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Abstract: In addition to its many functions, the placenta is characterized by the intensity and characteristics of its hormonal functions. It has been reported that the development and vascularization of the placenta with normal fetal growth is regulated by many factors such as growth factors, transcriptional factors, and homeobox proteins. This study was conducted to determine the distribution and possible physiological roles of HOXA11 and HLX in the uterus and placenta of pregnant cows. In this study, 27 pregnant Holstein breed cow uteri and placentas obtained from private slaughterhouses were used as tissue samples. In order to determine the pregnancy period of the obtained uterus, the ages of the fetuses were calculated according to the age calculation formula. The uteri were classified under three different groups as the first (69-89 days), second (99-178 days), and third (190-269 days) periods of pregnancy so that there are 9 animals in each group. Tissue samples were then subjected to immunohistochemistry to demonstrate the presence of HOXA11 and HLX proteins. It was observed that HOXA11 and HLX immunoreactions occurred prominently in the luminal and glandular epithelial cells of the uterus during pregnancy, and the maternal epithelial cells of the placenta and UTC and TGC/BNCs. In addition, it was determined that HOXA11 immunoreaction also formed in some vascular endothelial cells. Immunohistochemical findings in this study suggested that Hoxa11 and HLX may affect the proliferation and differentiation of cells in the uterus and placenta during pregnancy in cows, and that these proteins may contribute to normal uterine physiology, placentation formation, and continuity of pregnancy in cows. Key words: Hox proteins; HOXA11; HLX; placenta IMUNOLOKALIZACIJA PROTEINOV HOXA11 IN HLX V POSTELJICI KRAVE MED BREJOSTJO Izvleček: Za posteljico so poleg številnih funkcij značilne intenzivnost in karakteristike njenih hormonskih funkcij. Poročali so,da razvoj in ožiljenost posteljice pri normalni rasti ploda uravnavajo številni dejavniki, kot so rastni dejavniki, transkripcijski djavniki in homeoboks proteini. Ta študija je bila izvedena z namenom določiti porazdelitev in morebitno fiziološko vlogo protnov HOXA11 in HLX v maternici in posteljici brejih krav. Za vzorce tkiv smo uporabili maternice in posteljice 27 krav pasem Holstein, pridobljenih v zasebnih klavnicah. Obdobje brejosti smo izračunali na podlagi starosti plodov po formuli. Maternice smoglede na obdobje brejosti uvrstili v tri skupine po 9 vzorcev: 1 (69-89 dni), 2 (99-178 dni) in 3 (190-269 dni). Prisotnost proteinoHOXA11 in HLX v vzorcih tkiv smo preverili z imunohistokemijo. Proteini HOXA11 in HLX so bili prisotni predvsem v luminalnihin žleznih epitelijskih celicah maternice ter v epitelijskih celicah maternalne posteljice, UTC in TGC/BNC. Prisotnost HOXA1je bila zaznavna tudi v nekaterih žilnih endotelijskih celicah. Rezultati imunohistokemije v tej študiji kažejo na morebiten vplivproteinov HOXA11 in HLX na proliferacijo in diferenciacijo celic v maternici in posteljici krav med brejostjo in na morebitno vlogoteh proteinov pri normalni fiziologiji maternice, nastanku posteljice in vztrajanju brejosti pri kravah. Ključne besede: proteini Hox; HOXA11; HLX; posteljica

It is known that ghrelin, obestatin and leptin are involved in many biological processes such as appetite-energy regulation, growth hormone release and cell proliferation. In addition to these biological activities of in human and rodents, direct effects of these hormones on reproductive functions and regulation of smooth muscle contractions have gained importance in recent years. In mammals, gonadal functions are regulated by using a complex network of autocrine, paracrine and endocrine signals. The signals involved in the control of energy balance regulate reproductive functions by acting on different hypothalamic pituitary-gonadal axis. The underlying molecular mechanism of gonad is poorly understood and appears to be controlled on genetic, environmental, and hormonal factors. Ghrelin, obestatin and leptin, three recently discovered hormones, are especially coexpressed in endocrine cells. The purpose of this investigation was to examine the immunohistochemical expression and potential biological activity of bull and ram deferent ducts (DD) in relation to the tissue concentration of ghrelin, obestatin and leptin. Ghrelin, obestatin and leptin expression observed in the DD sections were restricted to the cytoplasm of the epithelial and muscle cells. Both ghrelin and leptin expressions were found in smooth muscle cells in muscular layer of DD and smooth muscle cells of blood vessels, and obestatin expression was found in the basal epithelial cells of the luminal epithelium. In conclusion, ghrelin, obestatin and leptin were expressed in varying concentrations in epithelial and muscle cells as well as blood vessels of the deferent ducts of both species. Given the multifunctional biological roles of these peptide hormones, we can suggest that these hormones function in the contraction and hypertrophy of smooth muscles, proliferation or apoptosis of epithelial and muscle cells in DD. Key words: deferent ducts; ghrelin; leptin; obestatin; ruminant; physiological function IZRAŽANJE IN BIOLOŠKA AKTIVNOST GRELINA, OBESTATINA IN LEPTINA V SEMENOVODU PRI BIKU IN OVNU Povzetek: Znano je, da so grelin, obestatin in leptin vključeni v številne biološke procese, kot so uravnavanje apetita, sproščanje rastnega hormona in spodbujanje delitev celic. Poleg omenjenih bioloških učinkov pri ljudeh in glodavcih so v zadnjih letih postali znani tudi njihovi neposredni učinki na delovanje spolnega sistema in uravnavanje kontrakcije gladkih mišic. Pri sesalcih je delovanje spolnih žlez uravnano z zapleteno mrežo avtokrinih, parakrinih in endokrinih sporočil. Sporočila, ki sodelujejo pri nadzoru energetskega ravnotežja v telesu pogosto vplivajo tudi na delovanje spolnega sistema z vplivom na hipotalamusno-hipofizno-spolno os. Osnovni molekularni mehanizem delovanja spolnih žlez je slabo raziskan, na njegovo delovanje pa vplivajo genetski, okoljski in hormonski dejavniki. Grelin, obestatin in leptin so trije hormoni, ki v telesu sodelujejo pri urejanju in ohranjanju energetskega ravnotežja. Namen raziskave je bil preučiti imunohistokemično izražanje in potencialno biološko aktivnost grelina, obestatina in leptina v semenovodu bikov in ovnov. Grelin, obestatin in leptin so izraženi v semenovodu in so bili omejeni na citoplazmo epitelijskih in mišičnih celic. Grelin in leptin sta bila izražena v gladkih mišičnih celicah mišične plasti semenovoda in gladkih mišičnih celicah krvnih žil. Obestatin je bil izražen v bazalnih epitelijskih celicah svetline semenovoda. Grelin, obestatin in leptin so torej izraženi v različnih koncentracijah v epitelnih in mišičnih celicah ter krvnih žilah v semenovodu pri biku in ovnu. Glede na raznolike biološke vloge teh peptidnih hormonov lahko predvidevamo, da ti hormoni morda sodelujejo pri krčenju in hipertrofiji gladkih mišic ter pri množenju ali apoptozi epitelijskih in mišičnih celic v semenovodu. Ključne besede: semenovod; grelin; leptin; obestatin; bik; oven; fiziološka funkcija