RESUMO
Animals evolve diverse pigment patterns to adapt to the natural environment. Countershading, characterized by a dark-colored dorsum and a light-colored ventrum, is one of the most prevalent pigment patterns observed in vertebrates. In this study, we reveal a mechanism regulating xanthophore countershading in zebrafish embryos. We found that csf1a and csf1b mutants altered xanthophore countershading differently: csf1a mutants lack ventral xanthophores, while csf1b mutants have reduced dorsal xanthophores. Further study revealed that csf1a is expressed throughout the trunk, whereas csf1b is expressed dorsally. Ectopic expression of csf1a or csf1b in neurons attracted xanthophores into the spinal cord. Blocking csf1 signaling by csf1ra mutants disrupts spinal cord distribution and normal xanthophores countershading. Single-cell RNA sequencing identified two col1a2+ populations: csf1ahighcsf1bhigh muscle progenitors and csf1ahighcsf1blow fibroblast progenitors. Ablation of col1a2+ fibroblast and muscle progenitors abolished xanthophore patterns. Our study suggests that fibroblast and muscle progenitors differentially express csf1a and csf1b to modulate xanthophore patterning, providing insights into the mechanism of countershading.
Assuntos
Pigmentação , Peixe-Zebra , Animais , Peixe-Zebra/metabolismo , Pigmentação/genética , Proteínas de Peixe-Zebra/genética , Proteínas de Peixe-Zebra/metabolismo , MúsculosRESUMO
PURPOSE: This study aimed to investigate the effect of the N6-methyladenosine (m6A) dependent ferroptosis on cisplatininduced Sertoli cell injury. MATERIALS AND METHODS: A cisplatin exposure mouse model was established by intraperitoneal injection of cisplatin in our study. TM4 cell lines was used for in vitro study. Ferroptosis was detected according to metabolomic analysis and a series of assays, including malondialdehyde, glutathione, and glutathione disulfide concentration detection, 2',7'-dichlorodihydrofluorescein diacetate and BODIPY 581/591 C11 probe detection, and transmission electron microscope imaging. Key ferroptosis-related genes were identified via transcriptomic analysis, western blot and immunohistochemistry. The m6A modification was demonstrated via m6A RNA immunoprecipitation and luciferase reporter assays. Immune cell infiltration was detected by mass cytometry, and verified by flow cytometry and immunofluorescence. RESULTS: Ferroptosis, but not other types of programmed cell death, is a significant phenomenon in cisplatin-induced testis damage and Sertoli cell loss. Ferroptosis induced by cisplatin in Sertoli cell/TM4 cell is GPX4 independent but is regulated by SLC7A11 and ALOX12. Both SLC7A11 and ALOX12 are regulated via m6A dependent manner by METTL3. Furthermore, overexpressed ALOX12-12HETE pathway may result in macrophage polarization and inflammatory response in cisplatin exposure testis. CONCLUSIONS: Cisplatin-induced Sertoli cell injury via ferroptosis and promoted ferroptosis in an m6A dependent manner. m6A modification of both SLC7A11 and ALOX12 mRNA could result in ferroptosis in our in vitro model. Further, overexpressed ALOX12 can cause more production of 12-HETE, which may be responsible for testis inflammation caused by cisplatin.
RESUMO
Bladder cancer (BC) is the second most common urological disease worldwide. Previous studies have reported that microRNA (miR)165p is associated with the development of BC, but whether miR165p regulates BC cell autophagy remains unknown. Thus, the aim of the present study was to investigate this issue. miR165p expression in BC cells was assessed by reverse transcriptionquantitative PCR. Cell viability and apoptosis were detected via Cell Counting Kit8 and flow cytometry assays, respectively. For cell autophagy detection, autophagic flux was detected using a mCherrygreen fluorescent proteinmicrotubuleassociated proteins 1A/1B light chain 3B (LC3) puncta formation assay, followed by determination of autophagyrelated protein markers. The targeting relationship between miR165p and caspase recruitment domain family member 10 (BIMP1) was confirmed using a dualluciferase reporter assay, followed by detection of the BIMP1/NFκB signaling pathway. The results showed that miR165p overexpression inhibited cell viability, whereas miR165p knockdown promoted cell viability in BC. Furthermore, miR165p overexpression induced autophagy, which was accompanied by increased autophagic flux and expression of the autophagyrelated proteins LC3II and beclin 1, as well as decreased p62 expression, whereas miR165p silencing led to an inhibition of autophagy in BC cells. Moreover, autophagy inhibitor 3methyladenine treatment inhibited cell autophagy and apoptosis in miR165poverexpressing cells. Mechanistic studies demonstrated that miR165p could inhibit the BIMP1/NFκB signaling pathway and this inhibition was achieved by directly targeting BIMP1. Furthermore, it was found that blockade of the BIMP1/NFκB signaling pathway inversed the inhibitory effects of miR165p knockdown on autophagy in BC cells. In vivo experiments further verified the tumorsuppressive effect on BC of the miR165p/BIMP1/NFκB axis. Therefore, the results of the present study indicated that miR165p promotes autophagy of BC cells via the BIMP1/NFκB signaling pathway, and an improved understanding of miR165p function may provide therapeutic targets for clinical intervention in this disease.
Assuntos
Proteínas Relacionadas à Autofagia/metabolismo , Autofagia , Regulação Neoplásica da Expressão Gênica , MicroRNAs/fisiologia , Transdução de Sinais , Neoplasias da Bexiga Urinária/genética , Neoplasias da Bexiga Urinária/metabolismo , Animais , Apoptose , Proteína Beclina-1/metabolismo , Biomarcadores Tumorais , Proteínas Adaptadoras de Sinalização CARD/metabolismo , Carcinoma/genética , Carcinoma/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular , Técnicas de Silenciamento de Genes , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Proteínas Associadas aos Microtúbulos/metabolismo , NF-kappa B/metabolismo , Proteínas de Ligação a RNA/metabolismoRESUMO
OBJECTIVE: Testicular mixed nonseminomatous germ cell cancer (TMNGCC) is rarely reported. This study aimed to explore the clinical symptoms, pathological characteristics and treatment methods of TMNGCC. METHODS: We analyzed the clinical data of 1 case of TMNGCC, observed its pathological characteristics under the light microscope by histology, cytochemistry, immunohistochemistry and immune marking, and investigated the clinical features of such tumors by reviewing the relevant literature. RESULTS: The patient presented with a chief complaint of painless testicular swelling for 3 years. Histopathological examinations revealed a tumor of papillary, fissural or adenoid structure, with large polygonal or columnar cells with one or more irregular vesicular nuclei, the nuclear membrane clear, the cytoplasm eosinophilic or basophilic, and the interstitium infiltrated by a few lymphocytes. Here are the immunohistochemical results: CD117 -, CK8-18 + +, CD30 + +, CK + + +, vimentin -, PLAP +/-, P53 +, AFP + and EMA + +. The tumor was pathologically diagnosed as teratogenic embryonic testicular cancer, and treated by radical surgery, followed by adjuvant chemotherapy according to the treatment of TMNGCC. One-year follow-up found the patient to be alive. CONCLUSION: TMNGCC is a rare malignant tumor, mostly with unobvious clinical symptoms. Its diagnosis primarily depends on physical examination, ultrasonography, CT, and measurement of serum tumor markers; its confirmation necessitates pathological examination, and its first-choice treatment is surgical resection.