Type of Article:  Original Research

Volume 7; Issue 3.3 (September 2019)

Page No.: 6929-6933

DOI: https://dx.doi.org/10.16965/ijar.2019.259


Mariyah Hidayat *1, Ihtisham Haider 2, Abdullah Khurram 3, Khalid P. Lone 4.

*1 Associate Professor, Department of Anatomy, Rahbar Medical and Dental College, Lahore, Pakistan.

2 Research student at University of Health Sciences, Lahore, Pakistan.

3 Teaching and research fellow, Department of Anatomy, University of Health Sciences, Lahore, Pakistan.

4 Professor, Department of Physiology & Cell Biology, University of Health Sciences, Lahore, Pakistan.

Address for correspondence: Dr Mariyah Hidayat, Associate Professor, Department of Anatomy, Rahbar Medical and Dental College, Lahore, Pakistan. E-Mail: drmariyah.hidayat@gmail.com


Objective: To immunohistochemically localize Bax proteins in pyramidal cells of cerebral cortex in hypothyroid neonates and to observe the effect of melatonin on these cells.

Study Design: Randomized controlled trial.

Place and duration of study: Animal House and Anatomy Department at University of Health Sciences, Lahore.

Material and Methods: Nine female wistar rats were randomly divided equally into three groups, including control(A), propylthiouracyl(B), melatonin treated propylthiouracyl (C) group and allowed to conceive. Medication was given throughout pregnancy and lactation. After delivery, 10 neonatal rats from each group were sacrificed on 22nd day of life and blood samples were immediately collected from the cardiac region for evaluating serum levels of TSH. The freshly extracted brains were immediately immersed in neutral buffered formalin for 3 weeks and later further processed. 3μm thick sections were sliced from cerebrum, deparaffinized and rehydrated, then washed in phosphate buffered saline and subsequently treated with primary and secondary antibodies.

Results:  PTU group had significant number of pyramidal neurons in cerebral cortex showing signs of apoptosis, whereas melatonin treated group showed reversal of these signs. Moreover, on serum analysis, the pups of dams taking PTU were severely hypothyroid whereas melatonin treated pups showed significant restoration of serum TSH levels.

Conclusion: Melatonin preserves the structure and function of pyramidal neurons of cerebral cortex of neonates if ingested by hypothyroid mothers during gestation and lactation.

Key words: Melatonin, Propylthiouracyl, hypothyroidism, thyroid stimulating hormone, anti-bax antibodies.


  1. Smallridge RC, Ladenson PW. Hypothyroidism in Pregnancy: Consequences to Neonatal Health, The Journal of Clinical Endocrinology & Metabolism, 2001;86(6):2349–2353. https://doi.org/10.1210/jcem.86.6.7577.
  2. Escobar GM, Obregón MJ, Rey FE. Maternal thyroid hormones early in pregnancy and fetal brain development, Best Pract. Res. Clin. Endocrinol. Metab. 2004;18(2):225-48.
  3. El Baba KA, Azar ST. Thyroid dysfunction in pregnancy. Int J Gen Med. 2012;5:227–230. doi:10.2147/IJGM.S27009.
  4. Cheng SY, Leonard JL, Davis PJ. Molecular aspects of thyroid hormone actions. Endocr. Rev. 2010;31(2):139–170. doi:10.1210/er.2009-0007.
  5. Bernal J. Thyroid Hormones in Brain Development and Function. [Updated 2015 Sep 2]. In: Feingold KR, Anawalt B, Boyce A, et al., editors. Endotext [Internet]. South Dartmouth (MA): MDText.com, Inc.; 2000-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK285549.
  6. Singh R, Upadhyay G and Godbole MM. Hypothyroidism alters mitochondrial morphology and induces release of apoptogenic proteins during rat cerebellar development. Journal of Endocrinology,2003;176:321–329.
  7. Kondo, Y. Tamura, J. W. Bawden, S. et al. The immunohistochemical localization of Bax and Bcl-2 and their relation to apoptosis during amelogenesis in developing rat molars. Arch Oral Biol. 2001;46(6): 557–568.
  8. Nita M and Grzybowski A. The Role of the Reactive Oxygen Species and Oxidative Stress in the Pathomechanism of the Age-Related Ocular Diseases and Other Pathologies of the Anterior and Posterior Eye Segments in Adults. Oxidative Medicine and Cellular Longevity, 2016; Article ID 3164734, 23 pages, 2016. https://doi.org/10.1155/2016/3164734.
  9. Ferreira et al., Melatonin: cell death modulator, Rev Assoc Med Bras. 2010;56(6):715-8.
  10. Rios ER. Melatonin: pharmacological aspects and clinical trends. Int J Neurosci. 2010 Sep;120(9):583-90. doi: 10.3109/00207454.2010.492921.
  11. Alonso-Alconada D, Alvarez A, Arteaga O, Martínez-Ibargüen A, Hilario E. Neuroprotective effect of melatonin: a novel therapy against perinatal hypoxia-ischemia. Int J Mol Sci. 2013;14(5):9379–9395. doi:10.3390/ijms14059379.
  12. Alghamdi BS. The neuroprotective role of melatonin in neurological disorders. J Neurosci Res. 2018;96(7):1136–1149. doi:10.1002/jnr.24220.
  13. Moriya T, Horie N, Mitome M, Melatonin influences the proliferative and differentiative activity of neural stem cells.J Pineal Res. 2007;42(4):411–418. doi: 10.1111/j.1600-079X.2007.00435.
  14. Srinivasan V, Spence DW, Pandi-Perumal SR, Brown GM, Cardinali DP. Melatonin in mitochondrial dysfunction and related disorders. Int J. Alzheimers Dis. 2011;326320.
  15. Tan DX, Manchester LC, Qin L, Reiter RJ. Melatonin: A Mitochondrial Targeting Molecule Involving Mitochondrial Protection and Dynamics. Int J Mol Sci. 2016;17(12):2124. Published 2016 Dec 16. doi:10.3390/ijms17122124.
  16. Xiao Q. Apoptosis in the developing cerebellum of the thyroid hormone deficient rat, Frontiers in Bioscience, 1998;3(1):A52-7
  17. Moog NK, Entringer S, Heim C, Wadhwa PD, Kathmann N, & Buss C. Influence of maternal thyroid hormones during gestation on fetal brain development. Neuroscience, 2015;342:68-100.
  18. Sousa AMM, Meyer KA, Santpere G, Gulden FO, Sestan N. Evolution of the Human Nervous System Function, Structure, and Development. Cell. 2017;170(2):226–247. doi:10.1016/j.cell.2017.06.036.
  19. Alkalby JM, Sarah JS. Effect Of Propylthiouracil-Induced Hypothyroidism On Reproductive EffIciency of Adult Male Rats. Bas.j.vet.Res. 2013;12(2).
  20. Lewinski A, Karbownik M. REVIEW. Melatonin and the thyroid gland. Neuro Endocrinol Lett. 2002;23 Suppl 1:73-8.
  21. Garcia-Marin, Fernandez-Santos, Morillo-Bernal, Gordillo-Martinez, Vazquez-Roman, Utrilla, Carrillo-Vico, Guerrero , Martin-Lacave. Melatonin in the thyroid gland: regulation by thyroid-stimulating hormone and role in thyroglobulin gene expression. J Physiol Pharmacol. 2015;66(5):643-52.
  22. Shomrat T, Nesher N. Updated View on the Relation of the Pineal Gland to Autism Spectrum Disorders. Front Endocrinol (Lausanne). 2019;10:37. doi:10.3389/fendo.2019.00037.
  23. Voiculescu, S.E. Zygouropoulos, N., Zahiu, CD, Zagrean, AM. Role of melatonin in embryo fetal development. J. Med. Life 2014;7(4):488–492.
  24. Reiter RJ, Tan DX, Korkmaz A, et al., Melatonin and stable circadian rhythms optimize maternal, placental and fetal physiology, Human Reproduction Update. 2014;2:293–307. https://doi.org/10.1093/humupd/dmt054.

Cite this article: Mariyah Hidayat, Ihtisham Haider, Abdullah Khurram, Khalid P. Lone. THE IMMUNOHISTOCHEMICAL LOCALIZATION OF BAX IN THE BRAIN OF HYPOTHYROID NEONATE DURING MATERNAL MELATONIN INTAKE. Int J Anat Res 2019;7(3.3):6929-6933. DOI: 10.16965/ijar.2019.259