ISSN 2415-3060 (print), ISSN 2522-4972 (online)
  • 43 of 50
Up
УЖМБС 2021, 6(1): 303–310
https://doi.org/10.26693/jmbs06.01.303
Biology

An Original Method for Removing and Fixing the Pituitary Gland of Sexually Mature Rats for Creating High-Quality Permanent Histological Preparations

Hryntsova N. B., Romanyuk A. M.
Abstract

The issue of high-quality creating histological preparations is an urgent issue of histology and pathological anatomy, experimental medicine and biology. The pituitary gland belongs to the smallest in mass and size of the endocrine glands in both humans and animals, it is an endocrine gland and occupies one of the central places in the endocrine regulation of the body's vital activity. The study of the features of the removal of the pituitary gland from Turkish saddle of the sphenoid bone of the skull of rats remains an important aspect of modern morphology. The purpose of the work was to develop an original technique for improving the first (removal) and second (material fixation) stages of creating histological preparations of the rat pituitary gland for morphological, morphometric, and immunohistochemical medico-biological experimental studies. Material and methods. The development of the methodology was carried out on 200 white sexually mature rats of different sex weighing 250-300 g at the age of 7-8 months in accordance with the National and European bioethical standards. Results and discussion. Modification of the technique of extracting the pituitary gland of rats includes the introduction of the animal into thiopental anesthesia, decapitation, separation of the skin and muscular integument of the head, resection of the occipital bone of the skull, exposure and removal of the brain, identification of the pituitary gland, followed by removal of the pituitary gland with a single complex (block) together with the pituitary fossa Turkish saddle and fragments of the sphenoid bone adjacent to it. The gland was fixed with a reduced concentration of a 5% solution of buffered neutral formalin. After 15-18 hours from the beginning of fixation, the complex was temporarily removed from the fixator, and the pituitary gland was removed from the pituitary fossa of Turkish saddle using eye scalpel and eye forceps. The organ had a well-fixed and compacted capsule, which prevented unnecessary trauma and fragmentation of the organ during removal. The removed pituitary gland was again immersed in a 5% solution of neutral buffered formalin for 2-3 hours. After the end of the fixation period, the pituitary gland had a well-fixed structure and was subject to the following standard stages of making permanent histological specimens

Keywords: pituitary gland, formalin, pituitary fossa, dissection

Full text: PDF (Ukr) 353K

References
  1. Adenomy gipofiza: klinika, diagnostyka, likuvannya [Pituitary adenomas: clinic, diagnosis, treatment]. Ed by BA Kadashev. M: OAO «Yzdatelstvo «Medytsyna»; 2007. 368 s. [Russian]
  2. Lutsik OD, Ivanova AY, Kabak KS, Chaykovskiy YuB. Gistologiya lyudini [Human histology]. K: Kniga plyus; 2003. 592 s. [Ukrainian]
  3. Zapadnyuk YP, Zapadnyuk VY, Zakharyya EA, Zapadnyu EA. Laboratornye zhyvotnye. Razvedenye, soderzhanye, yspolzovanye v eksperymente [Laboratory animals. Breeding, maintenance, use in the experiment]. K: Vyshcha shkola; 1983. 382s. [Russian]
  4. 3erbino DD, Bagriy MM, Bodnar YaYa. Patomorfologiya ta gistologiya (atlas) [Pathomorphology and histology (atlas)]. Vinnitsya: Nova Kniga; 2016. 799 s. [Ukrainian]
  5. Pykalyuk VS, Bessalova EYu. Vozmozhnosty makro-mykroanatomycheskykh metodov v yssledovanyy gypofyzov belykh krys [Possibilities of macro-micro-anatomical methods in the study of the pituitary glands of white rats]. Ukrayinskyy morfologichnyy almanakh. 2011; 3(9): 200-202. [Ukrainian]
  6. Chumachenko OYu, Redka OG. Anatomiya gipofiziv shchuriv u vikovomu aspekti v ekologichno nespriyatlivikh umovakh [Anatomy of rat pituitary glands in the age aspect in ecologically unfavorable conditions]. Pytannya bioindikatsiyi ta ekologiyi. 2015; 1(20): 187-193. [Ukrainian]
  7. Nozdrachev AD, Polyakov EL. Anatomyya krysy (Laboratornye zhyvotnye) [Anatomy of a Rat (Laboratory Animals]. Pod red akademyka AD Nozdracheva. SPb: Izdatelstvo «Lan»; 2001. 464 s. [Russian]
  8. Bobrysheva IV. Osobennosti stroeniya atsidofilnykh endokrinotsitov adenogipofiza krys posle primeneniya tsiklofosfamida [Features of the structure of acidophilic endocrinocytes of the adenohypophysis of rats after the use of cyclophosphamide]. Vestnik SAFU. Seriya «Mediko-biologicheskie nauki». 2016; 3: 59-65. [Russian] https://doi.org/10.17238/issn2308-3174.2016.3.59
  9. Kapytonova MYu, Ulla M, Kuznetsov SL, Khlebnykov VV, Nor-Ashykyn MNK, Akhmad A. Vozrastnaya morfofunktsyonalnaya kharakterystyka follykulo-zvezdchatykh kletok gypofyza krys pry stresse [Age morphofunctional characteristics of follicle-stellate cells of the pituitary gland of rats under stress]. Vestnyk RAMN. 2013; 11: 98–102. [Russian] https://doi.org/10.15690/vramn.v68i11.850
  10. Kashirina NK. Metodika identifikatsiyi ta vidilennya organiv endokrinnoyi sekretsiyi u mishey [Methods of identification and selection of endocrine secretion organs in mice]. Byuleten eksperimentalnoyi biologiyi i meditsini.1987; 5: 630-631. [Russian]
  11. Patent 81424 Ukraine, MPK A61B 17/00. Sposib preparuvannya gipofiza u shchuriv [The method of preparation of the pituitary gland in rats] / Kashchenko SA, Bobrysheva IV, Tatarenko DP, Bobrysheva AO. (UA); zayavnike i vlasniky patentu Kashchenko SA, Bobrysheva IV, Tatarenko DP, Bobrysheva AO. (UA). № u201301459; zayavl 07.02.13; opubl 25.06.13. Byul № 12. [Ukrainian]
  12. Patent 88650 Ukraine, MPK A61B 17/00. Modifikovaniy sposib preparuvannya gipofiza u bilikh laboratornikh shchuriv [Modified method of pituitary preparation in white laboratory rats] / Smirnov SM, Dubova GA, Dubova YuM, Tatarenko DP. (UA); zayavnike i vlasniky patentu Smirnov SM, Dubova GA, Dubova YuM, Tatarenko DP. (UA). № u201312254; zayavl 21.10.13; opubl 25.03.14. Byul № 6. [Ukrainian]
  13. Kalinin PL, Fomichov DV, Kadashev BA. Metodyka endoskopichnoyi endonazalnoyi transsfenoyidalnoyi adenomektomiyi [Methods of endoscopic endonasal transsphenoidal adenomectomy]. Pytannya neyrokhirurgiyi. 2007; 4: 42-45. [Russian]
  14. Kalinin PL, Kadashev BA, Fomichov DV, Kutin MA, Astaf'yeva LI. Khirurgichne likuvannya adenom gipofiza [Surgical treatment of pituitary adenomas]. Klinichni rekomendatsiyi. M; 2014. 3 s. [Russian]
  15. Hryntsova NB, Khomenko IV, Romanyuk AM, Bumeister VI, Kravtsova IA. Morphological and morphometric rearrangements of the rat adenohypophysial - thyroid system under the experimental extracellular dehydration. The world of medicine and biology. 2019; 2(68): 178-183. [Ukrainian] https://doi.org/10.26724/2079-8334-2019-2-68-174-179
  16. Kornyeykova IP. Korektsiya strukturnykh zmin gipofiza tvaryn riznogo viku, yaki vynykly za umov gipoosmolyarnoyi gipergidriyi [Correction of structural changes of the pituitary gland of animals of different ages, which arose under conditions of hypoosmolar hyperhydria]. Ukrayinskyy morfologichnyy almanakh. 2012; 4(15): 95-97. [Ukrainian]
  17. Bessalova YeYu. Morfologichni zmini organiv neyroendokrinnoyi sistemi samok ssavtsiv pri parenteralnomu vvedenni ksenogennoyi spinnomozkovoyi ridini [Morphological changes of organs in the neuroendocrine system and female savts with parenteral introduction of xenogenous spinal cord]. Naukoviy visnik Uzhgorodskogo universitetu. Seriya «Meditsina».2008; 33: 10-13. [Ukrainian]
  18. Fomyna KA. Organometrycheskye pokazately gypofyza posle dvukhmesyachnogo vozdeystvyya spyrtovoy nastoyky ekhynatsey [Organometric parameters of the pituitary gland after two months of exposure to alcoholic tincture of Echinacea] Visnyk morfologiyi. 2010; 2(16): 323-326. [Ukrainian]
  19. Elizarova ON, Zhidkova LV, Kochetkova TA. Posobie po toksikologii dlya laborantov [A guide to toxicology for laboratory assistants]. M: Meditsina; 1974. 157 s. [Russian]
  20. Mardar GI, Savchuk GG, Kavare VI, Kiptenko LI, Trybovska SV. Reaktsiya krovi, adenogipofiza ta kory nadnyrnykiv bilykh shchuriv na ionizuyuche oprominennya i preparat Ersol [The reaction of blood, adenohypophysis and adrenal cortex of white rats to ionizing radiation and the drug Ersol]. Naukovyy visnyk Chernivetskogo universytetu. Seriya «Biologiya». 2000; 77: 210-218. [Ukrainian]
  21. Sapozhnikov AG, Dorosevich AE. Gistologicheskaya i mikroskopicheskaya tekhnika. Rukovodstvo [Histological and microscopic techniques.] Smolensk: "SAU"; 2000; 475 s. [Russian]
  22. Afanasev YuY, Blanchuk VK, Vannykov LL, Donskykh NV. Osnovy gystologyy y gystologycheskoy tekhnyky [Fundamentals of Histology and Histological Technique]. Pod obshch red VG Elyseeva. M: Medytsyna; 1967. 267 s. [Russian]
  23. Kumar GL, Rudbeck L. Immunogistokhimicheskie metody: Rukovodstvo [Immunohistochemical Methods: Manual]. M; 2011. 224 s. [Russian]
  24. Patent 79002 Ukraine, MPK A61B 17/00. Preparuvalnyy stolyk dlya fiksatsiyi laboratornykh tvaryn [Dissection table for fixing laboratory twins] / Kashchenko SA, Tatarenko DP, Ivanov OS. (UA); zayavniky i vlasniky patentu Kashchenko SA, Tatarenko DP, Ivanov OS. (UA). № u201210848; zayavl 17.09.12; opubl 10.04.13. Byul № 7. [Ukrainian]
  25. Bagriy MM, Dibrova VA. Metodiki morfologichnikh doslidzhen [Methods of morphological research]. Vinnitsya: Nova Kniga; 2016. 295 s. [Ukrainian]
  26. Koveshnykov VG Tkachenko OYa, Shevchenko YV. K usovershenstvovanyyu metodyky gystologycheskoy obrabotky melkykh ob’ektov [To improve the technique of histological processing of small objects]. Ukrayinskyy medychnyy almanakh. 2001; 5(4): 62-64. [Ukrainian]