Beruflich Dokumente
Kultur Dokumente
6 AUTHORS, INCLUDING:
Ingrid Gracielle Martins da Silva
Rodrigo Gurgel-Gonalves
University of Braslia
University of Braslia
5 PUBLICATIONS 15 CITATIONS
SEE PROFILE
SEE PROFILE
Snia N Bo
University of Braslia
University of Braslia
SEE PROFILE
SEE PROFILE
Artculo de Original
1
2
Laboratory of Medical Parasitology and Vector Biology, Faculty of Medicine, University of Brasilia, Brazil.
Laboratory of Electron Microscopy, Department of Cell Biology, Institute of Biological Sciences, University of
Brasilia.
ABSTRACT
The mechanism of invasion of triatomine salivary glands by Trypanosoma rangeli remains little
known. To promote further information on the initial process of parasite invasion into glandular cells, an
ultrastructural investigation was conducted after in vitro and in vivo infection of Rhodnius robustus salivary
glands by a Brazilian strain of T. rangeli (SC-58). The Scanning Electron Microscopy images showed
that the flagellates (trypomastigotes and/or epimastigotes) adhere themselves in the basal membrane of
the glands mainly by the flagellum. It was also observed agglomerate of flagellates adhering in the basal
membrane near the glandular duct, mostly epimastigotes. After one hour of infection, multiple clusters
of flagellates spread out near to visible pores were recorded, an event not observed on the surface of the
control gland. In Transmission Electron Microscopy it was observed the presence of various cross-sections
and longitudinal figures of trypanosomes filling the gland lumen. The infected glands presented parasites
similar to epimastigotes and trypomastigotes adhered to the microvilli and spreaded in the lumen. Under
our experimental conditions flagellates (trypomastigotes and/or epimastigotes) appear to cause lesions in
the basal membrane, crossing through it and invading the cells of glandular epithelium, in most cases using
the tip of the flagellum.
Key words: Rhodnius robustus, Trypanosoma rangeli, Experimental infection, Salivary glands,
Scanning electron microscopy, Transmission electron microscopy.
31
BARRETO-SANTANA D. et al.
RESUMEN
Algunos aspectos del proceso de penetracin del Trypanosoma rangeli en glndulas salivales de
triatominos an no estn claros. Para promover informaciones adicionales de los eventos que transcurren
durante el pasaje del parasito a travs de estas clulas glandulares, una investigacin ultra-estructural
fue realizada luego despus de experimentos in vitro e in vivo de infeccin de las glndulas salivales de
Rhodnius robustus con la cepa SC-58 de T. rangeli. Las imgenes a microscopa electrnica de barrido
mostraron que los flagelados (tripomastigotes y/o epimastigotes) se adhieren a la membrana basal de las
glndulas por los flagelos. Fue tambin documentada a presencia de flagelados adheridos en la membrana
basal de la regin cercana al ducto glandular, y las formas mayoritarias eran epimastigotes. Con una hora
de infeccin, varias agrupaciones de flagelos fueron vistos junto a los poros hecho no observado en la
superficie de la glndula de control. En la microscopa electrnica de transmisin se observ la presencia
de tripanosomas esparcidos por el lumen de la glndula. En condiciones in vitro e in vivo los flagelados
producen lesiones en la membrana basal, cruzndola y siguiendo para las clulas del epitelio glandular, en
la mayora de los casos utilizando la punta del flagelo.
Palabras clave: Rhodnius robustus, Trypanosoma rangeli, Infeccin experimental, Glndulas salivales;
Microscopa electrnica de barrido, Microscopa electrnica de transmisin.
INTRODUCTION
Trypanosoma rangeli (Kinetoplastida, Trypanosomatidae) is a protozoan flagellate, which infects several species of blood-sucking insects (Hemiptera, Reduviidae, Triatominae) and mammals,
including man. T. rangeli is widely distributed in
South and Central America, often overlapping its
geographical distribution with that of Trypanosoma
cruzi, the etiologic agent of Chagas disease (Cuba
Cuba, 1998, Grisard et al, 1999).
Briefly, the life cycle of T. rangeli in triatomines
(mainly Rhodnius species) begins with the intake
of blood trypomastigotes from infected vertebrate
hosts. The parasites then colonize the digestive tract
of insects, adhere in the intestinal wall and differentiate into epimastigotes forms, which are able to
multiply and cross the intestinal epithelium. After
reaching the hemolymph, the parasites invade hemocytes, multiply and migrate infecting the bug
salivary glands (Cuba Cuba, 1998). Nevertheless the
developmental stages responsible for the penetration
into the gland and the mechanisms involved in this
process remain little known (Meirelles et al, 2005).
The invasion of the salivary glands of triatomines
by T. rangeli was first studied in level of optical
microscopy (Groot, 1952; Herrer, 1964; Watkins,
1971; Cuba Cuba, 1975). Few studies have been
pursued using electron microscopy, among them,
32
33
BARRETO-SANTANA D. et al.
Figure 1. Scanning Electron Microscopy (SEM) of the salivary gland uninfected Rhodnius robustus (X140). Arrows indicate small fractures in the basal lamina (X1.600).
Figure 2. SEM of R. robustus salivary gland after 30 minutes of infection by Trypanosoma rangeli (X110). In A, flagellates in the basal lamina near the duct (X3.000). In B,
flagellates present in the basal lamina near a slit (arrow) (X1.400).
Figure 3. T. rangeli covering the basal membrane of the salivary gland of R. robustus
after 24 hours pos-infection (X80). Indent highlighting the gland surface entirely covered
by flagellates (X190).
Figure 4-8. Scanning electron microscopy from the salivary glands of R. robustus infected by T. rangeli (SC-58), in
vivo infection.
Figure 4. Epimastigotes of T. rangeli adhered by flagellum (X3.300).
Figure 5. Trypomastigote adhered to the basal membrane. Flagellar pocket (Fp) undulating membrane (Um) (X8.500).
Figure 6. Salivary gland of R. robustus (X60). Highlighted the accumulation of flagellates (arrows) near the salivary
duct (X2.200).
Figure 7. Flagellates adhered to the basal
membrane by the middle part (arrow) of
the flagellum (X2.700).
Figure 8. Flagellates adhered to basal
membrane by the posterior part (arrow)
(X6.000). Flagellar pocket (Fp).
34
35
BARRETO-SANTANA D. et al.
36
the cells of the salivary gland for an unknown mechanism. After reaching the gland lumen, the parasites
appear as epimastigotes and continue to be attached
by the flagella to the microvilli of the salivary gland
cells. Similarly, in our study, it was observed flagellates with epimastigote characteristics, both in the
space between the basal lamina and the epithelium
of the salivary gland, and in the lumen of the gland,
the cells were adhered to the microvilli.
In TEM, Meirelles et al. (2005) demonstrated
that the flagellum of epimastigotes always appear
arranged in the basal lamina of salivary gland, penetrating individually through small holes. In contrast, in the in vitro infection, there was penetration
of various aggregates of flagellates by larger and
visible holes. Flagellates found penetrating in the
basal membrane exhibit characteristics of epimastigotes and those adhered in the membrane showed
characteristics of trypomastigotes. Apparently,
as suggested by Meirelles et al, (2005), epimastigotes produce a higher amount of a lytic molecule
consisting of a pore-forming protein, the so called
rangelysin allowing the passage of parasites by epithelial barriers. These interpretations cited by the
authors deserve further in-depth studies to define
more clearly the molecular or biochemical basis of
this important mechanism of glandular invasion.
In our evaluation under in vitro and in vivo experimental conditions flagellates (trypomastigotes and/
or epimastigotes) appear to cause lesions in the basal membrane, crossing through it and invading the
cells of glandular epithelium, in most cases using
the tip of the flagellum as an element of invasion.
REFERENCES
1. BASSERI HR, TEW IF, RATCLIFFE NA. 2002.
Identication and distribution of carbohydrate moieties
on the salivary glands of Rhodnius prolixus and their
possible involvement in attachment/invasion by Trypanosoma rangeli. Exp Parasitol. 100: 226-234.
2. BARRETO-SANTANA D, STARLING J, GURGELGONALVES R, CUBA CUBA CA. 2011. Biologia
comparativa e comportamento alimentar de Rhodnius
neglectus e Rhodnius robustus (Triatominae) sob
condies de laboratrio. Rev Soc Bras Med Trop.
44(4): 490-495.
3. CUBA CUBA CA. 1975. Estudo de uma cepa peruana de
Trypanosoma rangeli IV. Observaes sobre a evoluo
e morfognese do T. rangeli na hemocele e nas glndulas
salivares de Rhodnius ecuadoriensis. Rev Inst Med Trop.
17: 284-297.
4. CUBA CUBA CA. 1998. Revisin de los aspectos bio-
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
37