International Journal of Current Research and Review
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IJCRR - 7(14), July, 2015

Pages: 35-39

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HISTOLOGICAL AND ULTRA STRUCTURAL STUDIES OF THYMIC MYOID CELLS IN NANDANAM CHICKEN

Author: T.A. Kannan, Geetha Ramesh, S. Ushakumari, Sabiha Hayath Basha

Category: Healthcare

Abstract:Light and transmission electron microscopic studies of thymic myoid cells in Nandanam Chicken was done in various age groups ranging from day-old to forty weeks. Thymus is the central lymphoid organ in fowl, provides a complex environment essential for T-cell development and maturation. The thymic gland in chicken was covered by a thin connective tissue capsule. The connective tissue septa divided the gland into lobules. Thymic parenchyma consisted of an outer darker cortex and inner pale medulla. Under light microscope, numerous large cells with eosinophilic cytoplasm were observed in the medulla and also in the cortico-medullary junction, the myoid cells. Number of myoid cells increased as age advances. Under electron microscope, the cytoplasm of the myoid cell showed myofibrils as in skeletal muscle fibre. Cytoplasm also contained few mitochondria, free
ribosome and smooth endoplasmic reticulum. Small bundles of unmyelinated nerve fibres were observed in the thymic medulla

Keywords: Histology, Ultrastructure, Thymus, Myoid cell, Chicken

Full Text:

INTRODUCTION

Thymus is the central lymphoid organ occurs in all vertebrate species, situated on either side of the neck, close to the jugular vein (King, 1983). Bone marrow-derived T-cell precursors undergo differentiation, maturation eventually leading to migration of positively selected thymocytes to the peripheral lymphoid organs such as the spleen (Savino and Dardenne, 2000 and Varga et al., 2009) and GALT including the caecal tonsil and the lymph nodes (Ciriaco et al., 2003; Gail Pearse, 2006 and Karen Staines et al., 2013). Thymus is unique among the lymphoid organs in being an epithelial organ and well known for its cellular organization (Gail Pearse, 2006). Thymic epithelium shows subpopulation heterogenicity in both capsule and thymic parenchyma that form the three-dimensional framework of the thymus (Mohammad et al., 2007). Among these heterogenous cell population, there is an unique and interesting myoid cells of the thymus. Myoid cells are first described in Amphibians (Mayer, 1888) and they have been found to vary considerably due to cell turnover in the thymus in different vertebrates (Raviola and Raviola, 1967; Nakamura et al., 1986 and Geetha Ramesh and Vijayaragavan, 1997). In domestic fowl, they are identified as large cells in the thymic medulla and characterised by the presence of myofibrils in their cytoplasm similar to those found in skeletal muscle fibre (Gilmore and Bridges, 1974). Thymic myoid cells plays an important role of protecting thymocytes from apoptosis (Panse and Berrilh-Aknin, 2005). Although, fine structure of myoid cells has been described by Frazier (1973) in fowl, little details are available of their presence in Nandanam chicken of different age groups. Hence, an attempt has been made to study the histological and ultrastructural details of this unique cell type of thymic parenchyma of Nandanam chicken. It is a dual purpose, colored variety with good disease resistance and most popular among poultry farmers due to its adaptability to backyard farming. Nandanam strain was developed in Poultry Research Station, Tamil Nadu Veterinary and Animal Sciences University, Chennai.

MATERIALS AND METHODS

Over 36 specimens of thymic tissue were collected from six different age groups such as day-old, four, eight, twelve,twenty and forty weeks. Six birds were used in each age group. The thymus was removed immediately after high cervical dislocation and fixed for light and electron microscopy (Gilmore and Bridges, 1974). For light microscopy, tissue pieces were fixed in 10% neutral buffered formalin, dehydrated in alcohol, cleared in xylene and embedded in paraffin wax. Five micron thick sections were made and stained with Haematoxylin and Eosin (Bancroft and Stevens, 2007) and Masson’s trichrome for collagen and muscle fibres (Kiernan, 1981). For electron microscopy, small pieces of tissue were fixed for 2 hours in 3 % glutaraldehyde buffered to pH 7.4 with 0.1 M sodium cacodylate buffer at 40 C. The tissue was washed overnight in three changes of buffer at 40 C, post-fixed in 1 % osmium tetroxide in 0.1 M sodium cacodylate buffer at 40 C for 2 hours and washed overnight in buffer. Dehydration in ethanol and propylene oxide was followed by embedding in propylene oxide: epoxy resin mixture and embedded in Epon-araldite mixture after the method of Glauert (1967). Semi thin (1 micron) sections were stained by toluidine blue. Ultra thin sections (600 Ao to 900Ao ) were prepared on Leica ultracut microtome, mounted on uncoated copper grids and stained with saturated solution of uranyl acetate and lead citrate. Screening of ultra thin sections were made with transmission electron microscope (Phillips - Teknai-10) operated at 60-kilowatt ampere (KVA). The images were documented and processed in computer.

RESULTS

Light microscopy

In the present study, thymic gland was surrounded by a thin connective tissue capsule composed mainly of collagen and few elastic fibes. Fine trabeculae / septa extended from the capsule and divided the parenchyma into lobules. The parenchyma was observed to have a dark outer cortex and a pale inner medulla within the lobule in day-old, four weeks and ten weeks of age groups. The ratio of cortex to medulla of the thymus in day-old chick reversed in adult age groups. The cortex consisted of numerous small, medium and large lymphocytes lying within a meshwork of reticuloepithelial cells. Whereas the medulla contained predominantly retiucloepithelial cells with a fewer lymphocytes. In all the age groups, myoid cells were most numerous in the medulla, less in cortex. Also found in the cortico-medullary junction. The extramedullary location often found in young age groups rather than adult birds. The myoid cells were large and of varying shape in different age groups studied. From day-old to 20 weeks of age, they were seen as elongated, spindle shaped cells with striations (Fig. 1). In 40 weeks of age, the cells appeared ovoid or round. The nucleus was large and oval in shape with a single, discrete nucleolus. The cytoplasm appeared homogenous under light microscope and striations noticed in some cells. The cytoplasm was red in colour after Masson’s trichrome stain. The myoid cells were fewer in the thymic medulla on dayold and increased in number upto 40 weeks of age, in the present study

 Electron microscopy

Under transmission electron microscope, the cytoplasm of the myoid cell contained myofibrils of skeletal muscle (Fig.2). However, the overall arrangement of those myofibrils were not as in skeletal muscle fibre. The cytoplasm contained few mitochondria and smooth endoplasmic retiuclum and also free ribosomes and glycogen granules. The nucleus contained dispersed chromatin. In younger age groups, the myofibrils were oriented along the long axis and changes in the orientation was noticed in 40 weeks of age. The myoid cells of the chicken thymus were found mainly in the medulla in all the age groups studied. The number of myoid cells increased as age advances in the present study. Small bundles of unmyelinated nerve fibres were observed in the thymic medulla in the present study (Fig.3). However, no nerve fibres were seen to terminate directly on the surface of the myoid cell, although some fibres lay in close association with them.

DISCUSSION

Light microscopy

Though there are literatures available on the histoarchitecture of avian thymus, the detailed study about the thymic myoid cell in Nandanam chicken is scanty. Hence, the present study was therefore designed to observe the light and electron microscopic details of myoid cells in Nandanam chicken. In chicken, thymic gland was surrounded by a connective tissue capsule and fine trabeculae / septa extended from the capsule and divided the parenchyma into lobules (Hodges, 1974; Firth, 1977; Bhattacharya and Binaykumar, 1983; Gail Pearse, 2006; Leena et al., 2008). These septa are considered as important structure for immigration and emigration of T-lymphocyte precursor cells and distribution of antigens within the thymus. They are also thought to guide the invasion of interdigitating reticular cells, macrophages and act as a pathway for blood vessels and nerves to and from the thymus (Seifert and Christ, 1990 and Ritter and Crispe, 1992). In all the age groups, the parenchyma was observed to have a dark outer cortex and a pale inner medulla and the cellular composition of the cortex was in agreement with the findings of Hodges (1974), Gilmore and Bridges, (1974) in fowl and King and Mc Lelland (1981) in birds. The extramedullary location of the myoid cells in the present study supports the ontogenic similarities between lower and higher vertebrates (Kendall, 1980; Saad and Zapata, 1992; Bowden et al., 2005 and Mohamed et al., 2007). A similar finding was observed by Gilmore and Bridges (1974) and Kendall (1980) in chicken and Geetha Ramesh and Vijayaragavan (1997) in buffalo calves. But Robert et al. (1978) described that myoid cells were rare in human thymus. The change in shape of the myoid cells between age groups in the present study was assumed to be change in cell morphology to be an expression of degeneration corresponding to the degeneration of the thymus as postulated by Raviola and Raviola (1967) and van de Velde and Friedman (1967). The presence of striations in the cytoplasm recognised the presence of muscular elements (Itoh, 1983 and Leena et al., 2008). Increase in number of myoid cells from day-old to forty weeks of age in the present study leads to a postulation that these myoid cells directly or indirectly related with the involution of the organ, which is functional synchronization with the amount of sex hormone present in the blood (Vijayaragavan, 1988). Whereas Bockman (1968) assumed that the myoid cells were involved in the mechanism of tolerance to muscle self-antigen in human, but Mandel (1968a and b) opined that the thymic striated muscles may act as a local source of antigen for the self recognition of skeletal muscle in guinea pig.

Electron microscopy

Under transmission electron microscope, changes noticed in the cell organelles and orientation of myofibrils indicted the degenerating changes in involuting thymus (Van de Velde and Friedman, 1970 and Leena et al., 2012). The presence of myoid mainly in the medulla in all the age groups studied was similar to thymic tissue from humans, amphibians, reptiles, birds and various mammals (van de Velde and Friedman, 1966, 1967; Strauss et al.,1966; Henry,1968; Toro et al., 1969 and Bridges et al., 1970). However, Morris (1971) and Sugimura (1972) have reported their presence in the medulla of the ten to twelve month old Ox, and they are present in the normal adult human thymus (Henry, 1968). The myoid cells seen in the present study are structurally similar to those described in the frog thymus (Toro et al., 1969). Two main theories have been put forward to explain the presence of myoid cells within the thymus: (1) that aberrant mesodermal elements from the branchial arches become incorporated accidentally into the thymus during its embryological development (van de Velde and Friedman, 1966 and Frazier, 1973) and (2) they develop within the thymus as an intrinsic part of the organ (Kapa et al. 1968; Bockman and Winborn, 1969 and Toro et al., 1969). These latter authors suggested that the myoid cells develop from the epithelial cells of the thymic cytoreticulum. Toro and Olah (1967) and Toro et al. (1968) have also observed this process in the development of myoid cells during the in vitro culture of embryonic rat thymus tissue. The number of myoid cells increased as age advances in the present study does not support the first theory that they represent the outcome of an embryological accident. Hence, the present study supports the second theory that the cells develop as an integral part of the normal thymus and as such it is reasonable to suggest that they play a physiological role therein. The function of the myoid cells is still unknown. However, Raviola and Raviola (1967) and Rimer (1980) found that myoid cells play no physiological role because myofilaments are organized in random directions and myoid cells have no anchoring apparatus. But in the present study, myofilaments were shown to arrange regularly, and cross striations were clear. Hence, we are inclined to think that myoid cells push lymphatic cells to circulation around hatching as Toro et a1. (1969) suggested, though anchoring apparatus was not found in this study.

CONCLUSIONS

Thymic parenchyma was made up of an outer darker cortex and inner pale medulla. The myoid cells were numerous in the medulla and were large cells with eosinophilic cytoplasm. They were also observed in the cortico-medullary junction. Under electron microscope, the cytoplasm of the myoid cell showed myofibrils as in skeletal muscle fibre. Cytoplasm also contained few mitochondria, free ribosome and smooth endoplasmic reticulum. Small bundles of unmyelinated nerve fibres were observed in the thymic medulla. Number of myoid cells increased as age advances.

Conflict of Interests

The authors declare that they have no competing interests among them.

ACKNOWLEDGEMENT

Authors acknowledge the immense help received from the scholars whose articles are cited and included in references of this manuscript. The authors are also grateful to authors / editors / publishers of all those articles, journals and books from where the literature for this article has been reviewed and discussed. The authors also acknowledge the Professor and Head, Department of Animal Biotechnology and Professor and Head, Centralised Instrumentation Laboratory, Madras Veterinary College for providing facilities to carry out this work.

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A Study by Kumar S. et al. entitled "A Study on Clinical Spectrum, Laboratory Profile, Complications and Outcome of Pediatric Scrub Typhus Patients Admitted to an Intensive Care Unit from a Tertiary Care Hospital from Eastern India" is awarded Best Article for Vol 13 issue 09
A Study by Mardhiah Kamaruddin et al. entitled "The Pattern of Creatinine Clearance in Gestational and Chronic Hypertension Women from the Third Trimester to 12 Weeks Postpartum" is awarded Best Article for Vol 13 issue 08
A Study by Sarmila G. B. et al. entitled "Study to Compare the Efficacy of Orally Administered Melatonin and Clonidine for Attenuation of Hemodynamic Response During Laryngoscopy and Endotracheal Intubation in Gastrointestinal Surgeries" is awarded Best Article for Vol 13 issue 07
A Study by M. Muthu Uma Maheswari et al. entitled "A Study on C-reactive Protein and Liver Function Tests in Laboratory RT-PCR Positive Covid-19 Patients in a Tertiary Care Centre – A Retrospective Study" is awarded Best Article of Vol 13 issue 06 Special issue Modern approaches for diagnosis of COVID-19 and current status of awareness
A Study by Gainneos PD et al. entitled "A Comparative Evaluation of the Levels of Salivary IgA in HIV Affected Children and the Children of the General Population within the Age Group of 9 – 12 Years – A Cross-Sectional Study" is awarded Best Article of Vol 13 issue 05 Special issue on Recent Advances in Dentistry for better Oral Health
A Study by Alkhansa Mahmoud et al. entitled "mRNA Expression of Somatostatin Receptors (1-5) in MCF7 and MDA-MB231 Breast Cancer Cells" is awarded Best Article of Vol 13 issue 06
A Study by Chen YY and Ghazali SRB entitled "Lifetime Trauma, posttraumatic stress disorder Symptoms and Early Adolescence Risk Factors for Poor Physical Health Outcome Among Malaysian Adolescents" is awarded Best Article of Vol 13 issue 04 Special issue on Current Updates in Plant Biology to Medicine to Healthcare Awareness in Malaysia
A Study by Kumari PM et al. entitled "Study to Evaluate the Adverse Drug Reactions in a Tertiary Care Teaching Hospital in Tamilnadu - A Cross-Sectional Study" is awarded Best Article for Vol 13 issue 05
A Study by Anu et al. entitled "Effectiveness of Cytological Scoring Systems for Evaluation of Breast Lesion Cytology with its Histopathological Correlation" is awarded Best Article of Vol 13 issue 04
A Study by Sharipov R. Kh. et al. entitled "Interaction of Correction of Lipid Peroxidation Disorders with Oxibral" is awarded Best Article of Vol 13 issue 03
A Study by Tarek Elwakil et al. entitled "Led Light Photobiomodulation Effect on Wound Healing Combined with Phenytoin in Mice Model" is awarded Best Article of Vol 13 issue 02
A Study by Mohita Ray et al. entitled "Accuracy of Intra-Operative Frozen Section Consultation of Gastrointestinal Biopsy Samples in Correlation with the Final Histopathological Diagnosis" is awarded Best Article for Vol 13 issue 01
A Study by Badritdinova MN et al. entitled "Peculiarities of a Pain in Patients with Ischemic Heart Disease in the Presence of Individual Combines of the Metabolic Syndrome" is awarded Best Article for Vol 12 issue 24
A Study by Sindhu Priya E S et al. entitled "Neuroprotective activity of Pyrazolone Derivatives Against Paraquat-induced Oxidative Stress and Locomotor Impairment in Drosophila melanogaster" is awarded Best Article for Vol 12 issue 23
A Study by Habiba Suhail et al. entitled "Effect of Majoon Murmakki in Dysmenorrhoea (Usre Tams): A Standard Controlled Clinical Study" is awarded Best Article for Vol 12 issue 22
A Study by Ghaffar UB et al. entitled "Correlation between Height and Foot Length in Saudi Population in Majmaah, Saudi Arabia" is awarded Best Article for Vol 12 issue 21
A Study by Siti Sarah Binti Maidin entitled "Sleep Well: Mobile Application to Address Sleeping Problems" is awarded Best Article for Vol 12 issue 20
A Study by Avijit Singh"Comparison of Post Operative Clinical Outcomes Between “Made in India” TTK Chitra Mechanical Heart Valve Versus St Jude Mechanical Heart Valve in Valve Replacement Surgery" is awarded Best Article for Vol 12 issue 19
A Study by Sonali Banerjee and Mary Mathews N. entitled "Exploring Quality of Life and Perceived Experiences Among Couples Undergoing Fertility Treatment in Western India: A Mixed Methodology" is awarded Best Article for Vol 12 issue 18
A Study by Jabbar Desai et al. entitled "Prevalence of Obstructive Airway Disease in Patients with Ischemic Heart Disease and Hypertension" is awarded Best Article for Vol 12 issue 17
A Study by Juna Byun et al. entitled "Study on Difference in Coronavirus-19 Related Anxiety between Face-to-face and Non-face-to-face Classes among University Students in South Korea" is awarded Best Article for Vol 12 issue 16
A Study by Sudha Ramachandra & Vinay Chavan entitled "Enhanced-Hybrid-Age Layered Population Structure (E-Hybrid-ALPS): A Genetic Algorithm with Adaptive Crossover for Molecular Docking Studies of Drug Discovery Process" is awarded Best article for Vol 12 issue 15
A Study by Varsha M. Shindhe et al. entitled "A Study on Effect of Smokeless Tobacco on Pulmonary Function Tests in Class IV Workers of USM-KLE (Universiti Sains Malaysia-Karnataka Lingayat Education Society) International Medical Programme, Belagavi" is awarded Best article of Vol 12 issue 14, July 2020
A study by Amruta Choudhary et al. entitled "Family Planning Knowledge, Attitude and Practice Among Women of Reproductive Age from Rural Area of Central India" is awarded Best Article for special issue "Modern Therapeutics Applications"
A study by Raunak Das entitled "Study of Cardiovascular Dysfunctions in Interstitial Lung Diseas epatients by Correlating the Levels of Serum NT PRO BNP and Microalbuminuria (Biomarkers of Cardiovascular Dysfunction) with Echocardiographic, Bronchoscopic and HighResolution Computed Tomography Findings of These ILD Patients" is awarded Best Article of Vol 12 issue 13 
A Study by Kannamani Ramasamy et al. entitled "COVID-19 Situation at Chennai City – Forecasting for the Better Pandemic Management" is awarded best article for  Vol 12 issue 12
A Study by Muhammet Lutfi SELCUK and Fatma entitled "Distinction of Gray and White Matter for Some Histological Staining Methods in New Zealand Rabbit's Brain" is awarded best article for  Vol 12 issue 11
A Study by Anamul Haq et al. entitled "Etiology of Abnormal Uterine Bleeding in Adolescents – Emphasis Upon Polycystic Ovarian Syndrome" is awarded best article for  Vol 12 issue 10
A Study by entitled "Estimation of Reference Interval of Serum Progesterone During Three Trimesters of Normal Pregnancy in a Tertiary Care Hospital of Kolkata" is awarded best article for  Vol 12 issue 09
A Study by Ilona Gracie De Souza & Pavan Kumar G. entitled "Effect of Releasing Myofascial Chain in Patients with Patellofemoral Pain Syndrome - A Randomized Clinical Trial" is awarded best article for  Vol 12 issue 08
A Study by Virendra Atam et. al. entitled "Clinical Profile and Short - Term Mortality Predictors in Acute Stroke with Emphasis on Stress Hyperglycemia and THRIVE Score : An Observational Study" is awarded best article for  Vol 12 issue 07
A Study by K. Krupashree et. al. entitled "Protective Effects of Picrorhizakurroa Against Fumonisin B1 Induced Hepatotoxicity in Mice" is awarded best article for issue Vol 10 issue 20
A study by Mithun K.P. et al "Larvicidal Activity of Crude Solanum Nigrum Leaf and Berries Extract Against Dengue Vector-Aedesaegypti" is awarded Best Article for Vol 10 issue 14 of IJCRR
A study by Asha Menon "Women in Child Care and Early Education: Truly Nontraditional Work" is awarded Best Article for Vol 10 issue 13
A study by Deep J. M. "Prevalence of Molar-Incisor Hypomineralization in 7-13 Years Old Children of Biratnagar, Nepal: A Cross Sectional Study" is awarded Best Article for Vol 10 issue 11 of IJCRR
A review by Chitra et al to analyse relation between Obesity and Type 2 diabetes is awarded 'Best Article' for Vol 10 issue 10 by IJCRR. 
A study by Karanpreet et al "Pregnancy Induced Hypertension: A Study on Its Multisystem Involvement" is given Best Paper Award for Vol 10 issue 09

List of Awardees

A Study by Ese Anibor et al. "Evaluation of Temporomandibular Joint Disorders Among Delta State University Students in Abraka, Nigeria" from Vol 13 issue 16 received Emerging Researcher Award


A Study by Alkhansa Mahmoud et al. entitled "mRNA Expression of Somatostatin Receptors (1-5) in MCF7 and MDA-MB231 Breast Cancer Cells" from Vol 13 issue 06 received Emerging Researcher Award


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