Comparative analysis of the Down syndrome hippocampal non-coding RNA transcriptomes using a mouse model

Zhaowei Cai,Zhilan Xiao,Yufang Wang,Huazhen Liu,Kangdi Zhang,Xiaoning Zhen,Xiaoling Jiang 한국유전학회 2020 Genes & Genomics Vol.42 No.11

Background Down syndrome (DS), caused by trisomy 21, is the most common human chromosomal disorder. Hippocampalabnormalities have been believed to be responsible for the DS developmental cognitive deficits. Cumulative evidences indicatedthat non-coding RNAs (ncRNAs) participated in brain development and function. Currently, few was known whetherdysregulated ncRNAs existed in DS whether the dysregulated ncRNAs played important pathology roles in DS. Objective The purpose of this study was generating an overview map of the dysregulated ncRNAs in DS, including themicroRNA (miRNA), long ncRNA (lncRNA) and circular RNA (circRNAs). DS mouse models are invaluable tools forfurther mechanism and therapy studies. Methods The well-studied DS mouse model Dp(16)1/Yey was used in this study as it contains the trisomy of the whole humanchromosome 21 syntenic region on mouse chromosomes 16. Hippocampi were isolated from pups of seven-days-old. Librariesfor miRNA, lncRNA and circRNAs were constructed separately, and the next generation sequencing method was utilized. Results Differentially expressed (DE) miRNAs, lncRNAs and circRNAs were reported. Relative few regulating relationshipwere found between the DE miRNAs and DE mRNAs. LncRNAs originated from the trisomic regions expressed in clusters,but not all of them were 1.5-fold increased expressed. Dramatic DE circular RNAs were found in the DS hippocampus. The host genes of the DE circRNAs were enriched on functions which were well-known impaired in DS, e.g. long-termpotentiation,glutamatergic synapse, and GABAergic synapse. Conclusions We generated the first DS developmental hippocampal ncRNA transcriptome map. This work laid foundationsfor further investigations on role of ncRNAs in hippocampal functions.

Plumbagin inhibits proliferation and promotes apoptosis of ovarian granulosa cells in polycystic ovary syndrome by inactivating PI3K/Akt/mTOR pathway

Cai Zhaowei,He Shaojuan,Li Tao,Zhao Li,Zhang Kerong 한국통합생물학회 2020 Animal cells and systems Vol.24 No.4

Polycystic ovary syndrome (PCOS) is recognized as a general endocrine disease and reproductive disorder. Although evidence indicates that PCOS has a complex etiology and genetic basis, the pathogenic mechanisms and signal pathway in PCOS remain unclear. In this study, the normal structure of follicle and corpus luteum were observed, and no cyst nor hyperemia was observed under the light microscopic study with hematoxylin and eosin (H&E) staining. Eestosterone and progesterone were evaluated by radioimmunoassay in rat serum. The alterations of proliferative ability and cell cycle distribution of each group were assessed by Cell Counting Kit-8 (CCK8) assay and flow cytometry. The protein expression of p-mTOR/mTOR, p-PI3K/PI3K, p-AKT/AKT, and GAPDH were analyzed by western blotting. Both doses of PLB could benefit the ovarian morphology and polycystic property. PLBinduced a suppress effect on the proliferation of rat ovarian granulosa cells. In addition, PLB also induced concentrationdependent apoptosis in rat ovarian granulosa cells. The rat ovarian granulosa cells treated with PLB that the expression levels of p-AKT, p-mTOR, and p-PI3K were significantly decreased in a concentration-dependent manner. PLB not only plays a critical role in attenuating the pathology and polycystic property changes in the ovary but can also induce rat ovarian granulosa cell apoptosis through the PI3K/Akt/mTOR signal pathway. This study showed the innovative role of PLB in the pathogenesis of PCOS and provides a new therapeutic modality for the treatment of PCOS.

Effects of Castration on Androgen Receptor, IGF-I Ea, MGF and Myostatin Gene Expression in Skeletal Muscles of Male Pigs

Yao, Yuchang,Cai, Zhaowei,Zhang, Lifan,Zhao, Chunjiang,Wu, Keliang,Xu, Ningying,Liu, Gang,Wu, Changxin Asian Australasian Association of Animal Productio 2009 Animal Bioscience Vol.22 No.8

Castration of male pig produces significant negative effects on skeletal muscle development. The androgen receptor (AR), two splice variants of insulin-like growth factor-I (IGF-I Ea and MGF) and the myostatin gene may play important roles in this process. In the present study, the expression of AR, IGF-I Ea, MGF and myostatin genes in three skeletal muscles, the brachialis, longissimus and semitendinosus, were studied using real-time quantitative RT-PCR. Our experimental design used 14 pairs of male Landrace sire${\times}$Yorkshire dam piglets. The two piglets in each pair were full sibs, one of which was castrated at 21 d of age; the other remained intact. The study group was divided into subgroups of equal size. Animals in the first subgroup were slaughtered at 147 d and those of the second at 210 d of age. Carcass weight and lean meat yield were similar between boars and barrows at 147 d of age (p>0.05), whereas barrows had lower carcass weight and less lean meat yield at 210 d of age (p<0.05). Castration caused down-regulation of AR gene expression at both 147 and 210 d of age (p<0.05). The two splice variants of the IGF-I gene from porcine skeletal muscle were cloned using RT-PCR, and it was found that MGF differs from IGF-I Ea in having a 52-base insert in the last coding exon of the mRNA. Both splice variants were down-regulated by castration only at 210 d of age (p<0.05). No differences in expression of the myostatin gene were observed between boars and barrows at either 147 or 210 d of age (p>0.05). These results suggest that the downregulation of AR, IGF-I Ea and MGF gene expression following castration helps to explain the negative effect of castration on skeletal muscle development.

Multiple Tolerances and Dye Decolorization Ability of a Novel Laccase Identified from Staphylococcus Haemolyticus

Xingxing Li,Dongliang Liu,Zhaowei Wu,Dan Li,Yifei Cai,Yao Lu,Xin Zhao,Huping Xue 한국미생물·생명공학회 2020 Journal of microbiology and biotechnology Vol.30 No.4

Laccases are multicopper oxidases with important industrial value. In the study, a novel laccase gene (mco) in a Staphylococcus haemolyticus isolate is identified and heterologously expressed in Escherichia coli. Mco shares less than 40% of amino acid sequence identities with the other characterized laccases, exhibiting the maximal activity at pH 4.0 and 60oC with 2,2'-azino-bis (3- ethylbenzothiazoline-6-sulphonic acid) diammonium salt (ABTS) as a substrate. Additionally, the Mco is tolerant to a wide range of pH, heavy metal ions and many organic solvents, and it has a high decolorization capability toward textile dyes in the absence of redox mediators. The characteristics of the Mco make this laccase potentially useful for industrial applications such as textile finishing. Based on BLASTN results, mco is found to be widely distributed in both the bacterial genome and bacterial plasmids. Its potential role in oxidative defense ability of staphylococci may contribute to the bacterial colonization and survival.

Single-nucleus RNA sequencing reveals cell type-specific transcriptome alterations of Down syndrome hippocampus using the Dp16 mouse model

Zhou Zuolin,Zhi Chunchun,Chen Die,Cai Zhaowei,Jiang Xiaoling 한국유전학회 2023 Genes & Genomics Vol.45 No.10

Background Down syndrome (DS), the most frequently occurring human chromosomal disorder, is caused by trisomy 21. The exact molecular effects of trisomy on certain cell populations in the brain remain poorly understood. Objective The purpose of this study was to investigate the effects of trisomy on the transcriptomes of various types of neurons and nonneuronal cells in the hippocampus. Methods A total of 8993 nuclei from the WT and 6445 nuclei from the Dp16 hippocampus were analyzed by single-nucleus RNA sequencing (snRNA-seq). Cell clustering was achieved by the Seurat program. Results Hippocampal cells were grouped into multiple neuronal and nonneuronal populations. Only a limited number of trisomic genes were upregulated (q < 0.001) over 1.25-fold in a specific type of hippocampal cell. Specifically, deregulation of genes associated with synaptic signaling and organization was observed in multiple cell populations, including excitatory neurons, oligodendrocytes, and microglia. This observation suggests the potential importance of synapse deficits in DS. Interestingly, GO annotation of the upregulated genes suggested potential activation of the immune system by hippocampal excitatory neurons. Fewer trisomic genes were altered in nonneuronal cells than in neurons. Notably, microglial transcriptome analysis revealed significantly (q < 0.001) increased expression of C1qb and C1qc, which suggested potential involvement of complement-mediated synapse loss mediated by microglia in DS. Conclusion The trisomy-related hippocampal deficits should be driven by a small amount, not all, of the trisomic genes in a specific type of cell. Our work may help to narrow down both the molecular and cellular targets for future gene therapies in DS.