Abstract
The pituitary is involved in the regulation of endocrine homeostasis. Therefore, animal models of pituitary disease based on a thorough knowledge of pituitary anatomy are of great importance. Accordingly, we aimed to perform a qualitative and quantitative description of polypeptide hormone secreting cellular components of the Göttingen minipig adenohypophysis using immunohistochemistry and stereology. Estimates of the total number of cells immune-stained for adrenocorticotrophic hormone (ACTH), prolactin (PRL), and growth hormone (GH) were obtained with the optical fractionator technique using Stereo Investigator software. Moreover, 3D reconstructions of cell distribution were made. We estimated that the normal minipig adenohypophysis contains, on average, 5.6 million GH, 3.5 million PRL, and 2.4 million ACTH producing cells. The ACTH producing cells were widely distributed, while the PRL and GH producing cells were located in clusters in the central and lateral regions of the adenohypophysis. The morphology of the hormone producing cells also differs. We visualized a clear difference in the numerical density of hormone producing cells throughout the adenohypophysis. The relative proportions of the cells analyzed in our experiment are comparable to those observed in humans, primates, and rodents; however, the distribution of cells differs among species. The distribution of GH cells in the minipig is similar to that in humans, while the PRL and ACTH cell distributions differ. The volume of the pituitary is slightly smaller than that of humans. These data provide a framework for future large animal experimentation on pituitary function in health and disease.
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References
Amar AP, Weiss MH (2003) Pituitary anatomy and physiology. Neurosurg Clin N Am 14:11–23. https://doi.org/10.1016/s1042-3680(02)00017-7
Arafah BM (1986) Reversible hypopituitarism in patients with large nonfunctioning pituitary adenomas. J Clin Endocrinol Metab 62:1173–1179. https://doi.org/10.1210/jcem-62-6-1173
Arafah BM, Kailani SH, Nekl KE, Gold RS, Selman WR (1994) Immediate recovery of pituitary function after transsphenoidal resection of pituitary macroadenomas. J Clin Endocrinol Metab 79:348–354. https://doi.org/10.1210/jcem.79.2.8045946
Asa SL, Penz G, Kovacs K, Ezrin C (1982) Prolactin cells in the human pituitary—a quantitative immuno-cytochemical analysis. Arch Pathol Lab Med 106:360–363
Bargmann W (1949) The neurosecretory connection between the hypothalamus and the neurohypophysis. Z Zellforsch Mikrosk Anat 34:610–634
Bargmann W, Hild W, Ortmann R, Schiebler TH (1950) Morphologic and experimental studies of the hypothalamohypophyseal system. Acta Neuroveg (wien) 1:233–275
Bjarkam CR et al (2004) A MRI-compatible stereotaxic localizer box enables high-precision stereotaxic procedures in pigs. J Neurosci Methods 139:293–298. https://doi.org/10.1016/j.jneumeth.2004.05.004
Bjarkam CR, Glud AN, Orlowski D, Sorensen JC, Palomero-Gallagher N (2016) The telencephalon of the Gottingen minipig, cytoarchitecture and cortical surface anatomy. Brain Struct Funct. https://doi.org/10.1007/s00429-016-1327-5
Bjarkam CR, Glud AN, Orlowski D, Sorensen JCH, Palomero-Gallagher N (2017a) The telencephalon of the Gottingen minipig, cytoarchitecture and cortical surface anatomy. Brain Struct Funct 222:2093–2114. https://doi.org/10.1007/s00429-016-1327-5
Bjarkam CR, Orlowski D, Tvilling L, Bech J, Glud AN, Sørensen JCH (2017b) Exposure of the pig CNS for histological analysis: a manual for decapitation, skull opening and brain removal. J Vis Exp. https://doi.org/10.3791/55511
Bonthius DJ, McKim R, Koele L, Harb H, Karacay B, Mahoney J, Pantazis NJ (2004) Use of frozen sections to determine neuronal number in the murine hippocampus and neocortex using the optical disector and optical fractionator. Brain Res Brain Res Protoc 14:45–57. https://doi.org/10.1016/j.brainresprot.2004.09.003
Cao D, Ma X, Zhang WJ, Xie Z (2017) Dissection and coronal slice preparation of developing mouse pituitary gland. J vis Exp. https://doi.org/10.3791/56356
Catt KJ (1970) Pituitary function. Lancet 1:827–831. https://doi.org/10.1016/s0140-6736(70)92423-2
Christensen AB, Sorensen JCH, Ettrup KS, Orlowski D, Bjarkam CR (2018) Pirouetting pigs: a large non-primate animal model based on unilateral 6-hydroxydopamine lesioning of the nigrostriatal pathway. Brain Res Bull 139:167–173. https://doi.org/10.1016/j.brainresbull.2018.02.010
Console GM, Gomez Dumm CL, Brown OA, Ferese C, Goya RG (1997) Sexual dimorphism in the age changes of the pituitary lactotrophs in rats. Mech Ageing Dev 95:157–166. https://doi.org/10.1016/s0047-6374(97)01878-2
Console GM, Jurado SB, Oyhenart E, Ferese C, Pucciarelli H, Dumm CLAG (2001) Morphometric and ultrastructural analysis of different pituitary cell populations in undernourished monkeys. Braz J Med Biol Res 34:65–74. https://doi.org/10.1590/S0100-879x2001000100008
Cruz-Orive LM, Weibel ER (1990) Recent stereological methods for cell biology: a brief survey. Am J Physiol 258:L148-156
Cushing H (1912) The pituitary body and its disorders, clinical states produced by disorders of the hypophysis cerebri. vol x. J.B. Lippincott Company, Philadelphia
Dacheux F (1980) Ultrastructural immunocytochemical localization of prolactin and growth hormone in the porcine pituitary. Cell Tissue Res 207:277–286. https://doi.org/10.1007/BF00237812
Dacheux F (1981) Ultrastructural localization of corticotropin, beta-lipotropin, and alpha- and beta-endorphin in the porcine anterior pituitary. Cell Tissue Res 215:87–101. https://doi.org/10.1007/BF00236251
Dacheux F (1984) Differentiation of cells producing polypeptide hormones (ACTH, MSH, LPH, alpha- and beta-endorphin, GH and PRL) in the fetal porcine anterior pituitary. Cell Tissue Res 235:615–621. https://doi.org/10.1007/BF00226960
Dacheux F, Dubois MP (1976) Ultrastructural localization of prolactin, growth hormone and luteinizing hormone by immunocytochemical techniques in the bovine pituitary. Cell Tissue Res 174:245–260. https://doi.org/10.1007/BF00222162
Dada MO, Campbell GT, Blake CA (1984) Pars distalis cell quantification in normal adult male and female rats. J Endocrinol 101:87–94
de Lima AR et al (2007) Muscular dystrophy-related quantitative and chemical changes in adenohypophysis GH-cells in golden retrievers. Growth Horm IGF Res 17:480–491. https://doi.org/10.1016/j.ghir.2007.06.001
Deniz OG, Altun G, Kaplan AA, Yurt KK, von Bartheld CS, Kaplan S (2018) A concise review of optical, physical and isotropic fractionator techniques in neuroscience studies, including recent developments. J Neurosci Methods 310:45–53. https://doi.org/10.1016/j.jneumeth.2018.07.012
Ettrup KS et al (2011) Basic surgical techniques in the Gottingen minipig: intubation, bladder catheterization, femoral vessel catheterization, and transcardial perfusion. J vis Exp. https://doi.org/10.3791/2652
Filippa V, Mohamed F (2006b) Immunohistochemical study of somatotrophs in pituitary pars distalis of male viscacha (Lagostomus maximus maximus) in relation to the gonadal activity. Cells Tissues Organs 184:188–197. https://doi.org/10.1159/000099626
Filippa V, Mohamed F (2006a) ACTH cells of pituitary pars distalis of viscacha (Lagostomus maximus maximus): immunohistochemical study in relation to season, sex, and growth. Gen Comp Endocrinol 146:217–225. https://doi.org/10.1016/j.ygcen.2005.11.012
Filippa V, Mohamed F (2010) Morphological and morphometric changes of pituitary lactotrophs of viscacha (Lagostomus maximus maximus) in relation to reproductive cycle, age, and sex. Anat Rec (hoboken) 293:150–161. https://doi.org/10.1002/ar.21013
Francis SM, Venters SJ, Duxson MJ, Suttie JM (2000) Differences in pituitary cell number but not cell type between genetically lean and fat coopworth sheep. Domest Anim Endocrinol 18:229–239. https://doi.org/10.1016/s0739-7240(99)00081-8
Garcia-Navarro S, Malagon MM, Gracia-Navarro F (1988) Immunohistochemical localization of thyrotropic cells during amphibian morphogenesis: a stereological study. Gen Comp Endocrinol 71:116–123. https://doi.org/10.1016/0016-6480(88)90302-4
Glud AN et al (2011) Direct MRI-guided stereotaxic viral mediated gene transfer of alpha-synuclein in the Gottingen minipig CNS. Acta Neurobiol Exp (wars) 71:508–518
Glud AN, Bjarkam CR, Azimi N, Johe K, Sorensen JC, Cunningham M (2016) Feasibility of three-dimensional placement of human therapeutic stem cells using the intracerebral microinjection instrument. Neuromodul J Int Neuromodul Soc 19:708–716. https://doi.org/10.1111/ner.12484
Goodman S, Check E (2002) The great primate debate. Nature 417:684–687. https://doi.org/10.1038/417684a
Gundersen HJ (1986) Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William R. Thompson. J Microsc 143:3–45
Gundersen HJ, Jensen EB (1987) The efficiency of systematic sampling in stereology and its prediction. J Microsc 147:229–263
Gundersen HJ et al (1988) The new stereological tools: disector, fractionator, nucleator and point sampled intercepts and their use in pathological research and diagnosis. APMIS 96:857–881
Harris GW (1948) Neural control of the pituitary gland. Physiol Rev 28:139–179. https://doi.org/10.1152/physrev.1948.28.2.139
Heidelbaugh JJ (2016) Endocrinology update: hypopituitarism. FP Essentials 451:25–30
Heiman J (1938) The anterior pituitary gland in tumor-bearing rats. Am J Cancer 33:423–442. https://doi.org/10.1158/ajc.1938.423
Heiman ML, Surface PL, Mowrey DH, DiMarchi RD (1990) Sexual dimorphic porcine pituitary response to growth hormone-releasing hormone. Domest Anim Endocrinol 7:273–276. https://doi.org/10.1016/0739-7240(90)90033-v
Hofmann I et al (2020) Linkage between growth retardation and pituitary cell morphology in a dystrophin-deficient pig model of Duchenne muscular dystrophy. Growth Horm IGF Res 51:6–16. https://doi.org/10.1016/j.ghir.2019.12.006
Hong GK, Payne SC, Jane JA Jr (2016) Anatomy, physiology, and laboratory evaluation of the pituitary gland. Otolaryngol Clin N Am 49:21–32. https://doi.org/10.1016/j.otc.2015.09.002
Jensen KN, Deding D, Sorensen JC, Bjarkam CR (2009) Long-term implantation of deep brain stimulation electrodes in the pontine micturition centre of the Gottingen minipig. Acta Neurochir 151:785–794. https://doi.org/10.1007/s00701-009-0334-1
Jurado S, Console G, Gomez Dumm C (1998) Sexually dimorphic effects of aging on rat somatotroph cells. An immunohistochemical and ultrastructural study. J Vet Med Sci 60:705–711. https://doi.org/10.1292/jvms.60.705
Kasper RS, Shved N, Takahashi A, Reinecke M, Eppler E (2006) A systematic immunohistochemical survey of the distribution patterns of GH, prolactin, somatolactin, beta-TSH, beta-FSH, beta-LH, ACTH, and Alpha-MSH in the Adenohypophysis of Oreochromis Niloticus, the Nile Tilapia. Cell Tissue Res 325:303–313. https://doi.org/10.1007/s00441-005-0119-7
Kiki I, Altunkaynak BZ, Altunkaynak ME, Vuraler O, Unal D, Kaplan S (2007) Effect of high fat diet on the volume of liver and quantitative feature of Kupffer cells in the female rat: a stereological and ultrastructural study. Obes Surg 17:1381–1388. https://doi.org/10.1007/s11695-007-9219-7
Koeppen BM, Stanton BA, Levy MN, Berne RM (2018) Berne & Levy physiology
Kovacs K, Horvath E, Asa SL, Stefaneanu L, Sano T (1989) Pituitary cells producing more than one hormone human pituitary adenomas. Trends Endocrinol Metab 1:104–107. https://doi.org/10.1016/1043-2760(89)90012-x
Kuwahara S, Sari DK, Tsukamoto Y, Tanaka S, Sasaki F (2004) Age-related changes in growth hormone (GH) cells in the pituitary gland of male mice are mediated by GH-releasing hormone but not by somatostatin in the hypothalamus. Brain Res 998:164–173. https://doi.org/10.1016/j.brainres.2003.10.060
Lee J-S (2006) Comparative study of immunocytochemical patterns of somatotrophs, mammotrophs, and mammosomatotrophs in the porcine anterior pituitary vol 1875. Digital Repository @ Iowa State University. http://lib.dr.iastate.edu/. https://doi.org/10.31274/rtd-180813-1876
Lee JS, Jeftinija K, Jeftinija S, Stromer MH, Scanes CG, Anderson LL (2004) Immunocytochemical distribution of somatotrophs in porcine anterior pituitary. Histochem Cell Biol 122:571–577. https://doi.org/10.1007/s00418-004-0715-8
Lillethorup TP et al (2018) Nigrostriatal proteasome inhibition impairs dopamine neurotransmission and motor function in minipigs. Exp Neurol 303:142–152. https://doi.org/10.1016/j.expneurol.2018.02.005
Lind NM, Moustgaard A, Jelsing J, Vajta G, Cumming P, Hansen AK (2007) The use of pigs in neuroscience: modeling brain disorders. Neurosci Biobehav Rev 31:728–751. https://doi.org/10.1016/j.neubiorev.2007.02.003
Mai JRK, Paxinos G (2012) The human nervous system, 3rd edn. Elsevier Academic Press, Amsterdam
Meijer JC, Trudeau VL, Colenbrander B, Poot P, Erkens JH, Van de Wiel DF (1988) Prolactin in the developing pig. Biol Reprod 39:264–269. https://doi.org/10.1095/biolreprod39.2.264
Mikami S, Chiba S, Hojo H, Taniguchi K, Kubokawa K, Ishii S (1988) Immunocytochemical studies on the pituitary pars distalis of the Japanese long-fingered bat, Miniopterus schreibersii fuliginosus. Cell Tissue Res 251:291–299
Milosevic V, Nestorovic N, Terzic M, Ristic N, Ajdzanovic V, Trifunovic S, Sekulic M (2009) Pituitary ACTH cells in female rats after neonatal treatment with SRIH-14. Folia Histochem Cytobiol 47:479–484. https://doi.org/10.2478/v10042-009-0104-1
Mitrofanova LB, Konovalov PV, Krylova JS, Polyakova VO, Kvetnoy IM (2017) Plurihormonal cells of normal anterior pituitary: facts and conclusions. Oncotarget 8:29282–29299. https://doi.org/10.18632/oncotarget.16502
Molitch ME (2017) Diagnosis and treatment of pituitary adenomas: a review. JAMA 317:516–524. https://doi.org/10.1001/jama.2016.19699
Musumeci G et al (2015) A journey through the pituitary gland: development, structure and function, with emphasis on embryo-foetal and later development. Acta Histochem 117:355–366. https://doi.org/10.1016/j.acthis.2015.02.008
Naik DR, Shirasawa N, Nogami H, Ishikawa H (1991) Immunohistochemistry of the pituitary pars distalis of the musk shrew, Suncus murinus. Gen Comp Endocrinol 84:27–35
Nakane PK (1970) Classifications of anterior pituitary cell types with immunoenzyme histochemistry. J Histochem Cytochem 18:9–20. https://doi.org/10.1177/18.1.9
Nishimura S, Okano K, Yasukouchi K, Gotoh T, Tabata S, Iwamoto H (2000) Testis developments and puberty in the male Tokara (Japanese native) goat. Anim Reprod Sci 64:127–131. https://doi.org/10.1016/s0378-4320(00)00197-4
Norgaard Glud A et al (2010) Direct gene transfer in the Gottingen Minipig CNS Using stereotaxic lentiviral microinjections. Acta Neurobiol Exp (wars) 70:308–315
Orlowski D, Glud AN, Palomero-Gallagher N, Sorensen JCH, Bjarkam CR (2019) Online histological atlas of the Gottingen minipig brain. Heliyon 5:e01363. https://doi.org/10.1016/j.heliyon.2019.e01363
Orstrup LH et al (2019) Towards a Gottingen minipig model of adult onset growth hormone deficiency: evaluation of stereotactic electrocoagulation method. Heliyon 5:e02892. https://doi.org/10.1016/j.heliyon.2019.e02892
Ozone C et al (2016) Functional anterior pituitary generated in self-organizing culture of human embryonic stem cells. Nat Commun 7:10351. https://doi.org/10.1038/ncomms10351
Peter B, De Rijk EP, Zeltner A, Emmen HH (2016) Sexual maturation in the female Gottingen minipig. Toxicol Pathol 44:482–485. https://doi.org/10.1177/0192623315621413
Raus Balind S, Manojlovic-Stojanoski M, Milosevic V, Todorovic D, Nikolic L, Petkovic B (2016) Short- and long-term exposure to alternating magnetic field (50 Hz, 0.5 mT) affects rat pituitary ACTH cells: stereological study. Environ Toxicol 31:461–468. https://doi.org/10.1002/tox.22059
Rosendal F et al (2010) Defining the intercommissural plane and stereotactic coordinates for the Basal Ganglia in the Gottingen minipig brain. Stereotact Funct Neurosurg 88:138–146. https://doi.org/10.1159/000303526
Sasaki F, Iwama Y (1988) Sex difference in prolactin and growth hormone cells in mouse adenohypophysis: stereological, morphometric, and immunohistochemical studies by light and electron microscopy. Endocrinology 123:905–912. https://doi.org/10.1210/endo-123-2-905
Sauleau P, Lapouble E, Val-Laillet D, Malbert CH (2009) The pig model in brain imaging and neurosurgery. Anim. Int. J. Anim. Biosci. 3:1138–1151. https://doi.org/10.1017/S1751731109004649
Sterio DC (1984) The unbiased estimation of number and sizes of arbitrary particles using the disector. J Microsc 134:127–136
Tan JH, Sasaki F (2000) Effect of age on immunocytochemical staining characteristics of adenohypophyseal cells in Mongolian pony mares and stallions. Am J Vet Res 61:826–831. https://doi.org/10.2460/ajvr.2000.61.826
Trifunovic S, Manojlovic-Stojanoski M, Ajdzanovic V, Nestorovic N, Ristic N, Medigovic I, Milosevic V (2014) Effects of genistein on stereological and hormonal characteristics of the pituitary somatotrophs in rats. Endocrine 47:869–877. https://doi.org/10.1007/s12020-014-0265-3
Trifunovic S et al (2016) Effects of prolonged alcohol exposure on somatotrophs and corticotrophs in adult rats: stereological and hormonal study. Acta Histochem 118:353–360. https://doi.org/10.1016/j.acthis.2016.03.005
Trifunovic S, Manojlovic-Stojanoski M, Nestorovic N, Ristic N, Sosic-Jurjevic B, Pendovski L, Milosevic V (2018) Histological and morphofunctional parameters of the hypothalamic–pituitary–adrenal system are sensitive to daidzein treatment in the adult rat. Acta Histochem 120:129–135. https://doi.org/10.1016/j.acthis.2017.12.006
Trouillas J, Guigard MP, Fonlupt P, Souchier C, Girod C (1996) Mapping of corticotropic cells in the normal human pituitary. J Histochem Cytochem 44:473–479. https://doi.org/10.1177/44.5.8627004
Vanputten LJA, Vanzwieten MJ, Mattheij JAM, Vankemenade JAM (1988) Studies on prolactin-secreting cells in aging rats of different strains. 1 Alterations in pituitary histology and serum prolactin levels as related to aging. Mech Ageing Dev 42:75–90. https://doi.org/10.1016/0047-6374(88)90064-4
Vidal S, Roman A, Moya L (1997) Description of two types of mammosomatotropes in mink (Mustela vison) adenohypophysis: changes in the population of mammosomatotropes under different physiological conditions. Acta Anat (basel) 159:209–217. https://doi.org/10.1159/000147986
Wang JF et al (2014) Establishment and characterization of dairy cow growth hormone secreting anterior pituitary cell model. In Vitro Cell Dev Biol Anim 50:103–110. https://doi.org/10.1007/s11626-013-9664-7
Watanabe H, Andersen F, Simonsen CZ, Evans SM, Gjedde A, Cumming P, DaNe XSG (2001) MR-based statistical atlas of the Gottingen minipig brain. Neuroimage 14:1089–1096. https://doi.org/10.1006/nimg.2001.0910
West MJ (2012) Basic stereology for biologists and neuroscientists. Cold Spring Harbor Laboratory Press
West MJ (2013) Tissue shrinkage and stereological studies. Cold Spring Harb Protoc. https://doi.org/10.1101/pdb.top071860
West MJ, Slomianka L, Gundersen HJ (1991) Unbiased stereological estimation of the total number of neurons in thesubdivisions of the rat hippocampus using the optical fractionator. Anat Rec 231:482–497. https://doi.org/10.1002/ar.1092310411
Willems C, Fu Q, Roose H, Mertens F, Cox B, Chen J, Vankelecom H (2016) Regeneration in the pituitary after cell-ablation injury: time-related aspects and molecular analysis. Endocrinology 157:705–721. https://doi.org/10.1210/en.2015-1741
Wolpert SM, Molitch ME, Goldman JA, Wood JB (1984) Size, shape, and appearance of the normal female pituitary gland. AJR Am J Roentgenol 143:377–381. https://doi.org/10.2214/ajr.143.2.377
Acknowledgements
This research was conducted at the Center for Experimental Neuroscience (CENSE), Department of Neurosurgery, Aarhus University Hospital. The authors sincerely thank the Department of Biomedicine, Aarhus University, for access to stereological facilities and acknowledge with gratitude the skillful assistance of Mrs. Trine W. Mikkelsen, Mrs. Lise M. Fitting, Mrs. Majken Sand, and Mrs. Anne Sofie Møller Andersen as well as the staff at Paaskehoejgaard. We would also express our gratitude to Lundbeck Foundation for funding this project.
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All authors contributed to the study conception and design, which was managed by ANG and JCS. JCS, HZ, LT, and DO collected the pituitary samples. Material preparation, histological processing, and data collection were planned and performed by LT, MW, and DO. LT, MW, CRB performed data analysis. The first draft of the manuscript was written by LT and DO, which was commented on and edited by MW, ANG, and CB. All authors read and approved the final version of the manuscript.
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Tvilling, L., West, M., Glud, A.N. et al. Anatomy and histology of the Göttingen minipig adenohypophysis with special emphasis on the polypeptide hormones: GH, PRL, and ACTH. Brain Struct Funct 226, 2375–2386 (2021). https://doi.org/10.1007/s00429-021-02337-1
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DOI: https://doi.org/10.1007/s00429-021-02337-1