Univocal identification of retinal ganglion cells(RGCs) is an essential prerequisite for studying their degeneration and neuroprotection. Before the advent of phenotypic markers, RGCs were normally identified using re...Univocal identification of retinal ganglion cells(RGCs) is an essential prerequisite for studying their degeneration and neuroprotection. Before the advent of phenotypic markers, RGCs were normally identified using retrograde tracing of retinorecipient areas. This is an invasive technique, and its use is precluded in higher mammals such as monkeys. In the past decade, several RGC markers have been described. Here, we reviewed and analyzed the specificity of nine markers used to identify all or most RGCs, i.e., pan-RGC markers, in rats, mice, and macaques. The best markers in the three species in terms of specificity, proportion of RGCs labeled, and indicators of viability were BRN3A, expressed by vision-forming RGCs, and RBPMS, expressed by vision-and non-vision-forming RGCs. NEUN, often used to identify RGCs, was expressed by non-RGCs in the ganglion cell layer, and therefore was not RGC-specific. γ-SYN, TUJ1, and NF-L labeled the RGC axons, which impaired the detection of their somas in the central retina but would be good for studying RGC morphology. In rats, TUJ1 and NF-L were also expressed by non-RGCs. BM88, ERRβ,and PGP9.5 are rarely used as markers, but they identified most RGCs in the rats and macaques and ERRβ in mice. However, PGP9.5 was also expressed by non-RGCs in rats and macaques and BM88 and ERRβ were not suitable markers of viability.展开更多
Objectives: To explore the role of the growth of new nerves (neo-neurogenesis) in the tumorogenesis of squamous cell carcinoma of tongue. Materials and Methods: 10 formalin-fixed specimens were gained from patients di...Objectives: To explore the role of the growth of new nerves (neo-neurogenesis) in the tumorogenesis of squamous cell carcinoma of tongue. Materials and Methods: 10 formalin-fixed specimens were gained from patients diagnosed with tongue squamous cell carcinoma. Animal models were made by subcutaneous injection in the dorsal midline with Tca-8113 cell line. Mice were sacrificed 2, 4, 6 weeks after cell injection, and tumor tissues were fixed in 4% paraformaldehyde, embedded in paraffin, sectioned. Detection of neo-neurogenesis was stained by Neurofila-ment-L antibody (NF-L) using immunohistochemistry method (IHC) in biopsy from both human body and animal model. Results: IHC staining of NF-L is positive in all 10 paraffins of tongue squamous cell carcinoma sections which suggest that newly formed nerves are observed in tumor microenvironment. NF-L staining is also positive in the paraffins from animal models indicating that the tongue cancer recruits newly formed nerves in its tumorogenesis. Conclusions: Tumor neo-neurogenesis may play an important role in the pathogenesis and development of tongue cancer. From a therapeutic perspective, further studies on the topic may provide new clinical opportunity.展开更多
Neurofilaments (NFs) and cytokeratins are both heteropolymers, which assemble into intermediate filaments (IPs) only when other proper IF subunit proteins are expressed simultaneously. To study the assembly property o...Neurofilaments (NFs) and cytokeratins are both heteropolymers, which assemble into intermediate filaments (IPs) only when other proper IF subunit proteins are expressed simultaneously. To study the assembly property of NFs, we constructed two recombinant adenovirus which could express NF-L or NF-M, fused with green fluorescent protein (GFP) respectively. Then they were introduced into vero cells, and expressed fusion protein. Double labels of GFP fluorescence and immunofluorescence staining indicated that NF-L-GFP or GFP-NF-M not only coassembled with endogenous vimentins, but also coassembled with keratins into a cytoplasmic network of filaments.展开更多
基金supported by the Spanish Ministry of Economy and Competitiveness(PID2019-106498GB-I0)Instituto de Salud Carlos III,Fondo Europeo de Desarrollo Regional“Una manera de hacer Europa”(PI19/00071)+2 种基金Fundación Séneca,Agencia de Ciencia y Tecnología Región de Murcia(19881/GERM/15)Spanish Ministry of Science and Innovation(PID 2019-106498 GB-I00)Intramural Research Program of the National Eye Institute,National Institutes of Health(NIH/NEI RO1 EY029087)。
文摘Univocal identification of retinal ganglion cells(RGCs) is an essential prerequisite for studying their degeneration and neuroprotection. Before the advent of phenotypic markers, RGCs were normally identified using retrograde tracing of retinorecipient areas. This is an invasive technique, and its use is precluded in higher mammals such as monkeys. In the past decade, several RGC markers have been described. Here, we reviewed and analyzed the specificity of nine markers used to identify all or most RGCs, i.e., pan-RGC markers, in rats, mice, and macaques. The best markers in the three species in terms of specificity, proportion of RGCs labeled, and indicators of viability were BRN3A, expressed by vision-forming RGCs, and RBPMS, expressed by vision-and non-vision-forming RGCs. NEUN, often used to identify RGCs, was expressed by non-RGCs in the ganglion cell layer, and therefore was not RGC-specific. γ-SYN, TUJ1, and NF-L labeled the RGC axons, which impaired the detection of their somas in the central retina but would be good for studying RGC morphology. In rats, TUJ1 and NF-L were also expressed by non-RGCs. BM88, ERRβ,and PGP9.5 are rarely used as markers, but they identified most RGCs in the rats and macaques and ERRβ in mice. However, PGP9.5 was also expressed by non-RGCs in rats and macaques and BM88 and ERRβ were not suitable markers of viability.
文摘Objectives: To explore the role of the growth of new nerves (neo-neurogenesis) in the tumorogenesis of squamous cell carcinoma of tongue. Materials and Methods: 10 formalin-fixed specimens were gained from patients diagnosed with tongue squamous cell carcinoma. Animal models were made by subcutaneous injection in the dorsal midline with Tca-8113 cell line. Mice were sacrificed 2, 4, 6 weeks after cell injection, and tumor tissues were fixed in 4% paraformaldehyde, embedded in paraffin, sectioned. Detection of neo-neurogenesis was stained by Neurofila-ment-L antibody (NF-L) using immunohistochemistry method (IHC) in biopsy from both human body and animal model. Results: IHC staining of NF-L is positive in all 10 paraffins of tongue squamous cell carcinoma sections which suggest that newly formed nerves are observed in tumor microenvironment. NF-L staining is also positive in the paraffins from animal models indicating that the tongue cancer recruits newly formed nerves in its tumorogenesis. Conclusions: Tumor neo-neurogenesis may play an important role in the pathogenesis and development of tongue cancer. From a therapeutic perspective, further studies on the topic may provide new clinical opportunity.
基金This work was supported by the National Science Found for Distinguished Young Scholars (Grant No. 39425009).
文摘Neurofilaments (NFs) and cytokeratins are both heteropolymers, which assemble into intermediate filaments (IPs) only when other proper IF subunit proteins are expressed simultaneously. To study the assembly property of NFs, we constructed two recombinant adenovirus which could express NF-L or NF-M, fused with green fluorescent protein (GFP) respectively. Then they were introduced into vero cells, and expressed fusion protein. Double labels of GFP fluorescence and immunofluorescence staining indicated that NF-L-GFP or GFP-NF-M not only coassembled with endogenous vimentins, but also coassembled with keratins into a cytoplasmic network of filaments.
