Cell Tiss. Res. t60, 539--549 (1975) 9 by Springer-Verlag 1975
Multiple Rough Endoplasmic Cisternae in the Endocrine Pancreas of the Adult Rat Dora Wassermann Department of Occupational Health, Hebrew University - Hadassah Medical School, Jerusalem, Israel Received January 30, 1975
Summary. Multiple rough endoplasmic cisternae were found in the parenchymatous cells of the endocrine pancreas of the adult rat (alpha, beta, D and intermediary cells) and were especially developed in beta cells. They are considered to be normal constituents of the parenchymatous cells of the endocrine pancreas. Their close proximity to Golgi dictyosomes and the accumulation of secretory material sometimes seen at the extremities of such cisternae, suggest that they may have a role in the secretory activity of these endocrine cells. Key words: Endocrine pancreas - - Adult rat - - Endoplasmic reticulum - - Electron microscopy. Introduction Multiple rough e n d o p l a s m i c cisternae (MREC), h a v e been r e p o r t e d u n d e r different n a m e s (Table 1) in some t u m o r a l a n d non t u m o r a l d i v i d i n g cells a n d a t i n t e r p h a s e in some cells generally c h a r a c t e r i z e d b y r a p i d g r o w t h a n d / o r f r e q u e n t m u l t i p l i c a t i o n , in a n i m a l tissues (Table 2) a n d in p l a n t s (Esau a n d Gill, 1971). M R E C h a v e been described as closely a p p o s e d cisternae of r o u g h e n d o p l a s m i c r e t i c u l u m ( R E R ) , s e p a r a t e d b y n a r r o w spaces. These spaces are in some cases electron lucent, in others, more electron dense t h a n t h e l u m e n of t h e cisternae (Kudo, 1971) or t h e s u r r o u n d i n g g r o u n d c y t o p l a s m (Leak et al., 1967; Szollosi, 1967). S o m e t i m e s t h e y p r e s e n t a central line of y e t higher electron d e n s i t y (Szollosi, 1967) or t w o such parallel lines (Odor, 1965; Szollosi, 1967). I n c e r t a i n cases n a r r o w connections e x t e n d between t h e limiting m e m b r a n e s . R i b o s o m e s are a t t a c h e d to t h e o u t e r m o s t cisternal m e m b r a n e s a n d are a b s e n t on the inner membranes of MREC. M R E C v a r y from less t h a n 1 tz to several microns in length. T h e y are linear or i r r e g u l a r l y curved, cylindrical or spiral structures. I n t h e l a t t e r cases, t h e central core of c y t o p l a s m i c m a t r i x sometimes contains o t h e r organellae (Epstein, 1961). I n m i t o t i c cells, M R E C can be seen on the surface of t h e c h r o m o s o m a l masses, a m i d t h e spindle fibers or a t a more d i s t a n t location. A t i n t e r p h a s e t h e y are f o u n d to be j u x t a n u c l e a r , when one of t h e c o n s t i t u e n t cisternae consists of a segment of t h e n u c l e a r m e m b r a n e a t which n u c l e a r pores are lacking, or in t h e rest of t h e c y t o p l a s m b e t w e e n the nuclear envelope a n d t h e cell limiting m e m b r a n e . These
Send o//print requests to: Dr. Dora Wassermann, Dept. of Occupational Health, Hebrew University - - Hadassah Medical School, P.O. Box 1172, Jerusalem, Israel.
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Table 1. Other names for "Multiple rough endoplasmic cisternae" References "Paired structures" "Remnants of nuclear envelope" "Spindle lamellae" "Apposition of two nuclear membranes" "Remnants (of the excess) of the nuclear membrane" "Paired double membranes", "Paired cisternae" and "Skeins or folded lamellac of quadruple membranes" "Pairs of double cytoplasmic membranes" "Aggregated endoplasmic reticulum" "Confronting cisternae of granular endoplasmic reticulum" "Paired cisternae" "Paired cisternae" or "Multilamellated arrays" "Parafusorial lamellae" "Multiple inner membrane pairs" "Ergastoplasmic lamellac" "Paired cisternae", "Doubled and tripled membranous lamellae" and "Membranous laminae" "Paired cisternae" and "Four layered lamellar (membranous) structures" "Subsurface" and "Intracytoplasmic confronting cisternae" "Paired cisternae" and "Double cytoplasmic membranes" "Tubular endoplasmic reticulum" "Paired double membranes" "Duplication of the nuclear envelope" "Pentalaminar membranous structures" and "Pentalaminar ribbons" "Spindle filaments" "Peculiar stacks of membranes" and "Membrane structures" "Twin cisternal complexes" and "Multilaminated cisternal membranes"
Andersen, H. et al. (1971) Brinkley, B. R. et al. (1967) Buck, R. C. et al. (1961) Cesarini, J. P. et al. (1968) Chang, J. P. and Gibley, C. W. (1968) Deane, H.W. (1964) Epstein, M. A. (1961) Esau, K. and Gill, R. H. (1971) Flickinger, C. J. (1969) Hruban, Z. et al. (1965) Hu, F. (1971) Jto, S. (1960) Johnson, F. R. and Roberts, K. ]~. (1964) Kaufmann, B. P. et al. (1956) Kelley, R. 0. (1970, 1971) Kudo, S. (1971) Kumegawa, M. et al. (1968) Leak, L. V. (1967) Locker, J. et al. (1968) Maul, G. G. (1970) Murray, R. G. et al. (1965) Napolitano, L. and Gagne, H. T. (1963) Porter, K. R. (1957) Procicchiani, G. et al. (1968) Szollosi, D. (1967)
cisternae m a y be finite a t their extremities, or continue with classical R E R , with a n n u l a t e lamellae (Procicchiani, 1968) or t h e Golgi a p p a r a t u s (Locker et al., 1968). M R E C h a v e been t h o u g h t to result a t p r o p h a s e from t h e b r e a k d o w n of t h e nuclear envelope. T h e y h a v e been considered to he, a t i n t e r p h a s e t h e residue of m e m b r a n e s which c o n t r i b u t e d to t h e f o r m a t i o n of t h e nuclear m e m b r a n e of d a u g h t e r nuclei, t h e residue of R E R d e g e n e r a t i o n (Chang a n d Gibley, 1968) or t h e p r o d u c t of t h e nuclear envelope ( H a n a o k a a n d F r i e d m a n , 1970) or of a g g r e g a t e d R E R cisternac. M R E C h a v e been t h o u g h t to c o n t r i b u t e to t h e r e s t o r a t i o n of the nuclear envelope in d a u g h t e r nuclei, or to be a source or R E R (Chang a n d Gibley, 1968;
Fig. 1. P a r t s of four ceils of a Langerhans islet in an adult rat. Multiple rough endoplasmic cisternae m a y be seen a t arrows in the alpha-cell (A), beta-cell (B) and D-cell (D). Also noted are: S secretory granules characteristic of each of these three kinds of cells, p beta-prosecretory granule, N nucleus, Ly lysosome, M mitochondria, L lumen of capillary and E endothelial cell. • 18810
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D. Wassermann Table 2
Multiple R E R eisternae were reported in
Human tissues
Tumoral
:Non tumoral
Animal tissues
Tumoral
:Non tumoral
in vivo
Phase of cell cycle Fibromyxosarcoma l~habdomyosarcoma 0steosarcoma l~eticulum cell sarcoma Giant ceil tumor of bone
Interphase Mitosis
in vitro
H-19 cells KB cells HeLa cells
Interphase ,, ,,
in vivo
Cells lining Rathke's pouch An erythroid cell in fetal liver Mesenchymal ceils
~Iitosis ,,
in vitro
Small lymphocyte Small lymphocyte
,, ,,
in vivo
Abdominal tumors from rats with Sr 8~ induced chloroleukemia The :Novikoff hepatoma of the rat The Morris 3683 hepatoma of the rat The Walker 256 carcinoma Yoshida aseites hepatoma ceils Rat endothelioma Solid hepatoma of rats
Interphase Mitosis
in vitro
Colcemid treated mouse BI~ melanoma
Interphase
in vivo
Testis of Drosophyla melanogaster Primary spermatocytes of Drosophyla virilis Epididymis
Mitosis
Primary oocytes Hamster oocytes of unilaminar follicles Rat thymocytes Hamster thymocytes A fibroblast Pigeon esophageal epithelium White adipose cells after prolonged low food intake hrcuate neuron The uterine glands of the pig
,,
in vitro
Chinese hamster Clone strain Don C.
Interphase
Mitosis ,, ,, Interphase ,, 7, Mitosis
H a n a o k a a n d F r i e d m a n , 1970; P r o c i e e h i a n i et al., 1968), t o be t h e e x p r e s s i o n of t e m p o r a r y i n a c t i v a t i o n of t h e s y n t h e t i c a c t i v i t y of R E R (Szollosi, 1967) or to r e p r e s e n t a f u n c t i o n a l s p e c i a l i s a t i o n of R E R (Szollosi, 1967). I n t h i s p a p e r we r e p o r t t h e o c c u r r e n c e of M R E C in t h e e n d o c r i n e p a n c r e a s , a t i n t e r p h a s e , as a n o r m a l c o n s t i t u e n t of its p a r e n c h y m a t o u s cells.
Multiple Rough Endoplasmie Cisternae in the Endocrine Pancreas
Observations
543
References Leak, L.V. et al. (1967) Hanaoka, H. and Friedman, B. (1970)
Not found in interphase r,
,,
A cloned cell line of melanotic melanoma A strain from an epidermoid carcinoma "Normal" HeLa cells
Maul, G. G. (1970) Kumegawa, M. et al. (1968) Epstein, M. A. (1961)
In the early development of the hypophysis
Andersen, H. et al. (1971) Fukuda, T. (1974) Kelley, R. O. (1970, 1971)
From limb bud of 15.5 mm embryo
Johnson, F. R. and Roberts, K. B. (1964) Proeicchiani, G. (1968)
Phytohaemaglutinine stimulated (peril. blood of healthy man) Multiple RER cisternae did not occur when the mitotic ra,te was reduced with actinomyein D or puromycin
Kelley, R. O. (1971) Hruban, Z. et al. (1965)
4-8 days after i.m. implantation in the leg of rats Cells of ascitic fluid of rats innoculated i.p. with ascitic fluid containing Yoshida hepatoma Transplantable hepatoma established after primary hepatoma induced by a chemical carcinogen
Buck, R. C. (1961) Locker, J. et al. (1968) Porter, K. g. (1957) Chang, J. P. and Gibley, C. W. J. (1968)
Almost exclusively in the pellet specimen
Hu, F. (1971)
Quoted by [14] First meiotic division In 17 and 22 days old rat fetuses, following the onset of androgen secretion at about 15 days In hamster, rat, mongolian gerbil and squirrel monkey
Kaufmann, B. P. et al. (1956) Ito, S. (1960) Flickinger, C. J. (1969)
14 days old hamsters In the lamina propria of the adult mouse gut In all the stages of mitosis In young male rats. Interpreted as adaptive altergtions in the metabolism of the cell From an ovariectomized female In the latter stages of pregnancy During colcemid synchronisation of metaphase
Szollosi, D. (1967) Odor, D. L. (1965) Murray, g. G. et al. (1965) Cesarini, J. P. et al. (1968) Deane, H. W. (1964) Kudo, S. (1971) Napolitano, L. and Gagne, H. T. (1963) King, J. C. and Williams, T. H. (1974) Crombie, P. R. (1972) Brinkley, B. R. et al. (1967)
Material and Method Six white, local strain, male rats, 3.5 and 14.5 months old, received a commercially available pellet food and water ad libitum. The animals were laparatomized under ether anesthesia. A fragment of the pancreas was immersed immediately after removal in 3.5% glutaraldehyde in phosphate buffer at pI-I 7.6 (PB), cut into fragments of less than 1 mm3
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Fig. 2. Parts of three beta-cells in the endocrine pancreas of an adult rat. Multiple rough endoplasmic cisternae (at arrows) are randomly distributed among the other organellae. i interstitial space, m microvilli, G Golgi complex, M mitochondrion, R E R rough endoplasmic reticulum, S secretory granules, P prosecretory granules. •
and kept for 11/2 hours at 4 ~ C. Afterwards they were washed in PB for 2 hours, postfixed in osmium tetroxide 1% in 0.28 M veronal buffer at pH 7.4 for 11/2hours, dehydrated in increasing concentrations of ethyl alcohol, followed by propylene oxide and imbedded in epon. Langerhans islets were identified in one ~z thick sections. Thin sections were stained with 1% w/v uranyl acetate, post stained with lead citrate (Reynolds, 1963) and examined under a Philips 300 electron microscope.
Results and Comments M R E C were seen in t h e p a r e n c h y m a t o u s cells of t h e endocrine p a n c r e a s of t h e n o r m a l a d u l t r a t s in our s t u d y . T h e y a p p e a r e d to r e p r e s e n t a n o r m a l c o n s t i t u e n t of alpha, beta, D (Figs. 1-5) a n d i n t e r m e d i a r y acino-endocrine cells (Figs. 6, 7). Their presence is m o s t f r e q u e n t in t h e b e t a cell where t h e y p r e s e n t g r e a t e r morphological v a r i a t i o n a n d q u a n t i t y . These M R E C comprise two a n d often more r o u g h endoplasmic cisternae, closely a p p o s e d one to another. The space s e p a r a t i n g t h e m is n a r r o w a n d uniform in width. The cisternal walls facing these spaces are d e v o i d of ribosomes while t h e opposite m e m b r a n e s of the cisternae, which face t h e open m a t r i x , are ribosome s t u d d e d . W h e n m o r e t h a n two cisternae are i n v o l v e d in such structures, only t h e o u t e r side of t h e p e r i p h e r a l cisternae are of t h e r o u g h t y p e . Most of t h e spaces e n c o u n t e r e d are more or less electron lucent, sometimes crossed b y fine electron dense lines (Fig. 3).
Fig. 3. Part of a beta-cell in the endocrine pancreas of an adult rat. At one extremity of multiple rough endoplasmie eisternae (thick arrows) prosecretory granules are formed (crossed arrows). The Golgi apparatus (G) is in close topographical relationship with a rough endoplasmic cisterna (thin arrow). M mitochondrion, R E R rough endoplasmic reticulum, S secretory granule, P prosecretory granule. • 56400
Fig. 4. Parts of two beta-cells in the endocrine pancreas of an adult rat. Multiple rough endoplasmic cisternae are closely apposed to Golgi apparatus (thin arrow). The space separating the two structures (arrow head) is narrow and resembles, more closely, the spaces between the components of the dictyosome, than the space of multiple rough endoplasmic cisternae (crossed arrow). Other multiple rough endoplasmic cisternae at double arrows. A dilated cisterna shows infolding of the smooth membranes of multiple rough endoplasmic cisternae (thick arrow). I interstitial space, M mitochondria, S secretory granule, P prosecretory granule, L lysosome. • 30800 Fig. 5. Part of a beta-cell of the endocrine pancreas of an adult rat. Multiple rough endoplasmic cisternae at arrows. The quadruple structure (at single arrow) is in close proximity to the Golgi apparatus. A space of the same width as the spaces which separate the dictyosome units, separates the top of the multiple rough endoplasmic cisternae from the outer strand of the Golgi dictyosome. R E R rough endoplasmic reticulum, M mitochondria, s secretory granule, P prosecretory granule, L lysosome, m microvilli. • 31350 Fig. 6. Part of an acinar beta-cell of the endocrine pancreas of an adult rat. The characteristics of the narrow spaces of multiple rough endoplasmic cisternae are maintained when these cisternae are infolded (arrows). R E R rough endoplasmic reticulum of the acinar type, Z zymogen granules, s secretory granule and p prosecretory granule of the beta-cell type, r ribosomes. • 46200 Fig. 7. Part of an acinar beta-cell of the endocrine pancreas of an adult rat. When multiple rough endoplasmic cisternae take a ring or spiral configuration (arrow), the characteristics of intercisternal spaces are preserved (thin arrow). R E R rough endoplasmic reticulum of the acinar type, rer rough endoplasmic reticulum of beta-cell type. I inclusion, containing a more electron dense matrix than that of the cell cytoplasm, a beta-secretory granule, ribosomes and a finely granulated material, M mitochondrion. • 46200
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Figs. 4 and 5
Multiple Rough Endoplasmic Cisternae in the Endocrine Pancreas
Figs. 6 and 7
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MREC are generally in c o n t i n u i t y with R E R (Fig. 1) b u t often the inner membranes are c o n t i n u o u s with the homolateral outer m e m b r a n e s at one or both extremities (Fig. 4). MREC m a y occupy a n y place between the nuclear envelope a n d the outer cell m e m b r a n e (Figs. 1-7). The narrow space between the closely apposed cisternae m a i n t a i n their u n i f o r m width even when these cisternae are dilated, vesicular, papillar (Fig. 6) circular a n d spiral (Fig. 7). These cisternae are seen f r e q u e n t l y in close p r o x i m i t y to the Golgi a p p a r a t u s (Figs. 2, 4, 5). Their morphology is in m a n y respects similar to the Golgi a p p a r a t u s (the presence of narrow spaces between the c o n s t i t u e n t s of both structures a n d the contiguous smooth membranes), b u t n o t identical. This is since the above m e n t i o n e d spaces are always larger in the M R E C t h a n i n Golgi dictyosomes (Figs. 5, 6) a n d all the m e m b r a n e s which make up the Golgi a p p a r a t u s are of the smooth type. These facts, together with the occasional presence of secretory material a t the extremities of cisternae belonging to MREC (Fig. 4), suggest t h a t these structures m a y have a role in the metabolic events which lead to the prod u c t i o n of secretory granules.
References Andersen, H., Billow, F. A., M~llgard, K. : The early development of the pars distalis of human foetal pituitary gland. Z. Anat. Entwickl.-Gesch. 185, 117-138 (1971) Brinkley, B. R., Stubblefield, E., Hsu, T. C. : The effects of colcemid inhibition and reversal on the fine structure of the mitotic apparatus of chinese hamster cells in vitro. J. Ultrastruct. Res. 19, 1-18 (t967) Buck, R. C.: Lamellae in the spindle of mitotic cells of Walker 256 carcinoma. J. biophys. biochem. Cytol. 11, 227-236 (1961) Cesarini, J. P., Benkoel, L., Bonneau, H. : Ultrastructure compar~e du thymus chez le hamster, jeune et adulte. C. R. Soc. Biol. (Paris) 162, 1975-1979 (1968) Chang, J. P., Gibley, C. W. J.: Ultrastructure of tumor cells during mitosis. Cancer Res. 28, 521-534 (1968) Crombie, P. R.: A new specialisation of the granular endoplasmic reticulum in the uterine glands of the pig in the latter stages of pregnancy. J. Cell Sci. 10, 693-704 (1972) Deane, H. W. : Some electron microscopic observations of the lamina propria of the gut with comments on the close association of macrophages, plasma cells and eosinophils. Anat. Rec. 149, 453474 (1964) Epstein, M. A. : Some unusual features of fine structure observed in the HeLa cells. J. biophys. biochem. Cytot. 10, 153-162 (1961) Esau, K., Gill, R. H. : Aggregation of endopiasmic reticulum and its relation to the nucleus in a differentiating sieve element. J. Ultrastruct. Res. 84, 144-158 (1971) Flickinger, C. J. : Fine structure of the Wolffian duct and cytodifferentiation of the epididymis in fetal rats. Z. Zellforsch. 96, 344-360 (1969) Fukuda, T.: Fetal Hemopoiesis. II Electron microscopic studies on human hepatic hemopoiesis. Virchows Arch. Abt. B 16, 249-270 (1974) Hanaoka, H., Friedman, B.: Paired cisternae in human tumor cells. J. Ultrastruct. Res. 32, 323-333 (1970) Hruban, Z., Swift, H., Rechcigl, M.: Fine structure of transplantable hepatoma of the rat. J. nat. Cancer Inst. 85,459 473 (1965) Hu, F. : Ultrastructural changes in the cell cycle of cultured melanoma cells. Anat. Rec. 170, 41-55 (1971) Ito, S. : The lamellar systems of cytoplasmic membranes in dividing spermatogenic cells of Drosophila virilis. J. hiophys, bioehem. Cytol. 7, 433442 (1960) Johnson, F. R., Roberts, K. B. : The growth and division of human small lymphocytes in tissue culture: an electron microscopic study. J. Anat. (Lond.) 98, 303-311 (1964)
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Kaufmann, B. P., Gay, H., De, D. N., Yoshida, Y., McDonald, M. R., Somme, R. : The nature of the materials of heredity. Ann. Rep., Carnegie Inst. of Wash., Yearbook 1956 Kelley, R. O.: An electron microscopic study of mcsenchyme during development of interdigital spaces in man. Anat. Rec. 168, 43-53 (1970) Kelley, R. O. : Ultrastructural comparisons of paired cisternae in leukemia and mesenchymal cells during mitosis. Anat. Rec. 171, 559-565 (1971) King, J. C., Williams, T. H. : Transformation of hypothalamic arcuate neurons. I. Changes associated with stages of the estrus cycle. Cell Tiss. Res. 153, 497-515 (1974) Kudo, S. : The "Paired Cisternae" in mitotic cells of the pigeon esophageal epithelium. Arch. hist. jap. 33, 3144 (1971) Kumegawa, M., Cattoni, M., Rose, G. G.: Electron microscopy of oral cells in vitro. II. Subsurface and intracytoplasmic confronting cisternae in strain KB ceils. J. Cell Biol. 36, 443452 (1968) Leak, L. V., Caulfield, J. B., Burke, J. F., McKhann, C. F. : Electron microscopic studies on a human fibromyosarcoma. Cancer Res. 27, 261-285 (1967) Locker, J., Goldblatt, P. J., Leighton, J.: Some ultrastructural features of Yoshida ascites hepatoma. Cancer Res. 28, 2039-2050 (1968) Maul, G. G. : On the relationship between the Golgi apparatus and annulate lamellae. J. Ultrastruct. Res. 30, 368-384 (1970) Murray, R. G., Murray, A. S., Pizzo, A. : The fine structure of mitosis in rat thymic lymphocytes. J. Cell Biol. 26, 601-619 (1965) Napolitano, L., Gagne, H. T. : Lipid depleted white adipose cells: an electron microscope study. Anat. Rec. 147, 273-294 (1963) Odor, D. L. : The ultrastructure of unilaminar follicles of the hamster ovary. Amer. J. Anat. 116, 493-522 (1965) Porter, K. R.: The submicroscopic morphology of protoplasm. Harvey Lect. Series 51, 175 (1957) Procicchiani, G., Miggiano, V., Arancia, G.: A peculiar structure of membranes in PHAstimulated lymphocytes. J. Ultrastruct. Res. 22, 195-205 (1968) Reynolds, E. S. : The use of lead citrate at high Ph as an electron-opaque stain in electron microscopy. J. Cell Biol. 17, 208-212 (1963) Szollosi, D. : Modification of the endoplasmic reticulum in some mammalian oocytes. Anat. Rec. 158, 59 74 (1967)
Multiple Rough Endoplasmic Cisternae in the Endocrine Pancreas of the Adult Rat Dora Wassermann Department of Occupational Health, Hebrew University - Hadassah Medical School, Jerusalem, Israel Received January 30, 1975
Summary. Multiple rough endoplasmic cisternae were found in the parenchymatous cells of the endocrine pancreas of the adult rat (alpha, beta, D and intermediary cells) and were especially developed in beta cells. They are considered to be normal constituents of the parenchymatous cells of the endocrine pancreas. Their close proximity to Golgi dictyosomes and the accumulation of secretory material sometimes seen at the extremities of such cisternae, suggest that they may have a role in the secretory activity of these endocrine cells. Key words: Endocrine pancreas - - Adult rat - - Endoplasmic reticulum - - Electron microscopy. Introduction Multiple rough e n d o p l a s m i c cisternae (MREC), h a v e been r e p o r t e d u n d e r different n a m e s (Table 1) in some t u m o r a l a n d non t u m o r a l d i v i d i n g cells a n d a t i n t e r p h a s e in some cells generally c h a r a c t e r i z e d b y r a p i d g r o w t h a n d / o r f r e q u e n t m u l t i p l i c a t i o n , in a n i m a l tissues (Table 2) a n d in p l a n t s (Esau a n d Gill, 1971). M R E C h a v e been described as closely a p p o s e d cisternae of r o u g h e n d o p l a s m i c r e t i c u l u m ( R E R ) , s e p a r a t e d b y n a r r o w spaces. These spaces are in some cases electron lucent, in others, more electron dense t h a n t h e l u m e n of t h e cisternae (Kudo, 1971) or t h e s u r r o u n d i n g g r o u n d c y t o p l a s m (Leak et al., 1967; Szollosi, 1967). S o m e t i m e s t h e y p r e s e n t a central line of y e t higher electron d e n s i t y (Szollosi, 1967) or t w o such parallel lines (Odor, 1965; Szollosi, 1967). I n c e r t a i n cases n a r r o w connections e x t e n d between t h e limiting m e m b r a n e s . R i b o s o m e s are a t t a c h e d to t h e o u t e r m o s t cisternal m e m b r a n e s a n d are a b s e n t on the inner membranes of MREC. M R E C v a r y from less t h a n 1 tz to several microns in length. T h e y are linear or i r r e g u l a r l y curved, cylindrical or spiral structures. I n t h e l a t t e r cases, t h e central core of c y t o p l a s m i c m a t r i x sometimes contains o t h e r organellae (Epstein, 1961). I n m i t o t i c cells, M R E C can be seen on the surface of t h e c h r o m o s o m a l masses, a m i d t h e spindle fibers or a t a more d i s t a n t location. A t i n t e r p h a s e t h e y are f o u n d to be j u x t a n u c l e a r , when one of t h e c o n s t i t u e n t cisternae consists of a segment of t h e n u c l e a r m e m b r a n e a t which n u c l e a r pores are lacking, or in t h e rest of t h e c y t o p l a s m b e t w e e n the nuclear envelope a n d t h e cell limiting m e m b r a n e . These
Send o//print requests to: Dr. Dora Wassermann, Dept. of Occupational Health, Hebrew University - - Hadassah Medical School, P.O. Box 1172, Jerusalem, Israel.
540
D. Wassermann
Table 1. Other names for "Multiple rough endoplasmic cisternae" References "Paired structures" "Remnants of nuclear envelope" "Spindle lamellae" "Apposition of two nuclear membranes" "Remnants (of the excess) of the nuclear membrane" "Paired double membranes", "Paired cisternae" and "Skeins or folded lamellac of quadruple membranes" "Pairs of double cytoplasmic membranes" "Aggregated endoplasmic reticulum" "Confronting cisternae of granular endoplasmic reticulum" "Paired cisternae" "Paired cisternae" or "Multilamellated arrays" "Parafusorial lamellae" "Multiple inner membrane pairs" "Ergastoplasmic lamellac" "Paired cisternae", "Doubled and tripled membranous lamellae" and "Membranous laminae" "Paired cisternae" and "Four layered lamellar (membranous) structures" "Subsurface" and "Intracytoplasmic confronting cisternae" "Paired cisternae" and "Double cytoplasmic membranes" "Tubular endoplasmic reticulum" "Paired double membranes" "Duplication of the nuclear envelope" "Pentalaminar membranous structures" and "Pentalaminar ribbons" "Spindle filaments" "Peculiar stacks of membranes" and "Membrane structures" "Twin cisternal complexes" and "Multilaminated cisternal membranes"
Andersen, H. et al. (1971) Brinkley, B. R. et al. (1967) Buck, R. C. et al. (1961) Cesarini, J. P. et al. (1968) Chang, J. P. and Gibley, C. W. (1968) Deane, H.W. (1964) Epstein, M. A. (1961) Esau, K. and Gill, R. H. (1971) Flickinger, C. J. (1969) Hruban, Z. et al. (1965) Hu, F. (1971) Jto, S. (1960) Johnson, F. R. and Roberts, K. ]~. (1964) Kaufmann, B. P. et al. (1956) Kelley, R. 0. (1970, 1971) Kudo, S. (1971) Kumegawa, M. et al. (1968) Leak, L. V. (1967) Locker, J. et al. (1968) Maul, G. G. (1970) Murray, R. G. et al. (1965) Napolitano, L. and Gagne, H. T. (1963) Porter, K. R. (1957) Procicchiani, G. et al. (1968) Szollosi, D. (1967)
cisternae m a y be finite a t their extremities, or continue with classical R E R , with a n n u l a t e lamellae (Procicchiani, 1968) or t h e Golgi a p p a r a t u s (Locker et al., 1968). M R E C h a v e been t h o u g h t to result a t p r o p h a s e from t h e b r e a k d o w n of t h e nuclear envelope. T h e y h a v e been considered to he, a t i n t e r p h a s e t h e residue of m e m b r a n e s which c o n t r i b u t e d to t h e f o r m a t i o n of t h e nuclear m e m b r a n e of d a u g h t e r nuclei, t h e residue of R E R d e g e n e r a t i o n (Chang a n d Gibley, 1968) or t h e p r o d u c t of t h e nuclear envelope ( H a n a o k a a n d F r i e d m a n , 1970) or of a g g r e g a t e d R E R cisternac. M R E C h a v e been t h o u g h t to c o n t r i b u t e to t h e r e s t o r a t i o n of the nuclear envelope in d a u g h t e r nuclei, or to be a source or R E R (Chang a n d Gibley, 1968;
Fig. 1. P a r t s of four ceils of a Langerhans islet in an adult rat. Multiple rough endoplasmic cisternae m a y be seen a t arrows in the alpha-cell (A), beta-cell (B) and D-cell (D). Also noted are: S secretory granules characteristic of each of these three kinds of cells, p beta-prosecretory granule, N nucleus, Ly lysosome, M mitochondria, L lumen of capillary and E endothelial cell. • 18810
542
D. Wassermann Table 2
Multiple R E R eisternae were reported in
Human tissues
Tumoral
:Non tumoral
Animal tissues
Tumoral
:Non tumoral
in vivo
Phase of cell cycle Fibromyxosarcoma l~habdomyosarcoma 0steosarcoma l~eticulum cell sarcoma Giant ceil tumor of bone
Interphase Mitosis
in vitro
H-19 cells KB cells HeLa cells
Interphase ,, ,,
in vivo
Cells lining Rathke's pouch An erythroid cell in fetal liver Mesenchymal ceils
~Iitosis ,,
in vitro
Small lymphocyte Small lymphocyte
,, ,,
in vivo
Abdominal tumors from rats with Sr 8~ induced chloroleukemia The :Novikoff hepatoma of the rat The Morris 3683 hepatoma of the rat The Walker 256 carcinoma Yoshida aseites hepatoma ceils Rat endothelioma Solid hepatoma of rats
Interphase Mitosis
in vitro
Colcemid treated mouse BI~ melanoma
Interphase
in vivo
Testis of Drosophyla melanogaster Primary spermatocytes of Drosophyla virilis Epididymis
Mitosis
Primary oocytes Hamster oocytes of unilaminar follicles Rat thymocytes Hamster thymocytes A fibroblast Pigeon esophageal epithelium White adipose cells after prolonged low food intake hrcuate neuron The uterine glands of the pig
,,
in vitro
Chinese hamster Clone strain Don C.
Interphase
Mitosis ,, ,, Interphase ,, 7, Mitosis
H a n a o k a a n d F r i e d m a n , 1970; P r o c i e e h i a n i et al., 1968), t o be t h e e x p r e s s i o n of t e m p o r a r y i n a c t i v a t i o n of t h e s y n t h e t i c a c t i v i t y of R E R (Szollosi, 1967) or to r e p r e s e n t a f u n c t i o n a l s p e c i a l i s a t i o n of R E R (Szollosi, 1967). I n t h i s p a p e r we r e p o r t t h e o c c u r r e n c e of M R E C in t h e e n d o c r i n e p a n c r e a s , a t i n t e r p h a s e , as a n o r m a l c o n s t i t u e n t of its p a r e n c h y m a t o u s cells.
Multiple Rough Endoplasmie Cisternae in the Endocrine Pancreas
Observations
543
References Leak, L.V. et al. (1967) Hanaoka, H. and Friedman, B. (1970)
Not found in interphase r,
,,
A cloned cell line of melanotic melanoma A strain from an epidermoid carcinoma "Normal" HeLa cells
Maul, G. G. (1970) Kumegawa, M. et al. (1968) Epstein, M. A. (1961)
In the early development of the hypophysis
Andersen, H. et al. (1971) Fukuda, T. (1974) Kelley, R. O. (1970, 1971)
From limb bud of 15.5 mm embryo
Johnson, F. R. and Roberts, K. B. (1964) Proeicchiani, G. (1968)
Phytohaemaglutinine stimulated (peril. blood of healthy man) Multiple RER cisternae did not occur when the mitotic ra,te was reduced with actinomyein D or puromycin
Kelley, R. O. (1971) Hruban, Z. et al. (1965)
4-8 days after i.m. implantation in the leg of rats Cells of ascitic fluid of rats innoculated i.p. with ascitic fluid containing Yoshida hepatoma Transplantable hepatoma established after primary hepatoma induced by a chemical carcinogen
Buck, R. C. (1961) Locker, J. et al. (1968) Porter, K. g. (1957) Chang, J. P. and Gibley, C. W. J. (1968)
Almost exclusively in the pellet specimen
Hu, F. (1971)
Quoted by [14] First meiotic division In 17 and 22 days old rat fetuses, following the onset of androgen secretion at about 15 days In hamster, rat, mongolian gerbil and squirrel monkey
Kaufmann, B. P. et al. (1956) Ito, S. (1960) Flickinger, C. J. (1969)
14 days old hamsters In the lamina propria of the adult mouse gut In all the stages of mitosis In young male rats. Interpreted as adaptive altergtions in the metabolism of the cell From an ovariectomized female In the latter stages of pregnancy During colcemid synchronisation of metaphase
Szollosi, D. (1967) Odor, D. L. (1965) Murray, g. G. et al. (1965) Cesarini, J. P. et al. (1968) Deane, H. W. (1964) Kudo, S. (1971) Napolitano, L. and Gagne, H. T. (1963) King, J. C. and Williams, T. H. (1974) Crombie, P. R. (1972) Brinkley, B. R. et al. (1967)
Material and Method Six white, local strain, male rats, 3.5 and 14.5 months old, received a commercially available pellet food and water ad libitum. The animals were laparatomized under ether anesthesia. A fragment of the pancreas was immersed immediately after removal in 3.5% glutaraldehyde in phosphate buffer at pI-I 7.6 (PB), cut into fragments of less than 1 mm3
544
D. Wassermann
Fig. 2. Parts of three beta-cells in the endocrine pancreas of an adult rat. Multiple rough endoplasmic cisternae (at arrows) are randomly distributed among the other organellae. i interstitial space, m microvilli, G Golgi complex, M mitochondrion, R E R rough endoplasmic reticulum, S secretory granules, P prosecretory granules. •
and kept for 11/2 hours at 4 ~ C. Afterwards they were washed in PB for 2 hours, postfixed in osmium tetroxide 1% in 0.28 M veronal buffer at pH 7.4 for 11/2hours, dehydrated in increasing concentrations of ethyl alcohol, followed by propylene oxide and imbedded in epon. Langerhans islets were identified in one ~z thick sections. Thin sections were stained with 1% w/v uranyl acetate, post stained with lead citrate (Reynolds, 1963) and examined under a Philips 300 electron microscope.
Results and Comments M R E C were seen in t h e p a r e n c h y m a t o u s cells of t h e endocrine p a n c r e a s of t h e n o r m a l a d u l t r a t s in our s t u d y . T h e y a p p e a r e d to r e p r e s e n t a n o r m a l c o n s t i t u e n t of alpha, beta, D (Figs. 1-5) a n d i n t e r m e d i a r y acino-endocrine cells (Figs. 6, 7). Their presence is m o s t f r e q u e n t in t h e b e t a cell where t h e y p r e s e n t g r e a t e r morphological v a r i a t i o n a n d q u a n t i t y . These M R E C comprise two a n d often more r o u g h endoplasmic cisternae, closely a p p o s e d one to another. The space s e p a r a t i n g t h e m is n a r r o w a n d uniform in width. The cisternal walls facing these spaces are d e v o i d of ribosomes while t h e opposite m e m b r a n e s of the cisternae, which face t h e open m a t r i x , are ribosome s t u d d e d . W h e n m o r e t h a n two cisternae are i n v o l v e d in such structures, only t h e o u t e r side of t h e p e r i p h e r a l cisternae are of t h e r o u g h t y p e . Most of t h e spaces e n c o u n t e r e d are more or less electron lucent, sometimes crossed b y fine electron dense lines (Fig. 3).
Fig. 3. Part of a beta-cell in the endocrine pancreas of an adult rat. At one extremity of multiple rough endoplasmie eisternae (thick arrows) prosecretory granules are formed (crossed arrows). The Golgi apparatus (G) is in close topographical relationship with a rough endoplasmic cisterna (thin arrow). M mitochondrion, R E R rough endoplasmic reticulum, S secretory granule, P prosecretory granule. • 56400
Fig. 4. Parts of two beta-cells in the endocrine pancreas of an adult rat. Multiple rough endoplasmic cisternae are closely apposed to Golgi apparatus (thin arrow). The space separating the two structures (arrow head) is narrow and resembles, more closely, the spaces between the components of the dictyosome, than the space of multiple rough endoplasmic cisternae (crossed arrow). Other multiple rough endoplasmic cisternae at double arrows. A dilated cisterna shows infolding of the smooth membranes of multiple rough endoplasmic cisternae (thick arrow). I interstitial space, M mitochondria, S secretory granule, P prosecretory granule, L lysosome. • 30800 Fig. 5. Part of a beta-cell of the endocrine pancreas of an adult rat. Multiple rough endoplasmic cisternae at arrows. The quadruple structure (at single arrow) is in close proximity to the Golgi apparatus. A space of the same width as the spaces which separate the dictyosome units, separates the top of the multiple rough endoplasmic cisternae from the outer strand of the Golgi dictyosome. R E R rough endoplasmic reticulum, M mitochondria, s secretory granule, P prosecretory granule, L lysosome, m microvilli. • 31350 Fig. 6. Part of an acinar beta-cell of the endocrine pancreas of an adult rat. The characteristics of the narrow spaces of multiple rough endoplasmic cisternae are maintained when these cisternae are infolded (arrows). R E R rough endoplasmic reticulum of the acinar type, Z zymogen granules, s secretory granule and p prosecretory granule of the beta-cell type, r ribosomes. • 46200 Fig. 7. Part of an acinar beta-cell of the endocrine pancreas of an adult rat. When multiple rough endoplasmic cisternae take a ring or spiral configuration (arrow), the characteristics of intercisternal spaces are preserved (thin arrow). R E R rough endoplasmic reticulum of the acinar type, rer rough endoplasmic reticulum of beta-cell type. I inclusion, containing a more electron dense matrix than that of the cell cytoplasm, a beta-secretory granule, ribosomes and a finely granulated material, M mitochondrion. • 46200
546
D. Wassermann
Figs. 4 and 5
Multiple Rough Endoplasmic Cisternae in the Endocrine Pancreas
Figs. 6 and 7
547
548
D. Wassermann
MREC are generally in c o n t i n u i t y with R E R (Fig. 1) b u t often the inner membranes are c o n t i n u o u s with the homolateral outer m e m b r a n e s at one or both extremities (Fig. 4). MREC m a y occupy a n y place between the nuclear envelope a n d the outer cell m e m b r a n e (Figs. 1-7). The narrow space between the closely apposed cisternae m a i n t a i n their u n i f o r m width even when these cisternae are dilated, vesicular, papillar (Fig. 6) circular a n d spiral (Fig. 7). These cisternae are seen f r e q u e n t l y in close p r o x i m i t y to the Golgi a p p a r a t u s (Figs. 2, 4, 5). Their morphology is in m a n y respects similar to the Golgi a p p a r a t u s (the presence of narrow spaces between the c o n s t i t u e n t s of both structures a n d the contiguous smooth membranes), b u t n o t identical. This is since the above m e n t i o n e d spaces are always larger in the M R E C t h a n i n Golgi dictyosomes (Figs. 5, 6) a n d all the m e m b r a n e s which make up the Golgi a p p a r a t u s are of the smooth type. These facts, together with the occasional presence of secretory material a t the extremities of cisternae belonging to MREC (Fig. 4), suggest t h a t these structures m a y have a role in the metabolic events which lead to the prod u c t i o n of secretory granules.
References Andersen, H., Billow, F. A., M~llgard, K. : The early development of the pars distalis of human foetal pituitary gland. Z. Anat. Entwickl.-Gesch. 185, 117-138 (1971) Brinkley, B. R., Stubblefield, E., Hsu, T. C. : The effects of colcemid inhibition and reversal on the fine structure of the mitotic apparatus of chinese hamster cells in vitro. J. Ultrastruct. Res. 19, 1-18 (t967) Buck, R. C.: Lamellae in the spindle of mitotic cells of Walker 256 carcinoma. J. biophys. biochem. Cytol. 11, 227-236 (1961) Cesarini, J. P., Benkoel, L., Bonneau, H. : Ultrastructure compar~e du thymus chez le hamster, jeune et adulte. C. R. Soc. Biol. (Paris) 162, 1975-1979 (1968) Chang, J. P., Gibley, C. W. J.: Ultrastructure of tumor cells during mitosis. Cancer Res. 28, 521-534 (1968) Crombie, P. R.: A new specialisation of the granular endoplasmic reticulum in the uterine glands of the pig in the latter stages of pregnancy. J. Cell Sci. 10, 693-704 (1972) Deane, H. W. : Some electron microscopic observations of the lamina propria of the gut with comments on the close association of macrophages, plasma cells and eosinophils. Anat. Rec. 149, 453474 (1964) Epstein, M. A. : Some unusual features of fine structure observed in the HeLa cells. J. biophys. biochem. Cytot. 10, 153-162 (1961) Esau, K., Gill, R. H. : Aggregation of endopiasmic reticulum and its relation to the nucleus in a differentiating sieve element. J. Ultrastruct. Res. 84, 144-158 (1971) Flickinger, C. J. : Fine structure of the Wolffian duct and cytodifferentiation of the epididymis in fetal rats. Z. Zellforsch. 96, 344-360 (1969) Fukuda, T.: Fetal Hemopoiesis. II Electron microscopic studies on human hepatic hemopoiesis. Virchows Arch. Abt. B 16, 249-270 (1974) Hanaoka, H., Friedman, B.: Paired cisternae in human tumor cells. J. Ultrastruct. Res. 32, 323-333 (1970) Hruban, Z., Swift, H., Rechcigl, M.: Fine structure of transplantable hepatoma of the rat. J. nat. Cancer Inst. 85,459 473 (1965) Hu, F. : Ultrastructural changes in the cell cycle of cultured melanoma cells. Anat. Rec. 170, 41-55 (1971) Ito, S. : The lamellar systems of cytoplasmic membranes in dividing spermatogenic cells of Drosophila virilis. J. hiophys, bioehem. Cytol. 7, 433442 (1960) Johnson, F. R., Roberts, K. B. : The growth and division of human small lymphocytes in tissue culture: an electron microscopic study. J. Anat. (Lond.) 98, 303-311 (1964)
Multiple Rough Endoplasmic Cisternae in the Endocrine Pancreas
549
Kaufmann, B. P., Gay, H., De, D. N., Yoshida, Y., McDonald, M. R., Somme, R. : The nature of the materials of heredity. Ann. Rep., Carnegie Inst. of Wash., Yearbook 1956 Kelley, R. O.: An electron microscopic study of mcsenchyme during development of interdigital spaces in man. Anat. Rec. 168, 43-53 (1970) Kelley, R. O. : Ultrastructural comparisons of paired cisternae in leukemia and mesenchymal cells during mitosis. Anat. Rec. 171, 559-565 (1971) King, J. C., Williams, T. H. : Transformation of hypothalamic arcuate neurons. I. Changes associated with stages of the estrus cycle. Cell Tiss. Res. 153, 497-515 (1974) Kudo, S. : The "Paired Cisternae" in mitotic cells of the pigeon esophageal epithelium. Arch. hist. jap. 33, 3144 (1971) Kumegawa, M., Cattoni, M., Rose, G. G.: Electron microscopy of oral cells in vitro. II. Subsurface and intracytoplasmic confronting cisternae in strain KB ceils. J. Cell Biol. 36, 443452 (1968) Leak, L. V., Caulfield, J. B., Burke, J. F., McKhann, C. F. : Electron microscopic studies on a human fibromyosarcoma. Cancer Res. 27, 261-285 (1967) Locker, J., Goldblatt, P. J., Leighton, J.: Some ultrastructural features of Yoshida ascites hepatoma. Cancer Res. 28, 2039-2050 (1968) Maul, G. G. : On the relationship between the Golgi apparatus and annulate lamellae. J. Ultrastruct. Res. 30, 368-384 (1970) Murray, R. G., Murray, A. S., Pizzo, A. : The fine structure of mitosis in rat thymic lymphocytes. J. Cell Biol. 26, 601-619 (1965) Napolitano, L., Gagne, H. T. : Lipid depleted white adipose cells: an electron microscope study. Anat. Rec. 147, 273-294 (1963) Odor, D. L. : The ultrastructure of unilaminar follicles of the hamster ovary. Amer. J. Anat. 116, 493-522 (1965) Porter, K. R.: The submicroscopic morphology of protoplasm. Harvey Lect. Series 51, 175 (1957) Procicchiani, G., Miggiano, V., Arancia, G.: A peculiar structure of membranes in PHAstimulated lymphocytes. J. Ultrastruct. Res. 22, 195-205 (1968) Reynolds, E. S. : The use of lead citrate at high Ph as an electron-opaque stain in electron microscopy. J. Cell Biol. 17, 208-212 (1963) Szollosi, D. : Modification of the endoplasmic reticulum in some mammalian oocytes. Anat. Rec. 158, 59 74 (1967)
