Vol 61, No 1 (2023)
Original paper
Published online: 2023-03-17

open access

Page views 2424
Article views/downloads 428
Get Citation

Connect on Social Media

Connect on Social Media

Expression of HIF1α, BNIP3, and beclin-1 in the brain of newborn and adult yaks (Bos grunniens)

Qian Zhang1, Yan Cui1, Sijiu Yu1, Junfeng He1, Yangyang Pan1, Meng Wang1, Gengquan Xu1
Pubmed: 36987743
Folia Histochem Cytobiol 2023;61(1):26-33.


Introduction. As a main consumer of energy, the brain is particularly susceptible to the effects of hypoxia. However, during long-term evolution, the brain of the plateau yak developed adaptive mechanisms enabling it to maintain normal physiological conditions.

Material and methods. A total of 20 male yaks belonging to two age groups [newborns (1–6 days old; n = 10) and adults (3–5 years old; n = 10)] were obtained, and the brain tissue was fixed and processed by standard methods. RT-qPCR, ELISA and IHC assays were used to investigate the expression and localization of HIF1α, BNIP3 and beclin-1 in the hippocampus, cerebral cortex, thalamus, medulla oblongata and cerebellum of newborn and adult yak brains and to explore their potential neuroprotective role.

Results. We found that the expression levels of HIF1α, BNIP3 and beclin-1 at the mRNA and protein levels varied in the different regions of yak brain, with the highest expression observed in the hippocampus, followed by the cerebral cortex, thalamus, medulla oblongata and the cerebellum. Moreover, the HIF1α, BNIP3 and beclin-1 expression were significantly higher in the newborn yaks’ brains than in the adult yak. The IHC results showed that HIF1α, BNIP3 and beclin-1 were mainly distributed in the neurons of the cerebral cortex, hippocampus, thalamus, medulla oblongata and cerebellum. In particular, HIF1α accumulated in the nucleus and cytoplasm. Furthermore, the immunoreactivity of BNIP3 and beclin-1 was concentrated in the cytoplasm.

Conclusions. The results indicate that the yak hippocampus and cerebral cortex may be more resistant to hypoxia than thalamus, medulla oblongata and cerebellum, and the expression of BNIP3 and beclin-1 may be regulated by HIF1α to serve a neuroprotective role in the yak’s brain to adaptation to hypoxia. Additionally, the brain of adult yaks may have a higher tolerance to hypoxia than the brain of newborn yaks.

Article available in PDF format

View PDF Download PDF file


  1. Wilson MH, Newman S, Imray CH. The cerebral effects of ascent to high altitudes. Lancet Neurol. 2009; 8(2): 175–191.
  2. Sharp FR, Bernaudin M. HIF1 and oxygen sensing in the brain. Nat Rev Neurosci. 2004; 5(6): 437–448.
  3. Zhou Y, Huang X, Zhao T, et al. Hypoxia augments LPS-induced inflammation and triggers high altitude cerebral edema in mice. Brain Behav Immun. 2017; 64: 266–275.
  4. Kim BH, Jeziorek M, Kanal HD, et al. Moderately inducing autophagy reduces tertiary brain injury after perinatal hypoxia-ischemia. Cells. 2021; 10(4).
  5. Fan P, Xie XH, Chen CH, et al. Molecular regulation mechanisms and interactions between reactive oxygen species and mitophagy. DNA Cell Biol. 2019; 38(1): 10–22.
  6. Yuan Y, Zhang X, Zheng Y, et al. Regulation of mitophagy in ischemic brain injury. Neurosci Bull. 2015; 31(4): 395–406.
  7. Wu X, Zheng Y, Liu M, et al. BNIP3L/NIX degradation leads to mitophagy deficiency in ischemic brains. Autophagy. 2021; 17(8): 1934–1946.
  8. Bellot G, Garcia-Medina R, Gounon P, et al. Hypoxia-induced autophagy is mediated through hypoxia-inducible factor induction of BNIP3 and BNIP3L via their BH3 domains. Mol Cell Biol. 2009; 29(10): 2570–2581.
  9. Tran S, Fairlie WD, Lee EF. BECLIN1: protein structure, function and regulation. Cells. 2021; 10(6).
  10. He Y, Yu S, Hu J, et al. Changes in the anatomic and microscopic structure and the expression of HIF-1α and VEGF of the yak heart with aging and hypoxia. PLoS One. 2016; 11(2): e0149947.
  11. Yang Bo, Yu S, Cui Y, et al. Histochemical and ultrastructural observations of respiratory epithelium and gland in yak (Bos grunniens). Anat Rec (Hoboken). 2010; 293(7): 1259–1269.
  12. Mawolo J, Du X, Liu X, et al. Expression and distribution of neuroglobin and hypoxia-inducible factor-1α in the diencephalon of young yaks. Medycyna Weterynaryjna. 2021; 77(04): 6515–2021.
  13. Wang DP, Li HG, Li YJ, et al. Hypoxia-inducible factor 1alpha cDNA cloning and its mRNA and protein tissue specific expression in domestic yak (Bos grunniens) from Qinghai-Tibetan plateau. Biochem Biophys Res Commun. 2006; 348(1): 310–319.
  14. Cao K, Dong YT, Xiang J, et al. Reduced expression of SIRT1 and SOD-1 and the correlation between these levels in various regions of the brains of patients with Alzheimer's disease. J Clin Pathol. 2018; 71(12): 1090–1099.
  15. Bergeron M, Yu AY, Solway KE, et al. Induction of hypoxia-inducible factor-1 (HIF-1) and its target genes following focal ischaemia in rat brain. Eur J Neurosci. 1999; 11(12): 4159–4170.
  16. Zhao TB, Ning HX, Zhu SS, et al. Cloning of hypoxia-inducible factor 1alpha cDNA from a high hypoxia tolerant mammal-plateau pika (Ochotona curzoniae). Biochem Biophys Res Commun. 2004; 316(2): 565–572.
  17. Görlach A, Camenisch G, Kvietikova I, et al. Efficient translation of mouse hypoxia-inducible factor-1alpha under normoxic and hypoxic conditions. Biochim Biophys Acta. 2000; 1493(1-2): 125–134.
  18. Chávez JC, Agani F, Pichiule P, et al. Expression of hypoxia-inducible factor-1alpha in the brain of rats during chronic hypoxia. J Appl Physiol (1985). 2000; 89(5): 1937–1942.
  19. Ruscher K, Isaev N, Trendelenburg G, et al. Induction of hypoxia inducible factor 1 by oxygen glucose deprivation is attenuated by hypoxic preconditioning in rat cultured neurons. Neurosci Lett. 1998; 254(2): 117–120.
  20. Chertok VM, Nevzorova VA, Zakharchuk NV. Comparative study of HIF-1α- and HIF-2α-immunopositive neurons and capillaries in rat cortex under conditions of tissue hypoxia. Bull Exp Biol Med. 2018; 165(4): 516–520.
  21. Bani Hashemi S, Braun J, Bernhardt WM, et al. HIF-1alpha subunit and vasoactive HIF-1-dependent genes are involved in carbon monoxide-induced cerebral hypoxic stress response. Eur J Appl Physiol. 2008; 104(1): 95–102.
  22. Field JT, Gordon JW. BNIP3 and Nix: Atypical regulators of cell fate. Biochim Biophys Acta Mol Cell Res. 2022; 1869(10): 119325.
  23. Zhang H, Bosch-Marce M, Shimoda LA, et al. Mitochondrial autophagy is an HIF-1-dependent adaptive metabolic response to hypoxia. J Biol Chem. 2008; 283(16): 10892–10903.
  24. Chinnadurai G, Vijayalingam S, Gibson SB. BNIP3 subfamily BH3-only proteins: mitochondrial stress sensors in normal and pathological functions. Oncogene. 2008; 27 Suppl 1(Suppl 1): S114–S127.
  25. Schmidt-Kastner R, Aguirre-Chen C, Kietzmann T, et al. Nuclear localization of the hypoxia-regulated pro-apoptotic protein BNIP3 after global brain ischemia in the rat hippocampus. Brain Res. 2004; 1001(1-2): 133–142.
  26. Balduini W, Carloni S, Buonocore G, et al. Protective role of autophagy in neonatal hypoxia-ischemia induced brain injury. Neurobiol Dis. 2008; 32(3): 329–339.
  27. Arruri V, Vemuganti R. Role of autophagy and transcriptome regulation in acute brain injury. Exp Neurol. 2022; 352: 114032.
  28. Livieri T, Cuttaia C, Vetrini R, et al. Old and promising markers related to autophagy in traumatic brain injury. Int J Mol Sci. 2022; 24(1).
  29. Zhu L, Qi B, Hou D. Roles of HIF1α- and HIF2α-regulated BNIP3 in hypoxia-induced injury of neurons. Pathol Res Pract. 2019; 215(4): 822–827.
  30. Lu Na, Li X, Tan R, et al. HIF-1α/Beclin1-Mediated autophagy is involved in neuroprotection induced by hypoxic preconditioning. J Mol Neurosci. 2018; 66(2): 238–250.
  31. Zhang J, Ney PA. Role of BNIP3 and NIX in cell death, autophagy, and mitophagy. Cell Death Differ. 2009; 16(7): 939–946.
  32. Fekadu J, Rami A. Beclin-1 deficiency alters autophagosome formation, lysosome biogenesis and enhances neuronal vulnerability of HT22 hippocampal cells. Mol Neurobiol. 2016; 53(8): 5500–5509.
  33. Erlich S, Shohami E, Pinkas-Kramarski R, et al. Closed head injury induces upregulation of Beclin 1 at the cortical site of injury. J Neurotrauma. 2005; 22(7): 750–762.
  34. Guo Y. Role of HIF-1a in regulating autophagic cell survival during cerebral ischemia reperfusion in rats. Oncotarget. 2017; 8(58): 98482–98494.
  35. Ostrowski RP, Zhang JH. The insights into molecular pathways of hypoxia-inducible factor in the brain. J Neurosci Res. 2020; 98(1): 57–76.