1.Deciphering primate retinal aging at single-cell resolution.
Si WANG ; Yuxuan ZHENG ; Qingqing LI ; Xiaojuan HE ; Ruotong REN ; Weiqi ZHANG ; Moshi SONG ; Huifang HU ; Feifei LIU ; Guoqiang SUN ; Shuhui SUN ; Zunpeng LIU ; Yang YU ; Piu CHAN ; Guo-Guang ZHAO ; Qi ZHOU ; Guang-Hui LIU ; Fuchou TANG ; Jing QU
Protein & Cell 2021;12(11):889-898
2.Expression and role of nitric oxide synthase in the testis and epididymis of Macaca fascicularis.
Li SUN ; Ya-Ping REN ; Wei JIANG ; Mei-Yan ZHANG ; Qiao-Yan HOU
National Journal of Andrology 2006;12(10):876-878
OBJECTIVETo investigate the expression and the role of nitric oxide synthase (NOS) in the testis and epididymis of macaca fascicularis.
METHODSThe immunohistochemical ABC method was used to observe the localization of nitric oxide synthase in the testis and epididymis of the macaca fascicularis.
RESULTS(1) nNOS immunoreactivity was found in the spermatogenic cells of seminiferous tubules, the epithelia of epididymal efferent ducts, sperm and the endothelia of blood vessels; (2) iNOS was expressed in the epididymal efferent duct, the sperm inside the duct, and the myoid cells and endothelia of blood vessels; (3) eNOS immunoreactivity was detected in the interstitial cells of the testis, the epididymal efferent duct, the sperm inside the duct, and the myoid cells and endothelia of blood vessels.
CONCLUSIONNOS is extensively expressed in the testis and epididymis of the macaca fascicularis and it may play an important role in such processes as spermatogenesis, sperm maturation and testosterone secretion.
Animals ; Epididymis ; metabolism ; Immunohistochemistry ; Macaca fascicularis ; Male ; Nitric Oxide Synthase ; biosynthesis ; physiology ; Testis ; metabolism
3.Generation of a Hutchinson-Gilford progeria syndrome monkey model by base editing.
Fang WANG ; Weiqi ZHANG ; Qiaoyan YANG ; Yu KANG ; Yanling FAN ; Jingkuan WEI ; Zunpeng LIU ; Shaoxing DAI ; Hao LI ; Zifan LI ; Lizhu XU ; Chu CHU ; Jing QU ; Chenyang SI ; Weizhi JI ; Guang-Hui LIU ; Chengzu LONG ; Yuyu NIU
Protein & Cell 2020;11(11):809-824
Many human genetic diseases, including Hutchinson-Gilford progeria syndrome (HGPS), are caused by single point mutations. HGPS is a rare disorder that causes premature aging and is usually caused by a de novo point mutation in the LMNA gene. Base editors (BEs) composed of a cytidine deaminase fused to CRISPR/Cas9 nickase are highly efficient at inducing C to T base conversions in a programmable manner and can be used to generate animal disease models with single amino-acid substitutions. Here, we generated the first HGPS monkey model by delivering a BE mRNA and guide RNA (gRNA) targeting the LMNA gene via microinjection into monkey zygotes. Five out of six newborn monkeys carried the mutation specifically at the target site. HGPS monkeys expressed the toxic form of lamin A, progerin, and recapitulated the typical HGPS phenotypes including growth retardation, bone alterations, and vascular abnormalities. Thus, this monkey model genetically and clinically mimics HGPS in humans, demonstrating that the BE system can efficiently and accurately generate patient-specific disease models in non-human primates.
Animals
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Disease Models, Animal
;
Female
;
Gene Editing
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Humans
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Lamin Type A/metabolism*
;
Macaca fascicularis
;
Progeria/pathology*
4.Single-nucleus transcriptomics reveals a gatekeeper role for FOXP1 in primate cardiac aging.
Yiyuan ZHANG ; Yandong ZHENG ; Si WANG ; Yanling FAN ; Yanxia YE ; Yaobin JING ; Zunpeng LIU ; Shanshan YANG ; Muzhao XIONG ; Kuan YANG ; Jinghao HU ; Shanshan CHE ; Qun CHU ; Moshi SONG ; Guang-Hui LIU ; Weiqi ZHANG ; Shuai MA ; Jing QU
Protein & Cell 2023;14(4):279-293
Aging poses a major risk factor for cardiovascular diseases, the leading cause of death in the aged population. However, the cell type-specific changes underlying cardiac aging are far from being clear. Here, we performed single-nucleus RNA-sequencing analysis of left ventricles from young and aged cynomolgus monkeys to define cell composition changes and transcriptomic alterations across different cell types associated with age. We found that aged cardiomyocytes underwent a dramatic loss in cell numbers and profound fluctuations in transcriptional profiles. Via transcription regulatory network analysis, we identified FOXP1, a core transcription factor in organ development, as a key downregulated factor in aged cardiomyocytes, concomitant with the dysregulation of FOXP1 target genes associated with heart function and cardiac diseases. Consistently, the deficiency of FOXP1 led to hypertrophic and senescent phenotypes in human embryonic stem cell-derived cardiomyocytes. Altogether, our findings depict the cellular and molecular landscape of ventricular aging at the single-cell resolution, and identify drivers for primate cardiac aging and potential targets for intervention against cardiac aging and associated diseases.
Aged
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Animals
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Humans
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Aging/genetics*
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Forkhead Transcription Factors/metabolism*
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Myocytes, Cardiac/metabolism*
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Primates/metabolism*
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Repressor Proteins/metabolism*
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Transcriptome
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Macaca fascicularis/metabolism*
5.Influence of Ovariectomy on Bone Turnover and Trabecular Bone Mass in Mature Cynomolgus Monkeys.
Jun IWAMOTO ; Azusa SEKI ; Masao MATSUURA ; Yoshihiro SATO ; Tsuyoshi TAKEDA ; Hideo MATSUMOTO ; James K YEH
Yonsei Medical Journal 2009;50(3):358-367
PURPOSE: To examine the influence of ovariectomy (OVX) on bone turnover and trabecular bone mass at the 3 clinically important skeletal sites in mature cynomolgus monkeys. MATERIALS AND METHODS: Six female cynomolgus monkeys, aged 17-21 years, were randomized into 2 groups by the stratified weight: the OVX and sham-operation groups (n = 3 in each group). The experimental period was 16 months. Lumbar bone mineral density (BMD) in vivo and serum and urinary bone turnover markers were longitudinally measured, and peripheral quantitative computed tomographic and bone histomorphometric analyses were performed on trabecular bone of the lumbar vertebra, femoral neck, and distal radius at the end of the experiment. RESULTS: OVX induced in a reduction in lumbar BMD compared with the sham controls and the baseline, as a result of increased serum levels of bone-specific alkaline phosphatase and urinary levels of cross-lined N- and C-terminal telopeptides of type I collagen. Furthermore, OVX induced reductions in trabecular volumetric BMD and trabecular bone mass compared with the sham controls, with increased bone formation rate at the lumbar vertebra, femoral neck, and distal radius. CONCLUSION: The results indicated that OVX in mature cynomolgus monkeys (17-21 years of age) increased bone turnover and induced trabecular bone loss at the three skeletal sites compared with the sham controls. Thus, mature cynomolgus monkeys could be utilized for preclinical studies to examine the effects of interventions on bone turnover and trabecular bone mass at the 3 clinically important skeletal sites.
Alkaline Phosphatase/blood
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Animals
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*Bone Density
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Collagen Type I/urine
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Female
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Femur Neck/metabolism
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Lumbar Vertebrae/metabolism
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Macaca fascicularis/*physiology
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Ovariectomy/*adverse effects
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Radius/metabolism
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Random Allocation
6.Expression in Pichia pastoris and properties of human serum albumin-interferon alpha2b chimera.
Shao-Hong CHANG ; Xin GONG ; Zhi-Yu YANG ; Tong-Ying WANG ; Guo-Chang MA ; Qing-Jun MA ; Jun WU
Chinese Journal of Biotechnology 2006;22(2):173-179
To reduce the serum clearance of interferon alpha2b, a chimeric gene encoding an human serum albumin(HSA)--human interferon alpha2b(IFNalpha2b) fusion protein was overexpressed in Pichia pastoris. After fermentation in a 5L bioreactor, the fusion protein, capable of cross-reacting with anti-IFN alpha and anti-HSA antibody, was purified from the culture of the recombinant yeast by ultrafiltration, blue Sepharose affinity, phenyl hydrophobic interaction and Q ion exchange chromatography. Its IFNa2b moiety exhibits antiviral activity similar to that of recombinant human IFNa2b. In Cynomolgus monkeys model, The fusion protein was detectable in plasma, even 336h after a single does of 90 microg/kg injection intravenously or subcutaneously. The elimination phase half-life of the fusion protein was 101h after intravenous injection and 68.2h after subcutaneous injection. Its Subcutaneous bioavailability was 67.9%. The enhanced pharmacokinetics of interferon a2b fused to human serum albumin suggest its promissing application in clinic medicine.
Animals
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Bioreactors
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microbiology
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Fermentation
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Humans
;
Interferon-alpha
;
biosynthesis
;
genetics
;
Macaca fascicularis
;
Pichia
;
genetics
;
metabolism
;
Recombinant Fusion Proteins
;
biosynthesis
;
genetics
;
pharmacokinetics
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Recombinant Proteins
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Serum Albumin
;
biosynthesis
;
genetics
7.A novel therapeutic anti-HBV antibody with increased binding to human FcRn improves in vivo PK in mice and monkeys.
Ciming KANG ; Lin XIA ; Yuanzhi CHEN ; Tianying ZHANG ; Yiwen WANG ; Bing ZHOU ; Min YOU ; Quan YUAN ; Chi-Meng TZENG ; Zhiqiang AN ; Wenxin LUO ; Ningshao XIA
Protein & Cell 2018;9(1):130-134
8.Intramanchette transport during primate spermiogenesis: expression of dynein, myosin Va, motor recruiter myosin Va, VIIa-Rab27a/b interacting protein, and Rab27b in the manchette during human and monkey spermiogenesis.
Shinichi HAYASAKA ; Yukihiro TERADA ; Kichiya SUZUKI ; Haruo MURAKAWA ; Ikuo TACHIBANA ; Tadashi SANKAI ; Takashi MURAKAMI ; Nobuo YAEGASHI ; Kunihiro OKAMURA
Asian Journal of Andrology 2008;10(4):561-568
AIMTo show whether molecular motor dynein on a microtubule track, molecular motor myosin Va, motor recruiter myosin Va, VIIa-Rab27a/b interacting protein (MyRIP), and vesicle receptor Rab27b on an F-actin track were present during human and monkey spermiogenesis involving intramanchette transport (IMT).
METHODSSpermiogenic cells were obtained from three men with obstructive azoospermia and normal adult cynomolgus monkey (Macaca fascicularis). Immunocytochemical detection and reverse transcription-polymerase chain reaction (RT-PCR) analysis of the proteins were carried out. Samples were analyzed by light microscope.
RESULTSUsing RT-PCR, we found that dynein, myosin Va, MyRIP and Rab27b were expressed in monkey testis. These proteins were localized to the manchette, as shown by immunofluorescence, particularly during human and monkey spermiogenesis.
CONCLUSIONWe speculate that during primate spermiogenesis, those proteins that compose microtubule-based and actin-based vesicle transport systems are actually present in the manchette and might possibly be involved in intramanchette transport.
Actins ; metabolism ; Adult ; Animals ; Biological Transport ; physiology ; Dyneins ; metabolism ; Humans ; Macaca fascicularis ; Male ; Microtubules ; metabolism ; Myosin Heavy Chains ; metabolism ; Myosin Type V ; metabolism ; Myosins ; metabolism ; Spermatids ; cytology ; metabolism ; Spermatogenesis ; physiology ; Testis ; cytology ; metabolism ; Transport Vesicles ; physiology ; Vesicular Transport Proteins ; metabolism ; rab GTP-Binding Proteins ; metabolism
9.Ferumoxide labeled Flk1+ CD31- CD34- human bone marrow mesenchymal stem cells and its in vivo tracing in the brains of Macaca Fascicularis.
Ming FENG ; Ren-Zhi WANG ; Hua ZHU ; Nan ZHANG ; Chang-Jun WANG ; Jun-Ji WEI ; Shan LU ; Qin LI ; Xiao-Ming YIN ; Qin HAN ; Wen-Bin MA ; Chuang QIN ; Chun-Hua ZHAO ; Yi-Hua AN ; Yan-Guo KONG
Acta Academiae Medicinae Sinicae 2008;30(5):559-563
OBJECTIVETo explore the method for labeling Flk1+ CD31- CD34- human bone marrow mesenchymal stem cells (hBMSCs) with ferumoxide-PLL and evaluate the feasibility of its tracing after transplantation into the brains of Macaca Fascicularis.
METHODSThe hBMSCs were incubated with ferumoxide-PLL. Trypan blue staining, Prussian blue staining, and transmission electron microscope were performed to show intracellular iron, marking efficiency, and the vigor of the labeled cells. After the hBMSCs were transplanted into the brains of cynomolgus monkeys by stereotaxis, magnetic resonance imaging (MRI) was performed to trace the cells in vivo. Cell survival and differentiation were studied with immunohistochemistry, Prussian blue staining, and HE staining.
RESULTSThe marking efficiency of the ferumoxide-PLL was 96%. Iron particles were found intracytoplasmic of the hBMSCs by Prussian blue staining and transmission electron microscopy. The relaxation rates of labeled cells in MRI were 4.4 and 4.2 times higher than those of the unlabeled cells. Hypointensity area was found by MRI three weeks after transplantation. Many hBMSCs and new vessels were found in the transplantation zone by pathological and immunofluorescence methods.
CONCLUSIONSFerumoxide-PLL can effectively label hBMSCs and thus increase its contrast in MRI results. The cells can survive in the brains of cynomolgus monkeys. The labeled hBMSCs can be traced in vivo by MRI.
Animals ; Antigens, CD34 ; analysis ; metabolism ; Bone Marrow Cells ; chemistry ; metabolism ; Bone Marrow Transplantation ; Brain ; blood supply ; metabolism ; Brain Chemistry ; Contrast Media ; chemistry ; Dextrans ; Ferrosoferric Oxide ; chemistry ; Humans ; Macaca fascicularis ; Magnetic Resonance Imaging ; methods ; Magnetite Nanoparticles ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; chemistry ; metabolism ; Platelet Endothelial Cell Adhesion Molecule-1 ; analysis ; metabolism ; Staining and Labeling ; methods ; Vascular Endothelial Growth Factor Receptor-2 ; analysis ; metabolism