Trunk and Paw Pad Skin Morphology of the African Giant Pouched Rat ( Cricetomys Gambianus , Waterhouse-1840)
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ORIGINAL ARTICLE Eur. J. Anat. 18 (3): 175-182 (2014) Trunk and paw pad skin morphology of the African giant pouched rat ( Cricetomys gambianus , Waterhouse-1840) Chikera S. Ibe* ,1 , Suleiman O. Salami 1 and Itopa E. Ajayi 2 1Department of Veterinary Anatomy, Ahmadu Bello University Zaria, Nigeria and 2Department of Veterinary Anatomy, University of Abuja, Abuja, Nigeria SUMMARY Key words : Gross morphology – Histology – Skin of trunk – Paw pads – African giant pouched rat The gross morphology and histology of the skin of the trunk and paw pads in the African giant pouched rat were investigated to evaluate their INTRODUCTION role in the adaptation of the rodent to its subterra- nean environment. Samples were stained with The African giant pouched rat ( Cricetomys gam- haematoxylin and eosin, Masson’s trichrome, Alci- bianus ) is a remarkable rodent in many respects. It an blue, Verhoeff's haematoxylin counterstained is one of the largest nocturnal rodents of the Muri- with Van Gieson, and Weigert’s haematoxylin dae family. Its habitat is distinct from other mem- counterstained with Picro-Ponceau stains. Dorsal- bers of the family: it lives in terrestrial burrows con- ly, fur covering the skin was loosely folded. Fur structed using its teeth and manus to loosen sub- covered the entire trunk to the level of the radio- strate and its pes to push excavated soil away carpal and talocrural joints of the forelimb and (Ewer, 1967). Although Rosevear (1969) and Ajayi hindlimbs, respectively. Skin of the dorsum was (1975) suggested that the pes maybe well adapted paler than its grey-coloured fur, while skin of the for digging, no detailed anatomical study has been ventrum was dirty white. There were more hair done to verify the suggestion. follicles dorsally then ventrally. The manus and The African giant pouched rat has been used to pes had five and six paw pads, respectively. detect land mines (Mott, 2004) and diagnose tu- Keratinocytes in the epidermis of the paw pads berculosis (Maggie, 2003) by their high acuity of decreased in number and lost their cellular con- odour perception. It is sequel to this importance tents as they migrated towards the stratum lucid- that the potential of the rat as an experimental um . Metatarsal pads had a significantly (P < 0.001) model for biomedical research is currently ex- thicker stratum corneum than metacarpal pads. plored. Documentation of the normal features of its Elastic fibres were observed in the metatarsal integumentary system will serve as a lead for fur- pads. Other results and additional information from ther physiologic and behavioural studies. The the literature were integrated to propose the effect adaptive morphology of the mystacial vibrissae of the structures on the adaptation of the African have been reported (Ibe et al., 2010). giant pouched rat to its subterranean environment The numeric and regional distribution of paw and tropical climate. pads differ among rodents. They provide a cush- ioning effect to the underlying skeletal structures, thus protecting them from dislocation (Brown and Yalden, 1973). In some animals, paw pads contain * Corresponding author: Chikera S. Ibe. Department of Veterinary Anatomy, Ahmadu Bello University Zaria, Nigeria. Tel: +2348032882105. E-mail: [email protected] Submitted: 15 February, 2013. Accepted: 14 February, 2014. 175 Rat trunk and paw pad skin morphology eccrine glands whose secretions confer a degree digital pads of the manus and pes, the metacarpal of friction between the pad and adjoining environ- and the metatarsal pads were measured and sam- ment (Haffner, 1998). Animals of the same genera ples taken. The skin samples were cut in such a can be differentiated by paw pad morphology; An- way that the epidermis, dermis, subcutis, deep derson (1999) noted significant morphometric dif- facia and underlying muscles were intact. Tissues ference in the manus paw pads of the thick-tailed were washed with saline and fixed in 10 % phos- galago ( Otolemur crassicaudatus ) and the Gar- phate-buffered formalin for 1 week. After fixation, nett's galago ( Otolemur garnenii ). Anatomy of paw the samples were washed in tap water and kept pads has been reported in different rodents therein for 24 hours. Finally, specimens were de- (Kimura et al., 1994; Krystufek and Macholan, hydrated in a graded series of alcohol (70 %, 80 1998; Musser and Durden, 2002; Wong et al., %, 90 %, 95 % and 100 %). Tissues were cleared 2006; Kimura et al., 2008) and used to explain ad- in xylene, paraffin embedded and labelled. aptation to a specific mode of life. Skin samples from the trunk were sectioned Animal fur is indicated in body insulation and pro- transversely at 5 µm using a Jung rotary micro- tection from the immediate environment. Sweat tome (Model 42339, Berlin, Germany). Repre- glands play pivotal role in cooling mechanism. sentative sections were mounted on glass slides, They have been reported to be sparsely distributed dried, deparaffinized, stained, and coverslipped in the naked mole-rat ( Heterocephalus glaber ) as a using DPX as the mountant. Alternate sections heat conservation mechanism, to compensate for were stained with haematoxylin and eosin (H/E), the absence of fur (Daly and Buffenstein, 1998). Masson’s trichrome (MT) and Alcian blue (AB) Furthermore, skin colour differs in animals and has stains. A paraffin section of the skin of the dorsal been associated with thermoregulation (Scharf, trunk was treated with the Masson-Fontana reduc- 2008). ing method to demonstrate melanin, using the The present study will be of value in future derm- method of Bancroft and Stevens (2008) by placing atoglyphic studies in the African giant pouched rat. the section in 10 % ammoniacal silver nitrate over- Since the rats are currently domesticated in Nige- night at room temperature in a dark room. The fol- ria as supplementary protein source (Olayemi et lowing day, it was washed in several changes of al., 2001) and in Europe as exotic pets (Cooper, distilled water and treated with 5 % sodium thiosul- 2008), there is a need to have a good knowledge phate. Following a tap water flush, it was counter of their adaptive morphology. This is a prerequisite stained with 1 % aqueous neutral red, dehydrated for an effective breeding and domestication pro- through a graded series of alcohols to xylene and gramme. Therefore, the aim of the study was to mounted in DPX. identify morphologic features of the skin of the Skin samples from the paw pads were sectioned trunk and paw pads in the African giant pouched transversely at 7 µm and processed as described rat that are related to its tropical climate and sub- for the trunk skin samples. Alternate sections were terranean environment and compare the results stained with H/E, Verhoeff's haematoxylin counter- with similar findings in other rodents. stained with Van Gieson’s (VVG) and Weigert’s haematoxylin counterstained with Picro-Ponceau MATERIALS AND METHODS (WHP). H/E was used as the conventional stain to study Seven healthy adult African giant pouched rats the general tissue architecture. VVG was used to weighing between 1.3 kg and 1.5 kg were captured demonstrate elastic fibres which stained black. from the wild in Kaduna state, Nigeria. They were WHP is very effective for visualising structures of given elephant grass ( Penicetum puperum ), the dermis and epidermis as well as for providing groundnut cake, potatoes and drinking water ad enhanced cellular detail (Hamrick, 1998). MT was libitum . Gross features were observed and record- indicated to demonstrate collagen fibres which ed via digital photography. Report of Brown and stained blue against a black (cell nuclei) and red Yalden (1973) was used as guide to describe ex- (cytoplasm, muscles or erythrocytes) background. ternal features of the paw. Paw length and width AB was used to demonstrate acid mucosubstance were defined using the method of Ade and Ziekur which stained blue against a pink background. The (1999). Following humane sacrifice, the skin of the Massons-Fontana staining displayed melanin as trunk, forelimbs, hindlimbs and paw pads was re- black colour against a red background. moved. The epidermis was divided into two broad layers Skin samples of 5 mm 2 each from shaved por- for clarity. These were the stratum corneum and tions of the dorsal trunk, at the level of the thoracic the non-cornified layer. The stratum lucidum , stra- spine, and the ventral trunk, at the level of the um- tum spinosum , stratum granulosum and stratum bilicus, were measured with a vernier calliper basale were grouped as the non-cornified layer. (MG6001DC, General Tools and Instruments The thickness of the stratum corneum and non- Company, New York; sensitivity: 0.01mm) and cut cornified layer of the metacarpal and metatarsal out with a scalpel blade. 5 mm 2 each, of the digital pads were obtained using a calibrated ocular mi- skin of digit 3 of the manus and pes, the 2 nd inter- crometer (Leitz Wetzlar, Germany) following ap- 176 C.S. Ibe et al. propriate calibration of a light microscope fur was grey in colour and covered the entire trunk (OLYMPUS ® - XSZ107BN, Hamburg, Germany) (Fig. 1a). It extended to the level of the radiocarpal with a stage micrometer (Graticules Ltd., London). and talocrural joints of the forelimbs and hindlimbs, For each pad, 15 points were randomly chosen respectively. The ventral fur was greyish white and and the mean thickness of the stratum corneum thinner than the dorsal fur (Fig. 1b). The ventral fur and non-cornified layer was utilized. The result terminated on the medial femoral skin from where was subjected to statistical analysis using stu- the grey-coloured fur continued to the talocrural dent’s t-test to determine the difference in thick- joint. The extent of the ventral fur in the forelimb ness of the two epidermal layers. Images of histo- was the same as the dorsal fur (to the radiocarpal logical sections were photographed with a digital joint).