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An Electron Microscope Study of the Trichomonas Criceti

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An Electron Microscope Study of the Trichomonas Criceti 320 Cytologia 26 An Electron Microscope Study of the Trichomonas criceti J. Chakraborty, N. N. Das Gupta and N. N. Ray Biophysics Division, Saha Institute of Nuclear Physics, Calcutta-9, and School of Tropical Medicine, Calcutta-12,India ReceivedJanuary 17, 1961 Introduction Trichomonad flagellates were studied under the optical microscopy by Kofoid and Swezy (1915), Wenrich (1921 and 1944), Sammuels (1941.and 1957), Buttrey (1954) and Ludvik (1954). Later on Anderson (1955a, b) and Anderson and Beams (1959) studied the ultrastructural organisation of these flagellates with the help of electron microscope and ultrathin sectioning tech niques. Inoki et al (1959) studied the ultrastructure of T. vaginalis. The present paper deals with some new findings in the ultrastructural organisation of Trichomonas criceti (Ray et al). Material and methods The organisms were obtained from the caecum of the hamster. They were fixed in 1% OsO4 solution buffered at pH 7.4 for 30 minutes. After fixation, dehydration, infiltration, embedding and sectioning were done as usual. The sections were examined under a Siemens' Elmiskop I at 60 K. V. Observations Anterior flagella: The flagellum is found to consist of a number of fibrils running longitudinally along its length, surrounded by a thin flagellar sheath. In transverse section of the flagella (Fig. 2) one central fibril is seen surrounded by nine peripheral fibrils, the average thickness of each of which is about 400A. In the longitudinal section through the flagella a maximum number of 3 to 4 fibrils can be very clearly counted (Figs. 1, 3 and 4). The flagellar sheath is thin, continuous and appears to be 230A thick. Basal apparatus of the anterior flagella: The fibrillar bundle of the flagella extends (1.2-1.4)ƒÊ into the body of the protozoon (Fig. 1). There is a double layered membrane, surrounding the base of the flagella, a por tion of which is clearly visible in Fig. 1. The outer layer of this membrane is found to be continuous but the inner one is possibly fibrillar as in the oblique section it appears as a dotted line. The total thickness of the two layers together is about 900A. At the base of the flagellum there is a double 1961 An Electron Microscope Study of the Trichomonas criceti 321 walled filamentous structure about 1.4ƒÊ long (Figs . 1 and 3). The costa is attached to this structure (Fig. 3) . Parabasal body: The parabasal body of these organisms con sists of a number of thick walled, long la mellar structures, ar ranged in a parallel array (Figs. 5, 6 and 7). The membranes of these lamellae are con stricted in some places . The distance between two membranes is about 150A and the thickness of each of the membranes is about (250 to 300) A. The parabasal filament origi nates from the blepha roplastic region and is attached to the parabasal body (Fig. 6). Recurrent flagel la, "accessory fila Figs. 1-4. 1, oblique section of T. criceti showing the anterior ment" and undulating flagella (FA), basal body (B), membrane of the kinetosomal membrane: The re vacuole (KM), recurrent flagellum (FR), undulating membrane current flagella have (UM), sheet of the undulating membrane (UMS), accessory filament (AF) and mitochondria (M). •~6,600. 2, transverse an internal structure section of the flagellum (F) showing the peripheral (FFP) and similar to that of the central fibrils (FFC) and the flagellar sheath (FS). •~32,700, anterior one (Fig. 1). 3, section passing through the basal apparatus of the flagellum showing the fibrils of the flagellum (FF), filamentous structure The undulating mem at the base of the flagellum (Fl) and a portion of the costa brane is composed of (CO). •~13,600. 4, longitudinal section of the anterior flagella a sheet-like structure. (FA), showing the fibrils (FF) and the sub-fibrils (FFS'). Between the recurrent •~ 25,000. flagella and the sheet of the undulating membrane, there is a granular structure which is termed as the accessory filament. In Fig. 5, the components of the undulating membrane are clearly visible. The undulating sheet, recurrent flagellum and the accessory filament are surrounded by a 120A thick mem brane (Figs. 1 and 5). Axostyle and costa: Axostyle is a long, thick, rod-like structure, trav- 322 J. Chakraborty, N. N. Das Gupta and H. N. Ray Cytologia 26 ersing throughout the entire length of the body of the protozoon (Fig. 9B). It has a thick double walled limiting membrane (Fig. 9B). There are small fibrillar connections between these two walls (Fig. 10). The thickness of this axostylar mem brane brane is about 600A. Inside the axostyle there are a number of rounded, filamentous or granular structures (Fig. 10). The costa is also a long curved structure (Fig. 11), originating from the blepharo plastic region of the flagellum (Fig. 3). It is composed of a number of double walled disk-like struc tures tures (•`320A in thickness) embedded in a homogeneous matrix (Fig. 11). They are separated from each other by a distance of 140A. Granular struc Figs. 5-8. 5, section passing through the sheet of the undulat ing membrane (USM). The recurrent flagellum (FR), accessory tures: Paracostal gran filament (AF) and the fine double walled membrane (UM) ules, para-axostylar surrounding them are clearly visible. The parabasal body granules and chromatic (PB) is also present. •~20,800. 6, section showing the lamella of the parabasal body (PB) and the parabasal filament (PF) granules are found •~ 26,700. 7, section passing through part of parabasal body scattered all over the (PB). The parabasal filament is absent in this figure. •~32,700. body of this protozoon 8, section showing flagellar extension inside the body of the (Figs. 11, 10 and 1). parasite (F). •~16,000. Besides these types of granules, there are also other granules, the exact nature of which is not yet known (Fig. 12). These granules are surrounded by a double walled 200A thick membrane. The limiting membrane is not smooth and is constricted or elevated at some places. A rounded closely packed dense network-like struc ture is seen inside these granules. Nucleus: The nucleus is a prominent elongated body, situated near-the anterior end. It is surrounded by a distinct double-walled membrane about 1961 An Electron Microscope Study of the Trich omonas criceti 323 450A thick. There is a small gap between the outer and inner nuclear memb ranes (Fig. 13). Both the outer as well as the inner nuclear membranes are interrupted by small pores (Figs. 13 and 14). The nucleus is filled up with the granular nucleo plasm (Fig. 14). Some times a few denser bodies are visible in the nucleoplasm (Fig. 10). They may be the sec tions passing through the chromosomes. Mitochondria: Besides the paracostal para-axostylar and chromatic granules some less dense rounded or oval bodies are scattered all over the body at random (Fig. 1), which are surrounded by a 200A thick smooth mem brane (Fig. 9B). They are the sections of mitochondria; some times very faint "mic rovilli" are also Figs. 9-11. 9A, section passing through the median and lateral visible inside them axostyle (MAX and LAX) and nucleus (N). •~5,000. 9B, (Fig. 9B). oblique section of axostyle (AX) showing the rounded (r) and fi lamentous (f) structure inside it. The mitochondria (M) with faint microvilli (MC) are also visible. •~26,700. 10, section Discussion showing the double walled membrane of the axostyle (AXM) and the fine fibrillar connections (AXF) between these two The fine structure membranes. The para-axostylar granules (PAG) and the sec of the flagella of tion of chromosomes (CH) are also clearly visible. •~16,000. Trichomonas criceti 11, section of costa (CO), showing the double walled disk-like shows the same structures (DK) and the paracostal granules (PCG). •~25,000. pattern, as observed in the flagella and cilia of other, protozoan cells. Chang (1956), Inoki et al (1957), Pyne (1958), Pyne and Chakraborty (1958) found one central fibril surrounded by nine peripheral fibrils in L. donovani. In the present investigation also one central and nine peripheral fibrils are cearly visible, thus agreeing with the observations of Anderson and Beams (1959) and Inoki et al (1959) in T. muris and T. vaginalis. The central as well as the peripheral fibrils extend into the body forming 324 J. Chakraborty, N. N. Das Gupta and H. N. Ray Cytologia 26 the basal body. No granular structure, which may be called as the basal granule was found in this species. A double-walled filamentous structure is visible at the base of the flagellum (Fig. 3) the exact nature of which is not yet un derstood. The pelli cular layer invaginates inside the body of the parasite near the base of the flagellum, form ing the membrane of a vacuole. A similar structure was also found by Pyne and Chakraborty in Lei shmania donovani (1958). The double-walled lamellae and thick walled vesicles in the parabasal body found in this organism, re semble those reported by Anderson and Figs. 12-13. 12, section passing through the granules (G) of Beams in T. muris unknown nature. These granules are surrounded by a double (1959). The parabasal walled limiting membrane (LM). Closely packed dense net filament also shows work like structures (DPN) are visible inside these granules. the same internal •~ 53,300, 13, section of a portion of nucleus (N) showing the double walled nuclear membrane (NM) and the pores (NMP). structure. •~ 53,300. The similarity in structure between the recurrent and the anterior flagella and the granular nature of the accessory filament found in this work also agree with the observations of Anderson and Beams in T.
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