Seasonal changes in the protein profile and enzyme activity of the epididymal luminal fluid in the lizard, Eutropis carinata (Schneider, 1801)

in Animal Biology
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The epididymis of the male reproductive system is known to be involved in sperm maturation via the production of polypeptides, glycoproteins, surface proteins, enzymes and other factors. During the annual reproductive cycle, the epididymis of the lizard Eutropis carinata undergoes dramatic changes, both morphologically and biochemically, that occur in a well-organized sequence. The present study reveals the sequential changes that occur in the production and concentration of proteins in the epididymal luminal fluid throughout the annual reproductive cycle. A one-dimensional electrophoretic profile of the epididymal luminal proteins revealed a total of 18 bands in the regenerative phase, 22 bands during breeding and 17 bands in the post-breeding as well as regressed phases of the reproductive cycle. By two-dimensional electrophoresis, the protein complexes that are unique to the breeding phase were further resolved based on their pI and the molecular weight of each protein of the protein complex was determined. This is the first study to observe that proteins that are present during the reproductively inactive phase disappear during the reproductively active phase. The Periodic Acid Schiff (PAS) test for protein profiles revealed the presence of proteins with a carbohydrate moiety. Certain enzymes, such as acid phosphatase, alkaline phosphatase, and α-glucosidase, are highly sensitive to seasonal changes and their activity parallels the production of the epididymal proteins. This study provides evidence for androgen-dependent cyclical changes in the pattern of protein profiles and enzyme activity of the epididymal lumen in the lizard E. carinata.

Seasonal changes in the protein profile and enzyme activity of the epididymal luminal fluid in the lizard, Eutropis carinata (Schneider, 1801)

in Animal Biology



AndòS.CiarciaG.PannoM.L.ImbrognoE.TarantinoG.BuffoneM.BeraldiE.AngeliniF. & BotteV. (1992) Sex steroids levels in the plasma and testis during the reproductive cycle of lizard Podarcis s. sicula Raf. Gen. Comp. Endocrinol.851-7.

AnupC.P.MelvinP.ShilpaN.GandhiM.N.JadhavM.AliH. & KiniK.R. (2015) Proteomic analysis of elicitation of downy mildew disease resistance in pearl millet by seed priming with β-aminobutyric acid and Pseudomonas fluorescens. J. Proteomics12058-74.

AranhaI.BhagyaM. & YajurvediH.N. (2006) Ultrastructural study of the epididymis and the vas deferens and electrophoretic profile of their luminal fluid proteins in the lizard Mabuya carinata. J. Submicrosc. Cytol. Pathol.3837-43.

AveralH.I.ManimekalaiM. & AkbarshaM.A. (1992) Differentiation along the ductus epididymis of the Indian garden lizard, Calotes versicolor (Daudin). Biol. Struct. Morphogen.453-57.

BedfordJ.M. (1975) Maturation, transport and fate of spermatozoa in the epididymis. In: E.D.W. Hamilton & R.O. Greep (Eds) Handbook of Physiology pp. 303-318. American Physiology SocietyWashington DC, USA.

BrooksD.E. (1981) Secretion of protein and glycoprotein by the rat epididymis. Biol. Reprod.251099-1117.

ChapmanD.A. & KillianG.J. (1984) Glycosidase activities in principal cells, basal cells, fibroblasts and spermatozoa isolated from the rat epididymis. Biol. Reprod.31627-636.

ChoiH.HanC.JinS.KwonJ.T.KimJ.JeongJ.KimJ.HamS.JeonS.YooY.J. & ChoC. (2015) Reduced fertility and altered epididymal and sperm integrity in mice lacking ADAM71. Biol. Reprod.9370-71.

CiarciaG.LancieriM.SuzukiH.ManzoC.VitaleL.Tornese BuonamassaD. & BotteV. (1986) A specific nuclear protein and poly(ADPribose) transferase activity in lizard oviduct during the reproductive cycle. Mol. Cell. Endocrinol.47235-241.

DacheuxJ.L.DacheuxF. & PaquignonM. (1989) Changes in sperm surface membrane and luminal protein fluid content during epididymal transit in the boar. Biol. Reprod.40635-651.

DacheuxJ.L.DruartX.FouchecourtS.SyntinP.GattiJ.L.OkamuraN. & DacheuxF. (1998) Role of epididymal secretory proteins in sperm maturation with particular reference to the boar. J. Reprod. Fertil. Suppl.5399-107.

DacheuxJ.-L.GattiJ.L. & DacheuxF. (2003) Contribution of epididymal secretory proteins for spermatozoa maturation. Microsc. Res. Tech.617-17.

DacheuxJ.-L.DacheuxF. & DruartX. (2016) Epididymal protein markers and fertility. Anim. Reprod. Sci.16976-87.

DepeigesA. & DufaureJ.P. (1980) Major proteins secreted by the epididymis of Lacerta vivipara. Isolation and characterization by electrophoresis of the central core. Biochim. Biophys. Acta628109-115.

DepeigesA. & DufaureJ.P. (1981) Major proteins secreted by the epididymis of Lacerta vivipara. Identification by electrophoresis of soluble proteins. Biochim. Biophys. Acta – Protein Struct.667260-266.

DepeigesA. & DufaureJ.-P. (1983) Binding to spermatozoa of a major soluble protein secreted by the epididymis of the lizard Lacerta vivipara. Gamete Res.7401-406.

DepeigesA. & DacheuxJ.L. (1985) Acquisition of sperm motility and its maintenance during storage in the lizard, Lacerta vivipara. J. Reprod. Fertil.7423-27.

DepeigesA.BetailG.CouletM. & DufaureJ.P. (1985) Histochemical study of epididymal secretions in the lizard, Lacerta vivipara. Localization of lectin-binding sites. Cell Tissue Res.239463-466.

DepeigesA.ForceA. & DufaureJ.P. (1987) Production and glycosylation of sperm constitutive proteins in the lizard Lacerta vivipara. Evolution during the reproductive period. Comp. Biochem. Physiol. B86233-240.

DosemaneD. & BhagyaM. (2015) In vitro study of the spermatozoa motility in the lizard Eutropis carinata. J. Zool. Res.1189.

DosemaneD. & BhagyaM. (2016) Study of the spermatozoa motility acquisition patterns in excurrent duct system of the lizard Eutropis carinata. Int. J. Sci. Tech.442-45.

DufaureJ.P. & ChambonM. (1978) Uptake of [3H]testosterone in several organs of the male viviparous lizard (Lacerta vivipara Jacquin) and selective retention by the epididymis. Gen. Comp. Endocrinol.3623-29.

DufaureJ.P.CourtyY.DepeigesA.MesureM. & ChevalierM. (1986) Evolution and testosterone content of the epididymis during the annual cycle of the lizard Lacerta vivipara. Biol. Reprod.35667-675.

DufaureJ.R. & Saint-GironsH. (1984) Histologie comparée de l’epididyme et des ses sécrétions chez les reptiles (lézards et serpentes). Arch. Anat. Microsc.7315-26.

EspondaP. & BedfordJ.M. (1987) Post-testicular change in the reptile sperm surface with particular reference to the snake, Natrix fasciata. J. Exp. Zool.241123-132.

EzerN. & RobaireB. (2002) Androgenic regulation of the structure and functions of the epididymis. In: B. Robaire & B.T. Hinton (Eds) The Epididymis and the Vas Deferens pp. 297-316. Kluwer Academic/Plenum Publ.New York, NY, USA.

FouchécourtS.MétayerS.LocatelliA.DacheuxF. & DacheuxJ.-L. (2000) Stallion epididymal fluid proteome: qualitative and quantitative characterization; secretion and dynamic changes of major proteins. Biol. Reprod.621790-1803.

Gigon-DepeigesA. & DufaureJ.P. (1977) Secretory activity of the lizard epididymis and its control by testosterone. Gen. Comp. Endocrinol.33473-479.

JonesR.C. (1987) Changes in protein composition of the luminal fluids along the epididymis of the tammar, Macropus eugenii. J. Reprod. Fertil.80193-199.

KoskimiesA.I. & KormanoM. (1975) Proteins in fluids from different segments of the rat epididymis. J. Reprod. Fertil.43345-348.

LaemmliU.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature227680-685.

LichtP. (1984) Reptiles. In: G.E. Lamming (Ed.) Marshall’s Physiology of Reproduction pp. 206-282. Churchill LivingstoneEdinburgh, UK.

LinhardtK. & WalterK. (1963) Phosphatases. In: H.U. Bergmeyer (Ed.) Methods of Enzymatic Analysis pp. 783-785. Academic PressNew York, NY, USA.

LoftsB. & TsuiH.W. (1977) Histological and histochemical changes in the gonads and epididymides of the male Soft-shelled turtle, Trionyx sinensis. J. Zool181(1) 57-68.

LowryO.H.RosebroughN.J.FarrA.L. & RandallR.J. (1951) Protein measurement with the Folin phenol reagent. J. Biol. Chem.193265-275.

MausfeldP.SchmitzA.BöhmeW.MisofB.VrcibradicD. & RochaC.F.D. (2002) Phylogenetic affinities of Mabuya atlantica Schmidt, 1945, endemic to the Atlantic Ocean archipelago of Fernando de Noronha (Brazil): necessity of partitioning the genus Mabuya Fitzinger, 1826 (Scincidae: Lygosominae). Zool. Anz.241281-293.

MediniR.BhagyaM. & SamsonS. (2018) Identification and characterisation of the epididymal proteins in the lizard Eutropis carinata (Reptilia, Squamata) (Schneider, 1801). Gen. Comp. Endocrinol.25976-84.

MesureM.ChevalierM.DepeigesA.FaureJ. & DufaureJ.P. (1991) Structure and ultrastructure of the epididymis of the viviparous lizard during the annual hormonal cycle: changes of the epithelium related to secretory activity. J. Morphol.210133-145.

NirmalB.K. & RaiU. (1997) Epididymal influence on acquisition of sperm motility in the gekkonid lizard, Hemidactylus flaviviridis. Arch. Androl.39105-110.

NirmalB.K. & RaiU. (2000) Epididymal protein secretion and its androgenic control in wall lizards Hemidactylus flaviviridis (Ruppell). Indian J. Exp. Biol.38720-726.

Orgebin-CristM.C.DanzoB.J. & DaviesJ. (1975) Endocrine control of the development and maintenance of sperm fertilizing ability in the epididymis. In: Handbook of Physiology pp. 319-338. American Physiological SocietyWashington DC, USA.

RamagliL.S. (1999) Quantifying protein in 2-D PAGE solubilization buffers. Methods Mol. Biol.11299-103.

RavetV.CourtyY.DepeigesA. & DufaureJ.P. (1987) Changes in epididymal protein synthesis during the sexual cycle of the lizard, Lacerta vivipara. Biol. Reprod.37901-907.

RheubertJ.L.SeverD.M.SiegelD.S. & TrauthS.E. (2015) Male reproductive anatomy: the gonadoduct, sexual segment of kidney and cloaca. In: G.M. Jamieson (Ed.) Reproductive Biology and Phylogeny of Lizards and Tuatara pp. 253-301. CRC PressLondon, UK.

RussellL.D.PetersonR.N.RussellT.A. & HuntW. (1983) Electrophoretic map of boar sperm plasma membrane polypeptides and localization and fractionation of specific polypeptide subclasses. Biol. Reprod.28393-413.

SamsonS.BhagyaM.Chandrashekar sagarB.K. & AranhaI. (submitted) Renal sex segment of the lizard Eutropis carinata: A light microscopic and ultrastructural seasonal study. J. Morphol.

SarkarH.B.D. & ShivanandappaT. (1984) Androgen control of the accessory reproductive organ in the skink, Mabuya carinata (Schn.). In: A. Gopala Krisna & A.K. Saxena (Eds) Recent Trends in Life Science pp. 217-246. Manu PublishersKanpur, India.

SawadaY.TsunoT.UekiH.YamamotoH.FukagawaY.OkiT. & PradimicinQ. (1993) A new pradimicin aglycone, with α-glucosidase inhibitory activity. J. Antibiotics46507-510.

SegrestJ.P. & JacksonR.L. (1972) Molecular weight determination of glycoproteins by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. Methods Enzymol2854-63.

ShivakumarG.R.SarkarH.B.D. & SekharappaB.N. (1979) Histochemical profile of testis and epididymis in the lizard Psammophilus dorsalis (Gray). Indian J. Exp. Biol.17826-830.

ShivakumarG.R. & SarkarH.B.D. (1980) Effect of cyproterone acetate on the testis and epididymis of the lizard Psammophilus dorsalis (Gray). Experientia36616-618.

ShivanandappaT. & SarkarH.B.D. (1979a) Seasonal lipid cycle and steroidogenic activity in the testis of the tropical skink, Mabuya carinata (Schneider): a histochemical study. Gen. Comp. Endocrinol.38491-495.

ShivanandappaT. & SarkarH.B.D. (1979b) Histochemical localization of steroidogenic enzyme in the reptilian epididymis. Curr. Sci.55175-178.

ShivanandappaT. & SarkarH.B.D. (1987) Androgenic regulation of epididymal function in the skink, Mabuya carinata (Schn.). J. Exp. Zool.241369-376.

SinghD.K.JenaN. & MohantyN. (2016) Seasonal variation in nucleic acids, protein and certain enzymes in testis and epididymis of common Indian rock lizard, Psammophilus blanfordanus. The Bioscan112137-2141.

SyntinP.DacheuxF.DruartX.GattiJ.L.OkamuraN. & DacheuxJ.L. (1996) Characterization and identification of proteins secreted in the various regions of the adult boar epididymis. Biol. Reprod.55956-974.

TurnerT.T. (1984) Resorption versus secretion in the rat epididymis. J. Reprod. Fertil.72509-514.

TurnerT.T.RileyT.A.VagnettiM.FlickingerC.J.CaldwellJ.A. & HuntD.F. (2000) Postvasectomy alterations in protein synthesis and secretion in the rat caput epididymidis are not repaired after vasovasostomy. J. Androl.21276-290.

VerderameM. (2014) The involvement of the androgen receptor in the secretion of the epididymal corpus in the lizard Podarcis sicula. Int. J. Zool.20141-6.

VerderameM.AngeliniF. & LimatolaE. (2012) Expression of the estrogen receptor alpha switches off the secretory activity in the epididymal channel of the lizard Podarcis sicula. Mol. Reprod. Dev.79107-117.

VoglmayrJ.K.FairbanksG.JackowitzM.A. & ColellaJ.R. (1980) Post-testicular developmental changes in the ram sperm cell surface and their relationship to luminal fluid proteins of the reproductive tract. Biol. Reprod.22655-667.

WangX.WangD.WangD.WangH.ChangL.YiX.PengM. & GuoA. (2012) Systematic comparison of technical details in CBB methods and development of a sensitive GAP stain for comparative proteomic analysis. Electrophoresis33296-306.

XieS.-W.LiG.-T.QuL.-J.CaoY.WangQ.ZhouJ.-Y.ZhongR.-H.GuoX.-J. & ZhuY. (2016) Identification of new epididymal luminal fluid proteins involved in sperm maturation in infertile rats treated by dutasteride using iTRAQ. Molecules21602. DOI:10.3390/molecules21050602.


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    Weight of the epididymis of the lizard, E. carinata during successive phases of the reproductive cycle.

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    Total protein content of the epididymal luminal fluid in the lizard, E. carinata during successive phases of the reproductive cycle.

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    SDS-PAGE profile of epididymal luminal fluid proteins in the lizard, E. carinata during the breeding, regenerative, regressive and post-breeding phases of the reproductive cycle. Abbreviations (as they appear from left to right on the gel): Sm, Standard marker; Br, breeding phase; Reg, regenerative phase; Regr, regressed phase; Pb, post-breeding phase.

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    Master gel image generated by PDQuest software representing protein spots detected upon analysis of 2D gels of protein derived from luminal fluid from the epididymis in the lizard, E. carinata during the breeding phase. The proteins were fractionated in the mass range of 97.4-14.2 kDa and in the pH range 3-11.

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    PAS-stained SDS-PAGE profile of epididymal luminal fluid proteins in the lizard, E. carinata. Abbreviations (as they appear from left to right on the gel): Sm, standard marker; +ve, positive sample (BSA); Br, breeding phase; Pb, post-breeding phase; Regr, regressed phase; Reg, regenerative phase; −ve, negative sample (trypsin).

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    Activities of the enzymes acid phosphatase, alkaline phosphatase and alpha glucosidase in the lumen of the epididymis in the lizard, E. carinata during successive phases of the reproductive cycle.


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