Journal of Endocrinology and Reproduction

Open Access Open Access  Restricted Access Subscription Access
Open Access Open Access Open Access  Restricted Access Restricted Access Subscription Access

Transforming Growth Factor β: Cloning and Expression Profiling in the Brain of Catfish Model


Affiliations
1 Department of Animal Biology, School of Life Sciences, University of Hyderabad, P.O. Central University, Hyderabad - 500046, Telangana, India
     

   Subscribe/Renew Journal


Transforming growth factor-beta (tgf-β) is implicated as a signalling molecule and considered a vital regulator for neuronal survival, development, ovarian growth, and many other biological processes. The present study was aimed to analyze the expression of tgf-β and its significance during the early brain and gonadal development. Initially, cloning of partial cDNA of tgf-β from the female brain was performed followed by expression analysis during various developmental stages using quantitative real-time PCR. Cloned tgf-β is approximately 210 bp, which encodes a putative protein of 69 amino acid residues, and showed high similarity with other counterparts. The tissue-specific expression of tgf-β was ubiquitous but significantly higher in brain as well as gonads of both male and female. Further, in ontogeny analysis, tgf-β mRNA levels were significantly greater in 75-, 100- and 125-days post-hatch during development. The pre-spawning stage showed higher expression of tgf-β when compared to all other reproductive stages. Immunolocalization study revealed the presence of Tgf-β protein in the preoptic area-hypothalamus which was well correlated with the mRNA expression in discrete regions of the catfish brain. Expression of tgf-β seems to have a role in the regulation of gonadal function through the brain-pituitary axis.

Keywords

Brain, Development, Expression, Signalling Molecule, Tgf-β
Subscription Login to verify subscription
User
Notifications
Font Size


  • Meyers EA, Kessler JA. TGF-β family signaling in neural and neuronal differentiation, development, and function. Cold Spring Harb Perspect Biol. 2017; 9(8):a022244. https://doi.org/10.1101/cshperspect.a022244. PMid:28130363 PMCid:PMC5538418.

  • Calp MK, Matsumoto JA, Van Der Kraak G. Activin and transforming growth factor-β as local regulators of ovarian steroid genesis in the goldfish. Gen Comp Endocrinol. 2003; 132(1):142-50. https://doi.org/10.1016/S0016-6480(03)00060-1.

  • Krieglstein K, Strelau J, Schober A, Sullivan A, Unsicker K. TGF-β and the regulation of neuron survival and death. J Physiol Paris. 2002; 96(1-2):25-30. https://doi.org/10.1016/S0928-4257(01)00077-8.

  • Zheng S, Long J, Liu Z, Tao W, Wang D. Identification and evolution of TGF-β signaling pathway members in twenty-four animal species and expression in Tilapia. Int J Mol Sci. 2018; 19(4):1154. https://doi.org/10.3390/ijms19041154. PMid:29641448 PMCid:PMC5979292.

  • MacDougall TM, Van Der Kraak G. Peptide growth factors modulate prostaglandin E and F production by goldfish ovarian follicles. Gen Comp Endocrinol. 1998; 110(1):46-57. https://doi.org/10.1006/gcen.1997.7047. PMid:9514845.

  • Mamta SK, Senthilkumaran B. GDNF family receptor α-1 in catfish: Possible implication to brain dopaminergic activity. Brain Res Bull. 2018; 140:270-80. https://doi.org/10.1016/j.brainresbull.2018.05.010. PMid:29758254.

  • Findlay JK, Drummond AE, Dyson ML, Baillie AJ, Robertson DM, Ethier JF. Recruitment and development of the follicle: The roles of the transforming growth factor-β superfamily. Mol Cell Endocrinol. 2002; 191(1):35-43. https://doi.org/10.1016/S0303-7207(02)00053-9.

  • Knight PG, Glister C. TGF-β superfamily members and ovarian follicle development. Reproduction. 2006; 132(2):191-206. https://doi.org/10.1530/rep.1.01074. PMid:16885529.

  • Krieglstein K, Suter-Crazzolara C, Fischer WH, Unsicker K. TGF-beta superfamily members promote survival of midbrain dopaminergic neurons and protect them against MPP+ toxicity. The EMBO J. 1995; 14(4):736-42. https://doi.org/10.1002/j.1460-2075.1995.tb07052.x. PMid:7882977 PMCid:PMC398139.

  • Krieglstein K, Henheik P, Farkas L, Jaszai J, Galter D, Krohn K, Unsicker K. Glial cell line-derived neurotrophic factor requires transforming growth factor-β for exerting its full neurotrophic potential on peripheral and CNS neurons. J Neurosci. 1998; 18(23):9822-34. https://doi.org/10.1523/JNEUROSCI.18-23-09822.1998. PMid:9822741 PMCid:PMC6793316.

  • Schober A, Hertel R, Arumäe U, Farkas L, Jaszai J, Krieglstein K, Saarma M, Unsicker K. Glial cell line-derived neurotrophic factor rescues target-deprived sympathetic spinal cord neurons but requires transforming growth factor-β as cofactor in vivo. J Neurosci. 1999; 19(6):2008-15. https://doi.org/10.1523/JNEUROSCI.19-06-02008.1999. PMid:10066254 PMCid:PMC6782553.

  • Massague J. TGF-β signal transduction. Annu Rev Biochem. 1998; 67(1):753-91. https://doi.org/10.1146/annurev.biochem.67.1.753. PMid:9759503. 13. Burt DW, Law AS. Evolution of the transforming growth factor-beta superfamily. Prog Growth Factor Res. 1994; 5(1):99-118. https://doi.org/10.1016/0955-2235(94)90020-5.

  • Laing KJ, Pilström L, Cunningham C, Secombes CJ. TGF-β3 exists in bony fish. Vet Immunol Immunopathol. 1999; 72(1-2):45-53. https://doi.org/10.1016/S0165-2427(99)00116-6.

  • Tan B, Pascual A, de Beus A, Cooper K, Hull E. TGFβ (transforming growth factor β) and keratocyte motility in 24 h zebrafish explant cultures. Cell Biol Int. 2011; 35(11):1131-9. https://doi.org/10.1042/CBI20110063. PMid:21729005.

  • Carrella S, Barbato S, D’Agostino Y, Salierno FG, Manfredi A, Banfi S, Conte I. TGF-β controls miR-181/ERK regulatory network during retinal axon specification and growth. PLoS One. 2015; 10(12):e0144129. https://doi.org/10.1371/journal.pone.0144129. PMid:26641497 PMCid:PMC4671616.

  • Jason JY, Barnes AP, Hand R, Polleux F, Ehlers MD. TGF-β signaling specifies axons during brain development. Cell. 2010; 142(1):144-57. https://doi.org/10.1016/j.cell.2010.06.010. PMid:20603020 PMCid:PMC2933408.

  • (a) Chalazonitis A, Kalberg J, Twardzik DR, Morrison RS, Kessler JA. Transforming growth factor β has neurotrophic actions on sensory neurons in vitro and is synergistic with nerve growth factor. Dev Biol. 1992; 152(1):121-32. https://doi.org/10.1016/0012-1606(92)90162-A.

  • (b) Chalazonitis A, Kessler JA, Twardzik DR, Morrison RS. Transforming growth factor α, but not epidermal growth factor, promotes the survival of sensory neurons in vitro. J Neurosci. 1992; 12(2):583-94. https://doi.org/10.1523/JNEUROSCI.12-02-00583.1992. PMid:1740693 PMCid:PMC6575603.

  • Oliveira MA, Martinez ER, Butzge AJ, Doretto LB, Ricci JM, Rodrigues MS, Vigoya AA, Gómez-Gonzalez NE, Stewart AB, Nobrega RH. Molecular characterization and expression analysis of anti-Mullerian hormone in common carp (Cyprinus carpio) adult testes. Gene Expr Patterns. 2021; 40:119169. https://doi.org/10.1016/j.gep.2021.119169. PMid:33667682.

  • Ingman WV, Robertson SA. The essential roles of TGFβ1 in reproduction. Cytokine Growth Factor Rev. 2009; 20(3):233-9. https://doi.org/10.1016/j.cytogfr.2009.05.003. PMid:19497778.

  • Raghuveer K, Senthilkumaran B. Identification of multiple dmrt1s in catfish: localization, dimorphic expression pattern, changes during testicular cycle and after methyltestosterone treatment. J Mol Endocrinol. 2009; 42(5):437-48. https://doi.org/10.1677/JME-09-0011. PMid:19293249.

  • Mamta SK, Sudhakumari C, Kagawa H, Dutta-Gupta A, Senthilkumaran B. Controlled release of sex steroids through osmotic pump alters brain GnRH1 and catecholaminergic system dimorphically in the catfish, Clarias gariepinus. Brain Res Bull. 2020; 164:325-33. https://doi.org/10.1016/j.brainresbull.2020.08.022.PMid:32860867.

  • Hardie LJ, Laing KJ, Daniels GD, Grabowski PS, Cunningham C, Secombes CJ. Isolation of the first piscine transforming growth factor β gene: analysis reveals tissue-specific expression and a potential regulatory sequence in rainbow trout (Oncorhynchus mykiss). Cytokine. 1998; 10(8):555-63. https://doi.org/10.1006/cyto.1997.0334.PMid:9722928.

  • Harms CA, Ottinger CA, Blazer VS, Densmore CL, Pieper LH, Kennedy-Stoskopf S. Quantitative polymerase chain reaction for transforming growth factor-beta applied to a field study of fish health in Chesapeake Bay tributaries. Environ Health Perspect. 2000; 108(5):447-52. https://doi.org/10.1289/ehp.00108447. PMid:10811572 PMCid:PMC1638039.

  • Tafalla C, Aranguren R, Secombes CJ, Castrillo JL, Novoa B, Figueras A. Molecular characterization of sea bream (Sparus aurata) transforming growth factor β1. Fish Shellfish Immunol. 2003; 14(5):405-21. https://doi.org/10.1006/fsim.2002.0444. PMid:12711274.

  • Maehr T, Costa MM, Vecino JL, Wadsworth S, Martin SA, Wang T, Secombes CJ. Transforming growth factor-β1b: A second TGF-β1 paralogue in the rainbow trout (Oncorhynchus mykiss) that has a lower constitutive expression but is more responsive to immune stimulation. Fish Shellfish Immunol. 2013; 34(2):420-32. https://doi.org/10.1016/j.fsi.2012.11.011. PMid:23178261.

  • Senthilkumaran B, Okuzawa K, Gen K, Ookura T, Kagawa H. Distribution and seasonal variations in levels of three native GnRHs in the brain and pituitary of perciform fish. J Neuroendocrinol. 1999; 11(3):181-6. https://doi.org/10.1046/j.1365-2826.1999.00304.x. PMid:10201813.

  • Joy KP, Singh MS, Senthilkumaran B, Goos HT. Pituitary-gonadal relationship in the catfish Clarias batrachus (L): a study correlating gonadotrophin-II and sex steroid dynamics. Zool Sci. 2000; 17(3):395-404. https://doi.org/10.2108/jsz.17.395. PMid:18494596.

  • Kirubagaran R, Senthilkumaran B, Sudhakumari CC, Joy KP. Seasonal dynamics in gonadotropin secretion and E2-binding in the catfish Heteropneustes fossilis. Fish Physiol Biochem. 2005; 31(2-3):183. https://doi.org/10.1007/s10695-006-0022-5. PMid:20035456.

  • Acharjee A, Chaube R, Joy KP. Molecular cloning and characterization of the gonadotropin subunits GPα, FSHβ, and LHβ genes in the stinging catfish Heteropneustes fossilis: phylogeny, seasonal expression and pituitary localization. J Exp Zool A Ecol Genet Physiol. 2015; 323(8):567-85. https://doi.org/10.1002/jez.1949. PMid:26205349.

  • Zapata A, Diez B, Cejalvo T, Gutierrez-de Frias C, Cortes A. Ontogeny of the immune system of fish. Fish Shellfish Immunol. 2006; 20(2):126-36. https://doi.org/10.1016/j.fsi.2004.09.005. PMid:15939627.


Abstract Views: 220

PDF Views: 0




  • Transforming Growth Factor β: Cloning and Expression Profiling in the Brain of Catfish Model

Abstract Views: 220  |  PDF Views: 0

Authors

Sajwan-Khatri Mamta
Department of Animal Biology, School of Life Sciences, University of Hyderabad, P.O. Central University, Hyderabad - 500046, Telangana, India
Narlagiri Anusha
Department of Animal Biology, School of Life Sciences, University of Hyderabad, P.O. Central University, Hyderabad - 500046, Telangana, India
Balasubramanian Senthilkumaran
Department of Animal Biology, School of Life Sciences, University of Hyderabad, P.O. Central University, Hyderabad - 500046, Telangana, India

Abstract


Transforming growth factor-beta (tgf-β) is implicated as a signalling molecule and considered a vital regulator for neuronal survival, development, ovarian growth, and many other biological processes. The present study was aimed to analyze the expression of tgf-β and its significance during the early brain and gonadal development. Initially, cloning of partial cDNA of tgf-β from the female brain was performed followed by expression analysis during various developmental stages using quantitative real-time PCR. Cloned tgf-β is approximately 210 bp, which encodes a putative protein of 69 amino acid residues, and showed high similarity with other counterparts. The tissue-specific expression of tgf-β was ubiquitous but significantly higher in brain as well as gonads of both male and female. Further, in ontogeny analysis, tgf-β mRNA levels were significantly greater in 75-, 100- and 125-days post-hatch during development. The pre-spawning stage showed higher expression of tgf-β when compared to all other reproductive stages. Immunolocalization study revealed the presence of Tgf-β protein in the preoptic area-hypothalamus which was well correlated with the mRNA expression in discrete regions of the catfish brain. Expression of tgf-β seems to have a role in the regulation of gonadal function through the brain-pituitary axis.

Keywords


Brain, Development, Expression, Signalling Molecule, Tgf-β

References





DOI: https://doi.org/10.18519/jer%2F2022%2Fv26%2F215540