Creation and Analysis of Related Genetic Characteristics of BALB/cA.Cg.SHJH hr Mice

doi:10.12300/j.issn.1674-5817.2023.055

Abstract

Abstract:

Objective To introduce the Hr gene of spontaneously mutated SHJH ^hr mice into BALB/cAShjh inbred mice with clear genetic background,and provide a basis for study on the molecular mechanism of Hr gene mutation-induced abnormal phenotype and the application of this model. Methods Using a backcross-intercross breeding method guided by phenotypic monitoring, mutant genes from SHJH ^hr mice bred by spontaneous mutation were introduced into inbred BALB/cAShjh mice by homozygous mutation introgression, and the mice were bred into BALB/cA.Cg.SHJH ^hr (abbreviated as C.Cg.SHJH ^hr ) mice after 10 generations. The genotypes of 90 single nucleotide polymorphism (SNP) detection sites were analyzed in C.Cg.SHJH ^hr mice by multiplex PCR library construction followed by next generation sequencing. Then 14 biochemical locus marker genes were detected in C.Cg.SHJH ^hr mice according to the method of GB/T 14927.1-2008. Finally, whole genome exon sequencing was utilized to detect the mutated genes in this mouse. Results From May 2018 to March 2022, a total of 10 generations of backcross-intercross were conducted to complete the construction of the C.Cg.SHJH ^hr mouse line. Among the 90 SNPs loci detected, except for rs13484115 and rs13484116, all the other loci had the same genotype as the recipient mice BALB/cAShjh. The results of biochemical marker gene detection showed that all the 14 loci of the mouse were the same as those of the recipient mouse. Whole genome exon sequencing found that the mouse had 109 site mutations compared with the recipient mouse strain, including 71 synonymous mutations, 1 stopgain, 37 missense mutations, and 20 genes involved in protein sequence alterations (including the reported Hr gene). Conclusion C.Cg.SHJH ^hr mice were created. Through exon sequencing and genetic analysis, three Hr mutated genes and associated mutated genes that mainly cause phenotypic variations were identified, which provides a basis for expanding the application of C.Cg.SHJH ^hr mice in biomedical research.

Key words: BALB/cA.Cg.SHJH ^hr mice, Exon sequencing, Mutation, Genetic characteristics

CLC Number:

R-332

Xiaoqian TAN, Hao YANG, Huiqing TANG, Wei QU, Liang LI, Zhen QIAN, Jianzhong GU, Ping XU, Junhua XIAO. Creation and Analysis of Related Genetic Characteristics of BALB/cA.Cg.SHJH ^hr Mice[J]. Laboratory Animal and Comparative Medicine, 2023, 43(4): 363-370.

Figures/Tables 6

References 24

1	AHMAD W, FAIYAZ UL HAQUE M, BRANCOLINI V, et al. Alopecia universalis associated with a mutation in the human hairless gene[J]. Science, 1998, 279(5351):720-724. DOI: 10.1126/science.279.5351.720 .
2	AHMAD W, RATTERREE M S, PANTELEYEV A A, et al. Atrichia with papular lesions resulting from mutations in the rhesus macaque (Macaca mulatta) hairless gene[J]. Lab Anim, 2002, 36(1):61-67. DOI: 10.1258/0023677021911777 .
3	BALE S J. Of hairless mice and men: the genetic basis of congenital alopecia universalis/congenital atrichia[J]. J Cutan Med Surg, 1999, 3(6):309-311. DOI: 10.1177/120347549900300607 .
4	BERGMAN R, SCHEIN-GOLDSHMID R, HOCHBERG Z, et al. The alopecias associated with vitamin D-dependent rickets type IIA and with hairless gene mutations: a comparative clinical, histologic, and immunohistochemical study[J]. Arch Dermatol, 2005, 141(3):343-351. DOI: 10.1001/archderm.141.3.343 .
5	PANTELEYEV A A, PAUS R, AHMAD W, et al. Molecular and functional aspects of the hairless (hr) gene in laboratory rodents and humans[J]. Exp Dermatol, 1998, 7(5):249-267. DOI: 10.1111/j.1600-0625.1998.tb00295.x-i1 .
6	SUNDBERG J P, PRICE V H, KING L E Jr. The hairless gene in mouse and man[J]. Arch Dermatol, 1999, 135(6):718-720. DOI: 10.1001/archderm.135.6.718 .
7	ZLOTOGORSKI A, HOCHBERG Z, MIRMIRANI P, et al. Clinical and pathologic correlations in genetically distinct forms of atrichia[J]. Arch Dermatol, 2003, 139(12):1591-1596. DOI: 10.1001/archderm.139.12.1591 .
8	LIU Y, SUNDBERG J P, DAS S, et al. Molecular basis for hair loss in mice carrying a novel nonsense mutation (Hr^rh-R) in the hairless gene (Hr)[J]. Vet Pathol, 2010, 47(1):167-176. DOI: 10.1177/0300985809352970 .
9	唐慧青, 常书福, 于志锋, 等. SHJH ^hr 小鼠部分生物学特性及衰老表型的测定与分析[J]. 实验动物与比较医学,2023, 43(1):44-52. DOI:10.12300/j.issn.1674-5817.2022.069 .
	TANG H Q, CHANG S F, YU Z F, et al. Investigation on biological characteristics and aging phenotype of SHJH ^hr mice[J]. Lab Anim Comp Med, 2023, 43(1): 44-52. DOI:10.12300/j.issn.1674-5817.2022.069 .
10	GAO J F, LI Y C, GUAN Y L, et al. The accelerated aging skin in rhino-like SHJH^hr mice[J]. Exp Dermatol, 2022, 31(10):1597-1606. DOI: 10.1111/exd.14632 .
11	FOSTER H L, DAVID SMALL J, FOX J G. The Mouse in biomedical research[M]. New York: Academic Press, 1981.
12	钱强, 徐园, 王亚恒, 等. 基于多重PCR靶向二代测序的近交系小鼠遗传质量监测方法建立[J]. 实验动物与比较医学, 2019, 39(2): 111-117. DOI: 10.3969/j.issn.1674-5817.2019.02.008 .
	QIAN Q, XU Y, WANG Y H, et al. Mouse genetic quality monitoring method establishment based on next-generation sequencing through multiplex PCR[J]. Lab Anim Comp Med, 2019, 39(2): 111-117. DOI: 10.3969/j.issn.1674-5817.2019.02.008 .
13	SNELL G D. Congenic resistant strains of mice[M]//Origins of inbred mice. Amsterdam: Elsevier, 1978:119-156. DOI: 10.1016/b978-0-12-507850-4.50016-0 .
14	POSEY J E, O'DONNELL-LURIA A H, CHONG J X, et al. Insights into genetics, human biology and disease gleaned from family based genomic studies[J]. Genet Med, 2019, 21(4):798-812. DOI: 10.1038/s41436-018-0408-7 .
15	LUPSKI J R. Clinical genomics: from a truly personal genome viewpoint[J]. Hum Genet, 2016, 135(6):591-601. DOI: 10.1007/s00439-016-1682-6 .
16	KURAMOTO T, KUWAMURA M, TAGAMI F, et al. Kyoto rhino rats derived by ENU mutagenesis undergo congenital hair loss and exhibit focal glomerulosclerosis[J]. Exp Anim, 2011, 60(1):57-63. DOI: 10.1538/expanim.60.57 .
17	CICHON S, ANKER M, VOGT I R, et al. Cloning, genomic organization, alternative transcripts and mutational analysis of the gene responsible for autosomal recessive universal congenital alopecia[J]. Hum Mol Genet, 1998, 7(11):1671-1679. DOI: 10.1093/hmg/7.11.1671 .
18	ZLOTOGORSKI A, AHMAD W, CHRISTIANO A M. Congenital atrichia in five Arab Palestinian families resulting from a deletion mutation in the human hairless gene[J]. Hum Genet, 1998, 103(4):400-404. DOI: 10.1007/s004390050840 .
19	HSIEH J C, SISK J M, JURUTKA P W, et al. Physical and functional interaction between the vitamin D receptor and hairless corepressor, two proteins required for hair cycling[J]. J Biol Chem, 2003, 278(40):38665-38674. DOI: 10.1074/jbc.M304886200 .
20	PANTELEYEV A A, CHRISTIANO A M. The Charles River hairless rat mutation is distinct from the hairless mouse alleles[J]. Comp Med, 2001, 51(1):49-55.
21	POTTER G B, BEAUDOIN G M, DERENZO C L, et al. The hairless gene mutated in congenital hair loss disorders encodes a novel nuclear receptor corepressor[J]. Genes Dev, 2001, 15(20):2687-2701. DOI: 10.1101/gad.916701 .
22	THOMPSON C C, BOTTCHER M C. The product of a thyroid hormone-responsive gene interacts with thyroid hormone receptors[J]. Proc Natl Acad Sci U S A, 1997, 94(16):8527-8532. DOI: 10.1073/pnas.94.16.8527 .
23	SUNDBERG J P, DUNSTAN R W, COMPTON J G. Hairless mouse, HRS/J^hr/hr [M]//Integument and mammary glands. Berlin, Heidelberg: Springer, 1989:192-197. DOI: 10.1007/978-3-642-83749-4_32 .
24	LI Y C, GAO J F, GUAN Y L, et al. Novel rhino-like SHJH ^hr mice with thyroid dysfunction[J]. Zool Res, 2021, 42(6):734-738. DOI: 10.24272/j.issn.2095-8137.2021.211 .

位点Loci	C.Cg.SHJH ^hr	BALB/cA
Car2	b	b
Ce2	a	a
Es1	b	b
Es3	a	a
Es10	a	a
Gpd1	b	b
Gpi1	a	a
Hbb	d	d
Idh1	a	a
Mod1	a	a
Akp1	b	b
Pgm1	a	a
Trf	b	b
Pep3	a	a

位点Loci	C.Cg.SHJH ^hr	BALB/cA
Car2	b	b
Ce2	a	a
Es1	b	b
Es3	a	a
Es10	a	a
Gpd1	b	b
Gpi1	a	a
Hbb	d	d
Idh1	a	a
Mod1	a	a
Akp1	b	b
Pgm1	a	a
Trf	b	b
Pep3	a	a

突变位置 Mutation position	碱基变化 Base alteration	基因 Gene	涉及疾病/细胞功能 Related diseases/cell functions	下游信号通路调控 Signal pathway regulation	PolyPhen-2计算得分 PolyPhen-2 scores
70559773	C→T	Hr	全身性脱毛，伴有丘疹性病变的心房颤动	是	1.000
73523427	A→G	Nudet15	造血干细胞和祖细胞急性损失	是	0.000
74744708	T→C	esd	甲醛分解代谢	是	0.505
74811692	T→C	Lrch1	细胞对趋化因子的反应	否	0.047
74813612	T→C
75031916	G→A	5031414D18Rik	自噬体成熟、糖原、脂质代谢过程的调节	否	0.022
75032403	C→A
75036145	A→G
75125178	A→G	Lrrc63	膜蛋白	否	-
75224148	C→T	Lcp1	肌动蛋白丝束组件	否	0.166
78510551	C→G	Akap11	体型缩小，活动能力减退	否	0.997
79401743	G→A	Mtrf1	线粒体翻译终止；耳聋，痉挛性共济失调	否	0.959
79401763	G→T	Mtrf1
79406608	A→G	Mtrf1
79427505	C→G	Kbtbd	蛋白酶体介导的泛素依赖蛋白的分解代谢	是	0.006
79452831	G→C	Kbtbd	蛋白酶体介导的泛素依赖蛋白的分解代谢	是	0.006
79573278	A→G	Elf1	RNA聚合酶Ⅱ转录的正调控	是	0.000
79595387	G→A	Sugt1	参与动粒功能	是	0.000
79637996	C→G	Lect1	内皮细胞增殖负调控	否	0.005
79656659	A→G
79656692	T→C
86966512	G→A	Diaph3	遗传性听神经病变，先天性造血障碍性贫血	是	0.958
96201334	G→T	Klhl1	肌动蛋白结合蛋白	否	0.001
88469236	C→G	Pcdh20	细胞黏附	否	0.000
99046045	G→C	Mzt1	微管蛋白复合体组成	是	-
99061539	G→C	Bora	蛋白激酶活性的激活，细胞周期过程的调控，蛋白质定位的调节	是	0.995
99068023	C→T	Bora
99068337	G→C
99068407	C→G
99068529	A→G
99068542	A→G
99072637	T→A
99072704	G→A
99084109	C→A	Dis3	CUT分解代谢，RNA加工，rRNA分解代谢	是	0.000
99099549	G→T	Dis3	CUT分解代谢，RNA加工，rRNA分解代谢	是	0.000
99101172	A→G	Pibf1	有丝分裂核分裂，细胞因子产生，蛋白质磷酸化	否	0.982

突变位置 Mutation position	碱基变化 Base alteration	基因 Gene	涉及疾病/细胞功能 Related diseases/cell functions	下游信号通路调控 Signal pathway regulation	PolyPhen-2计算得分 PolyPhen-2 scores
70559773	C→T	Hr	全身性脱毛，伴有丘疹性病变的心房颤动	是	1.000
73523427	A→G	Nudet15	造血干细胞和祖细胞急性损失	是	0.000
74744708	T→C	esd	甲醛分解代谢	是	0.505
74811692	T→C	Lrch1	细胞对趋化因子的反应	否	0.047
74813612	T→C
75031916	G→A	5031414D18Rik	自噬体成熟、糖原、脂质代谢过程的调节	否	0.022
75032403	C→A
75036145	A→G
75125178	A→G	Lrrc63	膜蛋白	否	-
75224148	C→T	Lcp1	肌动蛋白丝束组件	否	0.166
78510551	C→G	Akap11	体型缩小，活动能力减退	否	0.997
79401743	G→A	Mtrf1	线粒体翻译终止；耳聋，痉挛性共济失调	否	0.959
79401763	G→T	Mtrf1
79406608	A→G	Mtrf1
79427505	C→G	Kbtbd	蛋白酶体介导的泛素依赖蛋白的分解代谢	是	0.006
79452831	G→C	Kbtbd	蛋白酶体介导的泛素依赖蛋白的分解代谢	是	0.006
79573278	A→G	Elf1	RNA聚合酶Ⅱ转录的正调控	是	0.000
79595387	G→A	Sugt1	参与动粒功能	是	0.000
79637996	C→G	Lect1	内皮细胞增殖负调控	否	0.005
79656659	A→G
79656692	T→C
86966512	G→A	Diaph3	遗传性听神经病变，先天性造血障碍性贫血	是	0.958
96201334	G→T	Klhl1	肌动蛋白结合蛋白	否	0.001
88469236	C→G	Pcdh20	细胞黏附	否	0.000
99046045	G→C	Mzt1	微管蛋白复合体组成	是	-
99061539	G→C	Bora	蛋白激酶活性的激活，细胞周期过程的调控，蛋白质定位的调节	是	0.995
99068023	C→T	Bora
99068337	G→C
99068407	C→G
99068529	A→G
99068542	A→G
99072637	T→A
99072704	G→A
99084109	C→A	Dis3	CUT分解代谢，RNA加工，rRNA分解代谢	是	0.000
99099549	G→T	Dis3	CUT分解代谢，RNA加工，rRNA分解代谢	是	0.000
99101172	A→G	Pibf1	有丝分裂核分裂，细胞因子产生，蛋白质磷酸化	否	0.982

[1]	SHEN Yan, XU Wang-yang, ZHU Hou-bao. Research Progress on Pathogenesis of Hereditary Diseases Caused by Mutations of Oxidoreductase DHTKD1 and Related Mouse Models [J]. Laboratory Animal and Comparative Medicine, 2018, 38(6): 468-472.
[2]	MA Deng-lei, ZHANG Lan. P301S Mutant Tau Transgenic Mouse and Their Applications [J]. Laboratory Animal and Comparative Medicine, 2017, 37(6): 491-496.
[3]	YANG Jin-lian, LU Yan-min, YANG Fan, ZHENG Hong, QU Cheng-kui, WANG Si-ying. The Effects of Leukemia Related Gain-of-function Mutant SHP-2 on Murine Angiogenesis in vivo [J]. Laboratory Animal and Comparative Medicine, 2012, 32(4): 259-264.
[4]	CHEN Zhen-yue1,2,XU Xiao-lei2,WU Li-qun1,LU Guo-ping1. Premature Aging in Zebrafish Heterozygous breakdance Mutant [J]. , 2010, 30(6): 410-414.
[5]	LIU Yue-huan1，WU Jiu-sheng2, SA Xiao-ying1, SHI Zhang-kui1. The Coat Color Mutants of Mongolian Gerbil and Its Genetics [J]. , 2002, 22(3): 168-171.