Objective To explore the expression and the changes of microtubule, aquaporin-4 (AQP4) and potassium ion channel 4.1 (Kir4.1) after spinal cord injury in rats.Methods Ninety female adult Sprague-Dawley rats were randomly divided into sham operation group (n=30) and injury group (n=60). The injury group was divided into six hours, one day, three days, five days and seven days subgroups, with twelve rats in each subgroup. Spinal cord injury at T10 was established with modified Allen's method (20 g×25 mm) in the injury group. The water content of spinal cord was measured at each time point after injury. Then, the pathology was observed with HE staining, the expression of α-Tubulin, AQP4 and Kir4.1 was detected and analyzed with immunohistochemical staining and Western blotting.Results The water content of the spinal cord was higher in the injured group than in the sham operation group (P<0.05), and was highest on the fifth day. HE staining showed that the gray matter hemorrhage at six hours after injury; one day after injury, the gray matter bled seriously, and neuron swelling was aggravated; three days after injury, the area of gray matter necrosis increased, and the edema phenomenon was obvious; five days and seven days after injury, the gray matter necrosis and the edema phenomenon were more serious. Western blotting and immunohistochemistry showed that the expression of AQP4 gradually increased after injury, and raised at peak on the fifth day; the expression of α-Tubulin and Kir4.1 was similar, and the expression gradually decreased after injury, especially on the fifth day.Conclusion The expression of α-Tubulin and Kir4.1 is similar after spinal cord injury, and is contrary to the expression of AQP4. α-Tubulin, AQP4 and Kir4.1 may be related after injury and may participate in the formation of spinal cord edema.
CHEN Tie-ge
,
GUO Yong-qiang
,
WANG Ming
,
ZHANG Dong-liang
,
XIA Ya-yi
,
WANG Jing
,
WU Ya-min
,
DANG Yue-xiu
,
ZHANG Hai-hong
. Expression and Change of Microtubule, Aquaporins-4 and K+ Ion Channel Protein-4.1 after Spinal Cord Injury in Rats[J]. Chinese Journal of Rehabilitation Theory and Practice, 2018
, 24(10)
: 1151
-1158
.
DOI: 10.3969/j.issn.1006-9771.2018.10.007
[1] Jin BJ, Hua Z, Binder DK, et al.Aquaporin-4-dependent K+ and water transport modeled in brain extracellular space following neuroexcitation[J]. J Gen Physiol, 2013, 141(1): 119-132.
[2] Zeng XN, Sun XL, Gao L, et al.Aquaporin-4 deficiency down-regulates glutamate uptake and GLT-1 expression in astrocytes[J]. Mol Cell Neurosci, 2007, 34(1): 34-39.
[3] Hubbard JA, Binder DK.Unaltered glutamate transporter-1 protein levels in aquaporin-4 knockout mice[J]. ASN Neuro, 2017, 9(1): 1759091416687846.
[4] Janke C, Bulinski JC.Post-translational regulation of the microtubule cytoskeleton: mechanisms and functions[J]. Nat Rev Mol Cell Biol, 2011, 12(12): 773-786.
[5] Ingber DE. Tensegrity I.Cell structure and hierarchical systems biology[J]. J Cell Sci, 2003, 116(Pt 7): 1157-1173.
[6] Yan X, Liu J, Wang X, et al.Pretreatment with AQP4 and NKCC1 inhibitors concurrently attenuated spinal cord edema and tissue damage after spinal cord injury in rats[J]. Front Physiol, 2018, 9: 6.
[7] Li XQ, Fang B, Tan WF, et al.miR-320a affects spinal cord edema through negatively regulating aquaporin-1 of blood-spinal cord barrier during bimodal stage after ischemia reperfusion injury in rats[J]. BMC Neurosci, 2016, 17: 10.
[8] Borgens RB, Liusnyder P.Understanding secondary injury[J]. Q Rev Biol, 2012, 87(2): 89-127.
[9] Tran LV.Understanding the pathophysiology of traumatic brain injury and the mechanisms of action of neuroprotective interventions[J]. J Trauma Nurs, 2014, 21(1): 30-35.
[10] Hasegawa H, Ma T, Skach W, et al.Molecular cloning of a mercurial-insensitive water channel expressed in selected water-transporting tissues[J]. J Biol Chem, 1994, 269(8): 5497-5500.
[11] Hubbard JA, Hsu MS, Seldin MM, et al.Expression of the astrocyte water channel aquaporin-4 in the mouse brain[J]. Asn Neuro, 2015, 7(5): 1759091415605486.
[12] Nagelhus EA, Mathiisen TM, Ottersen OP.Aquaporin-4 in the central nervous system: cellular and subcellular distribution and coexpression with KIR4.1[J]. Neuroscience, 2004, 129(4): 905-913.
[13] Saadoun S, Papadopoulos MC.Aquaporin-4 in brain and spinal cord oedema[J]. Neuroscience, 2010, 168(4): 1036-1046.
[14] Oklinski MK, Skowronski MT, Skowronska A, et al.Aquaporins in the spinal cord[J]. Int J Mol Sci, 2016, 17(12): E2050.
[15] Olsen ML, Campbell SC, Mcferrin MB, et al.Spinal cord injury causes a wide-spread, persistent loss of Kir4.1 and glutamate transporter 1: benefit of 17ß-oestradiol treatment[J]. Brain , 2010, 133(4):1013-1025.
[16] Najafi E, Stoodley MA, Bilston LE, et al.Inwardly rectifying potassium channel 4.1 expression in post-traumatic syringomyelia[J]. Neuroscience, 2016, 317: 23-35.
[17] Janmey PA.The cytoskeleton and cell signaling: component localization and mechanical coupling[J]. Physiol Rev, 1998, 78(3): 763-781.
[18] Ruschel J, Hellal F, Flynn KC, et al.Axonal regeneration. Systemic administration of epothilone B promotes axon regeneration after spinal cord injury[J]. Science, 2015, 348(6232): 347-352.
[19] Ruschel J, Bradke F.Systemic administration of epothilone D improves functional recovery of walking after rat spinal cord contusion injury[J]. Exp Neurol, 2018, 306: 243-249.
[20] Crunkhorn S.CNS injury: microtubule stabilizer repairs spinal cord injury[J]. Nat Rev Drug Discov, 2015, 14(5): 310.
[21] Zelinski B, Müller N, Kierfeld J.Dynamics and length distribution of microtubules under force and confinement[J]. Phys Rev E Stat Nonlin Soft Matter Phys, 2012, 86(4 Pt 1): 041918.
[22] Akhmanova A, Steinmetz MO.Tracking the ends: a dynamic protein network controls the fate of microtubule tips[J]. Nat Rev Mol Cell Biol, 2008, 9(4):309-322.
[23] Zhao H, Yao X, Wang TX, et al.PKCαregulates vasopressin-induced aquaporin-2 trafficking in mouse kidney collecting duct cells in vitro via altering microtubule assembly[J]. Acta Pharmacol Sin, 2012, 33(2): 230-236.
[24] Conner MT, Conner AC, Brown JE, et al.Membrane trafficking of aquaporin 1 is mediated by protein kinase C via microtubules and regulated by tonicity[J]. Biochemistry, 2010, 49(5): 821-823.
[25] Vossenkämper A, Nedvetsky PI, Wiesner B, et al.Microtubules are needed for the perinuclear positioning of aquaporin-2 after its endocytic retrieval in renal principal cells[J]. Am J Physiol Cell Physiol, 2007, 293(3): 1129-1138.
[26] Sunshine JE, Dagal A, Burns SP, et al.Methylprednisolone therapy in acute traumatic spinal cord injury: analysis of a regional spinal cord model systems database[J]. Anesth Analg, 2017, 124(4): 1200-1205.