Shegedin A. Yu., Yashchenko A. M.

EXPOSURE OF GLYCOCONJUGATES IN POSTNATAL MORPHOGENESIS OF RAT TESTIS AS REVEALED BY PAS REACTION AND LECTIN HISTOCHEMISTRY

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About the author:	Shegedin A. Yu., Yashchenko A. M.
Heading	EXPERIMENTAL MEDICINE
Type of article	Scentific article
Annotation	The combination of carbohydrate domains of nuclear and cell membranes, glycosylated cytoplasmic and nuclear structures and extracellular matrix components are coined by the term of glycome. Rapid glycome restructuring occurs during embryonic development and postnatal morphogenesis. A large number of publications showed that extensive restructuring of carbohydrate determinants occurs during spermatogenesis, but we couldn’t find any published work that regarded to changes of rat testis’ glycome throughout successive stages of postnatal morphogenesis. By using lectin histochemistry and PAS reaction it is possible to study the changes of carbohydrate determinants of glycopolymers during postnatal development of testis. 15 females of Wistar rats weighting 180-200g each were used in current study, whose offspring at 20th prenatal day of development, 1st, 20th and 40th days of postnatal developments were analyzed. Animals were killed by narcosis overdose and testis of males were fixed in in Bouin’s solution. General morphology paraffin sections of 5-7 µm were stained with H&E. Tissue carbohydrates were revealed by PAS reaction with subsequent counterstaining with hematoxylin. Carbohydrate determinants of biopolymers were studied using a set of lectins (Con A, PSA, GNA, PNA, VAA, CNFA). We have detected that morphogenic processes are accompanied by intense rearrangement of glycoconjugates, in particular, lectin binding carbohydrates within structural components of the testis. Mannose-specific lectins can be recommended as a selective histochemical markers of fetal Leydig cells, galactose-specific lectins – for specific labeling of acrosomes of rat sperm cells. Postnatal testicular morphogenesis is accompanied by rapid remodeling of glycoconjugates specific to studied lectin. Mannose specific lectins PSA, GNA, and Con A served as selective histochemical markers of Leydig cells, galactose specific lectins PNA, VAA and CNFA - as markers of acrosomal systems of rat sperm cells. Glycoconjugates recognized by both lectin groups were expressed by leptotene and zygotene spermatocytes starting from 20th day of postnatal development. Broadening of the spectrum of studied carbohydrate determinants by means of lectins specific to terminal N-Acetyl-D-glucosamine, N-Acetyl-D-galactosamine, L-fucose and sialic acids. Investigation of the influence of experimental pathology (altered balance of maternal thyroid hormones, in particular) on testicular glycoconjugates in offsprings.
Tags	rat testis, postnatal morphogenesis, lectin histochemistry, glycome
Bibliography	Voloshin N.A. Lektinyi zhivotnogo i rastitelnogo proishozhdeniya: rol v protsesah morfogeneza / N. A. Voloshin, E. A. Grigoreva // Zhurnal AMN Ukrainyi. – 2005. – T.11, No. 2. – S. 223-237. Tabachniuk N. V. Lektynohistokhimichni doslidzhennia ta embriohenez / N. V. Tabachniuk, I. Yu. Oliinyk, L. P. Lavriv// Klinichna anatomiia ta operatyvna khirurhiia. – 2010. – T.9, No. 3. – S. 95-100. Lutsik A. D. Lektinyi v gistohimii/ A. D. Lutsik, E. S. Detyuk, M. D. Lutsik // Lvov: Vyischa shkola, - 1989. Antoniuk V. O. Lektyny ta yikh syrovynni dzherela / V. O. Antoniuk // Lviv: Kvart, - 2005. Topolenko T. A. Rozpodil hlikoproteiniv v yaiechkakh shchuriv pislia vvedennia zhinochykh statevykh hormoniv u druhomu ta tretomu periodakh vahitnosti / T. A. Topolenko // Visnyk problem biolohii i medytsyny. – 2011. – T.2, Vyp.2. – S.267-269. Synenko M. M. Fukozo- i manozohlikany u strukturnykh komponentakh yaiechka shchuriv na tli eksperymentalnoho hipotyreozu / M. M. Synenko, Y. A. Nakonechnyi, O. V. Smolkova [ta in.] // Visnyk morfolohii. – 2010. – T.16, No.2. – S.416-419. Varyvoda O. Yu. Nokaut hena pttg u myshei suprovodzhuietsia pidvyshchenniam rivnia eksponuvannia vuhlevodnykh determinant DGalNAc / O. Yu. Varyvoda, Ye. Z. Filiak, O. D. Lutsyk [ta in.] // Biopolymers and Cell. – 2012. – V.28, No.2. – S.129-133. Yashchenko A. M. Hipotetychna rol retseptoriv lektyniv yak markeriv apoptozu v klitynakh simianyka pry tyroidnii patolohii / A. M. Yashchenko, B.R.Stoika, O.V.Smolkova [ta in.] // Tavrycheskyi medyko-byolohycheskyi vestnyk. – 2003. – T.6, No.3. – S.153-155. Dan X. Development and applications of lectins as biological tools in biomedical research / X. Dan, W. Liu // Med.Res.Rev. – 2015. – Vol.36. – P.221-247. Gabius H.J. The sugar code. Fundamentals of glycoscience/ H.J.Gabius [et al.] // Weinheim: Wiley, - 2009. Gallo A. Glycobiology of reproductive processes in marine animals/ A.Gallo, M.Costantini// Mar. Drugs. – 2012. – V.10, No.12. – P. 2861-2892. Krinke G. J. The laboratory rat / G. J. Krinke [et al.] // Amsterdam: Elsevier-Academic Press, - 2000. Lutsyk A. Structural, functional, and lectin histochemical characteristics of rat ovaries and endometrium in experimental hyper- and hypothyroidism / A. Lutsyk, E. Sogomonian // Folia Histochem. Cytobiol. – 2012. – Vol.50, No.3. – P.331-339. Parker G. A. Atlas of histology of the juvenile rat / G.A. Parker, C. A. Picut [et al.] // Amsterdam: Elsevier-Academic Press, - 2016. Roth J. Lectins for histochemical demonstration of glycans / J. Roth // Histochem.Cell Biol. – 2011. – Vol.136. – P.117-130. Sharon N. Lectins: carbohydrate-specific reagents and biological recognition molecules / N. Sharon // J.Biol.Chem. – 2007. – Vol.282. – P.2753-2764. Seco-Rovira V. Identification of apoptotic cells by means of lectin histochemistry / V. Seco-Rovira, E. Beltran-Frutos, C. Ferrer [et al.] // J. Cytol.Histol. – 2015. – Vol.6, No.2. – 309 p. Varki A. Essentials of glycobiology / A. Varki, R. D. Cummings, J. D. Esko [et al.] // Cold Spring Harbor: Cold Spring Harbor Laboratory Press, - 2009.
Publication of the article	«World of Medicine and Biology» №2(60), 2017 year, 174-178 pages, index UDK 611.631-019-018:547547.96]-08