Primary dissociated neuronal cultures are widely used research tools to investigate of pathological mechanisms and to treat various central and peripheral nervous system problems including trauma and degenerative neuronal diseases. We introduced a protocol that utilizes hippocampal and cortical neurons from embryonic day 17 or 18 mice. We applied appropriate markers (GAP-43 and synaptophysin) to investigate whether neurite outgrowth and synaptogenesis can be distinguished at a particular period of time. GAP-43 was found along the neural processes in a typical granular pattern, and its expression increased proportionally as neurites lengthened during the early in vitro period. Unlike GAP-43, granular immunoreactive patterns of synaptophysin along the neurites were clearly found from day 2 in vitro with relatively high immunoreactive levels. Expression of synaptic markers from cortical neurons reached peak level earlier than that of hippocampal neurons, although neurite outgrowths of hippocampal neurons were faster than those of cortical neurons. The amount of peak synaptic markers expressed was also higher in cortical neurons than that in hippocampal neurons. These results strongly suggest the usefulness of primary cultured neurons from mice embryos for synaptic function and plasticity studies, because of their clear and typical patterns of morphology that establish synapses. Results from this study also suggest the proper amount of time in vitro according to neuronal types (cortical or hippocampal) when utilized in experiments related with synaptogenesis or synaptic activities.
Animals ; Embryonic Structures ; GAP-43 Protein ; Mice ; Neurites ; Neurons ; Peripheral Nervous System ; Plastics ; Synapses ; Synaptophysin

The genetically engineered mice require special husbandry care and are mainly housed in Individually Ventilated Cage (IVC) systems and Static Micro Isolator Cages (SMIC) to minimize the risk for spreading undesirable microorganisms. However, the static micro isolation cage housing like SMIC are being replaced with IVC systems in many facilities due to a number of benefits like a higher density housing in limited space, better protection from biohazards and allergens and decreased work load due to decreased frequency of cage changing required in this system. The purpose of this study was to examine the reproductive performance of genetically engineered mice housed in individually ventilated cages (IVC) and Static Micro Isolator Cages (SMIC). When the B6C3-Tg (APPswe, PSEN1dE9) 85Dbo/Mmjax transgenic mice were housed in these two housing systems, the number of litters per dam, number of pups born per dam and number of pups weaned per dam were found to be slightly higher in the IVC as compared to the SMIC but the difference was not significant (P<0.05). In case of Growth Associated Protein 43 (GAP-43) knockout mice, the number of litters born per dam and the number of pups born per dam were marginally higher in the IVC as compared to those housed in SMIC but the difference was not significant (P<0.05). Only the number of pups weaned per dam were found to be significantly higher as compared to those housed in the SMIC system at P<0.05.
Allergens ; Animals ; GAP-43 Protein ; Hazardous Substances ; Housing* ; Mice* ; Mice, Knockout ; Mice, Transgenic

Although growth associated protein-43 (GAP-43) is known to play a significant role in the regulation of axonal growth and the formation of new neuronal connections in the hippocampus, there is only a few studies on the effects of acute stress on GAP-43 mRNA expression in the hippocampus. Moreover, the effects of repeated citalopram treatment on chronic mild stress (CMS)-induced changes in GAP-43 mRNA expression in the hippocampus have not been explored before. To explore this question, male rats were exposed to acute immobilization stress or CMS. Also, citalopram was given prior to stress everyday during CMS procedures. Acute immobilization stress significantly increased GAP-43 mRNA expression in all subfields of the hippocampus, while CMS significantly decreased GAP-43 mRNA expression in the dentate granule cell layer (GCL). Repeated citalopram treatment decreased GAP-43 mRNA expression in the GCL compared with unstressed controls, but this decrease was not further potentiated by CMS exposure. Similar decreases in GAP-43 mRNA expression were observed in CA1, CA3 and CA4 areas of the hippocampus only after repeated citalopram treatment in CMS-exposed rats. This result indicates that GAP-43 mRNA expression in the hippocampus may differently respond to acute and chronic stress, and that repeated citalopram treatment does not change CMS-induced decreases in GAP-43 mRNA expression in the GCL.
Animals ; Axons ; Citalopram ; GAP-43 Protein ; Hippocampus ; Humans ; Immobilization ; Male ; Neurons ; Rats ; RNA, Messenger

OBJECTIVE: To verify the effect of low energy laser irradiation (LELI) on the regeneration of injured sciatic nerve of the rat by showing the functional improvement and the elevated immunoreactivities (IRs) of growth-associated protein 43 (GAP-43). METHOD: Twenty rats, which had standardized compression injuries to the sciatic nerves, received the calculated LELI therapy immediately after the nerve injury and four consecutive days. The functional status was evaluated by sciatic functional index (SFI), and GAP-43-IRs was evaluated by immunohistochemistry and RT-PCR. RESULTS: The SFI was recovered in LELI rats faster than in the control group. Although expression of GAP-43 in the injured sciatic nerve was increased both in the LELI and control groups, the intensities of GAP-43-IRs were much greater in LELI treated group at 1 and 3 weeks after nerve injury. Both SFI and GAP-43-IRs reached the same level at 5 weeks after the nerve injury. CONCLUSION: LELI enhanced the neural regeneration after experimentally induced sciatic nerve injury at the early stage of recovery. Considering the effect of LELI on nerve regeneration was not fully explained until now, this study could suggest the meaningful explanation on the mechanism of LELI effectiveness on neural regeneration.
Animals ; GAP-43 Protein ; Immunohistochemistry ; Nerve Regeneration ; Peripheral Nerve Injuries ; Rats* ; Regeneration* ; Sciatic Nerve*

OBJECTIVETo investigate whether there is neogenesis of myelin sheath and neuron after transplantation of Schwann cells into cerebral hemorrhage lesion.

METHODSSchwann cells were expanded, labeled with BrdU in vitro and transplanted into rat cerebral hemorrhage with blood extracted from femoral artery and then injected into the basal nuclei. Double immunohistochemistry staining and electron microscopy were used to detect the expression of BrdU/MBP and BrdU/GAP-43 and remyelination.

RESULTSBrdU/MBP double positive cells could be seen at 1 week up to 16 weeks after transplantation of Schwann cells. Thin remyelination was observed under electron microscope. GAP-43 positive cells appeared after 12 weeks and were found more in Hippocamp.

CONCLUSIONSGrafted Schwann cells participate in remyelination and promoter nerve restore in rat cerebral hemorrhage.

Animals ; Cerebral Hemorrhage ; metabolism ; therapy ; GAP-43 Protein ; metabolism ; Rats ; Rats, Wistar ; Schwann Cells ; metabolism ; transplantation ; Sciatic Nerve ; cytology ; embryology

OBJECTIVE: To observe the effect of moxibustion at "Huantiao" (GB 30) on the expression of growth-associated protein-43 (GAP-43) in the sciatic nerve trunk and ventral horn of spinal cord (L METHODS: A total of 48 healthy male SD rats were randomly divided into a normal group, a sham operation group, a model group and a moxibustion group, 12 rats in each group. The rat model of primary sciatic pain was established by chronic constriction injury (CCI) of the sciatic nerve in the model group and the moxibustion group. On the 8th day of the experiment, moxibustion was adopted at "Huantiao" (GB 30) in the moxibustion group for 5-10 min, once a day for 14 consecutive days. Sciatic nerve function index (SFI) was measured and compared in each group at day 1, 7, 14 and 21. On the 21st day of the experiment, HE staining was used to observe the morphology of ventral horn of rat spinal cord and sciatic nerve trunk. Immunohistochemical method and real-time PCR were used to detect mRNA and protein expressions of GAP-43 in the spinal cord and sciatic nerve trunk of rats. RESULTS: On day 7, 14 and 21, there was no statistical difference in SFI between the sham operation group and the normal group ( CONCLUSION Moxibustion at "Huantiao" (GB 30) could improve the sciatic nerve function in rats with primary sciatica and its mechanism may be related to improving the expression of GAP-43 and enhancing the self-repair ability of the sciatic nerve after injury.
Animals ; Electroacupuncture ; GAP-43 Protein/genetics* ; Male ; Moxibustion ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; Sciatica/therapy* ; Spinal Cord

OBJECTIVETo investigate the effects of gestational isoflurane exposure on postnatal memory and learning and growth-associated protein-43 (GAP-43), neuropeptide Y(NPY) expression in the hippocampus of pups.

METHODSTwelve maternal Sprague-Dawley rats at gestation d 18(E18) were randomly divided into isoflurane group (n=6) and control group (n=6). Rats in isoflurane group were exposed to 1.3 % isoflurane for 6 h. For control group, animals breathed in 30 % oxygen and air mixed gas at the same condition. Spatial learning and memory of the offspring were determined with the Morris Water Maze(MWM) after postnatal 4 weeks. The changes of GAP-43 and NPY expression in the hippocampal CA1 region of the pups were determined by immunohistochemistry.

RESULTSIn MWM training, the escape latency to platform of the pups in isoflurane group was significantly longer, and the time spent in the third quadrant and times of original platform crossing were less than those of control animals (P<0.05). The number and optical density of GAP-43 and NPY positive neurons in the hippocampus of pups decreased significantly in the isoflurane group compared with the controls (P <0.01).

CONCLUSIONIsoflurane exposure in pregnant rats significantly impairs the spatial memory and learning of their pups at a juvenile age, which may be associated with the down-regulation of GAP-43 and NPY in the hippocampus.

Animals ; Female ; GAP-43 Protein ; metabolism ; Hippocampus ; drug effects ; metabolism ; Isoflurane ; pharmacology ; Maze Learning ; drug effects ; Neuropeptide Y ; metabolism ; Pregnancy ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the mechanisms of acupuncture treatment promoting the motor function recovery of neonate rats with cerebral palsy.

METHODSThe improved hypoxic-ischemic encephalopathy (HIE) means was performed to establish the model of neonate rats with cerebral palsy. All neonate rats were randomly divided into 3 groups: sham group, model group and acupuncture group (n = 20). We observed and scored motor function of rats, measured the levels of superoxide dismutase (SOD) and malondialdehyde (MDA) in serum, and also measured the expression of synaptophysin (SYP) and growth associated protein-43 (GAP-43) in the diseased region of cerebral tissue.

RESULTSThe motor function scores (11.3 +/- 0.29) and the serum level of SOD (147.1 +/- 12.7) U/ml in acupuncture treatment group were higher than those of model group ( P < 0.05). The serum level of MDA was lower in acupuncture treatment group than that of model group (P < 0.05). The expression of SYP and GAP-43 in the diseased region of cerebral tissue of acupuncture treatment group were higher than those of model group ( P < 0.05) .

CONCLUSIONAcupuncture-therapy could improve the motor function of neonate rats with cerebral palsy by decreasing the content of MDA in serum, increasing the contents of SOD in serum, and prolonging the upregulation of SYP and GAP-43 expressions in hmin tissue.

Acupuncture Therapy ; Animals ; Animals, Newborn ; Cerebral Palsy ; therapy ; Disease Models, Animal ; GAP-43 Protein ; metabolism ; Hypoxia-Ischemia, Brain ; therapy ; Malondialdehyde ; metabolism ; Rats ; Superoxide Dismutase ; metabolism ; Synaptophysin ; metabolism

Caveolin-1 (Cav-1) is a marker protein for caveolae, and acts as scaffolding protein to regulate the activities of signaling molecules. Previous studies indicate that Cav-1 mainly locates at the base of axonal and dendritic terminals of mouse primary hippocampal neurons and plays an active role in the regulation of injury-induced synaptic and terminal remodeling in central nervous system. The aim of this study was to identify the expression profile of Cav-1 protein in the brains of rats at different ages and to investigate the role of Cav-1 in Y-maze bright-dark discrimination learning (BDL). Firstly, the expressions of Cav-1 in the brains of young (1-month), adult (3-month) and aged (22-month) rats were observed by Western blot. Higher expression in the hippocampus and lower expression in the cortex were shown in the adult rats. It was also found that the score of BDL was related with the expression level of Cav-1. Secondly, using open-field test for spontaneous locomotor activities (SLA) and BDL, the role of Cav-1 in the learning and memory was observed. Compared with that in the control adult group, the Cav-1 protein expression in the hippocampus and prefrontal cortex of Y-maze trained adult rats significantly increased, while no marked changes in the cerebellum. These results suggest that Cav-1 protein is involved in BDL and plays an important role in the plasticity of central nervous system.
Age Factors ; Animals ; Brain Chemistry ; Caveolin 1 ; analysis ; physiology ; Discrimination Learning ; GAP-43 Protein ; analysis ; Male ; Maze Learning ; Rats ; Rats, Sprague-Dawley ; Synaptophysin ; analysis

OBJECTIVETo observe the effect of electroacupuncture on the neural plasticity in rats with cerebral infarction and investigate its mechanism.

METHODSSixty rats were randomly divided into a sham operation group, an acupuncture group and a non-acupuncture group, and each group was randomly divided into a 1-day subgroup, a 7-day subgroup and a 14-day subgroup. Cerebral infarction model was induced by the thread embolism method. The rats in the acupuncture group were treated by electroacupuncture at "Zusanli" (ST 36) and "Neiguan" (PC 6) for 20 minutes, once each day, while the rats in the sham operation group and the non-acupuncture group were just bound and fixed at the same time without acupuncture treatment. Neurological defects were assessed by Neurological Severity Score (NSS) and the changes of the expression of growth associated protein 43 (GAP-43) in peripheral nerve around cerebral infarction area were detected by immunohistochemistry technique.

RESULTSThe expression of the positive cells of GAP-43 around cerebral infarction area showed no significant distinction among the three groups at 1st day (P > 0.05), while the GAP-43 expression level of around cerebral infarction area in the acupuncture group (IOD:8. 990 1 +/- 0.098 7, 5.816 1 +/- 0.204 6) were significant higher than those in the sham operation group (IOD: 1.300 2 +/- 0.093 3, 1.362 6 +/- 0.216 6) and in the non-acupuncture group (IOD: 2.753 4 +/- 0.0875, 1.616 5 +/- 0.186 8) at 7th day and 14th day, respectively (all P < 0.01).

CONCLUSIONElectroacupuncture at "Zusanli" (ST 36) and "Neiguan" (PC 6) can improve the neural function and promote its remodeling in rats with cerebral infarction and the relevant mechanisms may be involved in enhancement of GAP-43 expressions around ischemic region.

Acupuncture Points ; Animals ; Cerebral Infarction ; genetics ; metabolism ; therapy ; Disease Models, Animal ; Electroacupuncture ; GAP-43 Protein ; genetics ; metabolism ; Humans ; Male ; Random Allocation ; Rats