In the present study,the effects of postnatal development on the number and distribution of skeletal muscle fibers of different types in hind leg of rat and rabbit were studied.The soleus muscles of rats aged 2 days,2...In the present study,the effects of postnatal development on the number and distribution of skeletal muscle fibers of different types in hind leg of rat and rabbit were studied.The soleus muscles of rats aged 2 days,2,4,6,8,and 10 weeks (body weight 10,32,95,190,280,and 320 g),and rabbits aged 2 days,2,4,8,12,16,20,and 24 weeks (body weight 100,220,400,750,1 200,1 600,2 100,and 2 500 g) were stained with succinic dehydrogenase.With an image analysis system,the X-Y coordinates of fibers were used to analyze the growth-related changes.The results of present study showed that three types of fibers were found in the soleus muscles of rat and rabbit,i.e.,type Ⅰ (slow oxidative),ⅡX (fast oxidative),and ⅡA (fast oxidative glycolytic).The type Ⅰ fibers were present throughout the muscle that had a uniform distribution and tended to increase in number with aging.Type ⅡX fibers were scattered throughout the muscle and decreased markedly in number with aging.Type ⅡA fibers were located at the central and deep regions,and showed a little or no change in number and distribution with aging.While be of age,type ⅡA and ⅡX fibers became restricted to the superficial region.No type ⅡB fibers were detected.Type ⅡA fibers had the largest diameter,type Ⅰ intermediate and type ⅡX the smallest.Mean cross-sectional area of each type fibers of rabbits was larger than that of rats.The present results indicate that the number and distribution of muscle fibers of different types in hind limb of rat and rabbit change with the process of postnatal growth.展开更多
Our previous studies showed that the early use of calpain inhibitors reduces calpain activity in multiple brain regions, and that postnatal treatment with calpeptin may lead to cerebellar motor dysfunction. However, i...Our previous studies showed that the early use of calpain inhibitors reduces calpain activity in multiple brain regions, and that postnatal treatment with calpeptin may lead to cerebellar motor dysfunction. However, it remains unclear whether postnatal calpeptin application affects hippocampus-related behaviors. In this study, Sprague-Dawley rats were purchased from the Animal Center of Anhui Medical University of China. For the experiments in the adult stage, rats were intraperitoneally injected with calpeptin, 2 mg/kg, once a day, on postnatal days 7–14. Then on postnatal day 60, the Morris water maze test was used to evaluate spatial learning and memory abilities. The open field test was carried out to assess anxiety-like activities. Phalloidin staining was performed to observe synaptic morphology in the hippocampus. Immunohistochemistry was used to count the number of NeuN-positive cells in the hippocampal CA1 region. DiI was applied to label dendritic spines. Calpeptin administration impaired spatial memory, caused anxiety-like behavior in adulthood, reduced the number and area of apical dendritic spines, and decreased actin polymerization in the hippocampus, but did not affect the number of NeuN-positive cells in the hippocampal CA1 region. For the neonatal experiments, neonatal rats were intraperitoneally injected with calpeptin, 2 mg/kg, on postnatal days 7 and 8. Western blot assay was performed to analyze the protein levels of Akt, Erk, p-Akt, p-Erk1/2, Erk1/2, SCOP, PTEN, mTOR, p-mTOR, CREB and p-CREB in the hippocampus. SCOP expression was increased, and the phosphorylation levels of Akt, mTOR and CREB were reduced in the hippocampus. These findings show that calpeptin administration after birth affects synaptic development in neonatal rats by inhibiting the Akt/mTOR signaling pathway, thereby perturbing hippocampal function. Therefore, calpeptin administration after birth is a risk factor for neurodevelopmental defects.展开更多
The organotypic retinal explant culture has been established for more than a decade and offers a range of unique advantages compared with in vivo experiments and cell cultures.However,the lack of systematic and contin...The organotypic retinal explant culture has been established for more than a decade and offers a range of unique advantages compared with in vivo experiments and cell cultures.However,the lack of systematic and continuous comparison between in vivo retinal development and the organotypic retinal explant culture makes this model controversial in postnatal retinal development studies.Thus,we aimed to verify the feasibility of using this model for postnatal retinal development studies by comparing it with the in vivo retina.In this study,we showed that postnatal retinal explants undergo normal development,and exhibit a consistent structure and timeline with retinas in vivo.Initially,we used SOX2 and PAX6 immunostaining to identify retinal progenitor cells.We then examined cell proliferation and migration by immunostaining with Ki-67 and doublecortin,respectively.Ki-67-and doublecortin-positive cells decreased in both in vivo and explants during postnatal retinogenesis,and exhibited a high degree of similarity in abundance and distribution between groups.Additionally,we used Ceh-10 homeodomain-containing homolog,glutamate-ammonia ligase(glutamine synthetase),neuronal nuclei,and ionized calcium-binding adapter molecule 1 immunostaining to examine the emergence of bipolar cells,Müller glia,mature neurons,and microglia,respectively.The timing and spatial patterns of the emergence of these cell types were remarkably consistent between in vivo and explant retinas.Our study showed that the organotypic retinal explant culture model had a high degree of consistency with the progression of in vivo early postnatal retina development.The findings confirm the accuracy and credibility of this model and support its use for long-term,systematic,and continuous observation.展开更多
文摘In the present study,the effects of postnatal development on the number and distribution of skeletal muscle fibers of different types in hind leg of rat and rabbit were studied.The soleus muscles of rats aged 2 days,2,4,6,8,and 10 weeks (body weight 10,32,95,190,280,and 320 g),and rabbits aged 2 days,2,4,8,12,16,20,and 24 weeks (body weight 100,220,400,750,1 200,1 600,2 100,and 2 500 g) were stained with succinic dehydrogenase.With an image analysis system,the X-Y coordinates of fibers were used to analyze the growth-related changes.The results of present study showed that three types of fibers were found in the soleus muscles of rat and rabbit,i.e.,type Ⅰ (slow oxidative),ⅡX (fast oxidative),and ⅡA (fast oxidative glycolytic).The type Ⅰ fibers were present throughout the muscle that had a uniform distribution and tended to increase in number with aging.Type ⅡX fibers were scattered throughout the muscle and decreased markedly in number with aging.Type ⅡA fibers were located at the central and deep regions,and showed a little or no change in number and distribution with aging.While be of age,type ⅡA and ⅡX fibers became restricted to the superficial region.No type ⅡB fibers were detected.Type ⅡA fibers had the largest diameter,type Ⅰ intermediate and type ⅡX the smallest.Mean cross-sectional area of each type fibers of rabbits was larger than that of rats.The present results indicate that the number and distribution of muscle fibers of different types in hind limb of rat and rabbit change with the process of postnatal growth.
基金supported by the National Natural Science Foundation of China,No.81601181,81673716(to GZ)the Natural Science Foundation of Anhui Province of China,No.1808085J15(to GZ)the Natural Science Research Project of Anhui Province of China,No.KJ2016A417(to GZ)
文摘Our previous studies showed that the early use of calpain inhibitors reduces calpain activity in multiple brain regions, and that postnatal treatment with calpeptin may lead to cerebellar motor dysfunction. However, it remains unclear whether postnatal calpeptin application affects hippocampus-related behaviors. In this study, Sprague-Dawley rats were purchased from the Animal Center of Anhui Medical University of China. For the experiments in the adult stage, rats were intraperitoneally injected with calpeptin, 2 mg/kg, once a day, on postnatal days 7–14. Then on postnatal day 60, the Morris water maze test was used to evaluate spatial learning and memory abilities. The open field test was carried out to assess anxiety-like activities. Phalloidin staining was performed to observe synaptic morphology in the hippocampus. Immunohistochemistry was used to count the number of NeuN-positive cells in the hippocampal CA1 region. DiI was applied to label dendritic spines. Calpeptin administration impaired spatial memory, caused anxiety-like behavior in adulthood, reduced the number and area of apical dendritic spines, and decreased actin polymerization in the hippocampus, but did not affect the number of NeuN-positive cells in the hippocampal CA1 region. For the neonatal experiments, neonatal rats were intraperitoneally injected with calpeptin, 2 mg/kg, on postnatal days 7 and 8. Western blot assay was performed to analyze the protein levels of Akt, Erk, p-Akt, p-Erk1/2, Erk1/2, SCOP, PTEN, mTOR, p-mTOR, CREB and p-CREB in the hippocampus. SCOP expression was increased, and the phosphorylation levels of Akt, mTOR and CREB were reduced in the hippocampus. These findings show that calpeptin administration after birth affects synaptic development in neonatal rats by inhibiting the Akt/mTOR signaling pathway, thereby perturbing hippocampal function. Therefore, calpeptin administration after birth is a risk factor for neurodevelopmental defects.
基金supported by the National Natural Science Foundation of China,Nos.81901156(to ZZ),82271200(to ZZ),82171308(to XC)the Fundamental Research Funds for the Central Universities,No.xzy012022035(to ZZ)+1 种基金the Natural Science Foundation of Shaanxi Province,Nos.2021JM-261(to QK),2023-YBSF-303(to ZZ)Traditional Chinese Medicine Project of Shaanxi Province,No.2019-ZZ-JC047(to QK)。
文摘The organotypic retinal explant culture has been established for more than a decade and offers a range of unique advantages compared with in vivo experiments and cell cultures.However,the lack of systematic and continuous comparison between in vivo retinal development and the organotypic retinal explant culture makes this model controversial in postnatal retinal development studies.Thus,we aimed to verify the feasibility of using this model for postnatal retinal development studies by comparing it with the in vivo retina.In this study,we showed that postnatal retinal explants undergo normal development,and exhibit a consistent structure and timeline with retinas in vivo.Initially,we used SOX2 and PAX6 immunostaining to identify retinal progenitor cells.We then examined cell proliferation and migration by immunostaining with Ki-67 and doublecortin,respectively.Ki-67-and doublecortin-positive cells decreased in both in vivo and explants during postnatal retinogenesis,and exhibited a high degree of similarity in abundance and distribution between groups.Additionally,we used Ceh-10 homeodomain-containing homolog,glutamate-ammonia ligase(glutamine synthetase),neuronal nuclei,and ionized calcium-binding adapter molecule 1 immunostaining to examine the emergence of bipolar cells,Müller glia,mature neurons,and microglia,respectively.The timing and spatial patterns of the emergence of these cell types were remarkably consistent between in vivo and explant retinas.Our study showed that the organotypic retinal explant culture model had a high degree of consistency with the progression of in vivo early postnatal retina development.The findings confirm the accuracy and credibility of this model and support its use for long-term,systematic,and continuous observation.
