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Annulate lamellae in human tumor cells
TISSUE
&CELL
1979 II (I)
P~hli.shrcl br Longntm
139-146
Group Ltd. Printd
in Grcwt Britcrin
C. N. SUN and H. J. WHITE
ANNULATE CELLS
LAMELLAE
IN HUMAN
TUMOR
ABSTRACT. Annulate lamellae have been found in a primitive neuroectodermal tumor, a metastatic cerebellar tumor, a testicular seminoma, a retinoblastoma and three melanomas. These annulate lamellae are arranged in stacked parallel arrays in the cytoplasm of tumor cells. The number of annulate lamellae observed to comprise a single stack varies from 2-4 in the seminoma tumor to 5-18 in the cerebellar tumor. Although the functional significance of annulate lamellae is still unknown, in many instances they have been found to be continuous with rough-surfaced cisternae of the endoplasmic reticulum and ribosomes have been demonstrated on the surface of annulate lamellae. This may suggest that annulate lamellae participate in protein synthesis.
Introduction first electron microscopic observation on annulate lamellae (AL) were made by McCulloch ( 1952) and Lansing et al. (I 952) in oocytes of marine animals. Palade (1955) described this organelle as fenestrated cisternae in the rat spermatocyte. The term ‘annulate lamellae’ was first used by Swift (I 956) to describe this organelle in the pancreatic acinar cells of the larval salamander, oocytes of snail and clam, and rat spermatids. The occurrence of the organelle has been documented in the oocytes of several different species of animals (McCulloch, 1952; Afzelius, 1955; Palade, 1955; Swift, 1956, 1968; Rebhun, 1956; Pasteels etal., 1958; Merriam, 1959; Gross et al., 1960; Kane, 1960; Hsu, 1963; Kessel, 1963, 1964, 1965, 1966), in spermatids and Sertoli cells (Ruthmann, 1958; Kaye et a/., 1961; Barer et al., 1960; Swift, 1956; Bawa, 1963; Nagano, 1966; Smith and Berlin, 1977; Sun et ul., 1977), and in cells of a variety of malignant tumors (Wessel and Bernhard, 1957; Epstein, 1957, 1963 ; I 959 ; Hoshino, I961 ; Binggeli, Chambers and Weiser, 1964 ; Svo boda, 1964;
THE
Veterans Administration of Arkansas for Medical Arkansas 72206. Received 31 March 1978. Revised 30 August 1978.
Hospital and University Sciences, Little Rock,
Ma and Webber, 1966; Locker cl Al., 1969: Ghadially and Parry, 1974). Annulate lamellae seem to have a widespread occurrence in animal tumors; however, their presence in human neoplastic material has been reported in only a few cases (Epstein, 1957; Kumegawa et al., 1961; Leak and Bensch, 1971: et al., 1967; Kadin Kovacs ef al., 1975, 1977). Recently, we found thes: structures in szvzral tumors. The increasing number of reports in neoplastic cells of annulate lamellae may suggest that the annulate lamellae have an unknown role in some neoplastic cells. Materials and Methods Biopsy specimens from a primitive neuroectodermal tumor, a metastatic cerebellar tumor, a testicular seminoma, a retinoblastoma and three melanomas were cut into small pieces about I mm3 and immediately fixed in 4% glutaraldehyde in phosphate buffer fat 2 hr, then post-fixed in 1 ‘A buffered osmium tetroxide for 1 hr. After dehydration, tissues were embedded in Epon 812. Thin sections were stained with uranyl acetate and lead citrate (Reynolds, 1963). Observations The 139
ultrastructural
features
of
annulate
140
SUN
lamellae from the above materials constitute the subject of the present study; other findings will not be presented here. Annulate lamellae are smooth-walled, paired membranes arranged in parallel stacks. The number of annulate lamellae observed to comprise a single stack varies with tissue examined (Figs. l-3 and Table 1). The membranes show regularly spaced discontinuities. By appropriate section, ‘pores’ on the membranes are seen which have an outside diameter of approximately 1000-1600 A. The center of the pores may appear empty, but sometimes a small central granule may be shown in tangential section (Fig. 3a). Granular and fibrillar material is present within the ‘pores’, especially in the vicinity of the periphery (Figs. 1, 3). We found in one case (p.e. tumor) that the annulate lamellae are in contact with the nuclear envelope (Fig. 4). More commonly, the annulate lamellae are in contact with the rough endoplasmic reticulum (Figs. 2, 3). In one of the melanomas, a peculiar membranous body was found in the cytoplasm of the tumor cell. In some profiles, this structure consists of stacks of braided canaliculate elements or channels which pursue a tortuous course and cross one
Fig.
1. Annulate
Table
AND
WHITE
I. Tissue examined No. of AL in
Tumor
a stack
Melanoma Seminoma Retinoblastoma Cerebellar tumor Primative neuroectodermal
tumor
8-10 2-4 t-8 5-18 4-7
another (Fig. 5). In many of the sections, profiles showed many ‘pores’ with central granules (Fig. 6). Here, too, channels are connected to the rough endoplasmic reticulum (Figs. 5, 6). A portion of the profile shows a similarity to the annulate lamellae (Fig. 6) of the myocardial cell in the chick embryo (Merkow and Leighton, 1966) and annulate lamellae of Sertoli cells in azoospermic human testis (Livni et al., 1977). These membranous structures may possibly represent modified annulate lamellae. Tn the cytoplasm of the primitive neuroectodermal tumor, there are many instances of membranous structures which may be precursors of the annulate lamellae (Fig. 7). These membranous structures are quite similar to the Golgi apparatus and sometimes
lamellae (AL) in retinoblastoma.
P, pore; N, nucleus.
x 50,000.
Fig. 2. Annulate lamellae (AL) in a nuclear pocket in melanoma. reticulum; G, Golgi apparatus; N, nucleus. x 16,000.
ER, endoplasmic
Fig. 3. Annulate lamellae reticulum. x 30,000.
ER, endoplasmic
(AL) in testicular
seminoma.
P, pore;
Fig. 3a. Tangential section of annuli: a small dense granule is sometimes in the center of the annulus ( +). ER, endoplasmic reticulum. x 30,000.
observed
Fig. 4. A stack of annulate lamellae is in contact with the nuclear envelope of a primitive neuroectodermal tumor (+). lntranuclear lamellae are also present (IAL). N, nucleus. x 43,000. Fig. 5. Peculiar configuration of agranular reticulum of braided channels in a melanoma tumor cell (+). ER, endoplasmic reticulum; G, Golgi apparatus. x 40,000. Fig. 6. Many of the profiles of the peculiar configuration show pores with central granules (+). ER, endoplasmic reticulum; G, Golgi apparatus. x 40,000. Fig. 7. Membranous structure may be a precursor annulate lamellae; G, Golgi apparatus. x 44,000.
of the annulate
lamellae (+).
AL,
SUN
144
AND
WHITE
Table 2. Some previous observations of annulate lamellae in human tumor cells No. of AL in a stack
Tumor
HeLa cells KB cells Pheochromocytoma Melanoma Apocrine hidrocytoma Warthin’s tumor Fibromyxosarcoma Adenomatous pituitary glands
4-8 4 5 4 3 6 2-l
are closely associated with the Golgi apparatus. Discussion Annulate lamellae occur frequently in rapidly growing and differentiating cell systems under normal and abnormal conditions (Porter, 1961). They are also occasionally associated with cell degeneration. An increase in the number of annulate lamellae has been noted in virus-infected animal (Koestner et al., 1966), and plant cells (Steinkamp and Hoefert, 1977), as well as in cells treated with a variety of chemicals (Hruban et al., 1965b, c). Since annulate lamellae are a constituent of many different types of cells, we may infer that the organelles have multiple functions that may differ depending upon the cell type. Among their suggested functions are protein synthesis (Merkow and Leighton, 1966; Wischnizer, 1970), nucleocytoplasmic exchanges (Kessel, 1968), nucleation sites for tubulin synthesis (De Brabander and Rogers, 1975), storage sites (Fanke, 1974), or as a disposal mechanism (Kessel, 1973). As for their origin, Afzelius (1955) first thought that this lameller system represented fragments of the nuclear membrane remaining in the cytoplasm after mitosis. Rebhum (1956) and Swift (1956) have suggested that annulate lamellae appeared to be synthesized as sheets on the surface of the nuclear envelope. In several cell systems, such as sea urchin and dragonfly oocytes, there is evidence that basophilic masses derived from the nucleus become associated with annulate lamellae (Kessel, 1968; Kessel and Beams, 1969). After the stacks of annulate lamellae have completely formed, rough endoplasmic reticulum extends from each of the annulate lamellae in a stack (Sun et al., 1977). Although many authors believe that annulate lamellae take their origin from the
Authors In In In In In
vitro vitro vitro vitro vivo In vivo In duo In vivo
Epstein (1961) Kumegawa et al. (1967)
Kadin and Bensch (1971) Maul (1970) Gross (1965) Tandler (1966) Leak et al. (1967) Kovacs (1975)
nuclear envelope, the precise origin of the annulate lamellae has not been firmly established. As early as 1955, Palade considered them to be a local differentiation of endoplasmic reticulum. Other authors (Kovacs et al., 1975) also believe they originate from endoplasmic reticulum. Hruban et al. (1965a) suggested that annulate lamellae may represent an intermediate stage in the formation of more typical endoplasmic reticulum, possibly containing only the messenger RNA before addition of free ribosome, or ribosomal RNA before the addition of protein. In the human tumor cells, there are very few instances of annulate lamellae and nuclear envelope association. Our study suggests that in tumors, annulate lamellae may possibly be formed from transformed cisternae of endoplasmic reticulum or the Golgi apparatus. Supporting this contention are reports by Maul (1970) and Scheer and Franke (1969). Hruban et al. (1965a), in a study of transplantable hepatomas of rats, reported their frequent occurrence particularly in rapidly growing tumors probably related to intense protein synthesis. Porter also noted their frequent occurrence in actively proliferating, embryonic, undifferentiated cells (Porter, 1961). By contract, in human tumors, AL are reported infrequently and paradoxically. Sometimes they even occur in slowly growing benign neoplasms (Tandler, 1966) (Table 2). More research is needed to answer the question of how the annulate lamellae relate to growth or protein metabolism. Acknowledgements This work was supported by Veterans Administration Research Funds. The technical assistance of Joe Meador and Bettye Stallings and the secretarial assistance of Diane Butler are greatly appreciated.
ANNULATE
LAMELLAE
IN TUMOR
CELLS
14.5
References AFZELIUS, B. A. 1955.The ulstrastructure of the nuclear membrane of the sea urchin oocyte as studied with the electron microscope. Expl Cell Res., 8, 147-158. BARER, R., JOSEPH, S. and MEEK, G. A. 1960. The origin and fate of the nuclear membrane in meiosis. Proc. R. Sot. B, 152, 353-366. BAWA, S. R. 1963. Fine structure of the Sertoli cell of the human testis. J. Ulrrastruct. Res., 9. 459-474. BINCGELI, M. F. 1959. Abnormal intranuclear and cytoplasmic formations associated with a chemically induced transplantable chicken sarcoma. J. biophys. biochem. Cytol., 5, 143-152. CHAMBERS, V. C. and WEISER, R. S. 1964. Annulate lamellae in sarcoma 1 cells. J. Cell Biol., 21, 133-I 39.
D~BRABANDER,M. and ROGERS, M. 1975. The formation of annulated lamellae induced by the disintegration of microtubules. J. Cell Sci., 19, 331-340. EPSTEIN, M. A. 1957. The fine structure of the cells in mouse sarcoma 37 axcitic fluids. J. biophvs. biochem. Cytol., 3, 567-576. EPSTEIN, M. A. 1961. Some unusual Cyrol., 10, 153-162.
features
of fine structure
observed
in HeLa cells. J. biophys. hiochem.
FRANKE. W. W. 1974. Structure, biochemistry and functions of the nuclear envelope. Int. Rev. Gyro/. (Suppl. 4), 71-236. GHADIALLY. F. N. and PARRY, E. W. 1974. lntranuclear annulate lamellae in Ehrilich ascites tumour cell. Virchorus Arch. path. Anat. Physiol., 15, 131-137. Gross. B. G. 1965. The apocrine hidrocystoma, an electron microscope study. Fedn Proc., 24, 432. GROSS, P. R.. PHILPOTT, D. E. and NASS, S. 1960. Electron microscopy of the centrifuged sea urchin egg, with a note on the structure of the ground cytoplasm. J. biophys. biochem. Cytol., 7, 135-141. HOSHINO, M. 1963. Submicroscopic characteristics of four strains of Yoshida ascites hepatoma of rats: a comparative study. Cancer Res., 23, 209-216. HRUBAN, 2.. SWIFT, H. and RECHCIGL, M. 1965a. Fine structure of transplantable hepatomas of the rat. J. natn. Cancer Inst.. 35, 459495. HI~UBAN,Z., SWIFT, H., DUNN, F. W. and LEWIS, D. E. 1965b. Effects of fl-3-furylalanine on the ultrastructure of the hepatocytes and pancreatic acinar cells. Lab. Invest., 14, 70-80. HI<~BAN. Z.. SWIFT. H. and SLESERS,A. 1965~. Effect of azoserine on the fine structure of the liver and pancreatic acinar cells. Cancer Res., 25, 708-723. Hsu, W. S. 1963. The nuclear envelope in the developing oocytes of the tunicate Boltenia Villosa. Z. Zellforsch. mikrosk.
Anat.. 58, 660-678.
KADIN, M. E. and BENSCH, K. G. 1971. Comparison of pheochromocytes with ganglion cells and neuroblasts grown in vitro. Cancer, 27, 1148-l 160. KANT, R. E. 1960. The effect of partial protein extraction on the structure of the eggs of the sea urchin. Arbacia punctulata.
J. biophys. biochem.
Cytol., 7, 21-26.
KAYF, G. r., PAPPAS, G. D., YASLJZUMI,G. and YAMAMOTO,H. 1961. The distribution of the endoplasmic reticulum during spermatogenesis in the crayfish, Camabroides japonicus. Z. Zellforsch. mikrosk. Anar., 53. 159-171. KFSSEL, R. G. 1963. Electron microscope J. C~,ll Biol., 19, 391-414.
Kksst~., R. G. 1964. Intranuclear mihrosk.
annulate
studies on the origin of annulate lamellae in oocytes
lamellae in oocytes of Necfrrrus.
of the tunicate,
Sryela partita. Z. Zellforvch.
Anat.. 63, 37-51.
Kksst L. R. G. 1965. Intranuclear and cytoplasmic annulate lamellae in tunicate oocytes. J. Cell Eiol.. 24. 471-487. KI SS~L, R. G. 1966. Ultrastructure and relationship of ooplasmic components in tunicates. Acra Embr.vo/. Morphol. Exp., 9, l-24. R. G. 1968. Annulate lamellae.
J. Ultrastrurf. Res. Suppl., 10, l-82. KISSEL, R. G. and BEAMS, H. W. 1969. Annulate lamellae and ‘yolk nuclei’ in oocytes of the dragonfly, Lebellula prrlchella. J. Cell Biol., 42, 185-201. K~ss~L. R. G. 1973. Structure and function of the nuclear envelope and related cytomembranes. Prop. Swg. Ktw~,
Membr. Sri., 6, 243-329. KOF,STNER,A., KASZA, L. and KINDIG, 0. 1966. Electron microscopy polioencephalomyelitis virus. Am. J. Path., 48, 129-148.
KOVA(.S. K., HORVATH, E. and BILBAO, J. M. 1975. Annulate glands. Arto Anat., 93, 249.-256.
IO
of tissue cultures
lamellae
in adenomas
infected with porcine of human
pituitary
146
SUN
AND
WHITE
KOVACS, K., HORVATH, E., BILBAO, J. M. and LISE, R. G. 1977. Annulate lamellae in spontaneous prolactin cell adenomas of the rat pituitary. Anat. Am., 141, 59-65. KUMEGAWA, M., CATTONI, M. and ROSE, G. G. 1967. Electron microscopy of oral cells in vitro. I. Annulate lamellae observed in strain KB cells. J. Cell Biol., 34, 897-1967. LANSING, A. I., HILLIER, J. and ROSENTHAL,T. B. 1952. Electron microscopy of some marine egg inclusions. Biol. Bull., 103, 294. LEAK, L. V., CANEFIELD,J. B., BURKE, J. F. and MEKHANN, C. F. 1967. Electron microscopic studies on a human fibromyxosarcoma. Cancer Res., 27, 261-285. Lrvwr, N., PALTIN, Z., SEGAL, S. and LAUFER, A. 1977. Fine structure of Sertoli and Leydig cells in a azoospermic human testis. Arch. Path. Lab. Med., 101, 442-445. LOCKER, J., GOLDBLATT, P. J. and LEIGHTON, J. 1969. Hematogenous metastasis of Yoshida ascites hepatoma in the chick embryo liver: ultrastructural changes in tumor cells. Cancer Res., 29, 1244. MA, M. H. and WEBBER,A. J. 1966. Fine structure of liver tumors induced in the rat by 3’-methyl-4-dimethylaminoazobenzene. Cancer Res., 26, 935. MAUL, G. 1970. On the relationship between Golgi apparatus and annulate lamellae. J. Uhastruct. Res., 30, 368-384. MCCULLOCH, D. 1952. Fibrous structures in the ground cytoplasm of the Arbacia egg. J. exp. Zool., 119, 47-59. MERKOW, L. and LEIGHTON, J. 1966. Increases numbers of annulate lamellae in myocardium of chick embryo incubated at abnormal temperatures. J. Cell Biol., 28, 127-137. MERRIAM, R. W. 1959. The origin and fate of annulate lamellae in maturing sand dollar eggs. J. biophys. biochem. Cytol., 5, 117-l 22. NAGONO, T. 1966. Fine structure of the Sertoli cell in the human testis. In Electron Microscopy (ed. R. Uyeda), Vol. 11, p. 661. Maruzen, Tokyo. PALADE, G. E. 1955. Studies on the endoplasmic reticulum. 11. Simple dispositions in cells in situ. J. biophys. biochem. Cytol., 1, 567-581. PASTEELS,J. J., CASTIAUX,P. and VANDERMURSSCHE,G. 1958. Cytochemical localizations and ultrastructure in the fertilized unsegmented egg of Panacentrotus lividus. .I. biophys. biochem. Cytol., 4, 575-578. PORTER, K. R. 1961. The ground substance: observations from electron microscopy. In The Cell (ed. A. E. Mirsky), Vol. II, p. 621. Academic Press, Inc., New York. REBHUN, L. I. 1956. Electron microscopy of basophilic structures of some invertebrate oocytes. J. biophys. biochem. Cytol., 2, 93-104. REYNOL.DS,E. S. 1963. The use of lead citrate at high pH as an electron-opaque stain in electron microscopy. .I. Cell Biol., 17, 208-212. RUTHMANN, A. 1958. Basiphilic lamellar systems in the crayfish spermatocyte. J. biophys. biochem. Cytol., 4, 267-273. SCHEER,U. and FRANKE, W. W. 1969. Negative staining and adenosine triphosphate activity of annulate lamellae of newt oocytes. J. Cell Biol., 42, 519-533. SMITH, F. E. and BERLIN, J. E. 1977. Cytoplasmic annulate lamellae in human spermatogenesis. Cell Tin. Res., 176, 234242. STEINKAMP, M. P. and HOEFERT, L. L. 1977. Annulate lamellae in phloem cells of virus-infected sonchus plants. J. Cell Biol., 74, 11 l-l 18. SUN, C. N., CHEW, E. C. and WHITE, H. J. 1976. lntranuclear annulate lamellae and intranuclear lamellae in the human spermatids. J. Cell Biol., 70, 4a. SUN, C. N. and WHITE, H. J. 1977. Annulate lamellae in human tumor cells. 35th A. Proc. Ekctron Microscopy Sot. Am., Boston, Mass., pp. 414-415. SUN,C. N., CHEW, E. C. and WHITE, H. J. 1977. Cytoplasmic annulate lamellae and intranuclear membranes in human spermatids and sperm. Cell Biol. Int. Rpts, 1, 345-351. SVORODA,D. J. 1964. Fine structure of nematomas induced in rats with p-dimethylaminoazobenzene. J. noln. Cancer Inst., 33, 315-338. SWIFT, H. 1956. The fine structure of annulate lamellae. J. biophys. biochem. Cytol., 2, 415-418. SWIFT, H. 1958. Cytoplasmic particulates and basophilia. In Symposium ofthe Chemical Basis ofDevelopmenf (eds. W. D. McElroy and B. Glass), pp. 174-213. Johns Hopkins Press, Baltimore, Maryland. TANDLER, B. 1966. Warthin’s tumor, electron microscopic studies. Arch. Otolor., 84, 68-76. WESSEL,W. and BERNHARD, W. 1957. Vergleichende elektroninmikroskopische Untersuchung von Ehrilchund Yoshida-Ascitestumorzellen. Z. Krebsforsch., 62, 140-162. WISCHNITZER, S. 1970. The annulate lamellae. lnt. Rev. Cytol., 27, 65-100.
&CELL
1979 II (I)
P~hli.shrcl br Longntm
139-146
Group Ltd. Printd
in Grcwt Britcrin
C. N. SUN and H. J. WHITE
ANNULATE CELLS
LAMELLAE
IN HUMAN
TUMOR
ABSTRACT. Annulate lamellae have been found in a primitive neuroectodermal tumor, a metastatic cerebellar tumor, a testicular seminoma, a retinoblastoma and three melanomas. These annulate lamellae are arranged in stacked parallel arrays in the cytoplasm of tumor cells. The number of annulate lamellae observed to comprise a single stack varies from 2-4 in the seminoma tumor to 5-18 in the cerebellar tumor. Although the functional significance of annulate lamellae is still unknown, in many instances they have been found to be continuous with rough-surfaced cisternae of the endoplasmic reticulum and ribosomes have been demonstrated on the surface of annulate lamellae. This may suggest that annulate lamellae participate in protein synthesis.
Introduction first electron microscopic observation on annulate lamellae (AL) were made by McCulloch ( 1952) and Lansing et al. (I 952) in oocytes of marine animals. Palade (1955) described this organelle as fenestrated cisternae in the rat spermatocyte. The term ‘annulate lamellae’ was first used by Swift (I 956) to describe this organelle in the pancreatic acinar cells of the larval salamander, oocytes of snail and clam, and rat spermatids. The occurrence of the organelle has been documented in the oocytes of several different species of animals (McCulloch, 1952; Afzelius, 1955; Palade, 1955; Swift, 1956, 1968; Rebhun, 1956; Pasteels etal., 1958; Merriam, 1959; Gross et al., 1960; Kane, 1960; Hsu, 1963; Kessel, 1963, 1964, 1965, 1966), in spermatids and Sertoli cells (Ruthmann, 1958; Kaye et a/., 1961; Barer et al., 1960; Swift, 1956; Bawa, 1963; Nagano, 1966; Smith and Berlin, 1977; Sun et ul., 1977), and in cells of a variety of malignant tumors (Wessel and Bernhard, 1957; Epstein, 1957, 1963 ; I 959 ; Hoshino, I961 ; Binggeli, Chambers and Weiser, 1964 ; Svo boda, 1964;
THE
Veterans Administration of Arkansas for Medical Arkansas 72206. Received 31 March 1978. Revised 30 August 1978.
Hospital and University Sciences, Little Rock,
Ma and Webber, 1966; Locker cl Al., 1969: Ghadially and Parry, 1974). Annulate lamellae seem to have a widespread occurrence in animal tumors; however, their presence in human neoplastic material has been reported in only a few cases (Epstein, 1957; Kumegawa et al., 1961; Leak and Bensch, 1971: et al., 1967; Kadin Kovacs ef al., 1975, 1977). Recently, we found thes: structures in szvzral tumors. The increasing number of reports in neoplastic cells of annulate lamellae may suggest that the annulate lamellae have an unknown role in some neoplastic cells. Materials and Methods Biopsy specimens from a primitive neuroectodermal tumor, a metastatic cerebellar tumor, a testicular seminoma, a retinoblastoma and three melanomas were cut into small pieces about I mm3 and immediately fixed in 4% glutaraldehyde in phosphate buffer fat 2 hr, then post-fixed in 1 ‘A buffered osmium tetroxide for 1 hr. After dehydration, tissues were embedded in Epon 812. Thin sections were stained with uranyl acetate and lead citrate (Reynolds, 1963). Observations The 139
ultrastructural
features
of
annulate
140
SUN
lamellae from the above materials constitute the subject of the present study; other findings will not be presented here. Annulate lamellae are smooth-walled, paired membranes arranged in parallel stacks. The number of annulate lamellae observed to comprise a single stack varies with tissue examined (Figs. l-3 and Table 1). The membranes show regularly spaced discontinuities. By appropriate section, ‘pores’ on the membranes are seen which have an outside diameter of approximately 1000-1600 A. The center of the pores may appear empty, but sometimes a small central granule may be shown in tangential section (Fig. 3a). Granular and fibrillar material is present within the ‘pores’, especially in the vicinity of the periphery (Figs. 1, 3). We found in one case (p.e. tumor) that the annulate lamellae are in contact with the nuclear envelope (Fig. 4). More commonly, the annulate lamellae are in contact with the rough endoplasmic reticulum (Figs. 2, 3). In one of the melanomas, a peculiar membranous body was found in the cytoplasm of the tumor cell. In some profiles, this structure consists of stacks of braided canaliculate elements or channels which pursue a tortuous course and cross one
Fig.
1. Annulate
Table
AND
WHITE
I. Tissue examined No. of AL in
Tumor
a stack
Melanoma Seminoma Retinoblastoma Cerebellar tumor Primative neuroectodermal
tumor
8-10 2-4 t-8 5-18 4-7
another (Fig. 5). In many of the sections, profiles showed many ‘pores’ with central granules (Fig. 6). Here, too, channels are connected to the rough endoplasmic reticulum (Figs. 5, 6). A portion of the profile shows a similarity to the annulate lamellae (Fig. 6) of the myocardial cell in the chick embryo (Merkow and Leighton, 1966) and annulate lamellae of Sertoli cells in azoospermic human testis (Livni et al., 1977). These membranous structures may possibly represent modified annulate lamellae. Tn the cytoplasm of the primitive neuroectodermal tumor, there are many instances of membranous structures which may be precursors of the annulate lamellae (Fig. 7). These membranous structures are quite similar to the Golgi apparatus and sometimes
lamellae (AL) in retinoblastoma.
P, pore; N, nucleus.
x 50,000.
Fig. 2. Annulate lamellae (AL) in a nuclear pocket in melanoma. reticulum; G, Golgi apparatus; N, nucleus. x 16,000.
ER, endoplasmic
Fig. 3. Annulate lamellae reticulum. x 30,000.
ER, endoplasmic
(AL) in testicular
seminoma.
P, pore;
Fig. 3a. Tangential section of annuli: a small dense granule is sometimes in the center of the annulus ( +). ER, endoplasmic reticulum. x 30,000.
observed
Fig. 4. A stack of annulate lamellae is in contact with the nuclear envelope of a primitive neuroectodermal tumor (+). lntranuclear lamellae are also present (IAL). N, nucleus. x 43,000. Fig. 5. Peculiar configuration of agranular reticulum of braided channels in a melanoma tumor cell (+). ER, endoplasmic reticulum; G, Golgi apparatus. x 40,000. Fig. 6. Many of the profiles of the peculiar configuration show pores with central granules (+). ER, endoplasmic reticulum; G, Golgi apparatus. x 40,000. Fig. 7. Membranous structure may be a precursor annulate lamellae; G, Golgi apparatus. x 44,000.
of the annulate
lamellae (+).
AL,
SUN
144
AND
WHITE
Table 2. Some previous observations of annulate lamellae in human tumor cells No. of AL in a stack
Tumor
HeLa cells KB cells Pheochromocytoma Melanoma Apocrine hidrocytoma Warthin’s tumor Fibromyxosarcoma Adenomatous pituitary glands
4-8 4 5 4 3 6 2-l
are closely associated with the Golgi apparatus. Discussion Annulate lamellae occur frequently in rapidly growing and differentiating cell systems under normal and abnormal conditions (Porter, 1961). They are also occasionally associated with cell degeneration. An increase in the number of annulate lamellae has been noted in virus-infected animal (Koestner et al., 1966), and plant cells (Steinkamp and Hoefert, 1977), as well as in cells treated with a variety of chemicals (Hruban et al., 1965b, c). Since annulate lamellae are a constituent of many different types of cells, we may infer that the organelles have multiple functions that may differ depending upon the cell type. Among their suggested functions are protein synthesis (Merkow and Leighton, 1966; Wischnizer, 1970), nucleocytoplasmic exchanges (Kessel, 1968), nucleation sites for tubulin synthesis (De Brabander and Rogers, 1975), storage sites (Fanke, 1974), or as a disposal mechanism (Kessel, 1973). As for their origin, Afzelius (1955) first thought that this lameller system represented fragments of the nuclear membrane remaining in the cytoplasm after mitosis. Rebhum (1956) and Swift (1956) have suggested that annulate lamellae appeared to be synthesized as sheets on the surface of the nuclear envelope. In several cell systems, such as sea urchin and dragonfly oocytes, there is evidence that basophilic masses derived from the nucleus become associated with annulate lamellae (Kessel, 1968; Kessel and Beams, 1969). After the stacks of annulate lamellae have completely formed, rough endoplasmic reticulum extends from each of the annulate lamellae in a stack (Sun et al., 1977). Although many authors believe that annulate lamellae take their origin from the
Authors In In In In In
vitro vitro vitro vitro vivo In vivo In duo In vivo
Epstein (1961) Kumegawa et al. (1967)
Kadin and Bensch (1971) Maul (1970) Gross (1965) Tandler (1966) Leak et al. (1967) Kovacs (1975)
nuclear envelope, the precise origin of the annulate lamellae has not been firmly established. As early as 1955, Palade considered them to be a local differentiation of endoplasmic reticulum. Other authors (Kovacs et al., 1975) also believe they originate from endoplasmic reticulum. Hruban et al. (1965a) suggested that annulate lamellae may represent an intermediate stage in the formation of more typical endoplasmic reticulum, possibly containing only the messenger RNA before addition of free ribosome, or ribosomal RNA before the addition of protein. In the human tumor cells, there are very few instances of annulate lamellae and nuclear envelope association. Our study suggests that in tumors, annulate lamellae may possibly be formed from transformed cisternae of endoplasmic reticulum or the Golgi apparatus. Supporting this contention are reports by Maul (1970) and Scheer and Franke (1969). Hruban et al. (1965a), in a study of transplantable hepatomas of rats, reported their frequent occurrence particularly in rapidly growing tumors probably related to intense protein synthesis. Porter also noted their frequent occurrence in actively proliferating, embryonic, undifferentiated cells (Porter, 1961). By contract, in human tumors, AL are reported infrequently and paradoxically. Sometimes they even occur in slowly growing benign neoplasms (Tandler, 1966) (Table 2). More research is needed to answer the question of how the annulate lamellae relate to growth or protein metabolism. Acknowledgements This work was supported by Veterans Administration Research Funds. The technical assistance of Joe Meador and Bettye Stallings and the secretarial assistance of Diane Butler are greatly appreciated.
ANNULATE
LAMELLAE
IN TUMOR
CELLS
14.5
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