Informatics Publishing Limited

P. Sekar ¹, G. Vengatesh ¹, M. Kathiresh Kumar ¹, S. Balaji ¹, M. A. Akbarsha ², M. M. Aruldhas ³

Affiliations
1 Department of Endocrinology, Dr. ALM Post-Graduate Institute of Basic Medical Sciences, University of Madras, Taramani Campus, Chennai - 600 113, IN
2 Department of Animal Science, Bharathidasan University, Tiruchirappalli - 620 024, IN
3 Department of Endocrinology, Dr. ALM Post-Graduate Institute of Basic Medical Sciences, University of Madras Taramani Campus, Chennai - 600 113, IN

Abstract

Reproductive and embryonic toxicity of hexavalent chromium (CrVI) is known, and adult testis is one of its vulnerable targets. However, it is not known if gestational and lactational exposure to excess Cr affects development and functions of Leydig cells during postnatal life. It is hypothesized that gestational/lactational exposure to CrVI may affect Leydig cell development and differentiation and its functions during postnatal life extending into adulthood. Pregnant [gestational days 9 to 21] and lactating [postnatal days (PND) 1 to 21] rats were exposed to 50ppm and 100ppm CrVI (K₂Cr₂O₇) through drinking water, and testis was collected on PND 30, 60, 90 and 120, and subjected to light and transmission electron microscopic analysis. Serum testosterone and estradiol were determined adopting RIA. Histological evaluation of testes revealed hypertrophy and vacuolation of Leydig cells of CrVI-exposed rats; transmission electron micrographs (TEM) showed lipid accumulation, swollen mitochondria and disorganized smooth endoplasmic reticulum. Lactational exposure to CrVI led to decrease in the number of mitochondria and collapse of mitochondrial cristae. In general, the changes were obvious in PND 30 rats, and became less pronounced by PND 60 to become normal by PND 90. Serum testosterone and estradiol levels showed a general trend of opposite response to CrVI exposure. Gestational exposure to CrVI caused increase in testosterone level in prepuberal rats, but the trend was reversed by PND 60, and by PND 120 its level was more than in coeval controls. A similar trend was noticed in rats which had lactational exposure to CrVI but for a consistent increase in both steroids in PND 30 and PND 60 old rats which were exposed to 50ppm CrVI. By PND 90, testosterone remained elevated or normal, but by PND 120 its level was increased due to lactational exposure to CrVI. On the contrary, serum estradiol in these rats was low by PND 90 and became normal by PND 120. The findings partially support the hypothesis proposed and it is concluded that the fetal type Leydig cells are the major targets for the toxic effects of CrVI exposure during gestational and lactational periods where in lactational exposure may have a persistent effect leading to increased testosterone: estradiol ratio. Nevertheless, the effects of CrVI on testosterone and estradiol are reversible, as the adult type Leydig cells are unaffected.

Keywords

CrVI, Estradiol, Leydig Cells, Sertoli Cells, Testicular Toxicity, Testosterone.

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L. Divya ¹, M. A. Akbarsha ², M. M. Aruldhas ³, O. V. Oommen ¹

Affiliations
1 Dept of Zoology, University of Kerala, Kariavattom, Thiruvananthapuram 695581, IN
2 Department of Animal Science, Bharathidasan University, Tiruchirappalli 620024, IN
3 Department of Endocrinology, Dr ALM PGIBMS, University of Madras, Taramani campus, Chennai 600113, IN

Abstract

Reactive oxygen species (ROS) are known to influence molecular and biochemical processes and signal transduction pathways, affecting cellular proliferation, differentiation and death in a variety of organisms. Amphibian metamorphosis encompasses all these three events within a short span of time. In the frog Clinotarsus curtipes development is largely akin to the post-embryonic development in mammals, displaying increased levels of ROS under in vivo physiological conditions. Scavenging ROS with an antioxidant revealed a serendipitous finding of turning to a pro-oxidant and a novel thyroid hormone mimetic with potential effects on neurotransmitter functions. Further, confirmatory studies, both conventional binding assays combined with in- silico approaches, revealed the ability of the compound to bind to human thyroid receptors thereby to mimic the thyroid hormone activity and thus function as potent endocrine disrupting chemical. Thus our study also cautions against the indiscriminate use of supplementary molecules without proper validation. Our studies on amphibian (Clinotarsus curtipes) development are valuable in examining the role of ROS in post embryonic development.

Keywords

Neurotransmitters, Metamorphosis, ROS, Oxidant.

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A. Faridha ¹, K. Faisal ¹, M. A. Akbarsha ¹

Affiliations
1 Department of Animal Science, Bharathidasan University, Tiruchirappalli - 620 024, IN

Abstract

In the background of the decreasing trend of male reproductive health in men and animals, the present study was aimed at finding gravimetric, histopathologoical and histometric changes in the testis of Swiss mouse in response to treatment of aflatoxin B₁ (AFB₁) in a chronic toxicity testing over different periods of time and at finding recovery, if any, after cessation of the treatment. AFB₁ suspended in corn oil and ethanol, was administered through intraperitoneal route to 90 day old Swiss mouse at a daily dose of 50mg/kg body weight for 7, 15, 35, 45 days. In the recovery group the mice were kept, after cessation of the treatment, under observation for 35 or 70 days. The testicles and seminal vesicles of the animals were subjected to histopathological analysis adopting paraffin/resin embedding and light microscopy. Computer assisted histometric analysis of several parameters was also made. In general there was little impact of the treatment in the animals treated for 7 days. In the animals treated for 15, 35 and 45 days, there was duration-dependent regression of the testis and seminal vesicles. Histopathological changes were observed in both the spermatogenic and androgenic compartments of the testis. Fragmentation of chromatin of pachytene spermatocytes, generation of uni- and multinucleate gaint cells, and premature loss of spermatids and spermatocytes were observed. Whereas the seminiferous tubules regressed, the Leydig cells underwent hypertrophy and distortion of shape of the nucleus. Generation of multiple micronucleate gaint cells and extensive loss of germ cells from the seminiferous epithelium were observed. Fertility of the mice was severely hampered. The various changes are discussed in relation to the possible effects of AFB₁ on the microtubules of the spindle fibers and the DNA of chromatin. Since spermatogonia and Sertoli cells are not vulnerable targets to AF toxicity, the treated animals recovered over a period of time.

Keywords

Aflatoxin B1, Male Reproductive Toxicity, Testis, Seminal Vesicles.

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R. S. Beyo ¹, L. Divya ¹, K. Ramachandran ¹, O. V. Oommen ¹, M. A. Akbarsha ²

Affiliations
1 Department of Zoology, University of Kerala, Kariavattom, Thiruvananthapuram, 695585, IN
2 Derpartment of Animal Science, Bharathidasan University, Thiruchirappalli, 620 024, IN

Abstract

Corpus luteum is a transient and well known endocrine gland that develops from the ruptured follicles in the female vertebrate ovaries, immediately after ovulation. A major source of progesterone, this gland is concerned with gestation in the viviparous animals and retention of eggs in the oviduct in the oviparous species. Caecilians are a group of amphibians with several aspects of reproductive anatomy and physiology unique. In as much as corpus lutem has been described in a few caecilian species, it has not been described from ultrastructural perspectives. Here in we describe the ultrastructural features of corpus luteum of a single specimen of Gegeneophis ramaswamii, which was captured immediately after it laid the eggs. The corpus luteum is formed by three types of cells, namely granulosa lutein cells, thecal lutein cells and phagocytic cells. Both the types of lutein cells are characterized by abundant mitochondria, endoplasmic reticulum and Golgi apparatus. The granulose lutein cells abound with discrete granules. The phagocytic cells engage in phagocytosing cell debris. The cellular features are discussed in relation to information from oviparous as well as viviparous vertebrates. Thus, the paper describes, for the first time the corpus luteum of a caecilian from ultrastructural perspectives.

Keywords

Corpus Luteum, Endothelium, Theca, SER, Follicle Cell.

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M. A. Akbarsha ¹, M. Manimekalai ²

Affiliations
1 Department of Animal Science, School of Life Sciences, Bharathidasan University, Tiruchirappalli - 620 024, IN
2 Department of Zoology, Periyar EVR College, Tiruchirappalli - 620 023, IN

Abstract

Histological analysis of the male reproductive tract of the peninsular flap-shelled turtle Lissemys p. punctata revealed that several minute ductuli efferentes reach the epididymis to form into a large thin-walled duct which probably, forms a temporary storage region of sperm arriving from the testis. Originating from this duct, the single long ductus epididymidis takes a highly tortuous course when it differentiates along its length, in terms of diameter and epithelial organization, into four regions, viz., the initial segment, caput, corpus and cauda. Turtle ductus epididymidis differs from that of lizards in the extensive pattern of folding of the epithelium of caput, corpus and cauda regions. The differentiation along the ductus epididymidis of the turtle signifies different functional attributes to the different regions. The epithelium of the initial segment of the turtle epididymis secretes large glycoprotein granules (4-8 μm) and that of the caput secretes minute granules (1-2 μm). The large granules possess a central core and a peripheral coat, whereas the minute granules are uniformly dense. Thus, turtle ductus epididymidis differentiates into initial segment, caput, corpus and cauda regions, and the initial segment and caput secrete seminal proteins in the lacertilian pattern as discrete granules.

Keywords

Epididymis, Glycoprotein Granules, Seminal Protein.

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O. V. Oommen ¹, M. Smita ¹, R. S. Beyo ¹, M. G. Jancy ², R. Girija ², M. A. Akbarsha ²

Affiliations
1 Department of Zoology, University of Kerala, Kariavattom, 695 581, Thiruvananthapuram, IN
2 Department of Animal Science, Bharathidasan University, Thinjchirappalli- 620 024, IN

Abstract

Caecilians, of the order Gymnophiona in the Lissamphibia, are circumtropically distributed across most moist tropical regions of the world. Gymnophiona are among the least understood terrestrial vertebrates and have received lesser attention than the other two orders of lissamphibians, the Urodela and Anura. Spermiogenesis, known as spermateleosis in lower vertebrates, is the transformation of the round spermatid into a highly specialized spermatozoon with a species-specific structure. Spermateleosis and sperm morphology of two species of caecilians, Ichthyophis tricolor and Uraeotyphlus cf. narayani, from the Western Ghats of Kerala, India, were studied using light and scanning electron microscopy. The cyst morphology is described in ten steps. We also report the sperm transporting course in this enigmatic amphibian in addition to localizing sperm in the kidney. This confirms the transport of sperm through the kidney.

Keywords

Amelwid Cells, Caecilians, Connective Tissue Strand, Cyst, Sertoli Cells, Spermatids, Spermatozoa, Sperm Transport.

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M. Smita ¹, O. V. Oommen ¹, M. G. Jancy ², M. A. Akbarsha ²

Affiliations
1 Department of Zoology, University of Kerala, Kariavottom 695 581, Trivandrum, Kerala, IN
2 Department of Animal Sciences, Bharathidasan University, Thiruchirappalli 620 024, Tamil Nadu, IN

Abstract

The Gymnophiona are the least wellknown amphibian order, an unfortunate situation because these animals can clarify many aspects of amphibian natural history, vertebrate evolution and the physiology of viviparity and fossorial life.

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M. G. Jancy ¹, M. Smita ², M. M. Ibrahmsha ³, O. V. Oommen ², M. A. Akbarsha ¹

Affiliations
1 Department of Animal Science, Bharathidasan Univeristy, Tiruchirappalti 620 024, IN
2 Department of Zoology, University of Kerala, Kariavattom 695 581, IN
3 Department of Zoology, Jamal Mohamed College, Tiruchirappalli 620 020, IN

Abstract

The retention of Mullerian duct in the adult male caecilians has been an intriguing aspect of reproductive biology of the vertebrates, as it defies the established knowledge of sex differentiation. It is hypothesized that this retention must have a bearing on the unique aspects of their reproduction namely internal fertilization and partial- to total viviparity, particularly in the secretion of a seminal plasma in which sperm should be suspended during ejaculation.

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B. Kadalmani ¹, C. Raghavan ¹, T. Thirunalasundari ¹, G. Subramanian ², M. A. Akbarsha ¹

Affiliations
1 Department of Animal Sciences, Bharathidasan University, Thiruchirappallli 620 024, IN
2 National Facility for Marine Cyanobacteria, Bharathidasan University, Thiruchirappalli 620 024, IN

Abstract

Cyanobacteria are prokaryotic aquatic organisms. Their potential to produce varied and extensive secondary metabolites, both intracellularly and more importantly extracellularly, has come to light, and attempts are afoot to find the application of these compounds as nutritional supplements, food additives, natural'colorants, pharmaceuticals, etc. The marine cyanobacteria are an example for diversity biomass and productivity. To find the potential for application of the marine cyanobacteria in human and animal welfare, the Department of Biotechnology, Govt, of India, has established the National Facility for Marine Cyanobacteria (NFMC), the only one of its kind in whole country, at Bharathidasan University. ICMR, Government of India, has funded a research project to find the application of marine cyanobacteria in male contraception and the results are promising.

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P. Sekhar ¹, S. Subramanian ², N. S. Venkatesh ², A. M. J. Sashi ², N. Kalpana ², J. Anbalagan ², G. Vengatesh ², P. Govindarajulu ², M. A. Akbarsha ³, M. M. Aruldhas ²

Affiliations
1 Post Graduate Department of Zoology, Voorhees College, Vellore-632001, IN
2 Department of Endocrinology, University of Madras, Dr. ALM Post Graduate Institute of Basic Medical Sciences, Taramani Campus, Chennai-600113, IN
3 Department of Animal Science, Bharathidasan University, Tiruchirappalli-620024, IN

Abstract

Heavy metals and chemicals are known to cause male infertility. Chromium, an occupational heavy metal pollutant is a wellknown carcinogen with toxic effect on skin, lung and kidney. A few early reports point out the reproductive toxicity of chromium in experimental animals. An extensive study conducted in our laboratory in adult rats and monkeys established that altered prooxidant - antioxidant balance leading to free radical toxicity in the testis as the mechanism underlying the reproductive toxicity of chromium.

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A. M. J. Sashi ¹, N. S. Venkatesh ¹, G. Vengatesh ¹, P. Sekhar ¹, J. Anbalagan ¹, N. Kala ¹, P. Govindarajulu ¹, M. A. Akbarsha ², M. M. Aruldhas ¹

Affiliations
1 Department of Endocrinology, Dr. ALM Post Graduate Institute of Basic Medical Sciences, University of Madras, Taramani Campus, Chennai-600113, IN
2 Department of Animal Science, Bharathidasan University, Tiruchirappalli-620024, IN

Abstract

Recent demonstration of thyroid hormone receptors (TR) in epididymis by our laboratory and that of Del Rio point out that epididymis may also be a target for the direct effect of T₃. Recent studies emanating from our laboratory have shown the importance of foetal-onset hypothyroidism on the growth, structure and function of the epididymis in adult rats. Since there are temporal difference in the foetal differentiation of hypothalamo-hypophyseal-testicular axis and the differentiation of testis and epididymis, it is hypothesized "Transient maternal hypothyroidism modifies specific cell types of the epididymis during pre-puberal, puberal and adult period in a temporal manner".

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R. S. Beyo ¹, O. V. Oommen ¹, M. A. Akbarsha ²

Affiliations
1 Department of Zoology, University of Kerala, Kariavattom, Thiruvananthapuram 695581, IN
2 Department of Animal Science, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620 024, IN

Abstract

This study is the first to describe the major stages of follicles of a caecilian from ultrastructural perspectives. The female reproductive system of Ichthyophis tricolor is described. Assembly of the folhcles involves the origin of primary oocytes from oogonia and the pre-granulosa cells from ovarian stroma, each from a separate nest, and the arrival of pre-granulosa cells around the meiotic oocyte to become granulosa cells. Appearance of a perivitelline space between the oocyte and the granulosa cells marks the beginning of pre-vitellogenic follicles. Macrovilli arise from the granulosa cells and ramiiy in the perivitelline space. In the vitellogenic follicles the macrovilli are further more prominent, and microvilli arising from the oolemma intercalate between the macrovilli. Yolk platelets appear in the cortical ooplasm, which are initially small and scattered but later the smaller ones fuse, and lipid and glycogen are added on resulting in large yolk platelets. The material of the vitelline envelope appears in the perivitelline space, which later condenses into the vitelline envelope. Some of the follicles undergo atresia when the granulosa cells become atrophied, and amoeboid cells from the theca enter the ooplasm and phagocytose the yolk.

Keywords

Caecilian, Follicle, Granulosa Cells, Ichthyophis tricolor, Oocyte, Ovary.

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K. Faisal ¹, S. Parveen ², R. Rajendran ³, R. Girija ¹, V. S. Periasamy ¹, B. Kadalmani ¹, A. Puratchikody ⁴, K. Ruckmani ⁴, B. M. J. Pereira ², M. A. Akbarsha ¹

Affiliations
1 Department of Animal Science, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620 024, IN
2 Department of Bioscience and Biotechnology, Reproductive Biology Laboratory, Indian Institute of Technology Roorke 247 667, IN
3 Green Chem, Attibele Industrial Area, Bangalore - 562 107, IN
4 Department of Pharmaceutical Engineering & Technology Bharathidasan Institute of Technology, Tiruchirappalli - 620 024, IN

Abstract

Quassia amara is a medicinal plant with several pharmacological properties. More recently, it has come to be known in the context of its potential biopesticidal application and the antimalarial property in two of its secondary metabolites, quassin and neo-quassin. There are also preliminary reports implicating this plant in male reproductive toxic effects, to the extent that the plant could as well be tested for male contraceptive efficacy. Therfore, we subjected the methanolic extract of the bark of this tree for male reproductive toxicological evaluation. This paper reports several derangements in the Cauda epididymidal sperm including a hither - to unreported one.

Keywords

Neo-Quassin, Quassia amara, Quassin, Sperm Abnormalities.

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K. Faisal , R. Girija , M. A. Akbarsha

Abstract

Quassia amara is a Brazilian medicinal plant traditionally used as an herbal remedy for a variety of diseases. Quassin is the major secondary chemical compound in it, which is under investigation as an anti-malarial compound. There are preliminary studies implicating Q. amara in male reproductive toxicity. Hence, this elaborate study was undertaken to find the male reproductive toxic effects of Q. amara and quassin. Methanolic extract of Q. amara bark and quassin were administered to male mice for 35 days. The cauda epididymidal sperm, testis and epididymis were subjected to analysis adopting light and transmission electron microscopy. Sperm counts, motility and viability decreased. The incidence of sperm with abnormal morphologies increased. There was premature loss of germ cells from the seminiferous tubules, which correlated with Sertoli cell pathology. The Leydig cells were vacuolated, suggesting a hypo-androgen status. The epididymis indicated severe histopathological changes, including reflection of a mitogenic/tumorigenic effect of Q. amara. Thus, this study indicates that Q. amara, when applied as a therapeutic, could be severely toxic to male reproductive mechanisms and the principal mechanism of action appears to be through Leydig cells. Quassin, the major secondary metabolite responsible for the therapeutic potential of Q. amara, is also the one causative of the male reproductive toxic effects. Thus, this study substantiates the claim that Q. amara is toxic to male reproductive system.

Keywords

Epididymis, Histopathology, Sperm, Testis, Quassia amara, Quassin.

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K. Faisal ¹, M. A. Akbarsha ¹

Affiliations
1 Department of Animal Science, School of Life Sciences, Bharathidasan University, Tiruchirappalli - 620024, Tamil Nadu, IN

Abstract

Ubiquitination is believed to play a critical role in removal of dead and/or defective spermatozoa in normal and, more importantly, under circumstances when such spermatozoa are produced in large numbers due to genetic defects or toxic manifestations. Ubiquitination under such instances would involve specific gene expressions, many of which are not yet clearly understood. In an exhaustive study in Swiss mouse model to find the spermatotoxic effect of quassin, a diterpene compound isolated from Quassia amara, we found most of the spermatozoa to be abnormal in morphology and unviable. In the present study, we aimed at analysing the transcriptional profile of three selected genes, Ubb, Ube2c and Psmb8, involved in the ubiquitin proteolytic pathway in the testis and epididymal segments of Q. amara bark methanol extract treated mice adopting semi-quantitative RT-PCR and to study the level of DNA damage of the treated mouse spermatozoa. The results revealed that the treatment induced considerable damage to the sperm DNA. All the three genes studied showed marked increase in their levels of expression in the treated mice compared to the corresponding controls. Thus, this study shows that Q. amara methanol extract is causative of sperm DNA damage and defective spermatozoa and, in such cases, the expression of specific genes concerned with ubiquitination pathway is increased, implying that ubiquitination-proteosomal degradation is involved in the processing of dead/defective spermatozoa.

Keywords

Defective Sperm, Proteosomal Degradation, Quassin, Quassia amara, Spermatotoxicity, Ubiquitination.

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V. Saraswathy ¹, R. Dileepkumar ², Achuthsankar S. Nair ¹, M. A. Akbarsha ³, Oommen V. Oommen ¹

Affiliations
1 Centre for Venom Informatics, Department of Computational Biology and Bioinformatics, University of Kerala-Kariavattom Campus, Thiruvananthapuram – 695581, Kerala, IN
2 Indriyam Biologicals, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Poojapura, Thiruvananthapuram – 695012, Kerala, IN
3 National College (Autonomous), Tiruchirappalli – 620001, Tamil Nadu, IN

Abstract

Envenomation is a serious neglected health issue at the global level that affects millions of people every year. It is highly prevalent among farmers and rural natives and is mainly due to the bite from snakes, spiders, frogs, dogs, wasps, bees, ants, etc. Many plants with antidote potential grow around our backyard without proper recognition and, unfortunately, several of them are under threat of extinction due to human interference and other environmental factors. The sustainable utilization of those antidote herbs can benefit as a life saver to the needy patients. The herbs can also be farmed and used commercially for the pharmaceutical application and incorporated with biotechnology and bioinformatics with a vision of synthesizing antidote drugs with less or no side effects. The objective of the present work is to create awareness among the public for the wise use of wild and local herbs, and their sustainable utilization with a computational case study on laboratory experiments done in two traditional plant based antidotes, selected from literature. The investigation is focused on Daboia russelii venom neutralization via in silico approach which can significantly reduce the time, expense, labour and samples taken. Our approach will add to the conventional non-specific polyvalent anti-snake venom (ASV) with more specific plant-based antidotes.

Keywords

Antidote, Envenomation, Herbal Medicines, Sustainable Utilization, Venom Informatics.

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R. Dileepkumar ¹, K. Anaswara ¹, V. Navya ², S. Beena ³, A. Jacob ⁴, L. Divya ⁵, A. S. Vijayasree ⁶, V. Deepthi ², G. Renganayaki ², P. R. Shidhi ², M. A. Akbarsha ⁷, K. P. Laladhas ⁸, Achuthsankar S. Nair ², P. R. Sudhakaran ², O. V. Oommen ²

Affiliations
1 Indriyam Biologics Private Limited, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Thiruvananthapuram 695 001, IN
2 Department of Computational Biology and Bioinformatics, University of Kerala, Thiruvananthapuram 695 581, IN
3 OmicsGen Life Sciences Pvt Ltd, Cochin 682 301, IN
4 Zoological Garden, Thiruvananthapuram 695 033, IN
5 Department of Animal Sciences, Central University of Kerala, Kasaragod 671 316, IN
6 Department of Zoology, Fatima Mata National College, Kollam 691 001, IN
7 Department of Biotechnology, National College (Autonomous), Tiruchirappalli 620 001, IN
8 St Stephen’s College, Pathanapuram 689 695, IN

Abstract

This study shows that the dominant mutation of Extension locus in the recessive locus of the agouti Indian three-striped palm squirrel results in constituting an active or hyperactive receptor. This is not inhibited by the agouti antagonist or agouti signalling protein, resulting in melanism. To the best of our knowledge, there is no earlier report of a melanic variant (black) of a three-striped palm squirrel (Funambulus palmarum Linn.) from India. The colour change is due to mutation and is traced to melanocortin-1 receptor (MC1R) gene, where it is proved to be a sequence alteration causing a frame shift in the Extension locus of the wild type. This would have probably caused the constitutive activation of MC1R.

Keywords

Agouti Signalling Protein, Dominant Mutation, Funambulus palmarum, Melanism, Melanocortin-1 Receptor.

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