Distribution of hyperpolarization-activated cyclic nucleotide gated cation channels 4 positive cells in rat myocardial tissue at different developmental stages

doi:10.16098/j.issn.0529-1356.2018.06.010

Abstract

Abstract:

Objective To investigate the changes of hyperpolarization-activated cyclic nucleotide gated cation channels 4(HCN4) expression in the rat heart at different developmental stages and to provide morphological basis for further study of HCN4 on cardiac function. Methods Four groups of rats, 15 rats per group, which were separately born for 1 day, 1 month, 3 months and 6 months were used in this study. The hearts of each group were collected. Paraffin section and frozen section were used to observe the distribution and morphological characteristics of HCN4 positive cells in myocardium of rats at different developmental stages by immunohistochemistry and fluorescence techniques. The protein content of HCN4 in left and right ventricles was observed by Western blotting. Results The immunohistochemical and fluorescence results showed that HCN4 positive cells in 1 day after birth, 1 month, 3 months and 6 months of the vascular smooth muscle cells, endocardial and vascular endothelial cells, epicardial mesothelial cells, fibroblasts, a large number of atrial myocytes, a small amount of cardiomyocytes, and conduction of tissue cells were expressed, but the number of expressed cells was uneven in different parts. Western blotting results showed that the expression of ventricular HCN4 protein decreased with age. Conclusion The expression of HCN4 decreased gradually with the increase of age in the rat myocardium, but the cells with strong proliferative ability are sustained expression. These results suggest that HCN4 may regulate the survival of many cells in myocardium.

Key words: Heart, Hyperpolarization-activated cyclic nucleotide gated cation channels 4, Immunohistochemistry, Immunofluorescence, Western blotting, Rat

ZHANG Ya-nan REN Ming-fen GUO Zhi-kun. Distribution of hyperpolarization-activated cyclic nucleotide gated cation channels 4 positive cells in rat myocardial tissue at different developmental stages[J]. Acta Anatomica Sinica, 2018, 49(6): 752-758.

References

［1］ Ye B, Nerbonne JM. Proteolytic processing of HCN2 and co-assembly with HCN4 in the generation of cardiac pacemaker channels［J］. J Biol Chem, 2009, 284(38): 25 553-25 559.

［2］ Brioschi C, Micheloni S, Tellez JO, et al. Distribution of the pacemaker HCN4 channel mRNA and protein in the rabbit sinoatrial node［J］. J Mol Cell Cardiol,2009, 47(2): 221-227.

［3］ Liang X, Wang G, Lin L, et al. HCN4 dynamically marks the first heart field and conduction system precursors［J］. Cir Res, 2013, 113(4): 399-407.

［4］ Verkerk AO, Wilders R, van Borren MM, et al. Pacemaker current (If) in the human sinoatrial node［J］. Eur Heart J, 2007, 28(20):2472-2478.

［5］ Stieber J, Hofmann F, Ludwig A. Pacemaker channels and sinus node arrhythmia ［J］. Trend Cardiovasc Med,2004,14(1) ∶23-28.

［6］ Fern ández-Velasco M, Goren N, Benito G,et al. Regional distribution of hyperpolarization-activated current (If) and hyperpolarization-activated cyclic nucleotide-gated channel mRNA expression in ventricular cells from control and hypertrophied rat hearts ［J］. J Physiol, 2003, 553(2): 395-405.

［7］ Dobrzynski H, Nikolski VP, Sambelashvili AT, et al. Site of origin and molecular substrate of atrioventricular junctional rhythm in the rabbit heart ［J］. Cir Res, 2003, 93(11): 1102-1110.

［8］ Marionneau C, Couette B, Liu J, et al. Specific pattern of ionic channel gene expression associated with pacemaker activity in the mouse heart ［J］. J physiol, 2005, 562(1): 223-234.

［9］ Chandler NJ,Greener ID,Tellez JO,et al.Molecular architecture of the human sinus node: insights into the function of the cardiac pacemaker［J］. Circulation,2009, 119(12): 1562-1575.

［10］ Garcia-Frigola C, Shi Y, Evans SM. Expression of the hyperpolarization-activated cyclic nucleotide-gated cation channel HCN4 during mouse heart development［J］. Gene Exp Patt, 2003, 3(6): 777-783.

［11］ Stieber J, Herrmann S, Feil S, et al. The hyperpolarization-activated channel HCN4 is required for the generation of pacemaker action potentials in the embryonic heart［J］. Proc Nat Acad of Sci, 2003, 100(25): 15235-15240.

［12］ Zheng YJ. Unnecessity of HCN4 encoded If in pacemaker activity of fetal ventricular cardiomyocyte［D］. Huazhong University of Science and Technology, 2011. (in Chinese)

郑云洁. HCN4 编码的IF在胚胎心室起搏活动中的非必要性［D］. 华中科技大学, 2011.

［13］ Qiao AX, Jing Y, Cai YJ, et al. Expression patterns of hyperpolarization-activated cyclic nucleotide-gated cation 4,connexin 43 and podoplanin in the embryonic mouse heart and development of cardiac conduction system ［J］. Acta Anatomica Sinica, 2012，43(3): 405-411. (in Chinese)

乔爱秀, 景雅, 蔡玉瑾, 等. 超极化激活环核苷酸门控阳离子通道4, 连接蛋白 43 和平足蛋白在小鼠胚胎心的表达与心传导系的发生［J］. 解剖学报, 2012，43(3): 405-411.

［14］ Li CX, Lu X, Guo ZhK. Evidence of mitosis in cardiac myocytes in vitro［J］, Acta Anatomica Sinica，2015,46（1）：63-68. (in Chinese)

李辞霞，卢杏，郭志坤.体外培养状态下心肌细胞有丝分裂的证据，解剖学报，2015, 46 (1):63-68.

［15］ Chang Y, Guo K, Li Q, et al.Multiple directional differentiation difference of neonatal rat fibroblasts from six organs［J］.Cell Physiol Biochem, 2016, 39(1):157-171.

［16］ Bucchi A, Baruscotti M, Robinson RB, et al. Modulation of rate by autonomic agonists in SAN cells involves changes in diastolic depolarization and the pacemaker current［J］. J Mole Cell Cardiol, 2007, 43(1): 39-48.

［17］ Maltsev VA, Lakatta EG. The funny current in the context of the coupled-clock pacemaker cell system［J］,Heart Rhythm,2012,9(2):302-306

［18］ Zicha S, Fernández-Velasco M, Lonardo G, et al. Sinus node dysfunction and hyperpolarization-activated (HCN) channel subunit remodeling in a canine heart failure model［J］. Cardiovasc Res, 2005, 66(3): 472-481.

［19］ Xia S, Wang Y, Zhang Y, et al. Dynamic changes in HCN2, HCN4, KCNE1, and KCNE2 expression in ventricular cells from acute myocardial infarction rat hearts［J］. Biochem Biophysical Res Communic, 2010, 395(3): 330-335.

［20］ Stillitano F, Lonardo G, Zicha S, et al. Molecular basis of funny current (If) in normal and failing human heart［J］. J Mol Cell Cardiol, 2008, 45(2): 289-299.

［21］ Wang QZh, Bao W, Zhou L, et al. Bone marrow mesenchymal stem cell transplantation effect on expression of HCN2 and HCN4 gene following myocardial infarctio［J］. Chinese Journal of Tissue Engineering Research, 2010, 14(1): 86-90. (in Chinese)

王庆志, 包巍, 周立, 等. 大鼠骨髓间充质干细胞移植梗死心肌 HCN2及HCN4基因的表达［J］. 中国组织工程研究, 2010, 14(1): 86-90.

[1]	YANG Meng-yi NIU Shi-bo ZHANG Jing DANG Jie MA Zhan-bing LU Hong HUO Zheng-hao. Association between index finger and ring finger length ratios and polymorphism of homeobox A11 gene locus among Ningxia college students [J]. Acta Anatomica Sinica, 2024, 55(1): 62-66.
[2]	HONG Xiao-min LI San-qiang CUI Qin-yi ZHENG Run-yue YANG Meng-li LUO Ren-li LI Qian-hui. Nuclear factor-κB signaling pathway and gender differences in alcoholic liver fibrosis [J]. Acta Anatomica Sinica, 2024, 55(1): 55-61.
[3]	XIE Hui-lan FANG Fang LIN Yi. Mechanism of Chir99021 regulating Wnt/β-catenin signaling pathway inhibiting osteogenic differentiation of rat dental pulp stem cells [J]. Acta Anatomica Sinica, 2024, 55(1): 67-72.
[4]	WEI Xi-xi LI Chao-hong ZHAO Chen-lu TANG Jia-ping LIU Yu-zhen. Characterization of group Ⅰ metabotropic glutamate receptors in rat superior cervical ganglion and their changes following chronic intermittent hypoxia [J]. Acta Anatomica Sinica, 2024, 55(1): 3-9.
[5]	YUAN Lei YANG Zhi-wei YANG Hui LIU Zheng-hai HE Jie WAN Wei. Panax notoginseng saponin relieving the inflammatory pain caused by complete Freund’s adjuvant by inhibiting the activation of astrocytes in mice [J]. Acta Anatomica Sinica, 2024, 55(1): 25-31.
[6]	MA Ting-ting CHEN Jian-hong LIU Ai-cui LI Hai-ning. Role of inhibiting lncRNA TUG1 to down-regulate nucleotide binding oligomerization domain like receptor protein 1 inflammasome in delaying the progression of Alzheimer’s disease [J]. Acta Anatomica Sinica, 2024, 55(1): 32-42.
[7]	ZOU Cheng-gong FENG Hao CHEN Bing TANG Hui SHAO Chuan SUN Mou YANG Rong HE Jia-quan . Research on the dynamic changes of neurological dysfunction and cognitive function impairment in traumatic brain injury [J]. Acta Anatomica Sinica, 2024, 55(1): 43-48.
[8]	LÜ Hai-yan SHEN Xi-ya ZHAO Fu-sheng ZHU Mei . Effects of tricholoma matsutake polysaccharides on 1-methy-4-pehnyl-pyridine ion-induced PC12 cell damage [J]. Acta Anatomica Sinica, 2024, 55(1): 49-54.
[9]	ZHENG Li-ping LIU Xia DU Xiao-hua WU Ya-juan LIU Shan-shan . Expression and distribution of brain-derived neurotrophic factor in different cerebrum regions of yak and cattle [J]. Acta Anatomica Sinica, 2024, 55(1): 10-16.
[10]	SU Peng WANG Xing-yi LIANG Jing-yan XIONG Tian-qing. Optimization and identification of a low density and high purity method for primary hippocampal neuron culture from fetal rats [J]. Acta Anatomica Sinica, 2024, 55(1): 113-119.
[11]	HAO Yong-ming GUO Lei ZHOU Cheng-hui PEI Han-jun LI Li ZHAO Zhe . Establishment of a rat model of myocardial hypertrophy by a modified abdominal aortic coarctation method [J]. Acta Anatomica Sinica, 2024, 55(1): 120-124.
[12]	XU Ya-ping YU Yue-xin CHEN Jin-fu WANG Ke-ke GUO Zhikun . Structural distribution and mechanism of elastic fibers and collagen fibers in ventricular interstitium of aged rats [J]. Acta Anatomica Sinica, 2023, 54(6): 716-721.
[13]	LIU Li-ping ZHU Meng-ni XU Hui . Effect of interleukin-6 on nucleated erythrocytes in lipopolysaccharide induced preeclampsia rats [J]. Acta Anatomica Sinica, 2023, 54(6): 722-729.
[14]	ZHAO Wei-ming LI Xiao YU Guo-ying WANG Gai-ping CHANG Cui-fang . Effect of serine protease inhibitor Kazaltype 1 on the proliferation of hepatocellular carcinoma cell and its mechanism [J]. Acta Anatomica Sinica, 2023, 54(6): 695-702.
[15]	WANG Wen-juan DING Yu-tong SU Hao REN Jie SUN Yan-rong WANG Han-fei LU Jia-li ZHANG Lin-qian BAI Yu QIN Li-hua. Changes of Nogo-A expression in hippocampus and striatum of rats under low estrogen condition [J]. Acta Anatomica Sinica, 2023, 54(6): 620-627.