [Google Scholar]
  11. Dean R. T., Barrett A. J. Lysosomes. Essays Biochem. 1976;12:1–40. [PubMed] [Google Scholar]
  12. Dimino M. J., Malcom S. E., Elfont E. A. Changes in acid hydrolase activities of ovarian subcellular fractions of immature rats after after gonadotropic treatment. Biol Reprod. 1977 Dec;17(5):780–786. doi: 10.1095/biolreprod17.5.780. [DOI] [PubMed] [Google Scholar]
  13. Dimino M. J., Reece R. P. Effects of gonadotropic hormones on rat ovarian lysosomes. Biol Reprod. 1973 Jun;8(5):523–530. doi: 10.1093/biolreprod/8.5.523. [DOI] [PubMed] [Google Scholar]
  14. Edwards R. G. Maturation in vitro of mouse, sheep, cow, pig, rhesus monkey and human ovarian oocytes. Nature. 1965 Oct 23;208(5008):349–351. doi: 10.1038/208349a0. [DOI] [PubMed] [Google Scholar]
  15. Elfont E. A., Roszka J. P., Dimino M. J. Cytochemical studies of acid phosphatase in ovarian follicles: a suggested role for lysosomes in steroidogenesis. Biol Reprod. 1977 Dec;17(5):787–795. doi: 10.1095/biolreprod17.5.787. [DOI] [PubMed] [Google Scholar]
  16. Fell H. B. Role of biological membranes in some skeletal reactions. Ann Rheum Dis. 1969 May;28(3):213–227. doi: 10.1136/ard.28.3.213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gordon S., Cohn Z. A. The macrophage. Int Rev Cytol. 1973;36:171–214. doi: 10.1016/s0074-7696(08)60218-1. [DOI] [PubMed] [Google Scholar]
  18. Guerrier P., Moreau M., Dorée M. Inhibition de la réinitiation de la méiose des ovocytes de Xenopus laevis par trois antiprotéases naturelles, l'antipïne, la chymostatine et la leupeptine. C R Acad Sci Hebd Seances Acad Sci D. 1977 Jan 24;284(4):317–319. [PubMed] [Google Scholar]
  19. Gwatkin R. B., Andersen O. F. Hamster oocyte maturation in vitro: inhibition by follicular components. Life Sci. 1976 Aug 15;19(4):527–536. doi: 10.1016/0024-3205(76)90232-0. [DOI] [PubMed] [Google Scholar]
  20. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  21. Lindner H. R., Tsafriri A., Lieberman M. E., Zor U., Koch Y., Bauminger S., Barnea A. Gonadotropin action on cultured Graafian follicles: induction of maturation division of the mammalian oocyte and differentiation of the luteal cell. Recent Prog Horm Res. 1974;30(0):79–138. doi: 10.1016/b978-0-12-571130-2.50007-9. [DOI] [PubMed] [Google Scholar]
  22. Lopata A., Fonseca J. R., Szego C. M. Cinemicrographic analysis of cytoplasmic and nuclear events associated with germinal vesicle breakdown in rat oocytes exposed to LH in vitro. J Reprod Fertil. 1977 Jul;50(2):211–216. doi: 10.1530/jrf.0.0500211. [DOI] [PubMed] [Google Scholar]
  23. MOUDGAL N. R., LI C. H. An immunochemical study of sheep pituitary interstitial cell-stimulating hormone. Arch Biochem Biophys. 1961 Oct;95:93–98. doi: 10.1016/0003-9861(61)90113-8. [DOI] [PubMed] [Google Scholar]
  24. Mitra S., Rao C. V. Receptors for gonadotropins and prostaglandins in lysosomes of bovine corpora lutea. Arch Biochem Biophys. 1978 Jan 15;185(1):126–133. doi: 10.1016/0003-9861(78)90151-0. [DOI] [PubMed] [Google Scholar]
  25. Moyle W. R., Moudgal N. R., Greep R. O. Cessation of steroidogenesis in Leydig cell tumors after removal of luteinizing hormone and adenosine cyclic 3',5'-monophosphate. J Biol Chem. 1971 Aug 25;246(16):4978–4982. [PubMed] [Google Scholar]
  26. Nekola M. V., Smith D. M. Failure of gonadotropins to induce in vitro maturation of mouse oocytes treated with dibutyryl cyclic AMP. J Exp Zool. 1975 Dec;194(3):529–533. doi: 10.1002/jez.1401940309. [DOI] [PubMed] [Google Scholar]
  27. ODOR D. L. Electron microscopic studies on ovarian oocytes and unfertilized tubal ova in the rat. J Biophys Biochem Cytol. 1960 Jun;7:567–574. doi: 10.1083/jcb.7.3.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Okazaki T., Strauss J. F., 3rd, Flickinger G. L. Lysosomal phospholipase A activities of rat ovarian tissue. Biochim Biophys Acta. 1977 May 25;487(2):343–353. doi: 10.1016/0005-2760(77)90010-8. [DOI] [PubMed] [Google Scholar]
  29. Peaucellier G. Initiation of meiotic maturation by specific proteases in oocytes of the polychaete annelid Sabellaria alveolata. Exp Cell Res. 1977 Apr;106(1):1–14. doi: 10.1016/0014-4827(77)90235-x. [DOI] [PubMed] [Google Scholar]
  30. Pietras R. J., Szego C. M. Surface modifications evoked by estradiol and diethylstilbestrol in isolated endometrial cells: evidence from lectin probes and extracellular release of lysosomal protease. Endocrinology. 1975 Dec;97(6):1445–1454. doi: 10.1210/endo-97-6-1445. [DOI] [PubMed] [Google Scholar]
  31. ROBBINS E., GONATAS N. K. THE ULTRASTRUCTURE OF A MAMMALIAN CELL DURING THE MITOTIC CYCLE. J Cell Biol. 1964 Jun;21:429–463. doi: 10.1083/jcb.21.3.429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. ROBBINS E., MARCUS P. I., GONATAS N. K. DYNAMICS OF ACRIDINE ORANGE-CELL INTERACTION. II. DYE-INDUCED ULTRASTRUCTURAL CHANGES IN MULTIVESICULAR BODIES (ACRIDINE ORANGE PARTICLES). J Cell Biol. 1964 Apr;21:49–62. doi: 10.1083/jcb.21.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. SOTELO J. R., PORTER K. R. An electron microscope study of the rat ovum. J Biophys Biochem Cytol. 1959 Mar 25;5(2):327–342. doi: 10.1083/jcb.5.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Schuetz A. W. Role of hormones in oocyte maturation. Biol Reprod. 1974 Mar;10(2):150–178. doi: 10.1095/biolreprod10.2.150. [DOI] [PubMed] [Google Scholar]
  35. Smith E. R., Bowers C. Y., Davidson J. M. Circulating levels of plasma gonadotropins in 4- and 5-day cycling rats. Endocrinology. 1973 Sep;93(3):756–758. doi: 10.1210/endo-93-3-756. [DOI] [PubMed] [Google Scholar]
  36. Sorensen R. A. Cinemicrography of mouse oocyte maturation utilizing Nomarski differential-interference microscopy. Am J Anat. 1973 Mar;136(3):265–276. doi: 10.1002/aja.1001360302. [DOI] [PubMed] [Google Scholar]
  37. Szego C. M., Seeler B. J. Hormone-induced activation of target-specific lysosomes: acute translocation to the nucleus after administration of gonadal hormones in vivo. J Endocrinol. 1973 Mar;56(3):347–360. doi: 10.1677/joe.0.0560347. [DOI] [PubMed] [Google Scholar]
  38. Szego C. M. Steroid-protein binding: from circulating blood to target cell nucleus. Gynecol Invest. 1976;7(5):251–279. doi: 10.1159/000301388. [DOI] [PubMed] [Google Scholar]
  39. Szego C. M. The lysosome as a mediator of hormone action. Recent Prog Horm Res. 1974;30(0):171–233. doi: 10.1016/b978-0-12-571130-2.50009-2. [DOI] [PubMed] [Google Scholar]
  40. Szego C. M. The role of cyclic AMP in lysosome mobilization and their nucleotropic translocation in steroid hormonal target cells. Adv Cyclic Nucleotide Res. 1972;1:541–564. [PubMed] [Google Scholar]
  41. Temin H. M. Stimulation by serum of multiplication of stationary chicken cells. J Cell Physiol. 1971 Oct;78(2):161–170. doi: 10.1002/jcp.1040780202. [DOI] [PubMed] [Google Scholar]
  42. Tsafriri A., Channing C. P., Pomerantz S. H., Lindner H. R. Inhibition of maturation of isolated rat oocytes by porcine follicular fluid. J Endocrinol. 1977 Nov;75(2):285–291. doi: 10.1677/joe.0.0750285. [DOI] [PubMed] [Google Scholar]
  43. Tsafriri A., Kraicer P. F. The time sequence of ovum maturation in the rat. J Reprod Fertil. 1972 Jun;29(3):387–393. doi: 10.1530/jrf.0.0290387. [DOI] [PubMed] [Google Scholar]
  44. Tsafriri A., Lindner H. R., Zor U., Lamprecht S. A. In-vitro induction of meiotic division in follicle-enclosed rat oocytes by LH, cyclic AMP and prostaglandin E 2 . J Reprod Fertil. 1972 Oct;31(1):39–50. doi: 10.1530/jrf.0.0310039. [DOI] [PubMed] [Google Scholar]
  45. Vermeiden J. P., Zeilmaker G. H. Relationship between maturation division, ovulation and leuteinization in the female rat. Endocrinology. 1974 Aug;95(2):341–351. doi: 10.1210/endo-95-2-341. [DOI] [PubMed] [Google Scholar]
  46. Wassarman P. M., Josefowicz W. J., Letourneau G. E. Meiotic maturation of mouse oocytes in vitro: inhibition of maturation at specific stages of nuclear progression. J Cell Sci. 1976 Dec;22(3):531–545. doi: 10.1242/jcs.22.3.531. [DOI] [PubMed] [Google Scholar]
  47. Weimar V. L., Haraguchi K. H. Immediate changes in acid phosphatase staining following injury to corneal mesenchymal cells. Life Sci. 1969 Jun 1;8(11):627–631. doi: 10.1016/0024-3205(69)90024-1. [DOI] [PubMed] [Google Scholar]
  48. Zamboni L. Fine morphology of the follicle wall and follicle cell-oocyte association. Biol Reprod. 1974 Mar;10(2):125–149. doi: 10.1095/biolreprod10.2.125. [DOI] [PubMed] [Google Scholar]
  49. Zeilmaker G. H., Verhamme C. M. Observations on rat oocyte maturation in vitro: morphology and energy requirements. Biol Reprod. 1974 Sep;11(2):145–152. doi: 10.1095/biolreprod11.2.145. [DOI] [PubMed] [Google Scholar]
  50. Zelenin A. V. Fluorescence microscopy of lysosomes and related structures in living cells. Nature. 1966 Oct 22;212(5060):425–426. doi: 10.1038/212425a0. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES