Characterisation of germline progenitor cells in the testes of phylostomid bats: Artibeus jamaicensis and Sturnira lilium
Norma Moreno-Mendoza
A * , Iván Cabrera-Campos B , Noe Zacaula-Juárez C and Tania J. Porras-Gómez A
A
B
C
Abstract
A population of sperm progenitor cells, known as Asingle spermatogonia, has been described in mammalian testes. During division cycles in spermatogenesis, some cells will form part of the Asingle spermatogonia group, while others form primary spermatocytes. Thus, during spermatogenesis, spermatogonia are the progenitor cells of spermatozoa.
In this study, we characterise the spermatogonial stem cells (SSCs) in the testicles of Artibeus jamaicensis and Sturnira lilium bats. The knowledge generated from this will contribute to the understanding of the biology of germ cells and the mechanisms of spermatogenesis in mammals, generating information on wildlife species that are important for biodiversity.
Testes were analysed by light and electron microscopy. Likewise, the expression of specific factors of stem cells (Oct4 and C-kit), germ cells (Vasa), cell proliferation (pH3 and SCP1) and testicular somatic cells (MIS, 3βHSD and Sox9) was characterised by immunofluorescence and western blot.
The histological analysis enabled the location of type Asingle, Apaired and Aaligned spermatogonia in the periphery of the seminiferous tubules adjacent to Sertoli cells. The expression of genes of stem and germ cells made it possible to corroborate the distribution of the SSCs.
Results indicate that type Asingle spermatogonia were not randomly distributed, since proliferative activity was detected in groups of cells adjacent to the seminiferous tubules membrane, suggesting the localisation of spermatogonial niches in a specific region of testes.
This study provides evidence for the existence of SSCs in the testis of chiropterans that contribute to the renewal of germline progenitor cells to maintain the reproduction of the organisms.
Keywords: Artibeus jamaicensis, As spermatogonia, bats, germ cells, spermatogenesis, spermatogonia, spermatogonial stem cells, Sturnira lilium, testis.
References
Aeckerle N, Drummer C, Debowski K, Viebahn C, Behr R (2015) Primordial germ cell development in the marmoset monkey as revealed by pluripotency factor expression: suggestion of a novel model of embryonic germ cell translocation. Molecular Human Reproduction 21, 66-80.
| Crossref | Google Scholar |
Aguilar J, Gonzalvo MC, Clavero A, Ortiz A, González E, Ortiz-Galisteo JR, Peralta L, Maza M, Castilla JA (2004) Células madre espermatogonias. Revista Internacional de Andrología 2, 54-59.
| Google Scholar |
Altman E, Yango P, Moustafa R, Smith JF, Klatsky PC, Tran ND (2014) Characterization of human spermatogonial stem cell markers in fetal, pediatric, and adult testicular tissues. Reproduction 148, 417-427.
| Crossref | Google Scholar |
Andrade CS, Pletsch ÂA, Soares EM, Morielle-Versute E, Taboga SR, Souza CC, Beguelini MR (2023) Annual reproductive cycle of males of the great fruit-eating bat, Artibeus lituratus: testicular variations, abiotic regulation and sperm analysis. Tissue and Cell 83, 102131.
| Crossref | Google Scholar |
Arita WH, Ceballos González G (1997) Los mamíferos de México: distribución y estado de conservación. Revista Mexicana de Mastozoología (Nueva Época) 2, 33-71.
| Crossref | Google Scholar |
Bai Y, Feng M, Liu S, Wei H, Li L, Zhang X, Shen C, Zhang S, Ma N (2016) Differential gene expression in mouse spermatogonial stem cells and embryonic stem cells. International Journal of Molecular Medicine 38, 423-432.
| Crossref | Google Scholar |
Beguelini MR, Moreira PRL, Faria KC, Marchesin SRC, Morielle-Versute E (2009) Morphological characterization of the testicular cells and seminiferous epithelium cycle in six species of Neotropical bats. Journal of Morphology 270, 943-953.
| Crossref | Google Scholar |
Beguelini MR, Puga CCI, Taboga SR, Morielle-Versute E (2011) Ultrastructure of spermatogenesis in the white-lined broad-nosed bat, Platyrrhinus lineatus (Chiroptera: Phyllostomidae). Micron 42, 586-599.
| Crossref | Google Scholar |
Boitani C, Di Persio S, Esposito V, Vicini E (2016) Spermatogonial cells: mouse, monkey and man comparison. Seminars in Cell & Developmental Biology 59, 79-88.
| Crossref | Google Scholar |
Campos-Sánchez R, Kapusta A, Feschotte C, Chiaromonte F, Makova KD (2014) Genomic landscape of human, bat, and ex vivo DNA transposon integrations. Molecular Biology and Evolution 31, 1816-1832.
| Crossref | Google Scholar |
Chen S-R, Liu Y-X (2015) Regulation of spermatogonial stem cell self-renewal and spermatocyte meiosis by Sertoli cell signaling. Reproduction 149, R159-R167.
| Crossref | Google Scholar |
Chiarini-Garcia H, Raymer AM, Russell LD (2003) Non-random distribution of spermatogonia in rats: evidence of niches in the seminiferous tubules. Reproduction 126, 669-680.
| Crossref | Google Scholar |
Clarke JR (1981) Physiological problems of seasonal breeding in eutherian mammals. Oxford Reviews of Reproductive Biology 3, 244-312.
| Google Scholar |
Clermont Y (1966) Renewal of spermatogonia in man. American Journal of Anatomy 118, 509-524.
| Crossref | Google Scholar |
Clermont Y, Leblond CP (1959) Differentiation and renewal of spermatogonia in the monkey, Macacus rhesus. American Journal of Anatomy 104(2), 237-273.
| Crossref | Google Scholar |
De Felici M (2013) Origin, migration, and proliferation of human primordial germ cells. In ‘Oogenesis’. (Eds G Coticchio, D Albertini, L De Santis) pp. 19–37. (Springer: London) doi:10.1007/978-0-85729-826-3_2
de Rooij DG (2017) The nature and dynamics of spermatogonial stem cells. Development 144, 3022-3030.
| Crossref | Google Scholar |
de Rooij DG, Russell LD (2000) All you wanted to know about spermatogonia but were afraid to ask. Journal of Andrology 21, 776-798.
| Crossref | Google Scholar |
Diao L, Turek PJ, John CM, Fang F, Reijo Pera A (2022) Roles of spermatogonial stem cells in spermatogenesis and fertility restoration. Frontiers in Endocrinology 13, 895528.
| Crossref | Google Scholar |
Di Carlo AD, Travia G, De Felici M (2000) The meiotic specific synaptonemal complex protein SCP3 is expressed by female and male primordial germ cells of the mouse embryo. International Journal of Developmental Biology 44, 241-244.
| Google Scholar |
Dym M (1994) Spermatogonial stem cells of the testis. Proceedings of the National Academy of Sciences of the United States of America 91, 11287-11289.
| Crossref | Google Scholar |
Dym M, Kokkinaki M, He Z (2009) Spermatogonial stem cells: mouse and human comparisons. Birth Defects Research Part C: Embryo Today: Reviews 87(1), 27-34.
| Crossref | Google Scholar |
Ehmcke J, Schlatt S (2006) A revised model for spermatogonial expansion in man: lessons from non-human primates. Reproduction 132, 673-680.
| Crossref | Google Scholar |
Fayomi AP, Orwig KE (2018) Spermatogonial stem cells and spermatogenesis in mice, monkeys and men. Stem Cell Research 29, 207-214.
| Crossref | Google Scholar |
França LR, Hess RA, Dufour JM, Hofmann MC, Griswold MD (2016) The Sertoli cell: one hundred fifty years of beauty and plasticity. Andrology 4, 189-212.
| Crossref | Google Scholar |
Griswold MD (2016) Spermatogenesis: the commitment to meiosis. Physiological Reviews 96, 1-17.
| Crossref | Google Scholar |
Guo J, Sosa E, Chitiashvili T, Nie X, Rojas EJ, Oliver E, Donnor Connect, Plath K, Hotaling JM, Stukenborg J-B, Clark AT, Cairns BR (2021) Single-cell analysis of the developing human testis reveals somatic niche cell specification and fetal germline stem cell establishment. Cell Stem Cell 28, 764-778.e4.
| Crossref | Google Scholar |
Hara K, Nakagawa T, Enomoto H, Suzuki M, Yamamoto M, Simons BD, Yoshida S (2014) Mouse spermatogenic stem cells continually interconvert between equipotent singly isolated and syncytial states. Cell Stem Cell 14, 658-672.
| Crossref | Google Scholar |
Heinrich A, DeFalco T (2020) Essential roles of interstitial cells in testicular development and function. Andrology 8, 903-914.
| Crossref | Google Scholar |
Hermann BP, Sukhwani M, Simorangkir DR, Chu T, Plant TM, Orwig KE (2009) Molecular dissection of the male germ cell lineage identifies putative spermatogonial stem cells in rhesus macaques. Human Reproduction 24, 1704-1716.
| Crossref | Google Scholar |
Hickford DE, Frankenberg S, Pask AJ, Shaw G, Renfree MB (2011) DDX4 (VASA) is conserved in germ cell development in marsupials and monotremes. Biology of Reproduction 85, 733-743.
| Crossref | Google Scholar |
INEGI (2021) Aspectos Geográficos Morelos. (Instituto Nacional de Estadística y Geografía, INEGI) Available at https://www.inegi.org.mx/contenidos/app/areasgeograficas/resumen/resumen_17.pdf
Ishiguro K-I (2023) Mechanism of initiation of meiosis in mouse germ cells. In ‘Current topics in developmental biology. Vol. 151’. (Ed. F Cole) pp. 1–26. (Academic Press) doi:10.1016/bs.ctdb.2022.04.005
IUCN (2022) The IUCN Red List of Threatened Species. Version 2022-2. Available at https://www.iucnredlist.org [Accessed 10 October 2023]
Jin C, Wang Z, Li P, Tang J, Jiao T, Li Y, Ou J, Zou D, Li M, Mang X, Liu J, Ma Y, Wu X, Shi J, Chen S, He M, Lu Y, Zhang N, Miao S, Sun F, Wang L, Li K, Yu J, Song W (2023) Decoding the spermatogonial stem cell niche under physiological and recovery conditions in adult mice and humans. Science Advances 9(31), eabq3173.
| Crossref | Google Scholar |
Jones DL, Wagers AJ (2008) No place like home: anatomy and function of the stem cell niche. Nature Reviews Molecular Cell Biology 9, 11-21.
| Crossref | Google Scholar |
Kakegawa R, Teramura T, Takehara T, Anzai M, Mitani T, Matsumoto K, Saeki K, Sagawa N, Fukuda K, Hosoi Y (2008) Isolation and culture of rabbit primordial germ cells. Journal of Reproduction and Development 54, 352-357.
| Crossref | Google Scholar |
Kanatsu-Shinohara M, Shinohara T (2013) Spermatogonial stem cell self-renewal and development. Annual Review of Cell and Developmental Biology 29, 163-187.
| Crossref | Google Scholar |
Karnovsky MJ (1965) A formaldehyde-glutaraldehyde fixative of high osmolality for use in Electron Microscopy. The Journal of Cell Biology 27, 137A Available at http://www.jstor.org/stable/1604673.
| Google Scholar |
Kemper CH, Peters PWJ (1987) Migration and proliferation of primordial germ cells in the rat. Teratology 36, 117-124.
| Crossref | Google Scholar |
Kim JY, Jung HJ, Yoon MJ (2015) VASA (DDX4) is a putative marker for spermatogonia, spermatocytes and round spermatids in stallions. Reproduction in Domestic Animals 50, 1032-1038.
| Crossref | Google Scholar |
Kubota H, Avarbock MR, Schmidt JA, Brinster RL (2009) Spermatogonial stem cells derived from infertile Wv/Wv mice self-renew in vitro and generate progeny following transplantation. Biology of Reproduction 81, 293-301.
| Crossref | Google Scholar |
McIntyre A, Summersgill B, Grygalewicz B, Gillis AJM, Stoop J, van Gurp RJHLM, Dennis N, Fisher C, Huddart R, Cooper C, Clark J, Oosterhuis JW, Looijenga LHJ, Shipley J (2005) Amplification and overexpression of the KIT gene is associated with progression in the seminoma subtype of testicular germ-cell tumours of adolescents and adults. Cancer Research 65, 8085-8089.
| Crossref | Google Scholar |
Morais DB, de Paula TAR, Barros MS, Balarini MK, de Freitas MBD, da Matta SLP (2013) Stages and duration of the seminiferous epithelium cycle in the bat Sturnira lilium. Journal of Anatomy 222, 372-379.
| Crossref | Google Scholar |
Nakagawa T, Sharma M, Nabeshima Y-I, Braun RE, Yoshida S (2010) Functional hierarchy and reversibility within the murine spermatogenic stem cell compartment. Science 328, 62-67.
| Crossref | Google Scholar |
Oakberg EF (1971) Spermatogonial stem-cell renewal in the mouse. The Anatomical Record 169, 515-531.
| Crossref | Google Scholar |
Oatley JM, Avarbock MR, Telaranta AI, Fearon DT, Brinster RL (2006) Identifying genes important for spermatogonial stem cell self-renewal and survival. Proceedings of the National Academy of Sciences of the United States of America 103, 9524-9529.
| Crossref | Google Scholar |
Pesce M, Wang X, Wolgemuth DJ, Scholer HR (1998) Differential expression of the Oct-4 transcription factor during mouse germ cell differentiation. Mechanisms of Development 71, 89-98.
| Crossref | Google Scholar |
Phillips BT, Gassei K, Orwig KE (2010) Spermatogonial stem cell regulation and spermatogenesis. Philosophical Transactions of the Royal Society B: Biological Sciences 365, 1663-1678.
| Crossref | Google Scholar |
Saitou M, Yamaji M (2012) Primordial germ cells in mice. Cold Spring Harbor Perspectives in Biology 4(11), a008375.
| Crossref | Google Scholar |
Sawicka A, Seiser C (2012) Histone H3 phosphorylation – a versatile chromatin modification for different occasions. Biochimie 94, 2193-2201.
| Crossref | Google Scholar |
Schrans-Stassen BHGJ, van de Kant HJG, de Rooij DG, van Pelt AM (1999) Differential expression of c-kit in mouse undifferentiated and differentiating type A spermatogonia. Endocrinology 140, 5894-5900.
| Crossref | Google Scholar |
Sorrentino V, Giorgi M, Geremia R, Besmer P, Rossi P (1991) Expression of the c-kit protooncogene in the murine male germ cells. Oncogene 6, 149-151.
| Google Scholar |
Subash SK, Kumar PG (2021) Self-renewal and differentiation of spermatogonial stem cells. Frontiers in Bioscience, Landmark 26, 163-205.
| Crossref | Google Scholar |
Tan K, Wilkinson MF (2020) A single-cell view of spermatogonial stem cells. Current Opinion in Cell Biology 67, 71-78.
| Crossref | Google Scholar |
Unni SK, Modi DN, Pathak SG, Dhabalia JV, Bhartiya D (2009) Stage-specific localization and expression of C-kit in the adult human testis. Journal of Histochemistry & Cytochemistry 57, 861-869.
| Crossref | Google Scholar |
Wu G, Schöler HR (2014) Role of Oct4 in the early embryo development. Cell Regeneration 3, 3:7.
| Crossref | Google Scholar |
Yoshida S, Sukeno M, Nabeshima Y-I (2007) A vasculature-associated niche for undifferentiated spermatogonia in the mouse testis. Science 317, 1722-1726.
| Crossref | Google Scholar |
Zhang L, Tang J, Haines CJ, Feng H, Lai L, Teng X, Han Y (2011) c-kit and its related genes in spermatogonial differentiation. Spermatogenesis 1, 186-194.
| Crossref | Google Scholar |
