Trefoil factor family peptides TFF1 and TFF3 in the nervous tissues of developing mouse embryo


  • Tatjana Belovari Department of Histology and Embryology, Faculty of Medicine, University of Osijek, Osijek
  • Nikola Bijelić Department of Histology and Embryology, Faculty of Medicine, University of Osijek, Osijek
  • Maja Tolušić Levak Department of Histology and Embryology, Faculty of Medicine, University of Osijek, Osijek
  • Mirela Baus Lončar Department of Molecular Medicine, Institute Ruđer Bošković, Zagreb, Croatia



trefoil factor, nervous system, embryonic development, mouse, immunohistochemistry


Trefoil factor family peptides (TFF1, TFF2, and TFF3) are predominantly found in mucous epithelia of various organs. However, they have also been reported in the nervous tissue, particularly mouse, rat, porcine, and human brain. The aim of this research was to determine the presence of TFF1 and TFF3 in the nervous system of developing mouse embryo. Mouse embryos, at the stages E15 to E17 were isolated, fixed in 4% paraformaldehyde and embedded in paraffin blocks. Sagittal 6µm sections were made, processed for immunohistochemistry, and incubated with anti-TFF1 or anti-TFF3 primary polyclonal rabbit antibodies. Labeled streptavidin-biotin method was used for TFF detection. TFF1 and 3 were found in the cytoplasm of ganglion cell somata, while TFF3 staining was also visible in the cytoplasm of neurons in different areas and nuclei of brain and medulla oblongata. Neurons in the gray matter of spinal cord were also TFF1 and TFF3 positive, and signal for both peptides was found in the choroid plexus. TFF peptides might be involved in the complex processes of nervous system development and differentiation and brain plasticity.


Download data is not yet available.


Blin N. Cytoprotective trefoil peptides abound in new functions. Cell Mol Life Sci 2005;62:2907–9.

Hoffmann W, Jagla W, Wiede A. Molecular medicine of TFF-peptides: from gut to brain. Histol Histopathol 2001;16(1):319–34.

Madsen J, Nielsen O, Tornoe I, Thim L, Holmskov U. Tissue Localization of Human Trefoil Factors 1, 2, and 3. J Histochem Cytochem 2007;55:505–13.

Regalo G, Wright NA, Machado JC. Trefoil factors: from ulceration to neoplasia. Cell Mol Life Sci 2005;62(24):2910–5.

Ahmed ARH, Griffiths AB, Tilby MT, Westley BR, May FEB. TFF3 is a normal breast epithelial protein and is associated with differentiated phenotype in early breast cancer but predisposes to invasion and metastasis in advanced disease. Am J Pathol 2012;180(3):904–16.

Chaiyarit P, Utrawichian A, Leelayuwat C, Vatanasapt P, Chanchareonsook N, Samson MH, et al. Investigation of trefoil factor expression in saliva and oral mucosal tissues of patients with oral squamous cell carcinoma. Clin Oral Investig 2012;16(6):1549–56.

Dhar DK, Wang TC, Tabara H, Tonomoto Y, Maruyama R, Tachibana M, et al. Expression of trefoil factor family members correlates with patient prognosis and neoangiogenesis. Clin Cancer Res 2005;11(18):6472–8.

Kannan N, Kang J, Kong X, Tang J, Perry JK, Mohankumar KM, et al. Trefoil factor 3 is oncogenic and mediates anti-estrogen resistance in human mammary carcinoma. Neoplasia 2010;12(12):1041–53.

Kosriwong K, Menheniott TR, Giraud AS, Jearanaikoon P, Sripa B, Limpaiboon T. Trefoil factors: tumor progression markers and mitogens via EGFR/MAPK activation in cholangiocarcinoma. World J Gastroenterol 2011;17(12):1631–41.

Chaiyarit P, Chayasadom A, Wara-Aswapati N, Hormdee D, Sittisomwong S, Nakaresisoon S, et al. Trefoil factors in saliva and gingival tissues of patients with chronic periodontitis. J Periodontol 2012;83(9):1129–38.

Rösler S, Haase T, Claassen H, Schulze U, Schicht M, Riemann D, et al. Trefoil factor 3 is induced during degenerative and inflammatory joint disease, activates matrix metalloproteinases, and enhances apoptosis of articular cartilage chondrocytes. Arthritis Rheum 2010;62(3):815–25.

Wills-Karp M, Rani R, Dienger K, Lewkowich I, Fox JG, Perkins C, et al. Trefoil factor 2 rapidly induces interleukin 33 to promote type 2 immunity during allergic asthma and hookworm infection. J Exp Med 2012;209(3):607–22.

Wright NA, Poulsom R, Stamp G, Van Norden S, Sarraf C, Elia G, et al. Trefoil peptide gene expression in gastrointestinal epithelial cells in inflammatory bowel disease. Scand J Gastroenterol Suppl 1992;193:76–82.

Baus-Loncar M, Kayademir T, Takaishi S, Wang T. Trefoil factor family 2 deficiency and immune response. Cell Mol Life Sci 2005;62(24):2947–55.

Hoffmann W. Trefoil factors TFF (trefoil factor family) peptide-triggered signals promoting mucosal restitution. Cell Mol Life Sci 2005;62(24):2932–8.

Thim L, Madsen F, Poulsen SS. Effect of trefoil factors on the viscoelastic properties of mucus gels. Eur J Clin Invest 2002;32(7):519–27.

Hirota M, Awatsuji H, Sugihara Y, Miyashita S, Furukawa Y, Hayashi K. Expression of pS2 gene in rat brain. Biochem Mol Biol Int 1995;35(5):1079–84.

Hinz M, Schwegler H, Chwieralski CE, Laube G, Linke R, Pohle W, et al. Trefoil factor family (TFF) expression in the mouse brain and pituitary: changes in the developing cerebellum. Peptides 2004;25(5):827–32.

Hirota M, Awatsuji H, Furukawa Y, Hayashi K. Cytokine regulation of PS2 gene expression in mouse astrocytes. Biochem Mol Biol Int 1994;33(3):515–20.

Hoffmann W, Jagla W. Cell type specific expression of secretory TFF peptides: colocalization with mucins and synthesis in the brain. Int Rev Cytol 2002;213:147–81.

Paunel-Görgülü AN, Franke AG, Paulsen FP, Dünker N. Trefoil factor family peptide 2 acts pro-proliferative and pro-apoptotic in the murine retina. Histochem Cell Biol 2011;135(5):461–73.

Jagla W, Wiede A, Dietzmann K, Rutkowski K, Hoffmann W. Co-localization of TFF3 peptide and oxytocin in the human hypothalamus. FASEB J 2000;14(9):1126–31.

Probst JC, Skutella T, Müller-Schmid A, Jirikowski GF, Hoffmann W. Molecular and cellular analysis of rP1.B in the rat hypothalamus: in situ hybridization and immunohistochemistry of a new P-domain neuropeptide. Brain Res Mol Brain Res 1995;33(2):269–76.

Schwarzberg H, Kalbacher H, Hoffmann W. Differential behavioral effects of TFF peptides: injections of synthetic TFF3 into the rat amygdala. Pharmacol Biochem Behav 1999;62(1):173–8.

Schwarz H, Jagla W, Wiede A, Hoffmann W. Ultrastructural co-localization of TFF3-peptide and oxytocin in the neural lobe of the porcine pituitary. Cell Tissue Res 2001;305(3):411–6.

Shi H-S, Yin X, Song L, Guo Q-J, Luo X-H. Neuropeptide Trefoil factor 3 improves learning and retention of novel object recognition memory in mice. Behav Brain Res 2012;227(1):265–9.

Paterson RW, Bartlett JW, Blennow K, Fox NC, Alzheimer's Disease Neuroimaging Initiative, Shaw LM, et al. Cerebrospinal fluid markers including trefoil factor 3 are associated with neurodegeneration in amyloid-positive individuals. Transl Psychiatry 2014;4(7):e419.

Otto WR, Patel K. Trefoil factor family (TFF)-domain peptides in the mouse: embryonic gastrointestinal expression and wounding response. Anat Embryol (Berl) 1999;199(6):499–508.

Gorba T, Bradoo P, Antonic A, Marvin K, Liu D-X, Lobie PE, et al. Neural regeneration protein is a novel chemoattractive and neuronal survival-promoting factor. Exp Cell Res 2006;312(16):3060–74.

Bijelić N, Belovari T, Baus Lončar M. Trefoil factor family protein 3 (TFF3) is present in cartilage during endochondral ossification in the developing mouse fetus. Acta Histochem 2013;115(3):204–8.

Katusić A, Jurić-Lekić G, Jovanov-Milosević N, Vlahović M, Jezek D, Serman L, et al. Development of the fetal neural retina in vitro and in ectopic transplants in vivo. Coll Antropol 2008;32(1):201–7.

Trefoil factor family peptides TFF1 and TFF3 in the nervous tissues of developing mouse embryo


Additional Files





Molecular Biology


How to Cite

Trefoil factor family peptides TFF1 and TFF3 in the nervous tissues of developing mouse embryo. Biomol Biomed [Internet]. 2015 Feb. 1 [cited 2024 May 19];15(1):33-7. Available from: