Alpha-synuclein expression in GnRH neurons of young and old bovine hypothalami
Yvan Bienvenu Niyonzima A , Yuuki Asato A , Tomoaki Murakami B and Hiroya Kadokawa
A *
A
B
Abstract
Understanding of central nervous system mechanisms related to age-related infertility remains limited. Fibril α-synuclein, distinct from its monomer form, is implicated in age-related diseases and propagates among neurons akin to prions.
We compared α-synuclein expression in gonadotropin-releasing hormone-expressing neurons (GnRH neurons) in the pre-optic area, arcuate nucleus, and median eminence of healthy heifers and aged cows to determine its role in age-related infertility.
We analysed mRNA and protein expression, along with fluorescent immunohistochemistry for GnRH and α-synuclein, followed by Congo red staining to detect amyloid deposits, and confocal microscopy.
Both mRNA and protein expressions of α-synuclein were confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and western blots in bovine cortex, hippocampus, and anterior and posterior hypothalamus tissues. Significant differences in α-synuclein mRNA expression were observed in the cortex and hippocampus between young and old cows. Western blots showed five bands of α-synuclein, probably reflecting monomer, dimer, and oligomers, in the cortex, hippocampus, hypothalamus tissues, and there were significant differences in some bands between young and old cows. Bright-field and polarised light microscopy did not detect obvious amyloid deposition in aged hypothalami; however, higher-sensitive confocal microscopy unveiled strong positive signal of Congo red and α-synuclein in GnRH neurons in aged hypothalami. Additionally, α-synuclein expression was detected in immortalised GnRH neurons, GT1-7 cells.
Alpha-synuclein was expressed in GnRH neurons, and some differences were observed between young and old hypothalami.
Alpha-synuclein may play an important role in aging-related infertility.
Keywords: aging, arcuate nucleus, Congo red staining, cortex, GT1-7 cell, hippocampus, median eminence, pre-optic area, ruminant.
References
Ahmed J, Fitch TC, Donnelly CM, Joseph JA, Ball TD, Bassil MM, Son A, Zhang C, Ledreux A, Horowitz S, Qin Y, Paredes D, Kumar S (2022) Foldamers reveal and validate therapeutic targets associated with toxic α-synuclein self-assembly. Nature Communications 13, 2273.
| Crossref | Google Scholar | PubMed |
Araki K, Yagi N, Aoyama K, Choong C-J, Hayakawa H, Fujimura H, Nagai Y, Goto Y, Mochizuki H (2019) Parkinson’s disease is a type of amyloidosis featuring accumulation of amyloid fibrils of α-synuclein. Proceedings of the National Academy of Sciences of the United States of America 116, 17963-17969.
| Crossref | Google Scholar | PubMed |
Bieri G, Gitler AD, Brahic M (2018) Internalization, axonal transport and release of fibrillar forms of alpha-synuclein. Neurobiology of Disease 109(Pt B), 219-225.
| Crossref | Google Scholar | PubMed |
Bonefeld BE, Elfving B, Wegener G (2008) Reference genes for normalization: a study of rat brain tissue. Synapse 62, 302-309.
| Crossref | Google Scholar | PubMed |
Campbell RE, Gaidamaka G, Han S-K, Herbison AE (2009) Dendro-dendritic bundling and shared synapses between gonadotropin-releasing hormone neurons. Proceedings of the National Academy of Sciences of the United States of America 106, 10835-10840.
| Crossref | Google Scholar | PubMed |
Canever JB, Soares ES, de Avelar NCP, Cimarosti HI (2023) Targeting α-synuclein post-translational modifications in Parkinson’s disease. Behavioural Brain Research 439, 114204.
| Crossref | Google Scholar | PubMed |
Castellani C, Fedrigo M, Frigo AC, Barbera MD, Thiene G, Valente M, Adami F, Angelini A (2017) Application of confocal laser scanning microscopy for the diagnosis of amyloidosis. Virchows Archiv 470, 455-463.
| Crossref | Google Scholar | PubMed |
Choi ML, Chappard A, Singh BP, Maclachlan C, Rodrigues M, Fedotova EI, Berezhnov AV, De S, Peddie CJ, Athauda D, Virdi GS, Zhang W, Evans JR, Wernick AI, Zanjani ZS, Angelova PR, Esteras N, Vinokurov AY, Morris K, Jeacock K, Tosatto L, Little D, Gissen P, Clarke DJ, Kunath T, Collinson L, Klenerman D, Abramov AY, Horrocks MH, Gandhi S (2022) Pathological structural conversion of α-synuclein at the mitochondria induces neuronal toxicity. Nature Neuroscience 25, 1134-1148.
| Crossref | Google Scholar | PubMed |
Dean DN, Lee JC (2021) Linking Parkinson’s disease and melanoma: interplay between α-Synuclein and Pmel17 amyloid formation. Movement Disorders 36, 1489-1498.
| Crossref | Google Scholar | PubMed |
Deyell JS, Sriparna M, Ying M, Mao X (2023) The Interplay between α-Synuclein and Microglia in α-Synucleinopathies. International Journal of Molecular Sciences 24, 2477.
| Crossref | Google Scholar | PubMed |
Dharmadana D, Reynolds NP, Conn CE, Valéry C (2019) pH-dependent self-assembly of human neuropeptide hormone GnRH into functional amyloid nanofibrils and hexagonal phases. ACS Applied Bio Materials 2, 3601-3606.
| Crossref | Google Scholar | PubMed |
Epelbaum J, Terrien J (2020) Mini-review: aging of the neuroendocrine system: insights from nonhuman primate models. Progress in Neuro-Psychopharmacology and Biological Psychiatry 100, 109854.
| Crossref | Google Scholar | PubMed |
Fields CR, Bengoa-Vergniory N, Wade-Martins R (2019) Targeting alpha-synuclein as a therapy for Parkinson’s disease. Frontiers in Molecular Neuroscience 12, 299.
| Crossref | Google Scholar |
Fortin JS, Hetak AA, Duggan KE, Burglass CM, Penticoff HB, Schott HC, II (2021) Equine pituitary pars intermedia dysfunction: a spontaneous model of synucleinopathy. Scientific Reports 11, 16036.
| Crossref | Google Scholar | PubMed |
Fowler DM, Koulov AV, Alory-Jost C, Marks MS, Balch WE, Kelly JW (2006) Functional amyloid formation within mammalian tissue. PLoS Biology 4, e6.
| Crossref | Google Scholar | PubMed |
Gegg ME, Verona G, Schapira AHV (2020) Glucocerebrosidase deficiency promotes release of α-synuclein fibrils from cultured neurons. Human Molecular Genetics 29, 1716-1728.
| Crossref | Google Scholar | PubMed |
Gossner AG, Foster JD, Fazakerley JK, Hunter N, Hopkins J (2011) Gene expression analysis in distinct regions of the central nervous system during the development of SSBP/1 sheep scrapie. Veterinary Microbiology 147, 42-48.
| Crossref | Google Scholar | PubMed |
Gunes S, Hekim GNT, Arslan MA, Asci R (2016) Effects of aging on the male reproductive system. Journal of Assisted Reproduction and Genetics 33, 441-454.
| Crossref | Google Scholar | PubMed |
Hsu LJ, Sagara Y, Arroyo A, Rockenstein E, Sisk A, Mallory M, Wong J, Takenouchi T, Hashimoto M, Masliah E (2000) α-synuclein promotes mitochondrial deficit and oxidative stress. The American Journal of Pathology 157, 401-410.
| Crossref | Google Scholar | PubMed |
Inoue K, Hosono M, Oyama H, Hirooka H (2020) Genetic associations between reproductive traits for first calving and growth curve characteristics of Japanese Black cattle. Animal Science Journal 91, e13467.
| Crossref | Google Scholar | PubMed |
Jin M, Wang S, Gao X, Zou Z, Hirotsune S, Sun L (2024) Pathological and physiological functional cross-talks of α-synuclein and tau in the central nervous system. Neural Regeneration Research 19, 855-862.
| Crossref | Google Scholar | PubMed |
Kadokawa H, Pandey K, Nahar A, Nakamura U, Rudolf FO (2014) Gonadotropin-releasing hormone (GnRH) receptors of cattle aggregate on the surface of gonadotrophs and are increased by elevated GnRH concentrations. Animal Reproduction Science 150, 84-95.
| Crossref | Google Scholar | PubMed |
Kadokawa H, Kotaniguchi M, Kereilwe O, Kitamura S (2021) Reduced gonadotroph stimulation by ethanolamine plasmalogens in old bovine brains. Scientific Reports 11, 4757.
| Crossref | Google Scholar |
Kereilwe O, Kadokawa H (2020a) Anti-Müllerian hormone and its receptor are detected in most gonadotropin-releasing-hormone cell bodies and fibers in heifer brains. Domestic Animal Endocrinology 72, 106432.
| Crossref | Google Scholar | PubMed |
Kereilwe O, Kadokawa H (2020b) Decreased Anti-Müllerian hormone and Anti-Müllerian hormone receptor type 2 in hypothalami of old Japanese Black cows. Journal of veterinary medical science 82, 1113-1117.
| Crossref | Google Scholar | PubMed |
Kinnart I, Manders L, Heyninck T, Imberechts D, Praschberger R, Schoovaerts N, Verfaillie C, Verstreken P, Vandenberghe W (2024) Elevated α-synuclein levels inhibit mitophagic flux. Nature Partner Journals Parkinson’s disease 10, 80.
| Crossref | Google Scholar | PubMed |
Lamberts JT, Hildebrandt EN, Brundin P (2015) Spreading of α-synuclein in the face of axonal transport deficits in Parkinson’s disease: a speculative synthesis. Neurobiology of Disease 77, 276-283.
| Crossref | Google Scholar | PubMed |
Leshin LS, Rund LA, Crim JW, Kiser TE (1988) Immunocytochemical localization of luteinizing hormone-releasing hormone and proopiomelanocortin neurons within the preoptic area and hypothalamus of the bovine brain. Biology of Reproduction 39, 963-975.
| Crossref | Google Scholar | PubMed |
Liposits Z, Merchenthaler I, Wetsel WC, Reid JJ, Mellon PL, Weiner RI, Negro-Vilar A (1991) Morphological characterization of immortalized hypothalamic neurons synthesizing luteinizing hormone-releasing hormone. Endocrinology 129, 1575-1583.
| Crossref | Google Scholar | PubMed |
Malhi PS, Adams GP, Pierson RA, Singh J (2006) Bovine model of reproductive aging: response to ovarian synchronization and superstimulation. Theriogenology 66, 1257-1266.
| Crossref | Google Scholar | PubMed |
Marsh SE, Blurton-Jones M (2012) Examining the mechanisms that link β-amyloid and α-synuclein pathologies. Alzheimer’s Research & Therapy 4, 11.
| Crossref | Google Scholar | PubMed |
Melki R (2018) Alpha-synuclein and the prion hypothesis in Parkinson’s disease. Revue Neurologique 174, 644-652.
| Crossref | Google Scholar | PubMed |
Miyamoto Y, Skarzynski DJ, Okuda K (2000) Is tumor necrosis factor α a trigger for the initiation of endometrial prostaglandin F2α release at luteolysis in cattle? Biology of Reproduction 62, 1109-1115.
| Crossref | Google Scholar | PubMed |
Moreno-Gonzalez I, Edwards G, III, Morales R, Duran-Aniotz C, Escobedo G, Jr, Marquez M, Pumarola M, Soto C (2022) Aged cattle brain displays Alzheimer’s disease-like pathology and promotes brain amyloidosis in a transgenic animal model. Frontiers in Aging Neuroscience 13, 815361.
| Crossref | Google Scholar | PubMed |
Niyonzima YB, Asato Y, Kadokawa H (2024) Alpha-synuclein expression in oxytocin neurons of young and old bovine brains. Journal of Reproduction and Development 70, 213-222.
| Crossref | Google Scholar | PubMed |
Obrenovich ME, Monnier VM (2004) Glycation stimulates amyloid formation. Science of Aging Knowledge Environment 2004, pe3.
| Crossref | Google Scholar |
Osoro K, Wright IA (1992) The effect of body condition, live weight, breed, age, calf performance, and calving date on reproductive performance of spring-calving beef cows. Journal of Animal Science 70, 1661-1666.
| Crossref | Google Scholar | PubMed |
Perissinotto F, Stani C, De Cecco E, Vaccari L, Rondelli V, Posocco P, Parisse P, Scaini D, Legname G, Casalis L (2020) Iron-mediated interaction of alpha synuclein with lipid raft model membranes. Nanoscale 12, 7631-7640.
| Crossref | Google Scholar | PubMed |
Puladi B, Dinekov M, Arzberger T, Taubert M, Köhler C (2021) The relation between tau pathology and granulovacuolar degeneration of neurons. Neurobiology of Disease 147, 105138.
| Crossref | Google Scholar | PubMed |
Putteeraj M, Soga T, Ubuka T, Parhar IS (2016) A “Timed” kiss is essential for reproduction: lessons from mammalian studies. Frontiers in Endocrinology 7, 121.
| Crossref | Google Scholar | PubMed |
Raghavan R, de Kruijff L, Sterrenburg MD, Rogers BB, Hladik CL, White CL, III (2004) Alpha-synuclein expression in the developing human brain. Pediatric and Developmental Pathology 7, 506-516.
| Crossref | Google Scholar | PubMed |
Rech RR, Giaretta PR, Brown C, Barros CSL (2018) Gross and histopathological pitfalls found in the examination of 3,338 cattle brains submitted to the BSE surveillance program in Brazil. Pesquisa Veterinária Brasileira 38, 2099-2108.
| Crossref | Google Scholar |
Schultz SW, Nilsson KPR, Westermark GT (2011) Drosophila melanogaster as a model system for studies of islet amyloid polypeptide aggregation. PLoS ONE 6, e20221.
| Crossref | Google Scholar | PubMed |
Scivetti M, Favia G, Fatone L, Maiorano E, Crincoli V (2016) Concomitant use of Congo red staining and confocal laser scanning microscopy to detect amyloidosis in oral biopsy: a clinicopathological study of 16 patients. Ultrastructural Pathology 40, 86-91.
| Crossref | Google Scholar | PubMed |
Sergeeva AV, Galkin AP (2020) Functional amyloids of eukaryotes: criteria, classification, and biological significance. Current Genetics 66, 849-866.
| Crossref | Google Scholar | PubMed |
Shahmoradian SH, Lewis AJ, Genoud C, Hench J, Moors TE, Navarro PP, Castaño-Díez D, Schweighauser G, Graff-Meyer A, Goldie KN, Sütterlin R, Huisman E, Ingrassia A, Gier Y, Rozemuller AJM, Wang J, Paepe AD, Erny J, Staempfli A, Hoernschemeyer J, Großerüschkamp F, Niedieker D, El-Mashtoly SF, Quadri M, Van IJcken WFJ, Bonifati V, Gerwert K, Bohrmann B, Frank S, Britschgi M, Stahlberg H, Van de Berg WDJ, Lauer ME (2019) Lewy pathology in Parkinson’s disease consists of crowded organelles and lipid membranes. Nature Neuroscience 22, 1099-1109.
| Crossref | Google Scholar | PubMed |
Simpson J, Yates CM, Watts AG, Fink G (1988) Congo red birefringent structures in the hypothalamus in senile dementia of the Alzheimer type. Neuropathology and Applied Neurobiology 14, 381-393.
| Crossref | Google Scholar | PubMed |
Sui Y-T, Bullock KM, Erickson MA, Zhang J, Banks WA (2014) Alpha synuclein is transported into and out of the brain by the blood-brain barrier. Peptides 62, 197-202.
| Crossref | Google Scholar | PubMed |
Sulzer D, Edwards RH (2019) The physiological role of α-synuclein and its relationship to Parkinson’s disease. Journal of Neurochemistry 150, 475-486.
| Crossref | Google Scholar | PubMed |
Tanco VM, Whitlock BK, Jones MA, Wilborn RR, Brandebourg TD, Foradori CD (2016) Distribution and regulation of gonadotropin-releasing hormone, kisspeptin, RF-amide related peptide-3, and dynorphin in the bovine hypothalamus. PeerJ 4, e1833.
| Crossref | Google Scholar | PubMed |
Tong J, Wong H, Guttman M, Ang LC, Forno LS, Shimadzu M, Rajput AH, Muenter MD, Kish SJ, Hornykiewicz O, Furukawa Y (2010) Brain α-synuclein accumulation in multiple system atrophy, Parkinson’s disease and progressive supranuclear palsy: a comparative investigation. Brain: a Journal of Neurology 133, 172-188.
| Crossref | Google Scholar | PubMed |
Tripathi T, Chattopadhyay K (2019) Interaction of α-Synuclein with ATP Synthase: switching role from physiological to pathological. ACS Chemical Neuroscience 10, 16-17.
| Crossref | Google Scholar | PubMed |
Urbanski HF (1991) Monoclonal antibodies to luteinizing hormone-releasing hormone: production, characterization, and immunocytochemical application. Biology of Reproduction 44, 681-686.
| Crossref | Google Scholar | PubMed |
van den Beld AW, Kaufman J-M, Zillikens MC, Lamberts SWJ, Egan JM, van der Lely AJ (2018) The physiology of endocrine systems with ageing. The Lancet Diabetes & Endocrinology 6, 647-658.
| Crossref | Google Scholar | PubMed |
Wang X-L, Feng S-T, Wang Y-T, Yuan Y-H, Li Z-P, Chen N-H, Wang Z-Z, Zhang Y (2022) Mitophagy, a form of selective autophagy, plays an essential role in mitochondrial dynamics of Parkinson’s disease. Cellular and Molecular Neurobiology 42, 1321-1339.
| Crossref | Google Scholar | PubMed |
Yamada M, Kotani Y, Nakamura K, Kobayashi Y, Horiuchi N, Doi T, Suzuki S, Sato N, Kanno T, Matsui T (2006) Immunohistochemical distribution of amyloid deposits in 25 cows diagnosed with systemic AA amyloidosis. Journal of Veterinary Medical Science 68, 725-729.
| Crossref | Google Scholar | PubMed |
Yanamandra K, Gruden MA, Casaite V, Meskys R, Forsgren L, Morozova-Roche LA (2011) α-synuclein reactive antibodies as diagnostic biomarkers in blood sera of Parkinson’s disease patients. PLoS ONE 6, e18513.
| Crossref | Google Scholar | PubMed |
Yin W, Mendenhall JM, Monita M, Gore AC (2009) Three-dimensional properties of GnRH neuroterminals in the median eminence of young and old rats. Journal of Comparative Neurology 517, 284-295.
| Crossref | Google Scholar | PubMed |
Zhang S, Zhu R, Pan B, Xu H, Olufemi MF, Gathagan RJ, Li Y, Zhang L, Zhang J, Xiang W, Kagan EM, Cao X, Yuan C, Kim S-J, Williams CK, Magaki S, Vinters HV, Lashuel HA, Garcia BA, James Petersson E, Trojanowski JQ, Lee VM-Y, Peng C (2023) Post-translational modifications of soluble α-synuclein regulate the amplification of pathological α-synuclein. Nature Neuroscience 26, 213-225.
| Crossref | Google Scholar | PubMed |
Zhou S, Shen Y, Zang S, Yin X, Li P (2021) The epigenetic role of HTR1A antagonist in facilitating GnRH expression for pubertal initiation control. Molecular Therapy - Nucleic Acids 25, 198-206.
| Crossref | Google Scholar | PubMed |
Zussy C, Brureau A, Delair B, Marchal S, Keller E, Ixart G, Naert G, Meunier J, Chevallier N, Maurice T, Givalois L (2011) Time-course and regional analyses of the physiopathological changes induced after cerebral injection of an amyloid β fragment in rats. The American Journal of Pathology 179, 315-334.
| Crossref | Google Scholar | PubMed |
