MICROSCOPY RESEARCH AND TECHNIQUE 75:282–284 (2012)
Ultrastructural Aspects of Callithrix penicillata Lingual Papillae ˜ O PEREIRA,1 ANA RITA DE LIMA,1 ERIKA BRANCO,1* WASHINGTON LUIZ ASSUNC ¸A ´ LUIZ REZENDE FRANCIOLLI,2 ROSE ELI GRASSI RICI,2 MARIA ANGELICA MIGLINO,2 ANDRE ´ AUGUSTO PEREIRA CARNEIRO MUNIZ,3 AND ALINE IMBELONI3 JOSE 1 Institute of Animal Health and Production—ISPA Faculty of Veterinary Medicine, Federal Rural University of Amazonia (UFRA), Belem, Brazil 2 Department of Surgery of the University of Sa˜o Paulo, Faculty of Veterinary Medicine and Animal Science, Sa˜o Paulo, Brazil 3 Evandro Chagas Institute (IEC) Sector-National Primate Center (CENP), Ananindeua, Para, Brazil
KEY WORDS
morphology; lingual papillae; Callithrix penicillata
ABSTRACT Callithrix penicillata belongs to the family Callitrichidae, Callithrix genus. They are basically insectivorous, but they consume fruits. The mucosa of the tongue is composed of some papillary types, revealing different levels of expertise. The present study attempted to describe the morphological and ultrastructural aspects of the dorsal surface of the C. penicillata, describing the characteristics and distribution of papillae found. Five tongues of C. penicillata (two females and three males), obtained from breeding colonies of CENP-Ananindeua-PA, died from natural causes. The material was fixed partly in a buffer solution paraformaldehyde 10% and partly in modified Karnovsky solution, divided into apex, body, and root, and then the fragments were used in light microscopy and scanning electron microscopy. The average length of the tongue of the females was 22 mm and for males 20.5 mm. Three types of papillae were described: filiform (along all tissue extension with 154 lm of diameter), fungiform (along all tissue extension with 275 lm of diameter), and vallate (just three units in caudal (dorsal) portion with 672 lm of diameter). Data analysis indicates that the distribution and ultrastructural morphology of the C. penicillata lingual papillae are some similar to other primates. Microsc. Res. Tech. 75:282–284, 2012. V 2011 Wiley Periodicals, Inc. C
INTRODUCTION The Callithrix penicillata presents preauricular tufts long, black, brush-shaped, white spots on the forehead, face, and head until the shoulder blades black or dark brown. Its general color is dark gray with dorsal striatum and low-ringed tail. These small primates reach about 20 cm long, weighing between 350 and 500 g, and have a density of 39 persons per km2 in groups of two to nine, with an average of six (Auricchio, 1995; Rowe, 1996). Callithrix penicillata usually feed on fruits, flowers, buds, insects, small vertebrates, seeds, shellfish, bird eggs, and gum (resin released by some species of trees, composed of sugars and minerals) (Castro, 2003). Feeding patterns of each species correlate directly to the amount and texture of the tongue mucosa, which comprises specific types of papillae—filiform, fungiform, vallate, and foliate—which presents different types of specialization (Getty, 1986; Pfeifer et al., 2000). Papillae arrangement may vary among primates species, including men. However, we can generally find the filiform papillae in a major number and distributed in all tongue surface as well as in a minor amount—the fungiform and vallate papillae. Depending of the species, a couple of foliates papillae can be found (Chamorro et al., 1994; Getty, 1986; Kobayashi et al., 1997; Young and Heath, 2001). In the face of few studies dealing with the tongue morphology of C. penicillata, this study opens new venues for this species preservation with new data about digestory system and tongue behavior related to different type of nutrition used when the animals are raised C V
2011 WILEY PERIODICALS, INC.
in captive. Thus, this study aimed to describe the lingual papillae of this species by macroscopic evaluation, light microscopy, and scanning electron microscopy (SEM). MATERIALS AND METHODS The tongues of five C. penicillata were studied in two females and three males, obtained from breeding colonies of National Primate Center (CENP)—Ananindeua, PA, Brazil. The animals died from natural causes. Tongues were measured and divided into apex, body, and root and fixed by immersion in buffer solution paraformaldehyde 10% for light microscopy and in modified Karnovsky solution containing 2.5% glutaraldehyde and 2% paraformaldehyde in 0.1 M sodium phosphate buffer (pH 7.2) at 48C, for 36 h, according to Watanabe and Yamada (1983). The microscopy analysis was performed by conventional techniques. Briefly, the tongue material was collected and processed in paraffin blocks. Hematoxylin and eosin-stained sections were prepared for each specimen, and sections of 5-lm thick were cut and mounted on slides. One set of samples was postfixed in buffered solution of osmium tetroxide at 1% for 1 h at 48C. Samples were then rinsed in distillate water by 3 h and ´ rika Branco, Rod. Ma´rio Covas, 1400, bl 6 ap 204, *Correspondence to: E Coqueiro, Ananindeua, Para´, Brazil. E-mail:
[email protected] Received 16 May 2011; accepted in revised form 13 June 2011 Contract grant sponsor: CENP-Ananindeu-PA DOI 10.1002/jemt.21055 Published online 2 September 2011 in Wiley Online Library (wileyonlinelibrary.com).
LINGUAL PAPILLAE IN THE CALLITHRIX
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Fig. 1. Tongue of C. penicillata. A: Photomacrograph: vallate papillae (arrow) and median lingual sulcus (hollow arrow), (B) Photomicrograph: filiform papillae (FP), stratified squamous epithelium (e), lamina propria (Lp), and skeletal muscle (sm), and (C–E) scanning electron microscopy: filiform (dotted areas and *), fungiform papillae (arrowhead), median lingual sulcus (hollow arrow), and vallate papillae (arrow). [(A) scale bar 1 cm, (B) HE, scale bar 50 lm, (C) scale bar 1 nm, and (D,E) scale bar 100 lm].
immersed in aqueous solution containing 1% tannic acid at room temperature during 1 h (Murakami, 1974), following the dehydration in a crescent series of alcohols (60% to absolute), and dried at critical point Balzers CPD-030 or Balzers CPP-020, using liquid CO2. Samples were mounted on metal stubs, coated with gold using spray EMITECH k 550, and analyzed by scanning electron microscope LEO 435VP. RESULTS In a macroscopic view, the tongue of C. penicillata was triangular-shaped and elongated in craniocaudal direction with acute apex, presence of a median lingual groove and three papillary distributed dorsally: the filiform papillae in greater numbers, followed by the fungiform papillae, and finally, only three units of vallate papillae, which are arranged exclusively in the caudal portion of the tongue. The initial measurement showed average total length of 22 mm in females and 20.5 mm in males, average width of 8.7 mm in females, and 7.5 mm in males (Fig. 1A). The light microscopy revealed a thin-stratified squamous epithelium and typical lamina propria mucosae in the base of the papillae (Fig. 1B). SEM analysis demonstrated three types of papillae being the largest distribution on the dorsal surface composed by filiform, fungiform (Figs. 1C and 1D), and vallate (Fig. 1E). The median lingual groove could also be observed in SEM (Fig. 1C). Microscopy Research and Technique
The filiform papillae that regardless of sex were present in greater quantities revealed irregular surface and average diameter of 71.6 lm showing as the smallest papillae detected. The fungiform papillae, which are interspersed with filiform, had rounded surface and average diameter of 210 lm. Located in the caudal portion of the tongue of C. penicillata, vallate papillae found very evident, with an average diameter of 672 lm (Figs. 1A and 1E). DISCUSSION Comparing to other species, the dorsal surface of the tongue of C. penicillata had a median groove, which is described only in carnivores (Dyce et al., 2010). In general, the literature reports that in the most primate species, the tongue is composed of three or four types of papillae: filiform, fungiform, vallate, and foliate (not always present in some species). Filiform papilla is the most common (Chamorro et al., 1986; Chibugo, 1993; Dyce et al., 2010; Pfeifer et al., 2000; Schwarze and Schro¨der, 1970; Sisson and Grossman, 1959), except for the manatee, which has only a small taste of these clusters at the apex of the tongue (Levin, 1999). The fungiform papillae are the second most frequent, followed by the vallate, which can vary from a simple pair as in the donkey (Abd-Elnaeim et al., 2002) to 2–5 U, distributed as principal and accessory, such as in the horse (Guimara˜es et al., 2007) or in large numbers
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located in the final third of the tongue as seen in ruminants and manatee (Ferreira et al., 1998; Emura et al., 2000; Levin, 1999). Foliate papillae are fewer in number, usually a pair, and species-dependent (Dyce et al., 2010; Levin, 1999; Sisson and Grossman, 1959); however, these are absent in C. penicillata. Little is known about the lingual papillae in primates; however, Watanabe et al. (1995) and Branco et al. (2011) described the microscopic structure of the Macaca fuscata and Saimiri sciureus lingual papillae, reporting that the fungiform papillae are more present in the dorsal cranial portion of the tongue than in the dorsal medial and caudal portions, fact differentiated from the manatee, which has a large number of fungiform papillae only on the side of the middle portion of the tongue (Levin, 1999). Watanabe et al. (1995) reveal that in M. fuscata, the fungiform papillae have a diameter ranging from 250 to 350 lm, whereas in S. sciureus, these papillae have an average diameter of 272 lm (Branco et al., 2011), revealing in both cases fungiform papillae than those found in C. penicillata (210 lm). As in S. sciureus (Branco et al., 2011) and in the dwarf armadillo (pichiy) (Ciuccio et al., 2008), in C. penicillata, these papillae are distributed in a diffuse, unlike in the white-lipped peccary (Tayassu pecari), which presents these papillae on the middle third of the tongue (Watanabe et al., 2009). In black rhinoceros (Diceros bicornis) and in maned three-toed sloth (Bradypus torquatus), the fungiform papillae present a distribution quite different, because they are arranged not only at the dorsal surface of the tongue, but also bordering all the vertex of it (Benetti et al., 2009; Emura et al., 2000). In M. Fuscata (Watanabe et al., 1995), as in S. sciureus (Branco et al., 2011), the filiform papillae were distributed along the tongue, with no specific points of concentration, fact which goes against our findings. The diameter of these papillae in M. fuscata varied of 100–200 lm, resembling as described in S. sciureus (154 lm), however, in C. penicillata recorded smaller diameter (71.6 lm). As in S. sciureus (Branco et al., 2011), the C. penicillata showed only three units of vallate papillae, situated in the caudal portion of the tongue, but with smaller diameter, because, in S. sciureus, it enrolls an average diameter of 830 lm against 672 lm of C. penicillata leaving it with a similar measurement reported by Watanabe et al. (1995) in M. Fuscata (600–800 lm). The number of vallate papillae found in C. penicillata is not common, as usual in most species is the presence of one or two pairs, as in the case of white-lipped peccaries (Tayassu peccary) and domestic carnivores (Noden and De Lahunta, 1985; Watanabe et al., 2009), However, it has been reported, in some rodents, just a unique papilla in dorsocaudal surface and centrally arranged (Stangl and Pfau, 1994). Our observation was remarkably contrasted to the 60 vallate papillae found in the black rhinoceros tongue (Emura et al., 2000). The S. sciureus is the only species that has the same number of vallate papillae that C. penicillata (three) (Branco et al., 2011). Unlike what was reported in other primates (Branco et al., 2011; Watanabe et al., 1995), our findings do not reveal the presence of foliate papillae; however, this is common on manatees, donkey, and swine (AbdElnaeim et al., 2002; Dyce et al., 2010; Levin, 1999).
REFERENCES Abd-Elnaeim MM, Zayed AE, Leiser R. 2002. Morphological characteristics of the tongue and its papillae in the donkey (Equus asinus): A light and scanning electron microscopical study. Ann Anat 184:473–480. Auricchio P. 1995. Primatas do Brasil. Sa˜o Paulo: Terra Brasilis. 168 p. Benetti EJ, Pı´coli LC, Guimara˜es JP, Motoyama AA, Miglino MA, Watanabe IS. 2009. Characteristics of filiform, fungiform and vallate papillae and surface of interface epithelium-connective tissue of the maned sloth tongue mucosa (Bradypus torquatus, Iliger, 1811): Light and scanning electron microscopy study. Anat Histol Embryol 38:42–48. Branco E, Guimara˜es JP, Miglino MA, Lacreta JR ACC, Ishizaki MN, Gomes BD, Muniz JAPC, Imbeloni A, Fioretto ET, Lima AR. 2011. Ultrastructural aspects of lingual papillae in squirrel monkey (Saimiri sciureus). Microsc Res Techniq 74:484–487. Castro CSS. 2003. Tamanho da a´rea de vida e padra˜o de uso do espac¸o em grupos de sagu¨is, Callithrix jacchus (Linnaeus) (primates callitrichidae). Rev Bras Zoo 20:91–96. Chamorro CA, De Paz P, Sandoval J, Fernandez JG. 1986. Comparative electron-microscopic study of the lingual papillae in two species of domestic mammals (Equus caballus and Bos taurus). I. Gustatory papillae. Acta Anat (Basel) 125:83–87. Chamorro CA, Fernandez JG, De Paz P, Pelaez B, Anel L. 1994. Scanning electron microscopy of the wild boar and pig lingual papillae. Histol Histopathol 9:657–667. Chibugo GA. 1993. The tongue. In: Evans HE, editor. Miller’s anatomy of the dog. Saunders: Philadelphia. pp. 396–414. Ciuccio M, Estecondo S, Casanave EB. 2008. Scanning electron microscopy study of the dorsal surface of the tongue in Zaedyus pichiy (Mammalia, Xenarthra, Dasypodidae). Int J Morphol 26:13–18. Dyce KM, Sack WO, Wensing CJG. 2010. Tratado de anatomia veterina´ria,4nd ed. Rio de Janeiro: Elsevier. 856 p. Emura S, Tamada A, Hayakawa K, Cehn H, Shoumura S. 2000. Morphology of the dorsal lingual papillae in the black rhinoceros (Diceros bicornis). Anat Histol Embryol 29:371–374. Ferreira JR, Augusto JC, Ferreira JR. 1998. Contribuic¸a˜o ao estudo anatoˆmico da distribuic¸a˜o e situac¸a˜o das papilas valadas em lı´nguas de bovinos (Bos taurus, L. 1758). Ar Cieˆnc Sau´de Unipar 2:231–239. Getty R. 1986. Anatomia dos animais dome´sticos,5th ed. Rio de Janeiro: Interamericana. 2000 p. Guimara˜es GC, Machado MRF, Santos ALQ. 2007. Morfologia, topografia e distribuic¸a˜o das papilas valadas em equ¨inos (Equus caballus, LINNAEUS, 1758) sem rac¸a definida. Biosci J 23:105–110. Kobayashi K, Kumakura M, Yoshimura K. 1997. Stereo structural differences of lingual papillae and their connective tissue cores in three kinds of artiodactyla. Rec Adv Microsc Cells Tiss Org 7:357–361. Levin MJ. 1999. Gross and microscopic observations on the lingual structure of the West Indian Manatee (Trichechus manatus latirostris). Dissertation. Faculty of the Virginia Polytechnic Institute and State University. 134p. Murakami T. 1974. A revised tannin-osmium method for non-coated scanning electron microscope specimens. Arch Histol Jpn 36:189– 193. Noden DM, De Lahunta A. 1985. Embriologia de los animales dome´sticos. Zaragoza: Acribia. 399 p. Pfeifer CJ, Levin M, Lopes MAF. 2000. Ultrastructure of the horse tongue: further observations on the lingual integumentary architecture. Anat Histol Embryol 29:37–43. Rowe N. 1996. The pictorial guide to living primates. New York: Pogonias. 274 p. Schwarze E, Schro¨der L. 1970. Compendio de anatomia veterina´ria. Zaragoza: Acribia. 212 p. Sisson S, Grossman JD. 1959. Anatomı´a de los animales dome´sticos,4th ed. Barcelona: Salvat. 952 p. Stangl FB Jr, Pfau RS. 1994. Gross morphology and distribution patterns of lingual papillae in some geomyid and heteromyid rodents. Proc Okla Acad Sci 74:25–29. Watanabe I, Yamada E. 1983. The fine structure of lamellated nerve endings found in the rat gingiva. Arch Hist Jpn 46:173–182. Watanabe IS, Inokuchi T, Hamasaki M, Yamada E. 1995. Threedimensional organization of the epithelium-connective tissue interface of the tongue and soft palate in the Macaca fuscata: A SEM study. Acta Microsc 4:59–73. Watanabe IS, Guimara˜es JP, Boleta AS, Almeida SRY, Righeti MM, Santos TC, Miglino MA, Kfoury JR Jr. 2009. Nerve endings of filliform, fungiform and vallate papillae of dorsal tongue mucosa of White-lipped peccary (Tayassu pecari): Neurohistological observations. Pesq Vet Bras 29:281–285. Young B, Heath J. 2001. Wheater, histologia funcional-texto e atlas em cores,4th ed. Rio de Janeiro: Uanabara Koogan, 424 p.
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