OBJECTIVETo explore the role of microtubule-associated protein 2 (MAP-2) and nestin in the development of tongue muscles of human embryos and fetuses.

METHODSPV immunohistochemistry was used to detect the expressions of MAP-2 and nestin proteins in the tongue tissues of human embryos and fetuses at the second, third and fourth months of gestation.

RESULTSMAP-2 and nestin positivity was detected in the tongue muscles of human embryos at 2 to 4 months of gestation. In the embryos at the second month of gestation, no obvious MAP-2 positive cells were found in the tongue muscles; at 3 and 4 months, the number of MAP-2-positive cells in the tongue muscles was 24.14∓8.28 and 15.86∓3.89, with the expression intensity of 109.42∓11.62 and 124.27∓8.73, respectively. At 2, 3 and 4 months of gestation, the number of nestin-positive cells in the tongue muscles was 12.50∓3.17, 19.00∓7.63, and 22.80∓6.91, with expression intensity of 119.99∓24.02, 102.20∓11.76, and 98.24∓10.66, respectively. As the gestational age increased, the number of MAP-2-positive cell number continued to decline following a transient increase but the expression intensity kept increasing; nestin-positive cells increased continuously but the expression intensity kept decreasing in the embryonic or fetal tongue muscles.

CONCLUSIONMAP-2 and nestin proteins are involved in the regulation of the development of tongue muscles in human embryos and fetuses.

Fetus ; metabolism ; Humans ; Microtubule-Associated Proteins ; metabolism ; Nestin ; metabolism ; Tongue ; embryology ; growth & development ; metabolism

OBJECTIVETo observe the morphological changes during development of the ventricular zone (VZ) and subventricular zone (SVZ) of human fetus and the distribution pattern of neural stem cells in the VA and SVZ.

METHODSHuman fetuses at the gestational ages of 9-11 weeks, 14-16 weeks, 22-24 weeks and 32-36 weeks were collected, and the brain sections of the VZ/SVZ under the frontal lobe were examined for cytoarchitecture and distribution of nestin-positive cells with HE staining, immunohistochemistry or immunofluorescence.

RESULTSThe thickness of VZ underwent no significant changes at the gestational ages of 9-24 weeks (P>0.05) and became obviously thinner at 32 weeks (P<0.05), while the thickness of SVZ increased during 9-24 weeks (P<0.05) without obvious thinning at 32 weeks (P>0.05). VZ was thicker than SVZ at 9-11 weeks but became markedly thinner than SVZ after 14 weeks (P<0.05). The VZ contained denser cells than SVZ and showed a distinct boundary between the VZ and SVZ. Large numbers of nestin-positive cells were detected in the VZ and SVZ, and nestin immunoreactivity was found primarily in the cell processes and occasionally in the soma. Some nestin-positive cells in the SVZ had 1-3 processes. Nestin immunoreactivity in the VZ and SVZ gradually grew weak with development. The cells positive for both nestin and Ki67 were located mainly in the inner zone of the VZ and throughout the SVZ, where some nestin-positive but Ki67-negative cells were also found.

CONCLUSIONThe SVZ fully extends and the neural stem cells in the VZ/SVZ can be morphologically heterogeneous during the development of fetal human brain.

Fetus ; Frontal Lobe ; cytology ; embryology ; metabolism ; Humans ; Nestin ; metabolism ; Neural Stem Cells ; metabolism ; Neurons ; metabolism

Astrocytoma ; metabolism ; Glial Fibrillary Acidic Protein ; metabolism ; Humans ; In Vitro Techniques ; Nestin ; metabolism

OBJECTIVETo explore the role of microtubule-associated protein 2 (MAP-2) and nestin in the development of tongue muscles of human embryos and fetuses.

METHODSPV immunohistochemistry was used to detect the expressions of MAP-2 and nestin proteins in the tongue tissues of human embryos and fetuses at the second, third and fourth months of gestation.

RESULTSMAP-2 and nestin positivity was detected in the tongue muscles of human embryos at 2 to 4 months of gestation. In the embryos at the second month of gestation, no obvious MAP-2 positive cells were found in the tongue muscles; at 3 and 4 months, the number of MAP-2-positive cells in the tongue muscles was 24.14∓8.28 and 15.86∓3.89, with the expression intensity of 109.42∓11.62 and 124.27∓8.73, respectively. At 2, 3 and 4 months of gestation, the number of nestin-positive cells in the tongue muscles was 12.50∓3.17, 19.00∓7.63, and 22.80∓6.91, with expression intensity of 119.99∓24.02, 102.20∓11.76, and 98.24∓10.66, respectively. As the gestational age increased, the number of MAP-2-positive cell number continued to decline following a transient increase but the expression intensity kept increasing; nestin-positive cells increased continuously but the expression intensity kept decreasing in the embryonic or fetal tongue muscles.

CONCLUSIONMAP-2 and nestin proteins are involved in the regulation of the development of tongue muscles in human embryos and fetuses.

Humans ; Microtubule-Associated Proteins ; metabolism ; Muscle, Skeletal ; embryology ; Nestin ; metabolism ; Tongue ; embryology

OBJECTIVETo investigate the role of microtubule-associated protein 2 (MAP-2) and nestin in gastric development in human embryos and fetuses.

METHODSImmunohistochemistry was used to detect the expressions of MAP-2 and nestin proteins in the gastric cardia, pyloric and gastric tissues of human embryos and fetuses during the second, third and fourth month of development.

RESULTSIn the second to fourth months of gestation, MAP-2 and nestin expressions were detected in the neural cells and neural fibers of the intermuscular nerve plexus and submucosal plexus in the gastric cardia, pyloric and gastric tissues. As the gestational age increased, the number of MAP-2- and nestin-positive cells and the expression intensity all increased in the myenteric plexus, but MAP-2 and nestin expressions were negative in the glandular and mucosal tissues of human embryonic and fetal gastric cardia, pylorus or gastric walls.

CONCLUSIONMAP-2 and nestin participate in the regulation of the development of gastric tissues in human embryos.

Fetus ; metabolism ; Humans ; Intermediate Filament Proteins ; metabolism ; Microtubule-Associated Proteins ; metabolism ; Nerve Tissue Proteins ; metabolism ; Nestin ; Stomach ; embryology ; metabolism

Immunohistochemistry and double immunofluorescent labeling techniques combined with confocal laser scanning microscope analysis were used to investigate the characteristic spatial induction profile of nestin following a transient middle cerebral artery occlusion in adult rat brain. The results showed that nestin was induced in ischemic core at 1 day after reperfusion. In addition to ischemic core, the expression of nestin increased in peri-ischemic I, II and III regions at 3 days and 1 week, then it decreased and narrowed along the rim of ischemic core 2 weeks after reperfusion. Double immunofluorescent labeling showed that nestin positive cells were mostly co-stained with GFAP,a astrocyte marker, in peri-ischemic I region 3 days after reperfusion. At 2 weeks, however nestin cells showed a long process and the cells double stained with nestin and NSE,a neuonal specific marker,increased in the ischemic brain. The results suggest that cerebral ischemia induces nestin expression in damaged neurons which might favor the neuroprotection against ischemic damage.
Animals ; Brain ; metabolism ; pathology ; Brain Ischemia ; metabolism ; pathology ; Immunohistochemistry ; Infarction, Middle Cerebral Artery ; metabolism ; pathology ; Nestin ; metabolism ; Neurons ; metabolism ; Rats

Living organisms are exposed to the geomagnetic field (GMF) throughout their lifespan. Elimination of the GMF, resulting in a hypogeomagnetic field (HMF), leads to central nervous system dysfunction and abnormal development in animals. However, the cellular mechanisms underlying these effects have not been identified so far. Here, we show that exposure to an HMF (<200 nT), produced by a magnetic field shielding chamber, promotes the proliferation of neural progenitor/stem cells (NPCs/NSCs) from C57BL/6 mice. Following seven-day HMF-exposure, the primary neurospheres (NSs) were significantly larger in size, and twice more NPCs/NSCs were harvested from neonatal NSs, when compared to the GMF controls. The self-renewal capacity and multipotency of the NSs were maintained, as HMF-exposed NSs were positive for NSC markers (Nestin and Sox2), and could differentiate into neurons and astrocyte/glial cells and be passaged continuously. In addition, adult mice exposed to the HMF for one month were observed to have a greater number of proliferative cells in the subventricular zone. These findings indicate that continuous HMF-exposure increases the proliferation of NPCs/NSCs, in vitro and in vivo. HMF-disturbed NPCs/NSCs production probably affects brain development and function, which provides a novel clue for elucidating the cellular mechanisms of the bio-HMF response.
Animals ; Cell Proliferation ; physiology ; Female ; Magnetic Fields ; Male ; Mice ; Nestin ; metabolism ; Neural Stem Cells ; cytology ; metabolism ; SOXB1 Transcription Factors ; metabolism

Objectives: To study the effects of Porphyromonas gingivalis (Pg) injected through tail vein on the molecular expression levels of biomarkers of neural stem cells (NSC) and neurons in the hippocampus of wild-type adult rats, and the effects on hippocampal neurogenesis. Methods: Eighteen male Sprague-Dawley (SD) rats were randomly divided into 3 groups based on the table of random numbers (n=6 in each group). In low-intensity group and high-intensity group, rats were injected intravenously through tail vein with 200 μl Pg ATCC33277 [1.0×10³ and 1.0×10⁸ colony forming unit (CFU), respectively] 3 times per week for 8 weeks. In the sham group, 200 μl of phosphate buffer saline (PBS) was given instead. Behavioral tests: the navigation and the exploration tests using Morris water maze (MWM) were applied to evaluate learning and memory ability of rats. Immunohistochemistry was performed to detect cells positively expressing nestin, doublecortin (DCX) and neuronal nuclei (NeuN) in the subgranular zone (SGZ) of rats in each group. Western blotting was used to evaluate the expression levels of nestin, DCX and NeuN in rat hippocampus. Results: Learning and memory abilities: on day 5 of navigation test, the lagency time was 22.83 (16.00, 38.34) s in the high-intensity group, significantly longer than the sham group [5.59 (5.41, 6.17) s] (t=-11.17, P<0.001). There were no significant differences between the low-intensity group [9.85 (8.75, 21.01) s] and the sham group (t=-6.83, P=0.080). Results in the exploration test showed that, in the high-intensity group, the number of fime crossing over the previous platform area within 60 s was 1.50 (1.00, 2.00), significantly less than the sham group [4.00 (2.75, 4.00)] (t=9.75, P=0.003); no significant differences between the low-intensity group [2.50 (2.00, 3.00)] and the sham one (t=4.50, P=0.382). Immunohistochemistry showed that the nestin⁺ cell density in the low-intensity group [(35.36±4.32) cell/mm²] and high-intensity group [(26.51±5.89) cell/mm²] were significantly lower than the sham group [(59.58±14.15) cell/mm²] (t=24.21, P=0.018; t=33.07, P=0.005); as for the mean absorbance of DCX⁺ cells, the low-intensity group (0.007±0.002) and the high-intensity group (0.006±0.002) were significantly lower than the sham group (0.011±0.001) (t=0.004, P=0.018; t=0.006, P=0.005); compared with the sham group [(1.13±0.14)×10³ cell/mm²], the density of NeuN⁺ neurons in the high-intensity group [(0.75±0.08)×10³ cell/mm²] was significantly reduced (t=0.38, P=0.017), and was not significantly changed in the low-intensity group [(0.88±0.19)×10³ cell/mm²] (t=0.25, P=0.075). Western blotting results showed that, compared with the sham group, the expression levels of nestin, DCX, and NeuN were significantly reduced in the high-intensity group (t=0.74, P<0.001; t=0.18, P=0.014; t=0.35, P=0.008), but were not statistically changed in the low-intensity group (t=0.18, P=0.108; t=0.08, P=0.172; t=0.19, P=0.077). Conclusions: Pg injected through tail vein may reduce learning and memory abilities of wild-type rats, and may reduce the number of nestin, DCX, and NeuN-positive cells, and the protein expression levels of the above molecules in the hippocampus.
Animals ; Biomarkers/metabolism* ; Hippocampus/metabolism* ; Male ; Nestin/metabolism* ; Neural Stem Cells/metabolism* ; Neurons/metabolism* ; Porphyromonas gingivalis/metabolism* ; Rats ; Rats, Sprague-Dawley ; Tail/metabolism*

OBJECTIVETo introduce the eukaryotic expression vector pEGFP-C1-PDX-1 into nestin-positive cell derived from bone marrow stromal cells by nucleofection and optimize the conditions for transfection.

METHODSThe recombinant plasmid was transfected into bone marrow stromal cells-derived nestin-positive cells with varied DNA quantities or the serum concentration in the medium. The expression of PDX-1 gene in the transfected cells was detected by RT-PCR.

RESULTSSatisfactory efficiency of transfection was achieved with the DNA quantity of 2-10 microg and medium serum concentration of 20%. PDX-1 expression was detected in the transfected cells by RT-PCR.

CONCLUSIONThe optimized transfection conditions result in enhanced efficiency of PDX-1 gene transfection into nestin-positive cells derived from bone marrow stromal cells, which may serve as the seed cells in tissue-engineering.

Bone Marrow Cells ; metabolism ; Genetic Vectors ; Homeodomain Proteins ; genetics ; Humans ; Intermediate Filament Proteins ; metabolism ; Nerve Tissue Proteins ; metabolism ; Nestin ; Plasmids ; Stromal Cells ; metabolism ; Trans-Activators ; genetics ; Transfection ; methods

Brain Neoplasms ; metabolism ; pathology ; surgery ; Diagnosis, Differential ; Humans ; Infant ; Magnetic Resonance Imaging ; Male ; Neoplasm Recurrence, Local ; Nestin ; metabolism ; Neuroectodermal Tumors, Primitive ; metabolism ; pathology ; surgery ; Temporal Lobe ; Vimentin ; metabolism